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秋田県健康環境センター年報 第10号
ISSN 1881-6053 秋 田 県 健 康 環 境 セ ン タ ー 年 報 秋田県健康環境センター年報 第 10 号 平成 26 年度 ANNUAL REPORT OF AKITA RESEARCH CENTER FOR PUBLIC HEALTH AND ENVIRONMENT 第 10 No. 10 2014 号 平 成 26 年 度 （ 2014 ） 秋田県健康環境センター Ƹ ơ NJ ƴ ᅸဋჄͤࡍؾǻȳǿȸƸŴᲫᲲ࠰ࡇƴᘓဃᅹܖᄂᆮƱؾǻȳǿȸƕ ወӳƠƯᚨᇌƞǕŴʻ࠰ưᲫᲪ࠰ƷራႸǛᡇƑƯƍLJƢŵ ࢘ǻȳǿȸƷᢃփƸŴʻࢸƷ૾ӼࣱǛܭNJƨžɶᧈᚘဒſƴඝƬƯᡶNJƯƍLJƢ ƕŴჄൟƷܤμȷ࣎ܤƷᄩ̬ƱƍƏᙻໜƔǒŴžͤࡍᘮܹƷ᧸ഥſƱžؾƷ̬μſ ƴӼƚƨӕኵǛؕஜ૾ᤆƱƠƯಅѦǛᘍƠƯƍLJƢŵ φ˳ႎƴƸŴᘍሁƔǒƷ̔᫂౨௹ƱƠƯŴज़௨ၐǍԼᘓဃƴ̞ǔኬᓏȷǦǤȫ ǹ౨௹ǛƸơNJŴԼɶƷസသᠾᕤ౨௹Ŵඕ߷ȷฯකŴئ൦ሁƷ൦ឋ౨௹Ŵ્ݧ ᏡยܭŴٻൢȷٓםȷᬳ᪦ยܭሁ࠼ርưٶૠƷ౨௹ǛܱƠƯƍLJƢŵ ́ƤƯᛦ௹ᄂᆮưƸŴ؏עƷᘍᛢ᫆ǍȋȸǺƴႺኽƠƨȆȸȞǛӕǓɥƛƯƍǔ DŽƔŴƷज़௨ၐᄂᆮǍؾᄂᆮƱσӷᄂᆮƢǔƳƲμႎƳӕኵƴNjӋьƠƯ ƍLJƢŵƜƷ˂ŴჄൟǁƷऴإ੩̓ƱƠƯज़௨ၐႆဃѣӼǛᛦ௹Ƣǔज़௨ၐऴإǻȳ ǿȸƷᢃփŴᆰ᧓્ݧዴǛႳᙻƢǔȢȋǿȪȳǰȝǹȈƷᚨፗŴ2/ሁٻൢ൲௨ ཋឋƷࠝႳᙻƳƲƴǑǓӲදॖإǛႆˋƠƯƍǔDŽƔŴЎௌሁƷ২ᘐਦݰǍЈЭᜒ ࡈሁǛܱƠŴͤࡍʖ᧸Ǎ̬ؾμƷॖᜤƷᣱǍգႆƴѐNJƯƍLJƢŵ ମ࠰ƸŴȇȳǰ༏Ʒϋज़௨ƕኖᲱᲪ࠰ƿǓƴႆဃƠŴLJƨŴᙱǢȕȪǫᜂưƸ ǨȜȩЈᘉ༏ƕ්ٻᘍƠLJƠƨŵʻ࠰ƸŴ᪡Ʒ/'45dzȭȊǦǤȫǹƷज़௨ƕਘٻ ƠϋǁƷˡƕүूƞǕǔƳƲŴᡈ࠰ƷͤࡍүೞƕفƠƯƍLJƢŵƜƷƨNJ࢘ǻȳ ǿȸưƸŴɢɟዬ࣯ʙ७ƕႆဃƠƨئӳưNjᡆᡮƳᛦ௹ȷ౨௹ƕưƖǔǑƏƳ˳Сૢ ͳƴѐNJƯƍLJƢŵ ƜƷ࠰إƸŴɼƴᲬᲰ࠰ࡇƴ࢘ǻȳǿȸƕᘍƬƨᛦ௹ᄂᆮǍಅѦܱጚƷಒᙲƴ ƭƍƯƱǓLJƱNJƨNjƷưƢŵஜǛᡫơƯٶƘƷႏಮƴ࢘ǻȳǿȸƷѣǁƷࣂྸ ᚐƱ᧙࣎Ǜ᭗NJƯƍƨƩƚǕƹ࠳ƍưƢŵ ᲬᲱ࠰ᲫᲬஉ ᅸဋჄͤࡍؾǻȳǿȸᧈ ش ር ┠ ḟ Ϩ ᗣ⎔ቃࢭࣥࢱ࣮ࡢᴫせ 1. ἢ 2. ᗇ⯋ࡢᴫせ 3. ⤌ 4. ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 1 ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 1 ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 1 ⫋ဨྡ⡙ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 2 5. ᴗົෆᐜ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 3 6. せᶵჾ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 4 ϩ ⧊ ᴗົᐇ⦼ 1. ヨ㦂᳨ᰝᐇ⦼ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 5 2. ◊ಟ࣭Ꮫ➼ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 9 3. ◊✲ᴗົᐇ⦼ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 14 Ϫ ㄪᰝ◊✲ሗ࿌ ࣭ ࢧ ࣝ ࣔ ࢿ ࣛ ࡢ Multiplex PCR ἲ ࡼ ࡿ ⾑ Ύ ᆺ ู ἲ ࡢ ☜ ❧ ͐͐͐͐͐͐͐͐͐͐͐ 21 ࣭ ᑠඣ྾ჾᝈ⏤᮶᳨యࡽࡢཎయ᳨ฟ≧ἣ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 25 ࣭ ᙉ ẘ ᆺ ࠾ ࡼ ࡧ ᙅ ẘ ᆺ Orientia tsutsugamushi ࡢ ࣐ ࢘ ࢫ ⾑ ୰ ࠾ ࡅ ࡿ ᥎ ⛣ ͐͐͐͐͐ 29 ࣭ ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣࡘ࠸࡚͐͐͐͐͐͐͐͐͐ 32 ࣭ 㣗ရࡽࡢ࢘ࣝࢫ᳨ฟἲࡋ࡚ࡢࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ ࣭ ϫ 㠉 ⢭ ᗘ ྥ ୖ 㛵 ࡍ ࡿ ◊ ✲ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 37 ␆Ỉ⏘≀୰ࡢື≀⏝་⸆ရ୍ᩧศᯒἲࡢ᳨ウཬࡧጇᙜᛶホ౯ࡘ࠸࡚͐͐͐͐͐͐͐ 59 Ⓨ⾲ᴗ⦼ 1. ᏛⓎ⾲ 2. ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ ㄅᥖ㍕ㄽᩥ ͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐͐ 70 80 ! ! ! ! !؝ॊৃࠂࢧҫӣү̴Ѡޕᅃ ! ! ! ! ! ! ! ! ! ! ! 秋田県健康環境センター年報 第10号 2014 ἢ 㠉 ᖺ ᭶ ᫂35. 7 ᫂ᮎᮇ 28. 1 39. 4 ⾨⏕ヨ㦂ᡤࢆ⛅⏣ᕷ∵ᓥ⏫タ❧ࠋ ᗇ⯋ࢆ⛅⏣ᕷᅵᡭ㛗⏫⛣㌿ࠋ ⾨⏕◊✲ᡤᨵ⛠ࠋ ⾨⏕⛉Ꮫ◊✲ᡤᨵ⛠ࠋ ᗇ⯋ࢆ⛅⏣ᕷྂᕝᇼ⏫㸦⌧༓⛅᫂ᚨ ⏫㸧᪂⠏⛣㌿ࠋ 39. 6 㡯 45. 7 බᐖᢏ⾡ࢭࣥࢱ࣮ࢆ⛅⏣ᕷ䭉ᓥࡢᕤᴗヨ㦂ሙෆ タ❧ࠋ ᗇ⯋ࢆ⛅⏣ᕷඵᶫ᪂⠏⛣㌿ࠋ ⎔ቃᢏ⾡ࢭࣥࢱ࣮ᨵ⛠ࠋ 48. 7 56. 4 61. 8 ᖹᡂ12. 4 ᗇ⯋ࢆ⛅⏣ᕷ༓⛅ஂಖ⏣⏫᪂⠏⛣㌿ࠋ ⎔ቃࢭࣥࢱ࣮ᨵ⛠ࠋ ⛅⏣ᕷᒣ⋤ࡢ┴ᗇ➨ᗇ⯋⥲ົ⌜ཬࡧ┘ど㺃 ሗ⌜ࢆ⨨ࡃࠋ ඵᶫศᐊᩜᆅෆࢲ࢜࢟ࢩࣥ㢮ศᯒᲷࢆ᪂⠏ࠋ 14. 3 18. 4 ⾨⏕⛉Ꮫ◊✲ᡤ⎔ቃࢭࣥࢱ࣮ࢆ⤌⧊⤫ྜࡋ㸪ᗣ⎔ቃࢭࣥࢱ࣮ࡋ࡚Ⓨ㊊ࠋ ༓⛅ᗇ⯋⏬⟶⌮ᐊཬࡧಖ⾨⏕㒊ࢆ㸪ඵᶫᗇ⯋⎔ቃ㒊ࢆタ⨨ࠋ 21. 4 ඵᶫᗇ⯋ࡢ⎔ቃ㒊ࢆ༓⛅ᗇ⯋⛣㌿ࡋ㸪ᗇ⯋ࢆ⤫ྜࠋಖ⾨⏕㒊ࡢ⌮Ꮫ㒊㛛⎔ቃ㒊ࡢ Ꮫ≀㉁㒊㛛ࢆ⤫ྜࡋࡓ⌮Ꮫ⌜ࢆ⎔ቃ࣭⌮Ꮫ㒊ෆタ⨨ࠋ⤌⧊ࢆ⏬⟶⌮ᐊ㸪ಖ⾨ ⏕㒊ཬࡧ⎔ቃ㺃⌮Ꮫ㒊ࡍࡿࠋ 22. 4 ಖᡤࡢヨ㦂᳨ᰝㄢࢆ⤫ྜࠋಖ⾨⏕㒊ࡢᚤ⏕≀⌜ࢆ⣽⳦⌜࢘ࣝࢫ⌜⦅ࡋ㸪ᗣ ⛉Ꮫ⌜ࢆᗣ⛉Ꮫ࣭⟶⌮⌜ྡ⛠ኚ᭦ࠋ⎔ቃ࣭⌮Ꮫ㒊ࢆ⌮Ꮫ㒊⎔ቃಖ㒊⦅ࠋ ⌮Ꮫ㒊ࡣ㸪⌮Ꮫ⌜ࢆ⦅ࡋࡓ㣗ရ⌮Ꮫ⌜⎔ቃ⌮Ꮫ⌜ࢆタ⨨ࠋ⎔ቃಖ㒊ࡣ ⎔ቃㄪᰝ⌜ࢆྡ⛠ኚ᭦ࡋࡓ⎔ቃಖ⌜ࢆタ⨨ࠋ 24. 4 ⏬⟶⌮ᐊࡢ⥲ົ࣭⏬⌜ࢆ⦅ࡋࠊ⥲ົ⟶⌮⌜⏬ሗ⌜ࢆタ⨨ࠋಖ⾨⏕㒊ࡢᗣ ⛉Ꮫ࣭⟶⌮⌜ࢆᗫṆࠋ⌮Ꮫ㒊ࡢ㣗ရ⌮Ꮫ⌜⎔ቃ⌮Ꮫ⌜ࢆ⤫ྜࡋࠊ⌮Ꮫ⌜ࢆタ⨨ࠋ ᗇ⯋ࡢᴫせ 1㸧ᡤᅾᆅ 2㸧ᩜ ᆅ 3㸧ᘓ ≀ ⛅⏣ᕷ༓⛅ஂಖ⏣⏫ 6 ␒ 6 ྕ 867.75 ੍㸦ᘓ≀ᘓᗋ㠃✚㸧 㕲➽ࢥࣥࢡ࣮ࣜࢺ㐀 5 㝵ᘓ 㸬⤌⧊ ᘏᗋ㠃✚ 4,553.52 m2 ᡤ ⏬⟶⌮ᐊ㛗 㸦ව㸧ಖ⾨⏕㒊㛗 㸦ᮏົ ⥲ົ⟶⌮⌜ 㸦ᖹᡂ 27 ᖺ 4 ᭶ 1 ᪥⌧ᅾ㸧 㛗 ⏬ሗ⌜ ⌮Ꮫ㒊㛗 ⎔ቃಖ㒊㛗 ⌮Ꮫ⌜ ⎔ቃಖ⌜ ⏬⟶⌮ᐊ㛗㸧 ⣽⳦⌜ ࢘ࣝࢫ⌜ ᖿ 1 ௵◊✲ဨ 2 ୖᖍ◊✲ဨ 3 ୖᖍ◊✲ဨ 1 ௵◊✲ဨ 4 ୖᖍ◊✲ဨ 1 ᰝ 2 ᑓ㛛ဨ 1 ௵◊✲ဨ 1 ௵◊✲ဨ 2 ᑓ㛛ဨ 1 ௵◊✲ဨ 2 ᢏ⬟௵ 1 ◊✲ဨ 2 ◊✲ဨ 2 ◊✲ဨ 2 ᑓ㛛ဨ 2 ⮫⫋ဨ 1 㠀ᖖ⫋ဨ 1 ᢏᖌ 1 ◊✲ဨ 5 ⮫⫋ဨ 1 㠀ᖖ⫋ဨ 1 ⮫⫋ဨ 1 ィ 5 ィ 3 ィ 8 ィ 5 ィ ⥲⫋ဨᩘ 45 ྡ㸦ṇ⫋ဨ 36 ྡ, ᑓ㛛ဨ 4 ྡ㸪㠀ᖖ⫋ဨ 2 ྡ, ⮫⫋ဨ 3 ྡ㸧 - 1 - 9 ィ 11 秋田県健康環境センター年報 第10号 2014 㸬⫋ဨྡ⡙ 㸦ᖹᡂ 27 ᖺ 4 ᭶ 1 ᪥⌧ᅾ㸧 ⏬⟶⌮ᐊ ⥲ົ⟶⌮⌜ ⫋ ྡ Ặ ᡤ 㛗 㣤 ሯ ᨻ ᐊ 㛗 㰺 ⸨ ᚿಖᏊ ᖿ 㸦 ව 㸧 ⌜ 㛗 ᕤ ⸨ ṇ ᰝ ᮅ ⏤ె⌮ ᰝ ᖹ 㔝 㐍 ௵ ᅜ Ᏻ ຊ ௵◊✲ဨ㸦ව㸧⌜㛗 Ლ ㇂ ᫂ ᘯ ဨ ⸨ ㇂ 㝧 Ꮚ ဨ బ ⸨ ✨ ᢏ ⏬ሗ⌜ ⬟ ௵ ᑓ ಖ⾨⏕㒊 ⣽ ⳦ ⌜ ࢘ࣝࢫ⌜ ⌮ Ꮫ 㒊 ◊ ✲ 㛛 ⎔ቃಖ㒊 ⎔ቃಖ⌜ ⠊ ༤ 㸦ව㸧 㒊 㛗 㸦ᮏົ⏬⟶⌮ᐊ㛗㸧 ୖᖍ◊✲ဨ㸦ව㸧⌜㛗 㰺 ⸨ ᚿಖᏊ ᅭ Ꮚ 㝯 ಙ ୖ ᖍ ◊ ✲ ဨ ⏣ ୰ ㈗ Ꮚ ୖ ᖍ ◊ ✲ ဨ ⇃ ㇂ ඃ Ꮚ ௵ ◊ ✲ ဨ 㧘 ᶫ ᚿ ಖ ◊ ✲ ဨ 㔝 ㈗ அ ◊ ✲ ဨ ᶔ ᑿ ᣅ Ꮚ ୖᖍ◊✲ဨ㸦ව㸧⌜㛗 ᩪ ⸨ ༤ அ ௵ ◊ ✲ ဨ ⛅ 㔝 ⳹Ꮚ ௵ ◊ ✲ ဨ బ ⸨ ᐶ Ꮚ ◊ ✲ ဨ ᰘ ⏣ ࡕࡦࢁ ◊ ✲ ဨ బ ⸨ ⏤⾰Ꮚ 㛗 㕥 ᮌ ᛅ அ ௵◊✲ဨ㸦ව㸧⌜㛗 ᑠ ᯘ ㈗ ྖ ௵ ◊ ✲ ဨ ୰ ᮧ ῟ Ꮚ ௵ ◊ ✲ ဨ ᯇ ῲ ளᕼᏊ ௵ ◊ ✲ ဨ ᮧ ᒣ ຊ ๎ 㒊 ⌮Ꮫ⌜ ྡ ᑓ 㛛 ဨ బ ⸨ ᬕ ⨾ ◊ ✲ ဨ ᑠ ᕝ ༓ ◊ ✲ ဨ 㔝 ⚘ ᮁ ᢏ ᖌ బ ⸨ ⌮ ዉ 㒊 㛗 ῲ ᚿ ఙ ୖᖍ◊✲ဨ㸦ව㸧⌜㛗 㯮 ἑ ᪂ ௵ ◊ ✲ ဨ Ύ Ỉ ௵ ◊ ✲ ဨ ᡂ ⏣ ಟ ྖ ᑓ 㛛 ဨ 㧗 ᔱ ᑓ 㛛 ဨ ஂ ◊ ✲ ဨ ⋢ ⏣ ᑗ ᩥ ◊ ✲ ဨ 㕥 ᮌ ⣧ ᜨ ◊ ✲ ဨ బ ⸨ ◊ ✲ ဨ ⏕ 㨶 ◊ ✲ ဨ బ ⸨ Ύ 㝯 - 2 - ྖ 秋田県健康環境センター年報 第10号 2014 ᴗົෆᐜ 㸦ᖹᡂ 27 ᖺ 4 ᭶ 1 ᪥⌧ᅾ㸧 ࣭ே㸪᭹ົ ⥲ ົ ⟶ ⌮ ⌜ ࣭ண⟬㸪Ỵ⟬ ࣭ᗇ⯋⟶⌮㸪ᗢົ୍⯡ ⏬ ⟶ ࣭◊✲ࡢ⏬㸪ホ౯㸪㐍⾜⟶⌮ ⌮ ࣭ࢭࣥࢱ࣮୰㛗ᮇィ⏬ࡢ㐍⾜⟶⌮ ᐊ ⏬ ሗ ⌜ ࣭ᗈሗ㸪◊ಟ ᳨࣭ᰝᴗົ⟶⌮ ࣭⢭ᗘ⟶⌮ ࣭ឤᰁⓎ⏕ືྥㄪᰝࡶ࡞࠺ཎయ᳨ᰝᴗົ ࣭⣽⳦ឤᰁ㣗୰ẘࡢヨ㦂᳨ᰝཬࡧㄪᰝ◊✲ ࣭⤖᰾Ⓩ㘓⪅ሗㄪᰝ ⣽ ⳦ ⌜ ࣭ᗣ࡙ࡃࡾ㛵ࡍࡿㄪᰝ◊✲ ಖ ࣭⸆⪏ᛶ⳦㛵ࡍࡿㄪᰝ◊✲ ࣭་⸆ရ➼㛵ࡍࡿ᳨ᰝ ⾨ ࣭ཤ㣗ရཬࡧ⎔ቃ᳨య➼㛵ࡍࡿ⣽⳦᳨ᰝ ⏕ 㒊 ࣭ឤᰁⓎ⏕ືྥㄪᰝࡶ࡞࠺ཎయ᳨ᰝᴗົ ࢘ ࣝ ࢫ ⌜ ࣭࢘ࣝࢫឤᰁ㣗୰ẘࡢヨ㦂᳨ᰝཬࡧㄪᰝ◊✲ ࣭ࡘࡘࡀࡢᢠయ᳨ᰝཬࡧㄪᰝ◊✲ ࣭ឤᰁሗࢭࣥࢱ࣮ᴗົ ࣭㣗ရࡢᏳᛶಀࡿヨ㦂᳨ᰝཬࡧㄪᰝ◊✲ ࣭㣗ရᨺᑕ⬟ࡢ ᐃ ⌮ ࣭᭷ᐖᐙᗞ⏝ရヨ㈙᳨ᰝ ⌮ Ꮫ ⌜ Ꮫ ࣭ཤ㣗ရࡢ⌮Ꮫⓗ᳨ᰝ ࣭ᕤሙ㸪ᴗሙỈ୰ࡢᏛ≀㉁ࡢ᳨ᰝ 㒊 ࣭ᗫᲠ≀㛵ಀ᳨ᰝ ࣭⎔ቃ୰ࡢᏛ≀㉁㛵ࡍࡿㄪᰝ◊✲ ࣭බඹ⏝ỈᇦỈ㉁ㄪᰝ ࣭ᕤሙ㸪ᴗሙỈᇶ‽᳨ᰝ ࣭ᕤሙ㸪ᴗሙࡤ࠸↮ฟᇶ‽᳨ᰝ ⎔ ࣭ᗫᲠ≀㛵ಀ᳨ᰝ ቃ ࣭⏕ά⾨⏕㛵ಀ᳨ᰝ ಖ ⎔ ቃ ಖ ⌜ ࣭⎔ቃᨺᑕ⬟ࡢ ᐃ ࣭Ẽởᰁᖖ┘ど 㒊 ࣭⯟✵ᶵ㦁㡢ㄪᰝ ࣭㓟ᛶ㞵ㄪᰝ ࣭ࢫ࣋ࢫࢺ⎔ቃㄪᰝ ࣭⎔ቃಖ㛵ࡍࡿㄪᰝ◊✲ - 3 - 秋田県健康環境センター年報 第10号 2014 せᶵჾ 㸦ᖹᡂ 27 ᖺ 4 ᭶ 1 ᪥⌧ᅾ㸧 ᶵ ჾ ྡ ࢞ࢫࢡ࣐ࣟࢺࢢࣛࣇ つ ᱁ ࢪࣞࣥࢺ࣭ࢸࢡࣀࣟࢪ࣮ 7890A㸦FID㸧 ࠌ ࢪࣞࣥࢺ࣭ࢸࢡࣀࣟࢪ࣮ 7890A㸦FPD㸧 ࠌ ࢪࣞࣥࢺ࣭ࢸࢡࣀࣟࢪ࣮ 6890N㸦ECD㸧 ࢞ࢫࢡ࣐ࣟࢺࢢࣛࣇ㉁㔞ศᯒィ ᓥὠ GCMS-QP2010 Ultra ࠌ ᓥὠ GCMS-QP2010 Plus ࠌ ᓥὠ QP5050A ࠌ ᓥὠ QP5000 ࠌ ࢪࣞࣥࢺ࣭ࢸࢡࣀࣟࢪ࣮ 6890N/5973N ࢞ࢫࢡ࣐ࣟࢺࢢࣛࣇࢱࣥࢹ࣒ᆺ㉁㔞ศᯒィ ࢧ࣮ࣔࣇࢵࢩ࣮ࣕ TSQ QuantumGC 㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ ᪥❧〇సᡤ D-7000 ࠌ ᪥❧〇సᡤ L-7000 ࠌ ᪥❧〇సᡤ L-7000㸦GPC㸧 ࠌ ᪥ᮏ࢛࣮࢘ࢱ࣮ࢬ 2695 ࠌ ࢪࣞࣥࢺ࣭ࢸࢡࣀࣟࢪ࣮ 1200 㸦DAD࣭FLD㸧 ᾮయࢡ࣐ࣟࢺࢢࣛࣇࢱࣥࢹ࣒㉁㔞ศᯒィ AB ࢧ࢚ࢵࢡࢫ API4000 ࢜ࣥࢡ࣐ࣟࢺࢢࣛࣇ ᪥ᮏࢲ࢜ࢿࢡࢫ ICS-1100 ࠌ ᪥ᮏࢲ࢜ࢿࢡࢫ DX-320 ཎᏊ྾ගศගගᗘィ ࣂ࣭ࣜࣥࢸࢡࣀࣟࢪ࣮ࢬ AA-280FS ICP Ⓨගศගศᯒ⨨ ࢧ࣮ࣔࣇࢵࢩ࣮ࣕ iCAP 6300 Duo ࣀ࣐ࣝࣝ࣊࢟ࢧࣥ⮬ືᢳฟ⨨ ࣛ࣎ࢸࢵࢡ HX-1000-8 㧗㏿⁐፹ᢳฟ⨨ DIONEX ♫ ASE-200 㸪ASE-300 ࣮࢜ࢺࢼࣛࢨ࣮ ࣅ࣮࢚ࣝࢸࢵࢡ QuAAtro 2-HR ศ㞳⏝㉸㐲ᚰᶵ ᪥❧ᕤᶵ CP70MX 㟁Ꮚ㢧ᚤ㙾 ᪥ᮏ㟁Ꮚ JEM-1010 Ge ༙ᑟయ᳨ฟჾἼ㧗ศᯒ⨨ ࢭࢥ࣮ EG&G GEM20P㸪MCA7 ࢭࢥ࣮ EG&G GEM25-70㸪MCA7600 ࠌ PCR ࣉࣟࢲࢡࢺ᳨ฟᐃ㔞ࢩࢫࢸ࣒ ࣉࣛࢻࣂ࢜ࢩࢫࢸ࣒ࢬ ABI PRISM 7000 ⮬ື᰾㓟⢭〇⨨ ᪥ᮏࣟࢩ࣭ࣗࢲࢢࣀࢫࢸࢡࢫ MagNA Pure LC2.0 ࣔࢽࢱࣜࣥࢢ࣏ࢫࢺ ࣟ࢝ MAR-22 ✵㛫ᨺᑕ⥺㔞ࣔࢽࢱࣜࣥࢢࢩࢫࢸ࣒ ᮾⰪ SD22-T పࣂࢵࢡࢢࣛࣥࢻᨺᑕ⬟⮬ື ᐃ⨨ ࣟ࢝ LBC-4201B Ẽởᰁᖖ┘どࢸ࣓࣮ࣞࢱࢩࢫࢸ࣒ NEC ⯟✵ᶵ㦁㡢⮬ື ᐃ⨨ ࣜ࢜ࣥ NA-37 - 4 - ! ! ! ! ࣂ !؞Ⴡ૰౹ ! 秋田県健康環境センター年報 第10号 2014 㸬ヨ㦂᳨ᰝᐇ⦼ ಖ⾨⏕㒊⾜ᨻ౫㢗᳨ᰝ 㸦௳ᩘ㸧 㡯ࠉ┠ ᖺࠉᗘ ᖹᡂ24 ᖹᡂ25 ᖹᡂ26 ឤᰁⓎ⏕ືྥㄪᰝཎయ ࢘ࣝࢫศ㞳᳨ᰝ ᳨ูᰝᩘ ⣽⳦᳨ᰝ ࣥࣇ࢚ࣝࣥࢨឤᰁ※ㄪᰝ ឤᰁὶ⾜ண ㄪᰝ ᪥ᮏ⬻⅖ឤᰁ※ㄪᰝ ⫵⅖⌫⳦ឤᰁ※ㄪᰝ 861 1,052 1,438 100 70 䠉 1,070 100 70 0 䠉 2,699 4 4 155 155 133 133 䠉 755 3,282 1 0 134 134 124 124 䠉 496 1,798 1 1 122 122 147 150 㯞⑈PCR 㢼⑈PCR 2 23 3 0 0 0 0 0 0 㯞⑈࣭㢼⑈࣭Ⓨ⑈ᛶ࢘ࣝࢫ᳨ᰝ*2 ᪂ᆺࣥࣇ࢚ࣝࣥࢨ 䠉 29 276 6 218 䠉 0 䠉 0 䠉 289 0 SFTS࢘ࣝࢫ᳨ᰝ*4 ࢹࣥࢢ࢘ࣝࢫ᳨ᰝ*5 3㢮ឤᰁಀࢃࡿཎᚤ⏕≀᳨ᰝ 䠉 䠉 499 䠉 䠉 490 0 7 484 ࢝ࣥࣆࣟࣂࢡࢱ࣮㸦⸆ឤཷᛶヨ㦂㸧 ᆅ◊ ࣞࣇࣞࣥࢫࢭࣥࢱ࣮ᴗົ ࢪࣇࢸ࣭ࣜⓒ᪥တ࣭࣎ࢶࣜࢾࢫ ⤖᰾⳦RFLP᳨ᰝ㸪VNTR᳨ᰝ 61 223 96 87 37 159 67 105 53 52 81 104 䠉 407 ࣀࣟ࢘ࣝࢫ᳨ᰝ 㣗୰ẘ➼᳨ᰝ ⫶⭠⅖࢘ࣝࢫ᳨ᰝ㸦ࣀࣟ࢘ࣝࢫ➼㸧 ⣽⳦᳨ᰝ HIVᢠయ᳨ᰝ HIVᢠయ☜ㄆ᳨ᰝ ᛶჾࢡ࣑ࣛࢪᢠయ᳨ᰝ ᱵẘᢠయ᳨ᰝ Bᆺ⫢⅖ᢠཎ᳨ᰝ Cᆺ⫢⅖ᢠయ᳨ᰝ ⣽⳦࣭࢘ࣝࢫ➼ࡢヨ㦂᳨ᰝ 850 1,078 100 70 㯞⑈ᢠయ౯᳨ᰝ ᪂ᆺࣥࣇ࢚ࣝࣥࢨ➼྾ჾ࢘ࣝࢫ᳨ᰝ ᪂ᆺࣥࣇ࢚ࣝࣥࢨࢱ࣑ࣇࣝ⪏ᛶ᳨ᰝ *3 ࡘࡘࡀ⾑Ύ᳨ᰝ *1 826 467 404 1,013 190 3 32 32 953 290 3 32 16 902 293 3 25 16 බ⾗ᾎሙ➼ࣞࢪ࢜ࢿࣛᒓ⳦᳨ᰝ බඹ⏝ỈᇦỈ㉁⎔ቃㄪᰝ 56 243 121 47 103 47 Ỉ㉁ở⃮ᑐ⟇ ඵ㑻†Ỉ㉁ಖㄪᰝ ᕤሙ࣭ᴗሙỈᇶ‽᳨ᰝ 84 244 78 218 79 230 ᗫᲠ≀ᑐ⟇ ࣐ࢫ࣭ࢫࢡ࣮ࣜࢽࣥࢢ ⏘ᴗᗫᲠ≀➼ᇶ‽᳨ᰝ 20 7,362 5 20 2,617 3 20 828 䠉 627 775 0 0 0 0 389 120 19,660 0 0 13,028 䠉 䠉 䠉 䠉 䠉 䠉 8,542 ࡑࡢᚤ⏕≀Ꮫⓗ᳨ᰝ 㣗ရ⾨⏕ಀࡿ᳨ᰝ ⏕ά⾨⏕ಀࡿ᳨ᰝ ་⸆ရ➼┘どᣦᑟᴗົಀࡿ᳨ᰝ ᅜẸᗣ࣭ᰤ㣴ㄪᰝ 㣗ရཤ᳨ᰝ 㣗୰ẘ⳦ởᰁᐇែㄪᰝ ⢭ᗘ⟶⌮ බ⾗ᾎሙỈ㸪㐟Ὃࣉ࣮ࣝỈࡢ⭠⳦᳨ᰝ ㈚ࡋ࠾ࡋࡰࡾ᳨ᰝ ඛኳᛶ௦ㅰ␗ᖖ㸪ෆศἪᝈ*6 ་⸆ရ㸪་⸆㒊እရ㸪་⒪ᶵჾ㸦⣽⳦㸧 ᰤ㣴ㄪᰝ࣭ゎᯒ 㣗⏕ά≧ἣㄪᰝ࣭ゎᯒ ㌟య≧ἣㄪᰝ࣭ゎᯒ ⏕ά⩦័ㄪᰝ࣭ゎᯒ ⾑ᾮ᳨ᰝ ሷศ⃰ᗘ ᐃ ྜࠉࠉࠉࠉィ 䠉 3 *1 ⫶⭠⅖࢘ࣝࢫ᳨ᰝ㸦ࣀࣟ࢘ࣝࢫ➼㸧ࡘ࠸࡚ࡣ㸪ᖹᡂ25ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *2 㯞⑈࣭㢼⑈࣭Ⓨ⑈ᛶ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ,ᖹᡂ25ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *3 ᪂ᆺࣥࣇ࢚ࣝࣥࢨ➼྾ჾ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ,ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *4 SFTS࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ㸪ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋᖹᡂ25ᖺᗘ௨๓ࡢSFTS࢘ࣝࢫ᳨ᰝ௳ᩘࡣࡑࡢᚤ⏕≀ ᳨ᰝ䛻 ィୖ 䛧䛶䛔䜛䚹 *5 ࢹࣥࢢ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ㸪ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋᖹᡂ25ᖺᗘ௨๓ࡢࢹࣥࢢ࢘ࣝࢫ᳨ᰝ௳ᩘࡣࡑࡢᚤ⏕≀ ᳨ᰝィୖࡋ࡚࠸ࡿࠋ *6 ࣐ࢫ࣭ࢫࢡ࣮ࣜࢽࣥࢢࡘ࠸࡚ࡣ㸪ᖹᡂ25ᖺ8᭶ࡽእ㒊ጤク࡞ࡗࡓࠋ - 5 - 秋田県健康環境センター年報 第10号 2014 ಖ⾨⏕㒊୍⯡౫㢗᳨ᰝ 㸦௳ᩘ㸧 ࠉࠉࠉࠉࠉࠉࠉᖺࠉᗘ 㡯ࠉ┠ ᖹᡂ24 410 3 0 0 1 㸫 0 0 0 0 0 2 416 ⛅⏣ᕷಖᡤ౫㢗ศ㸦ᥖ㸧 ࢘ࣝࢫศ㞳➼᳨ᰝ 㣗୰ẘ㛵ಀ➼⫶⭠⅖࢘ࣝࢫ᳨ᰝ㸦ࣀࣟ࢘ࣝࢫ➼㸧 㯞⑈࣭㢼⑈࣭Ⓨ⑈ᛶ࢘ࣝࢫ᳨ᰝ*1 ᪂ᆺࣥࣇ࢚ࣝࣥࢨ ᪂ᆺࣥࣇ࢚ࣝࣥࢨ➼྾ჾ࢘ࣝࢫ᳨ᰝ*2 ⣽⳦࣭࢘ࣝࢫ➼ࡢヨ㦂᳨ᰝ ᪂ᆺࣥࣇ࢚ࣝࣥࢨࢱ࣑ࣇࣝ⪏ᛶ᳨ᰝ SFTS࢘ࣝࢫ᳨ᰝ*3 ࢹࣥࢢ࢘ࣝࢫ᳨ᰝ*4 ⣽⳦ᇵ㣴ྠᐃ᳨ᰝ ⣽⳦㑇ఏᏊゎᯒ᳨ᰝ HIVᢠయ☜ㄆ᳨ᰝ ྜࠉࠉࠉࠉィ ឤᰁⓎ⏕ືྥㄪᰝ㛵ࢃࡿ᳨ᰝ ᖹᡂ25 ᖹᡂ26 450 0 24 0 3 㸫 0 0 0 13 0 0 490 365 0 1 42 㸫 3 0 0 3 9 2 0 425 *1 㯞⑈࣭㢼⑈࣭Ⓨ⑈ᛶ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ,ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *2 ᪂ᆺࣥࣇ࢚ࣝࣥࢨ➼྾ჾ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ,ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *3 SFTS࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ㸪ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ *4 ࢹࣥࢢ࢘ࣝࢫ᳨ᰝࡘ࠸࡚ࡣ㸪ᖹᡂ26ᖺᗘࡽ᪂ࡓ㡯┠ࢆ㉳ࡇࡋࡓࠋ ሗᥦ౪ᴗົ 㸦௳ᩘ㸧 ࠉࠉࠉࠉࠉࠉࠉᖺࠉᗘ 㡯ࠉ┠ 㐌ሗ ᝈ⪅ሗ ᭶ሗ ᇶᖿ࣭ᆅ᪉ឤᰁሗࢭࣥࢱ࣮ 㸦ឤᰁⓎ⏕ືྥㄪᰝ౫㢗ᴗົ㸧 ཎయሗ 㞟 ሗ࿌ 㑏ඖ ゎᯒ ᥦ౪ 㞟 ሗ࿌ 㑏ඖ ゎᯒ ᥦ౪ 㞟 ࢘ࣝࢫ ሗ࿌ ⣽⳦ 㑏ඖ ゎᯒ ゎᯒホ౯ጤဨ㈨ᩱᥦ౪ ᭶ሗ ⤖᰾Ⓩ㘓⪅ሗㄪᰝ౫㢗ᴗົ ᝈ⪅ሗ ᖺሗ*1 ⰼ⢊ண㜵ᑐ⟇౫㢗ᴗົ *2 ࢫࢠⰼ⢊ணሗసᡂᥦ౪ ࢫࢠⰼ⢊ ᐃᩘ ࢫࢠ㞝ⰼⱆㄪᰝᩘ ⰼ⢊ᝈ⪅ㄪᰝ⚊ᩘ ྜࠉࠉࠉࠉィ *1 ᪂つ⤖᰾Ⓩ㘓ᝈ⪅ᩘ㸸98ேࠊᖺᮎ⤖᰾Ⓩ㘓⪅ᩘ㸸223ே㸦ᖹᡂ26ᖺ1᭶㹼12᭶㸧 *2 ⰼ⢊ண㜵ᑐ⟇౫㢗ᴗົࡣᖹᡂ24ᖺᗘ࡛⤊ࡋࡓࠋ - 6 - 㞟 ሗ࿌ 㑏ඖ ゎᯒ ᥦ౪ 㞟 ሗ࿌ 㑏ඖ ゎᯒ ᥦ౪ ᖹᡂ24 468 52 52 52 468 108 12 12 12 108 52 503 301 24 24 6 108 12 12 12 108 9 1 1 1 9 52 221 15 80 2,895 ᖹᡂ25 468 52 52 52 468 108 12 12 12 108 52 526 310 24 24 6 108 12 12 12 108 9 1 1 1 9 㸫 㸫 㸫 㸫 2,559 ᖹᡂ26 468 52 52 52 468 108 12 12 12 108 52 563 286 24 24 6 108 12 12 12 108 9 1 1 1 9 㸫 㸫 㸫 㸫 2,572 - 7 - 秋田県健康環境センター年報 第10号 2014 ⎔ቃಖ㒊⾜ᨻ౫㢗᳨ᰝ 㸦௳ᩘ㸧 㡯ࠉ┠ ᖺࠉᗘ ୍⯡⎔ቃẼ ᐃᒁ Ẽởᰁᖖ┘ど*1 Ẽởᰁᑐ⟇ ⮬ື㌴ฟ࢞ࢫ ᐃᒁ ᕤሙᒁ ࡤ࠸↮ฟᇶ‽᳨ᰝ ⚟ᓥཎᏊຊⓎ㟁ᡤᨾక࠺ ⥭ᛴ⎔ቃᨺᑕ⬟ㄪᰝ ࡑࡢ⥭ᛴ⎔ቃᨺᑕ⬟ㄪᰝ ᖹᡂ26 57 57 50 (477,729) (466,452) (422,880) 15 15 15 (129,750) (128,634) (128,966) 74 74 74 (584,893) (547,254) (530,999) 0 29 28 1,420 1,390 1,400 㓟ᛶ㞵ᐇែㄪᰝ ࢫ࣋ࢫࢺᑐ⟇ ▼⥥㣕ᩓㄪᰝ 36 44 60 ⺬ཱྀỈ 36 36 0 ⎔ቃヨᩱ 1,251 919 514 ␆⏘ヨᩱ 38 14 0 㝆ୗ≀ 30 0 0 ᾋ㐟ࡌࢇ 30 0 0 4,107 3,814 4,257 0 0 㻜 ⥭ᛴㄪᰝ 519 674 192 ᕤሙỈ 2,189 1,998 1,882 ỈᾎሙỈ 336 0 0 ᗏ㉁ㄪᰝ 36 36 36 ⥭ᛴㄪᰝ 158 0 0 336 317 317 240 240 240 0 0 0 ởᰁᅵተฎ⌮ᴗᡤ᳨ᰝ 36 46 69 㐟Ὃ⏝ࣉ࣮ࣝỈ㉁᳨ᰝ 16 24 24 බ⾗ᾎሙỈ㉁᳨ᰝ 64 32 32 㣗⫗⾨⏕᳨ᰝᡤ⮬᳨ᰝ 20 20 20 ᰾✀ศᯒ ᰾✀ศᯒ ⎔ቃㄪᰝ ᕤሙ࣭ᴗሙỈᇶ‽᳨ᰝ Ỉ㉁ở⃮ᑐ⟇ ඵ㑻†Ỉ㉁ಖᑐ⟇ㄪᰝ ᆅୗỈㄪᰝ ⋢ᕝ㓟ᛶỈᙳ㡪ㄪᰝ ༑⏣†Ỉ㉁ಖᑐ⟇ㄪᰝ ༑⏣†Ỉ㉁⏕ែ⣔ㄪᰝ ༑⏣†ὶධἙᕝㄪᰝ ⏕ά⾨⏕ಀࡿ᳨ᰝ ᖹᡂ25 㓟ᛶ㞵ㄪᰝ බඹ⏝ỈᇦỈ㉁ㄪᰝ ᅵተởᰁᑐ⟇ ᖹᡂ24 㦁㡢ᑐ⟇ ⯟✵ᶵ㦁㡢ㄪᰝ 758 714 722 Ꮫ≀㉁ᑐ⟇ Ꮫ≀㉁⎔ቃㄪᰝ 137 195 274 ⏘ᴗᗫᲠ≀➼ᇶ‽᳨ᰝ 543 529 505 1,322 1,221 992 0 0 0 13,658 12,292 11,564 ᗫᲠ≀ᑐ⟇ ⬟௦⏘ᴗᗫᲠ≀ฎ⌮ࢭࣥࢱ࣮㛵㐃ㄪᰝ ⥭ᛴㄪᰝ ྜࠉࠉࠉࠉィࠉ㸦Ẽởᰁᖖ┘どࢆ㝖ࡃ㸧 *1 Ẽởᰁᖖ┘どࡣ㸪 ᐃᑐ㇟㡯┠ᩘ㸦ᐇ ࢹ࣮ࢱᩘ㸧ࢆ⾲ࡍࠋ - 8 - 秋田県健康環境センター年報 第10号 2014 㸬◊ಟ࣭Ꮫ➼ ◊ಟ➼ཧຍ ᖺ᭶᪥ ◊ ಟ ྡ ཧຍ⪅ 㛤ദᆅ 26.05.16 ᖹᡂ 26 ᖺᗘཎయ➼ࡢໟ࣭㐠ᦙㅮ⩦ ᶔᑿᣅᏊ ᮾி㒔 26.06.12㹼27 ᶵჾศᯒ◊ಟ బ⸨ ᇸ⋢┴ బ⸨Ύ㝯 26.06.30 ࢡ࣐ࣟࢺᇶ♏ࢭ࣑ࢼ࣮2014 㔝⚘ᮁ ⛅⏣ᕷ ୰ᮧ῟Ꮚ 26.09.04 26.09.18㹼19 26.09.26 26.10.07㹼10 26.10.10 26.10.23㹼24 ᮾ㣗୰ẘ◊✲➨ 27 ᅇయ㆟࣭◊ಟ ಖ⾨⏕㒊ဨ ᖹᡂ 26 ᖺᗘᆅ᪉⾨⏕◊✲ᡤᅜ༠㆟ ᰘ⏣ࡕࡦࢁ ᾏ㐨࣭ᮾ࣭᪂₲ᨭ㒊ᚤ⏕≀◊✲㒊 ᶔᑿᣅᏊ ➨ 1 ᅇᨺᑕ⬟ởᰁᑐ⟇ࣉࣟࢪ࢙ࢡࢺጤဨ ⌋⏣ᑦಇ ⛅⏣ᕷ ⏣୰㈗Ꮚ ᮾி㒔 ᒾ㇂㔠ோ ᮾி㒔 ᮧᒣຊ๎ ᾏ㐨 㸦බ㈈㸧⤖᰾ண㜵⤖᰾◊ಟᡤ◊ಟ ᖹᡂ 26 ᖺᗘ⤖᰾⾜ᨻᢸᙜ⪅ࢥ࣮ࢫ ᖹᡂ 26 ᖺᗘ㣗ရ⾨⏕᳨ᰝタಙ㢗ᛶ☜ಖ㒊㛛㈐௵⪅◊ಟ ᖹᡂ 26 ᖺᗘᆅ᪉⾨⏕◊✲ᡤᅜ༠㆟ ᾏ㐨࣭ᮾ࣭ ᪂₲ᨭ㒊බ⾗⾨⏕ሗ◊✲㒊⥲࣭◊ಟ ⛅⏣ᕷ ᒣᙧ┴ ᖹᡂ 26 ᖺᗘᆅ◊ᾏ㐨࣭ᮾ࣭᪂₲ᨭ㒊 㔝⚘ᮁ ⾨⏕Ꮫ◊✲㒊⥲ ⌋⏣ᑦಇ 26.10.28㹼31 ᖹᡂ 26 ᖺᗘ⎔ቃᨺᑕ⬟ศᯒ◊ಟ బ⸨Ύ㝯 ༓ⴥ┴ 26.11.06㹼07 ᖹᡂ 26 ᖺᗘ⛅⏣┴㣗ရ⾨⏕┘どဨ➼◊ಟ 㔝⚘ᮁ ₲ୖᕷ 26.11.11㹼19 ⎔ቃᨺᑕ⬟ศᯒ◊ಟ ᑠᕝ༓ ༓ⴥ┴ ⌋⏣ᑦಇ ᒾᡭ┴ 26.10.23㹼24 26.11.28 ᒾᡭ┴⎔ቃಖ◊✲ࢭࣥࢱ࣮ ࠕI-RIEP ࢭ࣑ࢼ࣮࣭༴㝤ࢻࣛࢵࢢ◊ಟࠖ ᯇῲளᕼᏊ 㟷᳃┴ 27.01.16 ᖹᡂ 26 ᖺᗘᣦᐃ⸆≀ศᯒ◊ಟ㆟ࡘ࠸࡚ 27.01.26㹼28 ᖹᡂ 26 ᖺᗘ▼⥥┦ᕪ㢧ᚤ㙾ἲ◊ಟ బ⸨Ύ㝯 ᇸ⋢┴ 27.01.28 ༴㝤ࢻࣛࢵࢢ➼㐪ἲ⸆≀ศᯒࢯ࣮ࣜࣗࢩࣙࣥࢭ࣑ࢼ࣮ ⌋⏣ᑦಇ ᒾᡭ┴ 27.01.29㹼30 ᖹᡂ 26 ᖺᗘᏛ≀㉁⎔ቃᐇែㄪᰝ⎔ቃ⛉Ꮫࢭ࣑ࢼ࣮ 27.02.13㹼14 ᖹᡂ 26 ᖺᗘ⾨⏕⌮Ꮫศ㔝◊ಟཬࡧ㣗ရᏛศ㔝◊✲ ⪅⫱ᡂ◊ಟࢭ࣑ࢼ࣮ ⌋⏣ᑦಇ బ⸨ ⋢⏣ᑗᩥ 㔝⚘ᮁ ᮾி㒔 ᮾி㒔 ᮾி㒔 㔝㈗அ 27.02.17㹼18 ᖹᡂ 26 ᖺᗘᕼᑡឤᰁデ᩿ᢏ⾡◊ಟ బ⸨ᐶᏊ ᮾி㒔 ⇃㇂ඃᏊ 27.03.11 ᖹᡂ 26 ᖺᗘᨺᑕ⥺┘ど⤖ᯝ㞟ㄪᰝ᳨ウ 㔝⚘ᮁ ᮾி㒔 27.03.20 ᗫᲠ≀㛵㐃ヨᩱࡢศᯒ࣐ࢽࣗࣝฟ∧グᛕࢭ࣑ࢼ࣮ బ⸨ ᮾி㒔 - 9 - 秋田県健康環境センター年報 第10号 2014 Ꮫ➼ฟᖍ ᖺ᭶᪥ ฟᖍ⪅ Ꮫ ྡ 㸦ۑⓎ⾲⪅㸧 㛤ദᆅ 26.05.14㹼16 ➨ 23 ᅇ⎔ቃᏛウㄽ Ⳣۑཎ ๛ ி㒔ᗓ 26.05.15㹼16 ➨ 107 ᅇ᪥ᮏ㣗ရ⾨⏕ᏛᏛ⾡ㅮ₇ 㰺⸨ᚿಖᏊ ᮾி㒔 26.06.14㹼15 ➨ 55 ᅇ᪥ᮏ⮫ᗋ࢘ࣝࢫᏛ ᰘ⏣ࡕࡦࢁ ᾏ㐨 26.06.18㹼20 ➨ 88 ᅇ᪥ᮏឤᰁᏛᏛ⾡ㅮ₇ ۑ㔝㈗அ ⚟ᒸ┴ 㔝㈗அ 26.06.26㹼27 ⾨⏕ᚤ⏕≀ᢏ⾡༠㆟➨ 35 ᅇ◊✲ ඵᰗ ₶ ۑబ⸨ᐶᏊ ᮾி㒔 ᩪ⸨༤அ 26.08.15㹼17 ➨ 2 ᅇࢪ㣗ရᏳྜྠ㆟ The 2nd. AFSA Conference on Food Safety and Security ۑᩪ⸨༤அ ࣋ࢺࢼ࣒ 26.08.23 ➨ 6 ᅇ J ឤᰁไᚚࢿࢵࢺ࣮࣡ࢡࣇ࢛࣮࣒ࣛ ᩪ⸨༤அ ᐑᇛ┴ 26.09.17㹼19 ᖹᡂ 26 ᖺᗘẼ⎔ቃᏛ బ⸨ ឡ┴ 㧘ᶫᚿಖ 26.09.18㹼19 ➨ 35 ᅇ᪥ᮏ㣗ရᚤ⏕≀ᏛᏛ⾡⥲ ⛅ۑ㔝⳹Ꮚ ۑᩪ⸨༤அ 㜰ᗓ 㰺⸨ᚿಖᏊ 26.10.08 ᪥ᮏ⋇་බ⾗⾨⏕Ꮫ㸦ᮾᆅ༊㸧 㰺⸨ᚿಖᏊ ᐑᇛ┴ 26.10.16㹼17 ➨ 37 ᅇ㎰⸆ṧ␃ศᯒ◊✲ 㔝⚘ᮁ ᐑᇛ┴ ۑబ⸨ᐶᏊ ᒾᡭ┴ 26.10.18 ➨ 60 ᅇ᪥ᮏᐤ⏕Ꮫ࣭᪥ᮏ⾨⏕ື≀Ꮫ ᪥ᮏᨭ㒊ྜྠ ۑᑠᕝ༓ 26.11.08 ➨ 38 ᅇ⛅⏣┴་Ꮫ᳨ᰝᏛ 26.11.09 ➨ 26 ᅇ࢘ࣝࢫᛶୗ⑩◊✲Ꮫ⾡㞟 ᩪ⸨༤அ ᮾி㒔 26.11.10㹼12 ➨ 62 ᅇ᪥ᮏ࢘ࣝࢫᏛᏛ⾡㞟 ۑᩪ⸨༤அ ⚄ዉᕝ┴ ۑᶔᑿᣅᏊ ⛅⏣ᕷ ۑబ⸨ᐶᏊ 26.11.19 ➨ 12 ᅇ⛅⏣┴බ⾗⾨⏕ᏛᏛ⾡ ⏣ۑ୰㈗Ꮚ ⛅⏣ᕷ 㰺⸨ᚿಖᏊ 26.11.20㹼21 ➨ 51 ᅇᅜ⾨⏕Ꮫᢏ⾡༠㆟ᖺ ⏣⌋ۑᑦಇ ศ┴ 26.11.22 ➨ 368 ᅇ᪥ᮏ⓶⛉Ꮫ⛅⏣ᆅ᪉ బ⸨ᐶᏊ ⛅⏣ᕷ 26.11.25 ࠶ࡁࡓ⏘Ꮫᐁ㐃ᦠࣇ࢛࣮࣒ࣛ 2014 ۑᩪ⸨༤அ ᕝཎᚿ ⛅⏣ᕷ 㔝⚘ᮁ 26.12.03㹼05 ➨ 108 ᅇ᪥ᮏ㣗ရ⾨⏕ᏛᏛ⾡ㅮ₇ ۑᩪ⸨༤அ ▼ᕝ┴ 㰺⸨ᚿಖᏊ 26.12.05 ⛅⏣ᛂ⏝⏕⛉Ꮫ◊✲➨ 24 ᅇㅮ₇ ۑబ⸨ᐶᏊ ⛅⏣ᕷ 26.12.20㹼21 ➨ 21 ᅇࣜࢣࢵࢳ◊✲◊✲Ⓨ⾲ ۑబ⸨ᐶᏊ ᮾி㒔 27.02.13㹼15 ᖹᡂ 26 ᖺᗘ᪥ᮏ⋇་ᖌ⋇་Ꮫ⾡Ꮫᖺḟ 㰺⸨ᚿಖᏊ ᒸᒣ┴ 27.03.16㹼18 ➨ 49 ᅇ᪥ᮏỈ⎔ቃᏛ 27.03.19㹼22 ➨ 62 ᅇ᪥ᮏ⏕ែᏛ ᑠᯘ㈗ྖ ⏕㨶 ⏣⋢ۑᑗᩥ - 10 - ▼ᕝ┴ 㮵ඣᓥ┴ 秋田県健康環境センター年報 第10号 2014 ᗣ⎔ቃࢭࣥࢱ࣮ദ◊✲Ⓨ⾲ 㛤 ദ ᪥ 㸸 ᖹᡂ 26 ᖺ 7 ᭶ 9 ᪥㸦Ỉ㸧 㛤ദሙᡤ 㸸 ⛅⏣┴⥲ྜಖࢭࣥࢱ࣮ ₇ 1 2 㢟 ྡ Ⓨ ⾲ ⪅ ⛥࡞⾑Ύᆺ Agbeni ࡀྠᐃࡉࢀࡓ 3 ௳ࡢࢧࣝࣔࢿࣛឤᰁࡢゎᯒ 㔝㈗அ ⛅⏣┴࠾ࡅࡿ㣗⏝∵ࡢ⭠⟶ฟ⾑ᛶ⭠⳦ಖ⳦≧ἣศ㞳ᰴࡢ⣽⳦Ꮫⓗᛶ≧ 㛵ࡍࡿ◊✲㸫2001㹼2003 ᖺ 2012㹼2013 ᖺࡢㄪᰝᡂ⦼ ඵᰗ ₶ 3 ࢘ࢩ᪩ᮇึஙࡢ࢘ࣝࢫάຠᯝ㛵ࡍࡿ᳨ウ ᰘ⏣ࡕࡦࢁ 4 㞝≀ᕝὶᇦ࠾ࡅࡿ࢝ࢶࢶ࣒࢞ࢩ⏕ᜥ≧ἣㄪᰝ㸸2009 ᖺ㹼2012 ᖺ బ⸨ᐶᏊ 5 ࣐ࢶᯘᩓᕸࡉࢀࡓ㎰⸆ࡢᣲືࡘ࠸࡚ ᑠᯘ㈗ྖ 6 ⏣ἑ†ࡢỈ㉁ࡢኚ㑄 ⏕㨶 7 ᮍฎ⌮⋢ᕝ㓟ᛶỈࡢ୰ࡼࡿ pH ᨵၿࡑࢀక࠺ຠᯝ ᡂ⏣ಟྖ ࡑࡢࡢཱྀ㢌Ⓨ⾲ ᖺ᭶᪥ Ⓨ ⾲ ྡ ࣭ ₇ 㢟 ྡ Ⓨ⾲⪅ 㛤ദᆅ ᖹᡂ 26 ᖺᗘಖ⎔ቃᴗົ◊✲Ⓨ⾲ ࣭ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿᑠඣ྾ჾᝈࡽࡢཎయ᳨ฟ≧ἣ ᰘ⏣ࡕࡦࢁ 27.01.23 ࣭ᝈ⪅࠾ࡼࡧ࣐࢘ࢫࡢ⾑୰ Orientia tsutsugamushi ࡢᐃ㔞㔜㛵 బ⸨ᐶᏊ ₲ୖᕷ ࡍࡿ⪃ᐹ ࣭᭷ᶵࣜࣥ⣔㎰⸆ࡀΰධࡉࢀࡓ෭㣗ရࡢ᳨ᰝࡘ࠸࡚ 㔝⚘ᮁ ㅮᖌὴ㐵➼ ᢏ⾡ᨭ ᐇ᪥ 26.05.09㹼 09.04 ࡞ ෆ ᐜ ㅮᖌẶྡ ࢫ࣮ࣃ࣮ࢧ࢚ࣥࢫࣁࢫࢡ࣮ࣝ ᐇ㦂ヨᩱࡢศᯒ 26.05.30 ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢၟရ㛤 26.09.30 Ⓨ㛵ࡍࡿຓゝ 26.06.13㹼17 26.07.03㹼18 26.07.14㹼16 26.10.15 ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢᑟධ 㛵ࡍࡿຓゝ ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢᑟධ 㛵ࡍࡿຓゝ ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢᑟධ 㛵ࡍࡿຓゝ ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢᑟධ 㛵ࡍࡿຓゝ 26.12.03㹼04 ࡶࡳẆⅣࢆά⏝ࡋࡓࣇ࣮ࣝࢻ࡛ 27.02.18㹼20 ࡢỈ㉁ίヨ㦂ࡢࢻࣂࢫ 26.12.26 ࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢᑟධ 㛵ࡍࡿຓゝ ᑐ ᘏேᩘ ᶓᡭΎ㝠Ꮫ㝔 1 ྡ ᩪ⸨༤அ ࣓ࣝࢡ♫ 2 ྡ ᩪ⸨༤அ ዉⰋ┴ಖ◊✲ࢭࣥࢱ࣮ 2 ྡ ᩪ⸨༤அ ᯇᕷಖ⎔ቃ◊✲ᡤ 5 ྡ ᩪ⸨༤அ රᗜ┴すᐑಖᡤ 3 ྡ ᩪ⸨༤அ ⛅⏣ᕷಖᡤ 1 ྡ 8 ྡ 1 ྡ 23 ྡ 㧗ᔱ ྖ ᡂ⏣ಟྖ ᩪ⸨༤அ 㛗ᓮ┴⎔ቃಖ◊✲ࢭ ࣥࢱ࣮ ᒸᒣ┴ᩜಖᡤ ྜ - 11 - ㇟ ィ 秋田県健康環境センター年報 第10号 2014 ฟ๓ㅮᗙ ฟ๓ㅮᗙ ㅮᖌẶྡ ᐇᅇᩘ ᘏཧຍ⪅ᩘ Ἔ᩿࡛ࡁ࡞࠸⤖᰾ ⏣୰㈗Ꮚ 1 ᅇ 20 ྡ ⣽⳦ᛶឤᰁ࣭㣗୰ẘࡘ࠸࡚ 㧘ᶫᚿಖ 2 ᅇ 43 ྡ ࢘ࣝࢫᛶ㣗୰ẘࡘ࠸࡚ ᩪ⸨༤அ 2 ᅇ 197 ྡ బ⸨ᐶᏊ 2 ᅇ 100 ྡ ᩪ⸨༤அ 1 ᅇ 28 ྡ ᰘ⏣ࡕࡦࢁ 1 ᅇ 35 ྡ ⎔ቃᨺᑕ⬟ࡘ࠸࡚ ⌋⏣ᑦಇ 1 ᅇ 30 ྡ ⎔ቃ୰ࡢẼởᰁ≀㉁ࡘ࠸࡚ ῲᚿఙ 1 ᅇ 40 ྡ ΎỈ 1 ᅇ 20 ྡ 12 ᅇ 513 ྡ ࢘ࣝࢫᛶឤᰁࡘ࠸࡚ ྜ ィ ࡑࡢㅮᖌὴ㐵 ࡞ෆᐜ ᐇ᪥ ㅮᖌẶྡ ᗫᲠ≀ฎ⌮ࡘ࠸࡚ 26.05.20 ᑠᯘ㈗ྖ ⛅⏣┴❧Ꮫ 4 ྡ 㣗ရ୰ࡢṧ␃㎰⸆ࡘ࠸࡚ 26.05.27 ᑠᯘ㈗ྖ ⛅⏣┴❧Ꮫ 4 ྡ 26.06.02 ⏣୰㈗Ꮚ ⏘ᴗᢏ⾡ࢭࣥࢱ࣮ 50 ྡ ࢘ࣝࢫᛶ㣗୰ẘࡢண㜵ᑐ⟇ࡘ࠸࡚ 26.08.05 ᩪ⸨༤அ ⛅⏣ᕷಖᡤ 90 ྡ ≉Ṧ⅏ᐖಖᏳ 26.08.20 ᩪ⸨༤அ ᾘ㜵Ꮫᰯ 80 ྡ 26.09.26 㔝㈗அ 30 ྡ ಶయࡢᛂ 26.09.30 ඵᰗ 150 ྡ 㧗㱋⪅ࡢᗣឤᰁࡘ࠸࡚ 26.10.01 ᩪ⸨༤அ 140 ྡ ⏕ά⾨⏕㛵ಀႠᴗࡢឤᰁᑐ⟇ 26.10.27 ᩪ⸨༤அ 30 ྡ Orientia tsutsugamushi ᇵ㣴ᢏ⾡ྜྠ◊ಟ 26.10.30 3 ྡ 50 ྡ 70 ྡ 19 ྡ ⤖᰾➼㛵ࡍࡿሗཬࡧᗣ⟶⌮ୖࡢ ண㜵᪉ἲ➼ࡘ࠸࡚ ឤᰁࣛ࢝ࣝࢺ 㹼⣽⳦ࡽࡳࡓឤᰁ㹼 㹼31 ₶ ౫㢗ඖ ⛅⏣ᆅᇦ⯆ᒁ⚟♴⎔ ቃ㒊 ⛅⏣Ꮫ་Ꮫ⣔◊✲⛉ ⏤ᆅᇦ⯆ᒁ⚟♴⎔ ቃ㒊 ⛅⏣┴⏕ά⾨⏕Ⴀᴗᣦ ᑟࢭࣥࢱ࣮ బ⸨ᐶᏊ ⚟ᓥ┴⾨⏕◊✲ᡤ ⛅⏣┴❧Ꮫ Ꮫ㝔ࢭ࣑ࢼ࣮ 26.11.07 ⏕㨶 ⏕ά⾨⏕㛵ಀႠᴗࡢឤᰁᑐ⟇ 26.11.10 బ⸨ᐶᏊ ༴㝤ᛶ≀㉁➼ࡢᇶ♏▱㆑ 26.11.13 ᩪ⸨༤அ - 12 - ⛅⏣┴⏕ά⾨⏕Ⴀᴗᣦ ᑟࢭࣥࢱ࣮ ᾘ㜵Ꮫᰯ ཧຍ⪅ᩘ 秋田県健康環境センター年報 第10号 2014 ⮫ᗋ᳨ᰝࢆ᥈ồࡍࡿ㹼࣮ࣁࢺ࣮ࣈ 26.11.23 ඵᰗ ࡘࡘࡀࡘ࠸࡚ 26.11.25 బ⸨ᐶᏊ ༑⏣†ࡢỈ㉁ࡢ⌧ἣࡘ࠸࡚ 27.01.20 ಖሗᏛ ᪥⮫ᢏ᪥ᮏᨭ㒊་Ꮫ 50 ྡ ⛅⏣┴་ᖌ 12 ྡ ⏕㨶 ⛅⏣┴⏕ά⎔ቃ㒊 80 ྡ 27.01.22 ⏣୰㈗Ꮚ ⛅⏣Ꮫ་Ꮫ⣔◊✲⛉ 80 ྡ 㧗⃰ᗘࣜࣥฟỈࡢ㐺ᛂ➼ 27.01.24 ᡂ⏣ಟྖ ࠶ࡁࡓ⎔ቃ᠓ヰ 60 ྡ ⏕ά⾨⏕㛵ಀႠᴗࡢឤᰁᑐ⟇ 27.02.23 ᩪ⸨༤அ 40 ྡ ⏕ά⾨⏕㛵ಀႠᴗࡢឤᰁᑐ⟇ 27.03.02 ᩪ⸨༤அ 30 ྡ 1,072 ྡ ࡽ⮫ᗋ᳨ᰝࢆぢࡘࡵ┤ࡍ㹼 ⤖᰾ࡢሗ⟶⌮࣭⤫ィ ₶ ᳨ᰝ㒊 ⛅⏣┴⏕ά⾨⏕Ⴀᴗᣦ ᑟࢭࣥࢱ࣮ ⛅⏣┴⏕ά⾨⏕Ⴀᴗᣦ ᑟࢭࣥࢱ࣮ ྜィ 20 ᅇ どᐹ࣭ぢᏛ➼ཷධ ཧຍ⪅༊ศ ᖹᡂ 24 ᖺᗘ 2 ᑠ࣭୰Ꮫ⏕ ᖹᡂ 25 ᖺᗘ (2) ᖹᡂ 26 ᖺᗘ 0 0 㸫 ࣥࢱ࣮ࣥࢩࢵࣉ 11 (3) 10 (3) 17 (3) ࡑࡢࡢᏛ⏕ 37 (4) 53 (4) 28 (2) ୍⯡┴Ẹ 0 ᴗົ㛵ಀ⪅ 㸦་ᖌ⮫ᗋ◊ಟྵࡴ㸧 9 0 (3) 9 (1) ⛅⏣┴❧Ꮫ⏕≀㈨※⛉Ꮫ㒊ᛂ⏝ ⏕≀⛉Ꮫ⛉࣭⏕≀⏕⏘⛉Ꮫ⛉࣭⏕≀ ⎔ቃᏛ⛉㸦3 ᖺ⏕㸧㸪⛅⏣㧗ᑓ⎔ቃ 㒔ᕷᕤᏛ⛉࣭㟁ẼሗᕤᏛ⛉㸦3 ᖺ ⏕㸧㸪ᮏⲮ㧗➼Ꮫᰯ㸦2 ᖺ⏕㸧 ⛅⏣Ꮫ་Ꮫ㒊♫་Ꮫᐇ⩦㸦3 ᖺ ⏕㸧㸪⪷㟋ዪᏊ▷ᮇᏛᑓᨷㄢᗣ ᰤ㣴ᑓᨷ㸦2 ᖺ⏕㸧 0 㸫 0 㸫 ▼ᕝ┴⎔ቃಖ◊✲ࢭࣥࢱ࣮㸪㛗ᓮ┴ 10 ┴እ (7) 2 (1) 5 (4) ⎔ቃಖ◊✲ࢭࣥࢱ࣮㸪㯞ᕸᏛ⋇་ Ꮫ㒊⋇་Ꮫ⛉㸦5 ᖺḟ⏕㸧㸪ᒾᡭᏛ ㎰Ꮫ㒊ඹྠ⋇་Ꮫ⛉㸦3 ᖺḟ⏕㸧 ᅜእ ྜ ィ 3 (2) 2 (1) 2 (2) 72 (21) 76 (10) 52 (11) ྜྷᯘ┬ⓑᇛᕷ⎔ቃಖㆤᒁ㸪ྜྷᯘ┬๓ ᩔᕷ⎔ቃಖㆤᒁ ὀ㸧ᣓᘼෆࡢᩘᏐࡣᅋయᩘ ཷ㈹࣭⾲ᙲ➼ ཷ㈹᪥ 26.10.22 ⾲ ⏕ά⎔ቃᅜ ᙲ ྡ ཷ㈹⪅ ឤㅰ≧ ῲᚿఙ - 13 - ᤵ ᶵ ⏕ά⎔ቃᅜ 㛵 秋田県健康環境センター年報 第10号 2014 㸬◊✲ᴗົᐇ⦼ ࡁ࡞ࡗࡓ 21 ᰴࡘ࠸࡚㸪PCR ᆺูࢆ⾜ࡗࡓ ࡇࢁ 19 ᰴ࡛ᆺูྍ⬟࡛࠶ࡾ㸪PCR ᆺูࡢ᭷ ⣽⳦⌜ ⏝ᛶࡀ♧ࡉࢀࡓࠋ ࢝ࣥࣆࣟࣂࢡࢱ࣮ࡢᆺู᪉ἲࡢ᳨ウศ㞳 ⳦ᰴࡢ≉ᚩ㸫࢝ࣥࣆࣟࣂࢡࢱ࣮ࣞࣇࣞࣥ ࢫࢭࣥࢱ࣮ᴗົ㸫㸦ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ ຓ㔠ࠕᅜෆࡢཎయࢧ࣮࣋ࣛࣥࢫ㈨ࡍ ࡿᶵ⬟ⓗ࡞ࣛ࣎ࢿࢵࢺ࣮࣡ࢡࡢᙉ㛵ࡍ ࡿ◊✲ࠖ㸧 㸦 ᖹ ᡂ 2 5 ᖺ ᗘ 㹼 㸧 ࢧࣝࣔࢿࣛ⳦ࡢ㎿㏿࡞ྠᐃࡢࡓࡵࡢ᳨ฟἲ ཬࡧ⾑Ύᆺูἲࡢ☜❧ 㸦 ᖹ ᡂ 25 ᖺ ᗘ 㹼 ᖹ ᡂ 26 ᖺ ᗘ 㸧 ◊✲ᴫせ ࢧ ࣝ ࣔ ࢿ ࣛ ࡼ ࡿ 㣗 ୰ ẘ 㸪ឤ ᰁ ࡢ 㝿 ࡢ ㎿ ㏿ ࡞ ᳨ ᰝ య ไ ࡢ ᵓ ⠏ ࢆ ┠ ⓗ 㸪ࢧ ࣝ ࣔ ࢿ ࣛ ࡢ ◊✲ᴫせ ᖹ ᡂ 26 ᖺ ࡣ ┴ෆࡢᩓⓎୗ⑩ᝈ⪅⏤᮶ࡢ 47 㑇ఏᏊゎᯒἲࡼࡿ᳨ฟἲཬࡧ⾑Ύᆺูἲ ᰴ㸦C. jejuni 46 ᰴ㸪C. coli 1 ᰴ㸧ࡘ࠸࡚ Penner ࡢ☜❧ࢆ⾜ࡗࡓࠋ ἲ Lior ἲࡢẚ㍑᳨ウࢆ⾜ࡗࡓࠋࡲࡓ㸪ᆺู ᖹ ᡂ 26 ᖺ ᗘ ࡣ 㸪㣗 ⫗ 21 ᳨ య ࡘ ࠸ ࡚ ࣜ ⬟࡛࠶ࡗࡓᰴࡘ࠸࡚ࡣ㸪PCR ᆺูἲࡼࡾᆺ ࣝ ࢱ ࣒ PCR ἲ ࡼ ࡿ ࢧ ࣝ ࣔ ࢿ ࣛ ள ✀ I ࡢ ᳨ ูྍ⬟᳨ウࡋࡓࠋࡉࡽ㸪⸆⪏ᛶࡢഴྥ ฟ ࢆ ⾜ ࠸ 㸪ᇵ 㣴 ࡼ ࡿ ศ 㞳 ྠ ᐃ ᳨ ᰝ ࡢ ẚ ㍑ ࢆᢕᥱࡍࡿࡓࡵ㸪ࢸࢺࣛࢧࢡࣜࣥ㸪࢚ࣜࢫࣟ ࢆ ⾜ ࡗ ࡓ ࠋࡲ ࡓ 㸪ࢧࣝࣔࢿࣛࡢ⾑ΎᆺࢆỴᐃࡋ ࣐ࢩࣥ㸪ࢼࣜࢪࢡࢫ㓟㸪ࣀࣝࣇࣟ࢟ࢧࢩࣥ㸪 ࡚࠸ࡿせ࡞ O ᢠཎ 7 ✀㢮㸪H1 ᢠཎ 12 ✀㢮㸪 ࢜ࣇࣟ࢟ࢧࢩࣥ㸪ࢩࣉࣟࣇࣟ࢟ࢧࢩࣥࡢ 6 H2 ᢠཎ 7 ✀㢮ࡘ࠸࡚ Multiplex PCR ἲࢆ᳨ウ ࡘ࠸࡚ឤཷᛶヨ㦂ࢆᐇࡋࡓࠋ ࡋ㸪᳨ᰝἲࡢ┬ຊࢆ⾜ࡗࡓࠋ ྠᵝࡋ࡚㭜⫗ࡽศ㞳ࡋࡓ 13 ᰴ㸦C. jejuni ⤖ᯝ 12 ᰴ㸪C. coli 1 ᰴ㸧㸪㣗୰ẘࡽศ㞳ࡋࡓ ࢧࣝࣔࢿࣛࡀศ㞳ࡉࢀࡓࡢࡣ 21 ᳨య୰ 5 ᳨ C. jejuni 2 ᰴࡘ࠸࡚ࡶㄪᰝࡋࡓࠋ య࡛࠶ࡾ㸪⾑Ύᆺࡣ Infantis (O7:r:1,5) 1 ᳨య ⤖ᯝ ࡽࡣ Manhattan (O6,8:d:1,5)ࡶศ㞳ࡉࢀࡓࠋࣜ ᩓⓎୗ⑩ᝈ⪅⏤᮶ࡢ C. jejuni 46 ᰴࡢ࠺ࡕ㸪 ࣝࢱ࣒ PCR ࡛ࡣቑ⳦ᾮ 3 ᳨యࡽࢧࣝࣔࢿ Penner ἲ࡛༢୍ࡢ⾑Ύᆺᆺูྍ⬟࡛࠶ࡗࡓࡶ ࣛ㑇ఏᏊࡀ᳨ฟࡉࢀ㸪ḟቑ⳦ᾮ࡛ࡣࢧࣝࣔࢿ ࡢࡣ 28 ᰴ㸦60.9%㸧㸪Lior ἲ࡛ࡣ 22 ᰴ㸦47.8%㸧 ࣛࡀ ศ㞳 ࡉࢀࡓ 5 ᳨ య ࡣ࠸ ࡎࢀ ࡶ㝧 ᛶ ࡞ࡗ ࡛࠶ࡗࡓࠋC. coli ࡣ࠸ࡎࢀࡢᡭἲ࡛ࡶ༢୍ࡢ⾑ ࡓࠋ1 ᳨యࡘ࠸࡚ࡣ㸪ࢧࣝࣔࢿࣛศ㞳㝜ᛶ࡛ Ύᆺࡣᆺูࡉࢀ࡞ࡗࡓࠋࡲࡓ㸪ᩓⓎୗ⑩ ࠶ࡗࡓࡀ㸪ࣜࣝࢱ࣒ PCR ࡛ࡣ㝧ᛶ࡛࠶ࡗ ᝈ⪅⏤᮶ࡢ C. jejuni 42 ᰴ୰ 33 ᰴ㸦78.6%㸧ࡀ࢟ ࡓࠋࡇࡢ᳨ウࡼࡾ㸪ࣜࣝࢱ࣒ PCR ἲࡣ㸪 ࣀࣟࣥ⣔⸆⪏ᛶࢆ♧ࡋࡓࠋC. jejuni ࠾ࡅ ࢧࣝࣔࢿ᳨ࣛᰝࡢึᮇẁ㝵࡛࠶ࡿḟቑ⳦ࡲ ࡿ࢟ࣀࣟࣥ⣔⸆ࡢ⪏ᛶࡣ㸪௨๓ࡣ 30㹼40% ࡛⾜࠼ࡤ㸪㎿㏿ࢧࣝࣔࢿࣛࡢ᳨ฟࡀྍ⬟࡛࠶ ⛬ᗘ࡛࠶ࡗࡓࡀ㸪㏆ᖺࡑࡢྜࡀ㧗ࡃ࡞ࡗ࡚࠾ ࡿࡇࡀ♧၀ࡉࢀࡓࠋ ࡾ㸪⪏ᛶࡢ㐍⾜ࡀ༴ࡉࢀࡿࠋ୍᪉㸪➨୍㑅 ⾑Ύᆺูἲࡘ࠸࡚ࡣ㸪┴ෆ࡛ศ㞳ࡉࢀࡿ ᢥ⸆ࡢ࢚ࣜࢫ࣐ࣟࢩࣥࡘ࠸࡚ࡣ⪏ᛶᰴࡣ せ࡞ࢧࣝࣔࢿࣛࡘ࠸࡚ࡣ㸪Multiplex PCR ࡼ ᳨ฟࡉࢀ࡞ࡗࡓࠋ ࡾᐜ᫆⾑Ύᆺࢆ᥎ᐃࡍࡿࡇࡀྍ⬟࡞ࡗ 㭜⫗⏤᮶ᰴࡢ୍㒊࡛ࡣ㸪ᝈ⪅⏤᮶ᰴྠ୍ࡢ ࡓࠋPCR ἲࡼࡿᢠཎ㑇ఏᏊࡢ᳨ฟࡣ㸪㎿㏿࡛ ⾑Ύᆺࡀࡳࡽࢀࡓࠋ㞟ᅋឤᰁ࡛ࡣ㸪㭜⫗ࡀ ࠶ࡿࡇຍ࠼㸪ఱࡽࡢཎᅉ࡛ᢠཎࡀ⏘⏕ࡉ ཎᅉ㣗ရ᥎ᐃࡉࢀࡓࡶ࠶ࡗࡓࡇࡽ㸪 ࢀ࡞ࡃ࡞ࡗ࡚ࡋࡲࡗࡓኚ␗ᰴ➼࠾࠸࡚ࡶ᭷ 㭜⫗ࡣ࢝ࣥࣆࣟࣂࢡࢱ࣮ࡢឤᰁ※ࡋ࡚㔜せ ຠ࡛࠶ࡾ㸪ᚋࡢṇ☜࡞ឤᰁࡢࢧ࣮࣋ࣛࣥ ⪃࠼ࡽࢀࡿࠋ ࢫࡸ㣗୰ẘࡢཎᅉ✲᫂➼ᙺ❧ࡘ⪃࠼ࡽࢀ Penner ἲࡼࡿ⾑Ύᆺู࡛༢୍ࡢᆺỴᐃ࡛ ࡿࠋ - 14 - 秋田県健康環境センター年報 第10号 2014 㣗୰ẘ➼࡛ศ㞳ࡉࢀࡓ㯤Ⰽࣈࢻ࢘⌫⳦ ࡢ㑇ఏᏊᆺูࡘ࠸࡚㸦ཌ⏕ປാ⛉Ꮫ◊✲ ㈝⿵ຓ㔠ࠕ㣗ရ୰ࡢ㣗୰ẘ⳦➼ࡢ㑇ఏ≉ᛶ ཬࡧไᚚ㛵ࡍࡿㄪᰝࠖ㸧 ◊✲ᴫせ 㸦 ᖹ ᡂ 25 ᖺ ᗘ 㹼 ᖹ ᡂ 27 ᖺ ᗘ 㸧 ⒪㠃ࡽ⤖᰾ᑐ⟇ࢆᨭࡍࡿࡓࡵ㸪ศ㞳ᰴࡢ ᮏ┴ࡢ㧗㱋⪅⤖᰾ᑐ⟇ࡢ⾜ᨻᑐᛂᨭࢆ┠ ⓗࡋ࡚㸪⤖᰾⇞ࡢᏛⓗ࣭ศᏊᏛⓗ≉ ᚩ㛵ࡍࡿ▱ぢࢆ㞟✚ࡋ≉ᚩࢆ᫂ࡽࡍࡿࠋ ࣆࣛࢪࢼ࣑ࢻᑐࡍࡿ⪏ᛶࡢ᭷↓㸪ࢯࢽࢪ ࢻᑐࡍࡿ⪏ᛶࡢ᭷↓ࢆ㎿㏿࣭㧗⢭ᗘุᐃࡍ ◊✲ᴫせ ୍⯡ὶ㏻ࡋ࡚࠸ࡿ㣗ရ࠾ࡅࡿྛ✀㣗୰ ࡿ㑇ఏᏊデ᩿ᢏ⾡ࢆᑟධࡍࡿࠋࡲࡓ㸪㧗㱋⪅⤖ ẘ⳦ࡢởᰁ≧ἣཬࡧ⳦ᰴࡢᆺู➼ࡢᏛሗ ᰾ᑐ⟇ά⏝ྍ⬟࡞㧗㱋⪅ࡢ⏕άࣃࢱ࣮ࣥ ࡣ㣗୰ẘⓎ⏕ࡢཎᅉ✲᫂࠶ࡿ࠸ࡣ㣗ရ⾨⏕ ྜࢃࡏࡓᩍᮦ➼ࡢᣦᑟᣦ㔪ࢆసᡂࡍࡿࠋ ᣦᑟ➼㠀ᖖ᭷⏝⪃࠼ࡽࢀࡿࠋ㣗ရ᳨ᰝ࠶ ⤖ᯝ ࡿ࠸ࡣ㣗୰ẘ࠾࠸࡚ศ㞳ࡉࢀࡓ㯤Ⰽࣈ ⤖᰾⳦ศᏊᏛㄪᰝࡋ࡚㸪ศ㞳ᰴࡢ VNTR ࢻ࢘⌫⳦ࢆᑐ㇟㸪᭷⏝࡞ࢱࣆࣥࢢ᪉ἲࡘ ᳨ᰝࢆ㸪JATA-12 ἲࡼࡾᐇࡋࡓࠋ⤖᰾⳦⸆ ࠸᳨࡚ウࡍࡿࡓࡵ㸪ᖹᡂ 25 ᖺᗘࡣ㭜⫗ởᰁᐇ ឤཷᛶࡢ㑇ఏᏊデ᩿࡛ࡣ㸪ࢯࢽࢪࢻ⪏ᛶ ែㄪᰝ࡛ศ㞳ࡉࢀࡓ㯤Ⰽࣈࢻ࢘⌫⳦ࡘ࠸࡚ ࡘ࠸࡚㸪katG㸪mabA㸪ahpC 㑇ఏᏊࡢኚ␗ࡢ POT ἲ㸪MLVA ἲ㸪PFGE ἲࡼࡾ㑇ఏᏊᆺู ᭷↓ࢆ☜ㄆࡋࡓࠋ౪ヨᰴࡣᖹᡂ 24 ᖺ 7 ᭶㹼ᖹ ࢆᐇࡋࡓࠋࡲࡓࣈࢻ࢘⌫⳦࢚ࣥࢸࣟࢺ࢟ࢩࣥ ᡂ 25 ᖺ 6 ᭶ཷ㡿ᰴ 96 ᰴ࡛㸪ࡑࡢ࠺ࡕᇵ㣴ヨ㦂 ࡢ A㹼E,G,H,I ᆺ sea㹼see 㸪seg㸪seh㸪sei 㑇ఏᏊ ࡛ࢯࢽࢪࢻ⪏ᛶ࡛࠶ࡗࡓᰴ 8 ᰴ୰ 5 ᰴ⪏ ࡢಖ᭷≧ἣࡘ࠸࡚ PCR ἲࡼࡾ᳨ウࡋࡓࠋᖹ ᛶ㑇ఏᏊኚ␗ࡀ࠶ࡗࡓࠋࣆࣛࢪࢼ࣑ࢻ⪏ᛶ㑇ఏ ᡂ 26 ᖺᗘࡣ㸪ࡉࡽ㸪ᐇ㝿ࡢ㣗୰ẘ࠾ Ꮚ᳨ᰝࡣ 48 ᰴࡘ࠸࡚㸪pncA 㑇ఏᏊࡢኚ␗ࡢ ࡅࡿᛂ⏝ࡢྍ⬟ᛶࢆ᳨ウࡍࡿࡓࡵ㸪㯤Ⰽࣈࢻ࢘ ᭷↓ࢆ☜ㄆࡋࡓࠋPZase(-)ࡢ 1 ᰴࡶྵࡵ࡚㸪48 ⌫⳦㣗୰ẘ⏤᮶ᰴ㸪᭷ⱞࡸࡢཎ ᰴ࡚ pncA 㑇ఏᏊࡢኚ␗ࡣ࡞ࡗࡓࠋ ࡲࡓ㸪ᮏ┴ࡢ㧗㱋⪅⤖᰾ᑐ⟇ᨭࡢ୍⎔ࡋ ᅉ≀㉁᳨࡛ฟࡉࢀࡓ㯤Ⰽࣈࢻ࢘⌫⳦ࡘ ࠸࡚ྠᵝ᳨ウࡋࡓࠋ ࡚㸪ࠕ㧗㱋⪅⚟♴タ➼࠾ࡅࡿ⤖᰾ᑐᛂ࢞ ⤖ᯝ ࢻࣈࢵࢡࠖࡢసᡂྥࡅࡓࣥࢱࣅ࣮ࣗㄪᰝࢆ 㣗୰ẘ➼ 25 ⏤᮶ 75 ᰴࡣ POT ἲ࡛ ᐇࡋࡓࠋᖹᡂ 24㹼25 ᖺ⤖᰾ᝈ⪅ࡢⓎ⏕ࡀ 31 ✀㢮㸪SE ᆺ POT ᆺࡢ⤌ࡳྜࢃࡏ࡛ 35 ✀ ࠶ࡗࡓ≉ู㣴ㆤ⪁ே࣮࣒࣍㸪⪁ேಖタ㸪ᒃ 㢮㸪MLVA ἲ࡛ 36 ✀㢮ᆺูࡉࢀࡓࠋࡉࡽ㸪 Ꮿㆤᨭᴗᡤ➼ࡢ 10 ᡤࢆᑐ㇟㸪ᖹᡂ POT ἲ MLVA ἲࡢᆺูࡢホ౯ࡢࡓࡵ㸪POT 26 ᖺ 12 ᭶㹼ᖹᡂ 27 ᖺ 2 ᭶ㄪᰝࡋࡓࠋ⤖᰾ᝈ ᆺ 24 ✀㸦SE:POT ᆺ 27 ✀㸧㸪MLVA ἲ 27 ✀ ⪅Ⓨ⏕ࡢᏳࡸᅔࡗࡓࡣ㸪⤖᰾ࡢ▱㆑ࡸ⌮ ᆺูࡉࢀࡓ 53 ᰴࡘ࠸࡚ PFGE ᆺูࢆᐇࡋࡓ ゎ㊊ࡼࡿ㣗ჾᾘẘ㸪⾰㢮ᐷලࡢὙ℆➼ࡢ᪥ ࡇࢁ㸪26 ✀㢮ᆺูࡉࢀࡓࠋྠࡌ POT ᆺ㸪 ᖖⓗ࡞సᴗࡘ࠸࡚ࡢࡲ࠸࡛࠶ࡗࡓࠋ㐣ᗘ MLVA ᆺࡀ PFGE ἲ࡛⣽ศࡉࢀࡿ㸪㏫␗ ࡢᏳࢆᣢࡘ⫋ဨࡀ࠸ࡿ୍᪉࡛⤖᰾ࢆࡃ▱ ࡞ࡿ POT ᆺ㸪MLVA ᆺࡀྠ୍ࡢ PFGE ࣃࢱ࣮ࣥ ࡽࡎ༴ᶵព㆑ஈࡋ࠸⫋ဨࡶ࠸ࡓࠋ᥋ゐ⪅デ ࡞ࡿࡶࡳࡽࢀࡓࡀ㸪POT ἲ MLVA ἲ ࡛ࡣᐃᮇⓗ㛗ᮇⓗ㏣㊧ࡍࡿࡓࡵᑐᛂࡢ↹㞧 ࡼࡿᆺูࡣ PFGE ἲࡼࡿᆺูࡰྠ⛬ᗘࡢ ࡉᅔᝨࡋࡓタࡶከࡗࡓࠋࡋࡋ㸪ࡇࢀࡽ ゎᯒຊࢆ᭷ࡋ࡚࠸ࡿࡇࡀ☜ㄆࡉࢀࡓࠋࡲࡓ㸪 ࡢᏳゎᾘࡣಖᡤࡢᣦᑟຓゝࡀࡁ࠸ᙺ ⡆౽ᛶ࣭㎿㏿ᛶࡽࡶ POT ᆺูཬࡧ MLVA ᆺ ࢆᯝࡓࡋ࡚࠸ࡓࠋタཬࡧᴗᡤ࠾ࡅࡿ⤖ ูࡣ᭷⏝࡞ᆺูἲ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋ ᰾ᑐᛂ࢞ࢻࣈࢵࢡࡢᚲせᛶࡘ࠸࡚ࡣ㸪 タཬࡧᴗᡤࡀㄆ㆑ࡋ࡚࠸ࡓࠋᥖ㍕ෆᐜࡘ࠸ ࡚ࡣ⤖᰾ࡢṇࡋ࠸▱㆑㸪Ⓨ⏕ࡢලయⓗᑐᛂ㸪 㧗㱋⪅⤖᰾ᑐ⟇ᨭ⸆⪏ᛶ㎿㏿デ᩿ἲ ࡢᑟධ㛵ࡍࡿㄪᰝ◊✲ ᪥ᖖ⏕άୖࡢ୍⯡ⓗሗ㸪ಖᡤࡢ㐃ᦠ➼࡛ 㸦 ᖹ ᡂ 26 ᖺ ᗘ 㹼 ᖹ ᡂ 27 ᖺ ᗘ 㸧 ࡛⤖᰾ᑐᛂࡀศࡿࡶࡢࢆᕼᮃࡋ࡚࠸ࡓࠋ ࠶ࡾ㸪 ་Ꮫ⏝ ㄒࡀ ᑡ࡞ࡃ ⌮ゎࡋ ࡸࡍ࠸ ࡶࡢ 㸪1 - 15 - 秋田県健康環境センター年報 第10号 2014 ࢘ࣝࢫ⌜ ◊✲ᴫせ ᾏ㐨࣭ᮾ࣭᪂₲ࣈࣟࢵࢡࡣከࡃࡢ⾑Ύᆺ 㣗ရࡽࡢ࢘ࣝࢫ᳨ฟἲࡋ࡚ࡢࣃࣥࢯ ࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ⢭ᗘྥୖ㛵ࡍࡿ◊ ✲㸦ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠ࠕ㣗ရ୰ࡢ ཎ࢘ࣝࢫࡢ᳨ฟἲ㛵ࡍࡿ◊✲ࠖ㸧 ࡼࡿࡘࡘࡀࡸ⣚ᩬ⇕⩌ࣜࢣࢵࢳ࡞ 㸦 ᖹ ᡂ 25 ᖺ ᗘ 㹼 ᖹ ᡂ 27 ᖺ ᗘ 㸧 ἲࡘ࠸࡚㸪ࣈࣟࢵࢡෆᢏ⾡◊ಟࢆ⾜ࡗࡓࠋ◊ ከᵝ࡞ࣜࢣࢵࢳࡀⓎ⏕ࡍࡿ㧗ࣜࢫࢡᆅ ᇦ࡛࠶ࡿࠋᅇ㸪ࢃࢀࢃࢀࡣࡇࢀࡽᵝࠎ࡞ࣜࢣ ࢵࢳࡢ࠺ࡕ㸪ࡘࡘࡀࡢ⾑Ύᢠయ౯ ᐃ ಟ࠾࠸࡚ࡣ㸪ᐇ⩦ࢆ⾜࠺ඹヲ⣽࡞࣐ࢽࣗ ࣝࢆసᡂࡍࡿࡇ࡛㸪ᚋࡶᐃࡉࢀࡿᢸᙜ ◊✲ᴫせ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡣ㸪㣗ရ᳨యྵ ⪅ࡢ␗ື㓄៖ࡋࡓデ᩿యไࡢᐇᵓ⠏ࢆ ࡲࢀࡿ࢘ࣝࢫ⢏Ꮚࢆ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ⾲㠃 ┠ᣦࡋࡓࠋࡲࡓ㸪⛅⏣┴࠾ࡅࡿᴗࡋ࡚ྛ ྾╔ࡉࡏ࡚ᅇࡍࡿࡇࢆᇶᮏཎ⌮ࡋ࡚ ✀ ၨ Ⓨ ά ືࢆ ⾜ ࡗ ࡓ ࠋ ᙜ ┴ ࡣ 㧗 ཎ ᛶ ࡢ Kato ࠸ࡿࠋࡑࡢ྾╔ࡢࡓࡵࡢࠕ⢶ࠖࡢᙺࢆᯝࡓࡍ ᆺࡘࡘࡀᝈ⪅ࡀ⌧ᅾࡶᅜෆ࡛၏୍☜ㄆࡉ ᤕᤊᢠయࡋ࡚㸪ࡇࢀࡲ࡛ࡣ་⒪⏝ࡢ࣐࢞ࣥࢢ ࢀ࡚࠾ࡾ㸪ᖺᗘࡶ๓ᅇ㸦2008 ᖺ㸧ࡢᝈ⪅ឤᰁ ࣟࣈࣜࣥ〇ࢆ⏝࠸࡚ࡁࡓࠋࡋࡋ㸪་⸆ရ࡛ ᥎ᐃᆅྠ୍ᆅⅬ࡛ࡢᝈ⪅Ⓨ⏕ࡀ☜ㄆࡉࢀࡓࠋ ࠶ࡿࡇࡽ⸆ἲక࠺ไ⣙ࡀ࠶ࡾ㸪ᬑཬẁ ࡑࡢࡓࡵ㸪፹✀࢝ࢶࢶ࣒࢞ࢩࡢ⏕ᜥㄪᰝ⤖ 㝵࠾࠸࡚ࡣ୍⯡ࡢヨ⸆ࡋ࡚㉎ධ࣭⏝࡛ࡁ ᯝ㸦᪤ሗ㸧ࢆඖឤᰁ᥎ᐃᆅࡢ⮬య࠾ࡼࡧ㛵 ࡿᤕᤊᢠయ౪⤥※ࡀᮃࡲࢀࡓࠋᮏ◊✲࡛ࡣ㸪་ 㐃ᶵ㛵ᚋࡢᑐ⟇ࡘ࠸࡚ࡢㄝ᫂ࢆ⾜ࡗ ⸆ရ࡛ࡣ࡞࠸ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࡢᑟධ ࡓࠋࡉࡽ㸪᪩ᮇ⒪ࡀ㔜せࡉࢀࡿࡘࡘࡀ ࢆ᳨ウࡍࡿࡇ࡛᰿ᮏⓗ࡞ၥ㢟ゎỴࢆᅗࡗࡓࠋ ࡘ࠸࡚་⒪ᶵ㛵ࢆᑐ㇟ࡋࡓၨⓎࢆ⾜ࡗ ࡓࠋ⣚ᩬ⇕⩌ࣜࢣࢵࢳࡘ࠸࡚ࡣ㸪⚟ᓥ┴㸪 ḟ㸪ᬑཬ㝿ࡋ࡚㞀ᐖ࡞ࡿഇ㝧ᛶࡢၥ㢟 ᑐࡋ࡚ᑐ⟇ࢆ᳨ウࡋࡓࠋ㏆ᖺࡢ㑇ఏᏊゎᯒᢏ ᪂₲┴࡛ࡢ᪥ᮏ⣚ᩬ⇕ᝈ⪅Ⓨ⏕ࢆཷࡅ㸪ᆅᇦ ⾡ࡢ㧗ᗘక࠸㸪ᐇ㦂ᐊෆ࡛ PCR ቑᖜ⏘≀ࡀ ࡛ࡢᚋࡢᝈ⪅Ⓨ⏕ഛ࠼ࡿࡓࡵ㸪᳨ᰝయไᵓ ΰධࡋุ࡚ᐃ⤖ᯝᙳ㡪ࢆཬࡰࡍࣜࢫࢡࡀ㧗 ⠏ࡢࡓࡵࡢ‽ഛࢆ⾜ࡗࡓࠋ ࡲࡗࡓࡇࡽ㸪୍ᒙࡢᑐ⟇ᙉࡀồࡵࡽࢀࡓ ⤖ᯝ ᅇ㸪⚟ᓥ┴࣭㟷᳃┴ࡢྛᢸᙜ⪅ࡀ␗ື࡞ ࡢ࡛ᑐᛂ⟇ࢆ᳨ウࡋࡓࠋ ࡗࡓࡓࡵᗘࡢᢏ⾡◊ಟࢆᐇࡋࡓࡀ㸪ᮏࣈࣟ ⤖ᯝ NoV-GI 4㸪NoV-GI 6㸪NoV-GII 2㸪NoV-GII 4㸪 ࢵࢡෆ࠾ࡅࡿ᳨ᰝయไࢿࢵࢺ࣮࣡ࢡࢧ࣏ ཬࡧ NoV-GII 6 ࡢྛᆺࢆ⏝࠸࡚㸪ᕤᴗ⏝་⒪ ࣮ࢺయไࢆಖᣢࡍࡿୖ࡛㠀ᖖ᭷ព⩏࡛࠶ࡗ ⏝ࡢ࣐࢞ࣥࢢࣟࣈࣜࣥࢆ⏝࠸ࡓ㝿ࡢᅇ⋡ࡢ ࡓᛮࢃࢀࡿࠋࡉࡽ㸪⛅⏣┴࠾࠸࡚ࡣၨⓎ ẚ㍑ࢆ⾜ࡗࡓࡇࢁ㸪࡚࠾࠸࡚୧⪅ࡣྠ➼ 㛵ࡋᐇࡋࡓᑐᛂࢆࡿࡇࡀ࡛ࡁࡓࡀ㸪 ࡛࠶ࡗࡓࠋ ᚋࡶྛᆅ᪉ࡢᐇែᐇぢྜࡗࡓ᳨ᰝయไ ഇ 㝧 ᛶ 㜵 Ṇ ࡢ ࡓ ࡵ ࡢ ᪉ ⟇ ࡋ ࡚ 㸪 dUTP UNG (Uracil-N-Glycosylase)ࡢ⤌ࡳྜࢃࡏࡼࡿ ࡢᩚഛࢆ㐍ࡵࡿࡶ㸪ᵝࠎ࡞ࣜࢣࢵࢳ ࡢၨⓎࢆ⥅⥆ࡍࡿࡇࡀ⫢せ⪃࠼ࡽࢀࡓࠋ DNA ởᰁࡢ↓ຠ㸪2nd PCR real-time PCR ࢆ⏝࠸࡚㸪Ct ್ࢆࡶࡗุู࡚ࡍࡿ᪉ἲࡀ᭷ຠ࡛ ࠶ࡿࡇࢆ♧ࡍࡇࡀ࡛ࡁࡓࠋ ,3 ἲ⏝ࡍࡿᢠཎࡢ᭱㐺ྥࡅࡓ᳨ウ 㸦ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠ࠕࢲࢽ፹ᛶ ⣽⳦ឤᰁࡢデ᩿࣭⒪యไᵓ⠏ࡑࡢᇶ ┙࡞ࡿᢏ⾡࣭ሗࡢయ⣔㛵ࡍࡿ◊ ✲ࠖ㸧 㸦 ᖹ ᡂ 2 4 ᖺ ᗘ 㹼 ᖹ ᡂ 2 6 ᖺ ᗘ 㸧 ࣒ࣥࣉࢫ࢘ࣝࢫࡢὶ⾜ゎᯒ࡞ࡽࡧཎ ᛶⓎ⌧ࡢศᏊᶵᵓゎᯒ㸦ཌ⏕ປാ⛉Ꮫ◊✲ ㈝⿵ຓ㔠ࠕ࣡ࢡࢳࣥࡼࡾண㜵ྍ⬟࡞ᝈ ᑐࡍࡿண㜵᥋✀ࡢ⛉Ꮫⓗ᰿ᣐࡢ☜❧ཬࡧ ᑐ⟇ࡢྥୖ㛵ࡍࡿ◊✲ࠖ㸧 㸦 ᖹ ᡂ 24 ᖺ ᗘ 㹼 ᖹ ᡂ 26 ᖺ ᗘ 㸧 ◊✲ᴫせ ᡃࡀᅜ࡛ࡣ MMR ࣡ࢡࢳࣥࡢ୰Ṇ௨㝆࣒ࣥࣉ - 16 - 秋田県健康環境センター年報 第10号 2014 ࢫ࣡ࢡࢳࣥࡢ᥋✀⋡ࡀపୗࡋ㸪࠾ࡓࡩࡃࡐࡢ ࡁࡿࡶࡢࡢ㸪་⸆ရ࡛࠶ࡿࡇࡽᵝࠎ࡞ไ⣙ ὶ⾜ࡣ࠸ࡲࡔไᚚ࡛ࡁ࡚࠸࡞࠸ࠋࡇ࠺ࡋࡓ⌧ ࡀ࠶ࡿࠋᮏ◊✲࡛ࡣ㸪ไ⣙ࡢ࡞࠸ᤕᤊᢠయ౪⤥ ≧ࢆ㋃ࡲ࠼㸪࣒ࣥࣉࢫ࣡ࢡࢳࣥࡢᐃᮇ᥋✀ࡢ ※ࡘ࠸᳨࡚ウࡋ㸪㣗ရࡢ࢘ࣝࢫ᳨ᰝࡢᬑཬ ♫ⓗࢽ࣮ࢬࡀ㧗ࡲࡗ࡚࠸ࡿࠋ୍᪉㸪Ḣ⡿ඛ㐍 ᙺ❧࡚ࡿࡇࢆ┠ⓗࡍࡿࠋ ᅜ࠾࠸࡚ࡣ MMR ࣡ࢡࢳࣥ 2 ᅇ᥋✀ࡢᬑཬ ⤖ᯝ ࡶࢃࡽࡎ㸪࠾ࡓࡩࡃࡐࡢࡓࡧ㔜࡞ࡿ࢘ ࢺࣈࣞࢡࡀၥ㢟࡞ࡗ࡚࠸ࡿࠋ ࣀࣟ࢘ࣝࢫ㸦NoV㸧㸪ࢧ࣏࢘ࣝࢫ㸦SaV㸧㸪 A ᆺ⫢⅖࢘ࣝࢫ㸦HAV㸧㸪ࢹࣀ࢘ࣝࢫ 41 ᚋ࣡ࢡࢳࣥࡢᑟධࡼࡗ࡚࠾ࡓࡩࡃࡐ ᆺ㸦AdV41㸧ࡘ࠸࡚㣗ရὙᾮࡽࡢᅇࢆ ὶ⾜ࡢไᚚࢆ☜ᐇ࡞ࡶࡢࡍࡿࡓࡵࡣ㸪࣒ࣥ ヨࡳࡓࡇࢁ㸪㣗୰ẘࡢཎᅉ࢘ࣝࢫࡋ࡚᭱ ࣉࢫ࢘ࣝࢫࡢᅜෆ࠾ࡅࡿὶ⾜ືែࡢᢕᥱ㸪 ከࡢ NoV-GII 4 ᑐࡋ࡚ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜ ᢠཎᛶࡢゎᯒ㸪ཎᛶⓎ⌧ᶵᵓࡢゎ᫂࡞㸪࠸ ࣥࢆ⏝࠸ࡓሙྜࡢᅇ⋡ࡣ 52.0㸣࡛㸪་⒪⏝࢞ ࡃࡘࡢゎỴࡍࡁㄢ㢟ࡀ࠶ࡿࠋ࡞࡛ࡶ㸪ᅜ ࣐ࣥࢢࣟࣈࣜࣥ〇ࢆ⏝࠸ࡓሙྜࡣ 46.8㸣࡛࠶ ෆ࡛ࡢὶ⾜ືែࢆ⥙⨶ⓗ࣭ᣢ⥆ⓗ࡞ศᏊᏛࢹ ࡗࡓࠋ௨ୗ㸪NoV-GII 2 ᑐࡋ࡚ࡣ 38.6㸣/38.2㸣 ࣮ࢱࡢ㞟✚ࡀ㸪࣡ࢡࢳࣥຠᯝࢆⓗ☜ホ౯ࡋ⾜ 㸦ᕤᴗ⏝/་⒪⏝㸧㸪NoV-GII 6: 15.5㸣/13.3㸣㸪 ᨻୖࡢ⟇ᫎࡉࡏࡿୖ࡛ᚲ㡲࡛࠶ࡿࠋࡋ NoV-GI 4: 35.7 㸣 /37.6 㸣 㸪 NoV-GI 6: 12.1 㸣 ࡋ࡞ࡀࡽ㸪ࡑࢀࢆᐇ⌧ࡍࡿࡓࡵࡢᅜⓗ࡞ࢧ࣮ /6.1 㸣 㸪 SaV-GI 1: 24.3 㸣 /23.7 㸣 㸪 SaV-GII 3: ࣋ࣛࣥࢫࡢࢿࢵࢺ࣮࣡ࢡࢩࢫࢸ࣒ࡣ࠸ࡲࡔ↓ 18.1 㸣 /17.0 㸣 㸪 SaV-GIV 1: 29.7 㸣 /25.0 㸣 㸪 ࡃ㸪ࡑࡢᵓ⠏ࡣᛴົ࡞ࡗ࡚࠸ࡿࠋࡑࡇ࡛㸪ࢧ SaV-GV 1: 29.5㸣/24.8㸣㸪HAV: 17.8㸣/13.1㸣㸪 ࣮࣋ࣛࣥࢫࡢᇶ♏࡞ࡿࡁ㝔ࡸᆅ᪉⾨⏕ AdV41: 33.0㸣/32.7㸣࡛࠶ࡾ㸪ẚ㍑ࡋࡓ࡚࠾ ◊✲ᡤᅜ❧ឤᰁ◊ࢆ⧅ࡄࢿࢵࢺ࣮࣡ࢡࢆᵓ ࠸࡚୧⪅ࡣྠ➼࡛࠶ࡗࡓࠋ ⠏ࡋ㸪ࡑࢀࡼࡗ࡚࣒ࣥࣉࢫ࢘ࣝࢫࡢᅜෆὶ ࡲࡓ㸪ᐇ㝿ࡢởᰁ㣗ရࢆࣔࢹࣝࡋࡓẚ㍑ヨ ⾜≧ἣࡢᴫせࢆᢕᥱࡍࡿࡇࢆ┠ᣦࡋࡓࠋ 㦂࠾࠸࡚ࡶ㸪ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࢆ⏝࠸ ⤖ᯝ ࡓ ⣔ ࡣ ࣏ ࢸ ࢺ ࢧ ࣛ ࢲ ࡛ 40.6 㸣 㸪 ↝ ࡁ ࡑ ࡤ ࡛ ᖹᡂ 26 ᖺᗘᙜࢭࣥࢱ࣮࠾࠸᳨࡚ฟࡉࢀ 33.5 㸣 㸪 ་ ⒪ ⏝ ࡢ ࡑ ࢀ 㸦 ࣏ ࢸ ࢺ ࢧ ࣛ ࢲ ࡛ ࡓ࣒ࣥࣉࢫ࢘ࣝࢫ 9 ᰴࡘ࠸࡚㑇ఏᏊゎᯒࢆ 34.7㸣㸪↝ࡁࡑࡤ࡛ 32.4㸣㸧ྠ➼௨ୖࡢᅇ ⾜ࡗࡓࡇࢁ㸪࡚ Gw ᆺ࡛࠶ࡗࡓࠋ◊✲⌜ ⋡ࢆ♧ࡋࡓࠋࡇࢀࡽࡢࡇࡽ㸪⏝࠾࠸࡚ ᡤᒓࡍࡿ 20 ࣨᡤᆅ᪉⾨⏕◊✲ᡤయ࡛ 27 ᰴࡢ ≉ẁࡢไ⣙ࡢ↓࠸ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࢆ ゎᯒࢆ⾜ࡗࡓࡀ࡚ Gw ᆺ࡛࠶ࡗࡓࠋᖹᡂ 25 ヨ⸆ࡋ࡚ࣃࣥࢺࣛἲᑟධࡍࡿࡇࡣኚ ᖺᗘࡣᮾ᪥ᮏࡣ Ge ᆺ㸪す᪥ᮏࡣ Gw ᆺศ ྜ⌮ⓗ࡛࠶ࡾ㸪ỗ⏝ᛶࢆᢸಖࡍࡿព࡛ࡶ✚ᴟ ࡉࢀ࡚࠸ࡓࡀ㸪ᖹᡂ 26 ᖺᗘࡣ Gw ᆺࡢࡳࡀ᳨ฟ ⓗ࡞ά⏝ὶ㏻࣮ࣝࢺࡢᩚഛࡀᮃࡲࢀࡿࡶࡢ ࡉࢀࡓࠋ ⪃࠼ࡽࢀࡓࠋ 㣗ရࡢ࢘ࣝࢫ᳨ᰝ࠾ࡅࡿᤕᤊᢠయࡢ౪ ⤥※㛵ࡍࡿ◊✲㸦ྠ⏕ཌ⏕ᴗᅋࠕᆅ ᇦಖ⚟♴◊✲ຓᡂࠖຓᡂᴗ㸧 ஙඣႍᜥ㸪Ẽ⟶ᨭႍᜥࡢ࢘ࣝࢫ࣭⣽⳦ឤ ᰁ≧ἣࡢᢕᥱ㸦ᕷ❧⛅⏣⥲ྜ㝔ࡢඹྠ ◊✲㸧 㸦 ᖹ ᡂ 2 5 ᖺ ᗘ 㹼 ᖹ ᡂ 2 6 ᖺ ᗘ 㸧 㸦 ᖹ ᡂ 26 ᖺ ᗘ 㹼 ᖹ ᡂ 27 ᖺ ᗘ 㸧 ◊✲ᴫせ ஙඣႍᜥࡸẼ⟶ᨭႍᜥࡢᝈ⪅ࡽ᳨ฟࡉࢀ ◊✲ᴫせ 㣗ရ᳨య╔ࡋࡓ࢘ࣝࢫࡢ᳨ฟἲࡋ ࡿཎయ㸪⮫ᗋ⤒㐣ࡸணᚋࡢ㛵㐃ࢆㄪᰝࡋ㸪 ࡚㸪⏦ㄳ⪅ࡽࡼࡗ࡚㛤Ⓨࡉࢀࡓࣃࣥࢯࣝࣅ ⒪ᙺ❧࡚ࡿࡇࢆ┠ⓗࡍࡿࠋ⮫ᗋࢹ࣮ࢱ ࣭ࣥࢺࣛࢵࣉἲࡣ㸪㯤Ⰽࣈࢻ࢘⌫⳦ࡢ⾲㠃࢘ ྲྀᚓ⣽⳦᳨ᰝࡣᕷ❧⛅⏣⥲ྜ㝔ࡀ⾜࠸㸪ᙜ ࣝࢫ⢏Ꮚᢠయࡢ」ྜయࢆ྾╔ࡉࡏ࡚ᅇ ࢭࣥࢱ࣮࡛ࡣ࢘ࣝࢫࡢ᳨ฟࢆ⾜ࡗࡓࠋ ࡍࡿࡇࡀᇶᮏཎ⌮࡛࠶ࡿࠋᤕᤊᢠయࡢ౪⤥※ ⤖ᯝ ᇶ♏ᝈẼ⟶ᨭႍᜥࢆᣢࡘᑠඣ྾ჾឤ ࡋ࡚㸪་⒪⏝࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࡀ⏝࡛ - 17 - 秋田県健康環境センター年報 第10号 2014 ᰁᝈ⪅ࡽ᥇ྲྀࡋࡓ 91 ᳨య㸦᳨య᥇ ྲྀᮇ㛫㸸 ⤖ᯝ ᖹᡂ 25 ᖺ 12 ᭶㹼ᖹᡂ 27 ᖺ 2 ᭶㸧ࢆᑐ㇟㸪9 ㎰⸆ࡢ୍ᩧศᯒἲࡘ࠸࡚㸪ࢡ࢚ࢵࢳ࣮ࣕࢬ ✀ࡢཎయࡘ࠸᳨࡚ฟࢆヨࡳࡓࠋ⤖ᯝ㸪65 ᳨ ἲ ࢆ ᇶ ᪂ つ ศ ᯒ ἲ ࢆ ᳨ ウ ࡋ ࡓ ࠋ GC ཬ ࡧ యࡽఱࡽࡢཎయࡀ᳨ฟࡉࢀ㸪᳨ฟ⋡ࡣ LC-MS/MS ࡢྛ ᐃ⏝ࡢ 2 ἲࢆᵓ⠏ࡋ㸦௨ୗ GC 71.4㸣࡛࠶ࡗࡓࠋᮏ┴ࡢឤᰁⓎ⏕ືྥㄪᰝ ἲ㸪LC ἲࡍࡿ㸧㸪GC ἲ 275 㡯┠㸪LC ἲ 161 ࠾ࡅࡿ྾ჾ⏤᮶᳨యࡽࡢཎయ᳨ฟ⋡ 㡯┠ࡘ࠸࡚㸪୍ᚊᇶ‽ࣞ࣋ࣝࡢ ᐃࡀྍ⬟ 㸦p.25㹼p.28 ཧ↷㸧ẚ㍑ࡋ㸪ᅇ᳨ウᑐ㇟ ࡞ࡗࡓࠋ ࡋࡓᝈ⪅⩌ࡽࡢ᳨ฟ⋡ࡣ㧗⋡࡛࠶ࡾ㸪Ẽ⟶ᨭ ᪂つศᯒἲࡢጇᙜᛶホ౯ヨ㦂ࢆ㸪ཌ⏕ປാ┬ ႍᜥᝈ⪅࠾ࡅࡿ᫆ឤᰁᛶࡀ♧၀ࡉࢀࡓࠋ᳨ฟ ㏻▱ࡢ࢞ࢻࣛࣥᚑ࠸㸪8 㣗ရ㸦ࡾࢇࡈ㸪 ࡉࢀࡓཎయࡢෆヂࡣ㸪RS ࢘ࣝࢫ 39 ᳨య㸪 ࠺ࢀࢇࡑ࠺㸪⋞⡿㸪ࡡࡂ㸪ࡤࢀ࠸ࡋࡻ㸪࢟ࣕ ࣛࣀ࢘ࣝࢫ 18 ᳨య㸪࣎࢝࢘ࣝࢫ 5 ᳨య㸪 ࣋ ࢶ 㸪 ࣞ ࣔ ࣥ 㸪 ㇋ 㸧 ࢆ ᑐ ㇟ ᐇ ࡋ ࡓ ࠋ LC ࣃࣛࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫ 5 ᳨య㸦1 ᆺ 1 ᳨ ἲࡢࡡࡂ㸦6 㸧ࢆ㝖ࡁ㸪ྛἲ࡛⣙ 8 ࡢ㡯┠ య㸪2 ᆺ 3 ᳨య㸪3 ᆺ 1 ᳨య㸧㸪ࣄࢺࢥࣟࢼ࢘ ࡀホ౯ᇶ‽㐺ྜࡋࡓࠋ ࣝࢫ 1 ᳨య࡛࠶ࡗࡓ㸦㔜ឤᰁࢆྵࡴ㸧ࠋ LC ἲࡢࡡࡂࡣ㸪┿ᗘࡀホ౯ᇶ‽ࢆୗᅇࡾ㸪 ࡡ࡚ࡽẼ⟶ᨭႍᜥࡢ㛵㐃ࡀᣦࡉࢀ࡚࠸ 㐺ྜ࡞ࡗࡓࡶࡢࡀከࡗࡓࠋᑐ⟇ࡋ࡚⢭ ࡓ RS ࢘ࣝࢫ࠾ࡼࡧࣛࣀ࢘ࣝࢫࡢ᳨ฟࡀ 〇㔞ࢆ༙㔞ࡋ⢭〇ᗘྜࢆ㧗ࡵࡓࡇࢁ㸪ᅇ ✺ฟࡋ࡚ከࡗࡓࡇࡽ㸪ᨵࡵ࡚Ẽ⟶ᨭႍᜥ ⋡ࡀୖࡀࡾᨵၿࡍࡿࡇࡀ࡛ࡁࡓࠋ ᝈ⪅ᑐࡍࡿ୧࢘ࣝࢫࡢᙳ㡪ࡀ☜ㄆࡉࢀࡓࠋ ຍᕤ 㣗ရ 㸦෭ 㣯 Ꮚ㸪 ෭ ࢢ ࣮ࣜ ࣥࣆ ࣮ࢫ 㸪 㔝⳯ࢪ࣮ࣗࢫ㸧 GC ἲཬࡧ LC ἲࢆ㐺⏝ࡋ㸪 ጇᙜᛶホ౯ヨ㦂ྠࡌ᮲௳࡛ῧຍᅇヨ㦂ࢆ ⌮Ꮫ⌜ ⾜ࡗࡓࠋᅇ⋡㸪⢭ᗘࡶⰋዲ࡞⤖ᯝࡀᚓࡽ ࢀ㸪ᮏἲࡣຍᕤ㣗ရࡶ᭷⏝࡛࠶ࡗࡓࠋ 㣗ရ୰ࡢṧ␃㎰⸆ཬࡧṧ␃ື≀⏝་⸆ရࡢ ୍ᩧศᯒἲ㛵ࡍࡿ◊✲ ᳨ウࡋ㸪ࢭࢺࢽࢺࣜࣝ 0.1%ࢠ㓟Ỉࡢΰᾮ 㸦 ᖹ ᡂ 24 ᖺ ᗘ 㹼 ᖹ ᡂ 26 ᖺ ᗘ 㸧 ࡼࡿᢳฟཬࡧ㝖ࢱࣥࣃࢡ᧯సࢆ⾜࠸㸪C18 ᅛ┦ ື≀⏝་⸆ရ 115 㡯┠ࡢ୍ᩧศᯒἲࡘ࠸࡚ ࣒࡛࢝ࣛ⬺⬡ࡋࡓᚋ㸪LC-MS/MS ࡛ ᐃࡍࡿ᪉ ἲࢆᵓ⠏ࡋࡓࠋ7 㣗ရ㸦㭜⫗㸪∵⫗㸪㇜⫗㸪ࢧ ◊✲ᴫせ 㣗ရ୰ࡢṧ␃㎰⸆ཬࡧື≀⏝་⸆ရࡢṧ␃ᇶ ࢣ㸪㭜༸㸪∵ங㸪ࡣࡕࡳࡘ㸧ࢆᑐ㇟ጇᙜᛶホ ‽್ࡼࡿつไࡣ㸪࣏ࢪࢸࣈࣜࢫࢺ᪉ᘧࡀ᥇ࡽ ౯ヨ㦂ࢆᐇࡋࡓ⤖ᯝ㸪࡚ࡢ㣗ရ࡛⣙ 8 ௨ ࢀ࡚࠾ࡾ㸪ᙜࢭࣥࢱ࣮࡛ࡣ㸪⌧ᅾ㸪㎰⸆㸪ື≀⏝ ୖࡀホ౯ᇶ‽㐺ྜࡋࡓࠋ ་⸆ရࢆྜࢃࡏ㸪⣙ 240 㡯┠ࢆ GC ཬࡧ LC-MS/MS ࢆ⏝࠸࡚ศᯒࡋ࡚࠸ࡿࠋ ࢪ࢜࢟ࢧࣥศゎ⳦ࡢᇵ㣴㛗ᮇಖᏑ᮲ ௳ࡢ᳨ウ 㸦 ᖹ ᡂ 2 5 ᖺ ᗘ 㹼 ᖹ ᡂ 2 7 ᖺ ᗘ 㸧 ࡢ㣗ရࡢከᵝక࠸㸪⏝ࡉࢀࡿ㎰⸆➼ ࡢ✀㢮ࡀቑ࠼㸪ຍᕤ㣗ရࡢ᳨ᰝࡢᚲせᛶࡶ㧗ࡲࡗ ࡚࠸ࡿࡇࡽ㸪᭦࡞ࡿ᳨ᰝ㡯┠ࡢᣑ㸪ከᵝ ࡞㣗ရᑐᛂ࡛ࡁࡿ㎿㏿࡛㧗⢭ᗘ࡞ศᯒἲࡢ㛤 ◊✲ᴫせ ᗫỈࡽࡢ㝖ཤࡀ㞴ࡋ࠸ 1,4-ࢪ࢜࢟ࢧࣥࡘ Ⓨࡀᛴࡀࢀࡿࠋ ࡲࡓ㸪㣗ရ⾨⏕ἲࡢつ᱁ᇶ‽ࡢ㐺ྜࢆุ᩿ࡍ ࠸࡚㸪άᛶởἾ⏕≀ࡼࡿ࡞ฎ⌮᪉ᘧࡢᵓ ࡿࡓࡵ⏝࠸ࡿศᯒἲࡘ࠸࡚ࡣ㸪ཌ⏕ປാ┬ࡀ ⠏ࡀᚲせࡉࢀ࡚࠸ࡿࠋᮏ◊✲࡛ࡣ㸪≉␗ⓗ࡞ ㏻▱ࡋࡓ࢞ࢻࣛࣥᚑ࠸ጇᙜᛶホ౯ヨ㦂ࢆ ฎ⌮⬟ຊࢆ᭷ࡍࡿ 1,4-ࢪ࢜࢟ࢧࣥศゎ⳦ࡘ࠸ ⾜࠺ࡇࡀᚲ㡲࡞ࡗࡓࠋ ࡚ࡢ᭱㐺࡞ᇵ㣴᮲௳ࡢ᳨ウࢆ⾜࠸㸪⳦ࡗ࡚ ᮏ◊✲࡛ࡣ㸪᳨ᰝ㡯┠ᩘࡢᣑࢆᅗࡾ㸪ຍᕤ㣗 ࡢ᭱㐺࡞ ᗘࡸᚲせࡍࡿᰤ㣴※㸪㜼ᐖせᅉ࡞ ရࡶᛂ⏝࡛ࡁࡿ㎿㏿ࡘ㧗⢭ᗘ࡞ศᯒἲࡘ ࢆホ౯㸪⪃ᐹࡋ㸪ᐇ㝿ࡢᗫỈฎ⌮タࡢ⟶⌮ ࠸᳨࡚ウࡍࡿࠋࡲࡓ㸪᳨ウࡋࡓศᯒἲࡢጇᙜᛶホ ࡢࡓࡵࡢᣦᶆࢹ࣮ࢱࡍࡿࠋࡲࡓ㸪⳦ࡢ㛗ᮇಖ ౯ヨ㦂ࢆ⾜࠸㸪ࡼࡾಙ㢗ᛶࡢ㧗࠸᪉ἲࢆ☜❧ࡍࡿࠋ Ꮡ᪉ἲࢆᵓ⠏ࡍࡿࡇࡼࡾ✀ࡢ⥔ᣢࢆᅗࡾ㸪 - 18 - 㐠㌿᪉ἲࡸ⎔ቃࡢኚࡼࡾ⳦ࡀṚ⁛ࡍࡿ➼ ࢆᘬࡁ㉳ࡇࡍࡇࢆᐇ㦂ⓗ᫂ࡽࡋ࡚ࡁ ࡢ ࡢែഛ࠼ࡿࠋ ࡓࠋᖹᡂ 22 ᖺᗘࡽࡢ◊✲࡛ࡣ㸪ࡑࢀࡽࡢᑐ ⤖ᯝ ⟇ࡢᡭ❧࡚ࢆ᳨ウࡋ㸪ᮍฎ⌮ࡢᙉ㓟ᡂศࢆฎ⌮ ⏕≀ฎ⌮ᵴࡽ᥇ྲྀࡋࡓάᛶởἾࡘ࠸࡚㸪 ࡍࡿࡓࡵࡢ୰ᢏ⾡୍ᐃࡢ┠ฎࡀ❧ࡗࡓࠋࡑ ෭ࡼࡿ㛗ᮇಖᏑ᪉ἲࡢ᳨ウࢆ⾜ࡗࡓࠋ෭ ࡢᡂᯝࢆཷࡅ㸪ᮏ◊✲ㄢ㢟࡛ࡣ㸪pH ไᚚྍ⬟࡞ ๓ࡢヨᩱㄪ〇ࡣ㸪άᛶởἾヨᩱ 500 ml ࢆ㐲ᚰᶵ ୰ࡼࡾ㸪ୖグᮍฎ⌮ࡢᙉ㓟ᡂศࢆຠ⋡Ⰻࡃ ࡼࡾ ⬺Ỉᚋ 㸪 0㸪30㸪50㸣ࢢࣜࢭ ࣜࣥ ⁐ᾮ࡛ ᙅࡵࡿᢏ⾡ࢆ☜❧ࡍࡿࠋࡇࡢ୰క࠸㸪⏣ἑ ⨨ࡋ㸪ᗘ⬺Ỉࡍࡿࡇ࡛⾜࠸㸪-85Υࡢ෭ †ࡲ࡛ࡢὶᇦ࡛⏕ࡌ࡚࠸ࡿ※Ἠ୰ྵࡲࢀࡿ ᗜಖᏑࡋࡓࠋಖᏑᚋࡢศゎ⬟ヨ㦂ࡣ㸪άᛶở 㕲ࡸ࣑ࣝࢽ࣒࢘ࡢຍỈศゎᣲືࢆㄪᰝࡍࡿ Ἶヨᩱࢆ 1,4-ࢪ࢜࢟ࢧࣥ 2 mg/L㸪500 ml ⁐ᾮ ࡶ㸪⌧⾜୰ࡼࡿຍỈศゎ࢚ࣜࡢୖ ࡋ㸪ዲẼ᮲௳ୗ࡛᧠ᢾ㸪24 㛫ᚋࡢ 1,4-ࢪ࢜࢟ ὶഃࡢಁ㐍ࡘ࠸࡚ࡶేࡏ᳨࡚ウࡍࡿࠋ ࢧࣥ⃰ᗘࡼࡾῶᑡ㏿ᗘࢆ⟬ฟࡋ㸪ホ౯ࢆ⾜ࡗ ⤖ᯝ ࡓࠋ෭ಖᏑ 6 ࣨ᭶ᚋࡢῶᑡ㏿ᗘࡣ㸪ࢢࣜࢭࣜ ᖹᡂ 26 ᖺᗘࡣ㸪୰ฎ⌮タỈࡀᨺὶࡉࢀ ࣥ⃰ᗘ 0㸪30㸪50㸣ࡍ࡚࠾࠸࡚㸪෭๓ࡢ ࡿ⋢ᕝᨭὶࡢ㯮ᕝࡢྜὶࡍࡿᅗࡢᆅⅬյ ῶᑡ㏿ᗘࡰྠࡌ࡛࠶ࡾ㸪ᐜ᫆ಖᏑྍ⬟࡞ ࠾࠸࡚㸪MO 㓟ᗘҸ0㸦pH4.8 ௨ୖ㸧ࢆ┠ᣦࡋࡓ ࡇࢆ♧ࡍ⤖ᯝ࡞ࡗࡓࠋ෭ಖᏑヨ㦂ࡣ⥅⥆ ᐇドヨ㦂ࢆ 10 ᭶ 2 ᪥ᐇࡋ㸪ᅗࡢᆅⅬձ㹼 ୰࡛࠶ࡾ㸪ࡼࡾ㛗ᮇⓗ࡞ಖᏑ࡛ࡢάᛶ⥔ᣢࢆ☜ ո࠾࠸࡚ㄪᰝࢆ⾜ࡗࡓࠋୖグᮍฎ⌮ᙉ㓟ᡂศ ㄆࡋ࡚࠸ࡃࠋ 㸦pH ⣙ 2㸪ὶ㔞 20,000 L/min㸧ཬࡧ୰ฎ⌮Ỉ άᛶởἾ୰ࡢ 1,4-ࢪ࢜࢟ࢧࣥศゎ⳦ࢆᢕᥱࡍ ୰ࡢᮍฎ⌮㓟ᛶᡂศࢆ୰ࡍࡿࡓࡵ㸪ࣝ࢝ ࡿࡓࡵ㸪ḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ࡼࡾ㑇ఏᏊゎ ࣜᮦࡋ࡚ங≧ࡋࡓ㓟࢝ࣝࢩ࣒࢘㸦CaO㸧 ᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝ㸪 1,4-ࢪ࢜࢟ࢧࣥศゎ⳦ ࢆմࡢୖὶ㒊 4 㛫㐃⥆ᢞධࡋࡓ㸦ᢞධ㔞 ࡋ࡚▱ࡽࢀ࡚࠸ࡿ Mycobacterium ᒓࡸ Afipia ᒓ 300 kg/h㸧 ࠋ ➼ࡀ᳨ฟࡉࢀࡓࠋᚋࡶ㑇ఏᏊゎᯒࢆ⥅⥆ࡋ࡚ ⾜࠸㸪ศゎ⳦ࡢ༢㞳࣭ᇵ㣴ࡼࡾᛶ㉁ࢆ᥈ࡿண ᐃ࡛࠶ࡿࠋ ⎔ቃಖ⌜ ⋢ᕝୖὶ㒊࠾ࡅࡿ୰ᑐ⟇ࡢຠ⋡ S+ ࢆྵࡴỈ㉁ᨵၿࡢಁ㐍 㸦ᖹᡂ 25 ᖺᗘ㹼ᖹᡂ 27 ᖺᗘ㸧 ◊✲ᴫせ ᕷ࠶ࡿ⋢ᕝ Ἠࡢ※Ἠࡣ㸪ᖹᡂ 14 ᖺ ᅗ ⋢ᕝୖὶ㒊ࡢㄪᰝᆅⅬ ௨㝆㸪ᛴ⃭࡞⥲㓟ᗘࡢୖ᪼ࡀࡳࡽࢀࡓࠋࡇࡢኚ ᑐᛂࡍࡿࡓࡵ㸪ᅗࡢ୰ฎ⌮タ࡛ࡣ▼⅊ ୖグࡢ᮲௳࡛୰ฎ⌮ࡍࡿࡇࡼࡗ࡚㸪ࡍ ▼ࢆቑ㔞ࡋ㸪ฎ⌮┠ᶆ pH ࡛࠶ࡿ 3.5 ௨ୖࢆ⥔ᣢ ࡚ࡢㄪᰝᆅⅬ࡛ pH ࡀᨵၿࡋ࡚࠸ࡓࠋࡋࡋ ࡋ࡚୰ฎ⌮ࡋ࡚࠸ࡿࡶࡢࡢ㸪⏣ἑ†ࡢ pH ప ࡞ࡀࡽ㸪ᆅⅬյ࡛ࡣ㸪ヨ㦂๓ pH ⣙ 2.5 ࢆ pH ⣙ ୗࡀほ ࡉࢀ࡚࠸ࡿࠋࡇࡢ⌧㇟ࢆཷࡅ㸪ᙜࢭࣥ 4.4 ࡲ࡛ᨵၿࡍࡿࡇࡣ࡛ࡁࡓࡀ㸪MO 㓟ᗘҸ0 ࢱ࣮࡛ࡣ㸪ᖹᡂ 20 ࡽ 21 ᖺᗘࡢ◊✲ࡼࡗ࡚ ࡞ࡿ pH4.8 ࢆ㐩ᡂࡍࡿࡇࡣ࡛ࡁ࡞ࡗࡓࠋ ୰ฎ⌮タࢆ⤒⏤ࡋ࡞࠸ᙉ㓟ᡂศ୰ฎ ࡲࡓ㸪ᆅⅬո࠾ࡅࡿ 3 㛫ᚋࡢ pH ࡣ⣙ 3.8 ⌮タ࡛ฎ⌮࡛ࡁ࡞࠸㕲㸪࣑ࣝࢽ࣒࢘ࡢᡂศ ࡞ࡾ㸪ᖹᡂ 25 ᖺᗘ 2 ᅇᐇࡋࡓࡢ pH4.1 ࡀὶୗకࡗ࡚ຍỈศゎࡋ㸪H + ࢆᨺฟࡍࡿࡇ ࡣ฿㐩ࡋ࡞ࡗࡓࠋࡇࢀࡽࡢཎᅉࡣᆅⅬղࢆ ࡼࡾ㸪⋢ᕝὶᇦཬࡧ⏣ἑ†࠾ࡅࡿ pH పୗ ὶࢀࡿᕝࡢ⥲㓟ᗘࡀᖹᡂ 25 ᖺᗘẚ㍑ࡋ࡚⣙ - 19 - 秋田県健康環境センター年報 第10号 2014 3 ಸ⛬ᗘቑຍࡋ࡚࠸ࡓࡇࡼࡿ⪃࠼ࡽࢀࡿࠋ ୍᪉㸪⾲♧ࡍࡼ࠺ MO 㓟ᗘࡘ࠸࡚ᖹᡂ 25 ࡽ 26 ᖺᗘࡢ⤖ᯝࢆẚ㍑ࡋࡓࡇࢁ㸪ᆅⅬ յཬࡧոࡢ୧ᆅⅬ࡛ H26 ᖺࡢヨ㦂୰ࡢ MO 㓟ᗘ ࡀప࠸್ࡋ࡚ほ ࡉࢀ࡚࠸ࡿࡇࡽ㸪pH ࡣᫎࡋ࡞࠸㓟ᛶᡂศࡢῶᑡࡀᮏヨ㦂ࡢຠᯝ ࡋ࡚☜ㄆࡉࢀࡓࠋ ⾲ ୰ᐇドヨ㦂࠾ࡅࡿ 02 㓟ᗘࡢ ῶᑡᣲື MO 㓟 ᗘ 㸦mg/L㸧 ㄪᰝᖺᗘ ᖹᡂ 25 ᖺᗘ ᖹᡂ 26 ᖺᗘ ᆅⅬ յ ո յ ո ヨ㦂๓ 155 26 180 26 51 19 26 11 ヨ㦂୰ 3h ᚋ - 20 - ! ! ! ! ි !؟ਬड़ࢂ၅ ! 秋田県健康環境センター年報 第10号 2014 ࢧࣝࣔࢿࣛ⳦ࡢ㎿㏿࡞ྠᐃࡢࡓࡵࡢ᳨ฟἲཬࡧ⾑Ύᆺูἲࡢ☜❧㸦ᖹᡂ 25㹼26 ᖺᗘ㸧 ࢧࣝࣔࢿࣛࡢ 0XOWLSOH[3&5 ἲࡼࡿ⾑Ύᆺูἲࡢ☜❧ 㔝㈗அ 㧘ᶫᚿಖ ᶔᑿᣅᏊ ⇃㇂ඃᏊ ᖹᡂ 25 ᖺᗘࡽࡢㄪᰝ◊✲ᴗ࡛ࡣ㸪ࢧࣝࣔࢿࣛࡼࡿ㣗୰ẘ㸪ឤᰁⓎ⏕ࡢ㝿ࡢ㎿㏿࡞᳨ᰝయไ ࡢᵓ⠏ࢆ┠ⓗ㸪ࢧࣝࣔࢿࣛࡢ㑇ఏᏊゎᯒἲࡼࡿ᳨ฟἲཬࡧ⾑Ύᆺูἲࡢ᳨ウࢆ⾜ࡗࡓࠋᮏ◊✲ ࡼࡾ㸪⾑Ύᆺูࡘ࠸࡚ࡣ Multiplex PCR ἲࢆ⏝࠸࡚㸪せ࡞ᢠཎࡢ㑇ఏᏊࡢ᳨ฟࡀྍ⬟࡞ࡾ㸪㎿ ㏿ࡘṇ☜࡞⾑Ύᆺࡢྠᐃࡀྍ⬟࡞ࡗࡓࠋ ࡣࡌࡵ ࢆΰࡋ㸪≉␗ⓗ࡞จ㞟ᛂࢆ☜ㄆࡋࡓࠋ2 ࡘ ࢧࣝࣔࢿࣛࡣ㸪⣽⳦ᛶࡢ㣗୰ẘཬࡧឤᰁᛶ⫶ ┠ࡢ H ᢠཎࢆㄏᑟࡍࡿࡓࡵ㸪㝧ᛶ࡞ࡗࡓ H ᆺ ⭠⅖ࡢせ࡞ཎᅉ⳦ࡢࡦࡘ࡛࠶ࡿࠋࢧࣝࣔࢿ ࡢ⾑Ύࢆྵࡴ┦ㄏᑟᇵᆅ⳦ࢆ᥋✀ࡋ㸪⳦ࡢ㐠 ࣛࡣ㸪⳦య⾲㠃ࡢ O ᢠཎ 2 ࡘࡢࢇẟᢠཎ ືᛶࢆ☜ㄆࡋࡓࠋ┦ㄏᑟᇵᆅୖࡢ⳦ࢆ BHI ᇵᆅ ࡢ⤌ࡳྜࢃࡏ㸦O:H1:H2㸧ࡼࡗ࡚ 2,500 ✀㢮௨ ᥋✀ࡋ㸪ྠᵝࡋ࡚ 2 ┦┠ࡢ H ᢠཎࢆ≉ᐃࡋࡓࠋ ୖࡢ⾑ΎᆺࡀᏑᅾࡍࡿࠋ୰࡛ࡶ㸪ࣄࢺཎᛶ ࢆ♧ࡍ S.enterica ࡢள✀ I ࡣᅛ᭷ࡢྡ⛠ࡀ 3&5 ἲࡼࡿᢠཎ㑇ఏᏊࡢゎᯒ ⳦ᰴࡽࡢ DNA ᢳฟࡣ㸪ࣝ࢝ࣜຍ⇕ᢳฟ ࡉࢀ࡚࠸ࡿࠋࡑࡢࡓࡵ㸪ࢧࣝࣔࢿࣛ࠾ࡅࡿ⾑ Ύᆺࡣ㸪㔜せ࡞Ꮫⓗᣦᶆ࡞ࡗ࡚࠾ࡾ㸪㣗୰ ἲࡼࡾ⾜ࡗࡓࠋ O ᢠཎࡢ≉ᐃ㸪Franklin ࡽ ẘᑐ⟇ࡸឤᰁࡢࢧ࣮࣋ࣛࣥࢫᙺ❧ࡗ࡚࠸ ࡿࠋ 1) ཬࡧ Hong ࡽ 2) ࡢሗ࿌ࢆࡶ㸪Ex-taq HS㸦Takara㸧ࢆ⏝࠸࡚ ᙜࢭࣥࢱ࣮࠾࠸࡚ࡶ㸪㣗୰ẘࡢ᪩ᮇⓎぢࡸ ⾜ࡗࡓࠋO9 ⩌ࡢ≉ᐃࡣ Hirose ࡽ 3) 㸪O3 ⩌ࡢ ឤᰁࡢⓎ⏕ືྥࢆㄪᰝࡍࡿࡓࡵ㸪ࢧࣝࣔࢿࣛ ≉ᐃࡣ㸪Herrera-León ࡽ㸦2007㸧4) ࡢ primer ࡶ ࡢ⾑ΎᆺูࡼࡿᏛゎᯒࢆᐇࡋ࡚࠸ࡿࠋࡋ ⏝ࡋࡓࠋ H1 ᢠཎࡢ≉ᐃࡣ㸪Hong ࡽ ࡋ࡞ࡀࡽ㸪」ᩘᅇࡢᇵ㣴ࢆక࠺ᚑ᮶ࡢ⾑Ύᆺ 5) 6) 2) 㸪Herrera-León 7) ูἲࡣ㛫ࢆせࡋ㸪㏆ᖺ㸪ᚑ᮶ἲ࡛ࡣ⾑Ύᆺࢆ ࡽ㸦2004㸧 㸪 Levy ࡽ 㸪㔝 ࡢሗ࿌ࢆࡶ ≉ᐃ࡛ࡁ࡞࠸⳦ᰴࡶᩓぢࡉࢀࡿࡼ࠺࡞ࡗ࡚ࡁ Ex-taq HS㸦Takara㸧ࢆ⏝࠸࡚⾜ࡗࡓࠋ ࡓࠋࡑࡇ࡛㸪ᖹᡂ 25 ᖺᗘࡽࡢㄪᰝ◊✲ᴗ࡛ H2 ᢠཎࡢ≉ᐃࡣ㸪Hong ࡽ 2) ཬࡧ Echeita ࡽ 8) ࡣ㸪ࡼࡾ㎿㏿ࡘṇ☜࡞ࢧࣝࣔࢿ᳨ࣛᰝయไࡢ ࡢሗ࿌ࢆࡶ Ex-taq HS㸦Takara㸧ࢆ⏝࠸࡚⾜ ᵓ⠏ࢆ┠ⓗ㸪㑇ఏᏊゎᯒἲࡼࡿࢧࣝࣔࢿࣛ ࡗࡓࠋH2 ࡢ 1 complex ཬࡧ e,n complex ࡢヲ⣽ ࡢ⾑Ύᆺูἲࡢ᳨ウࢆ⾜ࡗ࡚ࡁࡓࠋ ゎᯒࡣ Echeita ࡽ 8) ࡢሗ࿌ࢆࡶ Standard taq ᮏሗ࡛ࡣ㸪ࢧࣝࣔࢿࣛࡢ⾑ΎᆺࢆỴᐃࡋ࡚࠸ 㸦ࢢࣛࢼ࣮㸧ࢆ⏝࠸࡚⾜ࡗࡓࠋ ࡿ 3 ࡘࡢᢠཎࡢྜᡂ㛵ࡍࡿ㑇ఏᏊࢆᑐ㇟ ࡋࡓ Multiplex PCR ἲࡼࡿ⾑Ύᆺูἲࡢ᳨ウ ᮏ◊✲࡛タィ࣭⏝ࡋࡓࣉ࣐࣮ࣛࡣ㸪⾲ 1 ࡢ ࠾ࡾࠋ ⤖ᯝࡑࡢ᭷⏝ᛶࡘ࠸࡚ሗ࿌ࡍࡿࠋ ⤖ᯝ⪃ᐹ ᪉ἲ 3&5 ἲࡼࡿ 2 ⩌ู Multiplex PCR ἲࡼࡾせ࡞ O ᢠཎ࡛࠶ࡿ ⾑Ύᆺู O ⩌ูࡣ㸪⾑ᾮᐮኳᇵᆅୖࡢ⳦ࢆᥙࡁྲྀࡾ㸪 O2 ⩌㸪O3 ⩌㸪O4 ⩌㸪O7 ⩌㸪O8 ⩌㸪O9 ⩌㸪 ࢫࣛࢻ࢞ࣛࢫୖ࡛ྛ O ⩌⾑Ύΰࡋ㸪≉␗ O13 ⩌ࡘ࠸࡚㸪ྠ᳨ฟࡀྍ⬟࡛࠶ࡗࡓ㸦ᅗ ⓗ࡞จ㞟ᛂࢆ☜ㄆࡋࡓࠋ 1A㸧ࠋࡓࡔࡋ㸪O2 ⩌ O9 ⩌ࡣඹ㏻ࡢ㑇ఏᏊࢆ H ᆺูࡣ㸪ቑ⳦ࡋࡓ BHI ᇵ㣴ᾮ 3%࣐࣍ࣝ ᑐ㇟ࡋ࡚࠾ࡾ㸪୧⪅ࡢศ㢮ࡣ Hirose ࡽ 3) ࡢ ࣜࣥຍ⏕⌮㣗ሷỈࢆຍ࠼ࡓᢠཎᾮྛ H ᆺ⾑Ύ O9 ⩌ࢆ᳨ฟࡍࡿ PCR ἲࡀ᭷ຠ⪃࠼ࡽࢀࡓࠋ - 21 - 秋田県健康環境センター年報 第10号 2014 ⾲ ⏝ࡋࡓ 3ULPHU ࡢ୍ぴ Mix Target atgggagcgtttgggttc cgcctctccactaccaacttc tgaaagaatatgtaattgtcagtgg tttcattatctctttgctctatcg atttgcccagttcggtttg ccataaccgacttccatttcc cgtcctataaccgagccaac ctgctttatccctctcaccg gatagcaacgttcggaaattc cccaatagcaataaaccaagc ctgaaaaatggtttagattg accattggatactgtaactg 2 min at 94°C/ 30 cycles (30 sec at 94°C, 30 sec at 55°C, 1 min at 72°C)/ 2 min at 72°C a/b/d a b d F R R R actcaggcttcccgtaacgc gaggccagcaccatcaagtgc gcttcatacagaccatctttagttg ggctagtattgtccttatcgg 2 min at 94°C/ 30 cycles (30 sec at 94°C, 30 sec at 55°C, 1 min at 72°C)/ 5 min at 72°C e,h/i/l,v/z10 e,h i l,v z10 F R R R R F R gcagatcaactctcagaccctggg aacgaaagcgtagcagacaag tagccatctttaccagttccc cctgtcactttcgtggttat gcactggcgttactcaatctc gtgatctgaaatccagcttcaag aagtttcgcactctcgtttttgg 2 min at 94°C/ 30 cycles (30 sec at 94°C, 30 sec at 55°C, 1 min at 72°C)/ 5 min at 72°C F R R R R actcaggcttcccgtaacgc attctgtttcgagtcggaat gatcaccagtaatagcagg gtcagtcgcatcagcaaagctt gagcatctttaacgctggca 2 min at 94°C/ 30 cycles (30 sec at 94°C, 30 sec at 55°C, 1 min at 72°C)/ 5 min at 72°C F R F R F R F R agaaagcgtatgatgtagtaag cagcaatttctgggtctgtaa agaaagcgtatgatgtgaaa attgtggttttagttgcgcc gtggggcaacmctcaatactg cctgccactttcgtggttgc taactggcgatacattgactg tagcaccgaatgatacagcc F R R R R F R F R R cttatgccrataatggtactacactg tttgaccaaykymgcgscatt ggttacagvagccgtaccag ctcctgtacttctgttttggttgta taatcgccatttttgtcgag gtggggcaacmctcaatactg cctgccactttcgtggttgc ggcaacccgacagtaactggcgatac ccatccttaaaggatacggc atcaacggtaacttcatatttg 1 min at 94°C/ 30 cycles (30 sec at 94°C, 20 sec at 58°C, 1 min at 72°C)/ 5 min at 72°C F R F R gaggaagggaaatgaagctttt tagcaaactgtctcccaccatac gaktataaattgtyagatattcttcttc ctragctatggcaatcacgc 2 min at 94°C/ 25 cycles (30 sec at 94°C, 30 sec at 55°C, 30 sec at 72°C)/ 2 min at 72°C F R F R taaagtatatggtgctgatttaacc gttaaaatgacagattgagcagag agccacgaacattccttaactatg gttattaataatacactaatcgccac 2 min at 94°C/ 25 cycles (30 sec at 94°C, 30 sec at 55°C, 30 sec at 72°C)/ 2 min at 72°C O4 O7 O⩌ O8 O3 O13 H1-2 G H1-3 c/r/k/y c r k y z6 1-complex H2c l,w e,n-comlex H2 1,2/5/6/7 1,2 1,5 1,6 1,7 l,w e,n,x/e,n,z15 e,n,x e,n,z15 O9 group PCR conditions F R F R F R F R F R F R O2/O9 H1-1 Primer Sequence (5' to 3') O9/O9,46 O9,46 O3,10/O1,3,19 O3 group O3,10 Fragment size (bp) Reference 624 2) 789 1) 341 2) 397 2) 281 2) 502 1) 423 551 763 200 272 330 422 532 6) 5) 2) 5) 2) 5) same as a/b/d 2 min at 94°C/ 30 cycles (30 sec at 94°C, 30 sec at 55°C, 1 min at 72°C)/ 5 min at 72°C 405 475 529 720 㻣㻕 399 This study 294 2) 241 8) 152 2) 390 99 291 191 241 8) modified from 8) 8) 55 135 - 22 - 614 3) 234 This study 345 4) 492 This study (C) (B) (A) (D) ᅗ 0XOWLSOH[3&5 ἲࡼࡿࢧࣝࣔࢿࣛࡢ⾑Ύᆺูἲ (A) lane M1: 100 bp DNA size ladder, P: Typhimurium (O4), Enteritidis (O9), Agbeni (O13), Hadar (O8), Infantis (O7), Anatum (O3). (B) lane M2: 50 bp DNA size ladder, a: Narashino, b: Paretyphi B, d: Livingstone, e,h: Newport, i: Typhimurium, l,v: Litchfield, z10: Mbandaka, G: Enteritidis, c: Choleraesuis, r: Infantis, k: Thompson, y: Bareilly. (C) P: Weltevreden (z6), Typhimurium (1-complex), Livingstone (l,w), Braenderup (e,n-complex). (D) 1,2: Typhimurium, 1,5: Infantis, 1,6: Anatum, 1,7: Bredeney, l,w: Livingstone, x: Hadar, z15: Braenderup. ࡼࡾ R ᆺኚ␗ࡋ㸪O ᢠཎࡢ≉␗ᛶࢆኻ࠺ࡇࡀ O9 ⩌ࡢ୰ࡣ㸪O9 O9,46 ཬࡧ O9,46,27 ࡀ Ꮡ ᅾ ࡍ ࡿ ࠋ O9 ࡛ ࡣ O ᢠ ཎ ྜ ᡂ 㑇 ఏ Ꮚ ⩌ ୰ ࡢ ࠶ࡿ 9) 10) ࠋPCR ἲࡼࡾ O ⩌ูࡀྍ⬟࡛࠶ࡗࡓ 2 H-rpt 㑇ఏᏊࡀḞᦆࡋ࡚࠸ࡿࡇࡽ 㸪ࡇࡢ㡿 ᰴࡢ࠺ࡕ㸪Sa2730 ࡣ Abony㸦O4:b:e,n,x㸧ࡢᇵ㣴 ᇦࢆᑐ㇟ PCR ἲࢆ⾜࠸㸪୧⪅ࡢ㚷ูࡀྍ⬟࡛ ୰ฟ⌧ࡋࡓ R ᆺ⳦࡛࠶ࡿࡀ㸪PCR ἲ࡛ࡣඖࡢ ࠶ࡿࡇࢆ☜ㄆࡋࡓ㸦ᅗ 2㸧ࠋ ⾑Ύᆺྠ୍࡛࠶ࡿࡇࡀ☜ㄆ࡛ࡁࡓࠋ ࢧ ࣝ ࣔ ࢿ ࣛ ࡢ ⾑ Ύ ᆺ ࡢ ୰ ࡣ 㸪 Typhi ࡸ O3 ⩌ࡢ୰ࡶ㸪O3,10 O1,3,19 ࡀᏑᅾࡍࡿࠋ ᮏ◊✲࡛ࡣ㸪wbaK 㑇ఏᏊࢆᑐ㇟ࡋࡓ PCR ἲ Paratyphi A ࡢࡼ࠺ࢳࣇࢫࡸࣃࣛࢳࣇࢫࡢ ࡼࡾ୧⪅ࡢ㚷ูࡀྍ⬟࡛࠶ࡿࡇࢆ☜ㄆࡋࡓ ཎᅉ࡞ࡾ㔜ࡋࡸࡍ࠸⾑ΎᆺࡶᏑᅾࡍࡿࠋ 㸦ᅗ 2㸧ࠋ Typhi ࡛ࡣ㸪vi ᢠཎࡤࢀࡿⳒ⭷ᵝᢠཎࡢᙳ O ᢠཎࡀ࡛᫂࠶ࡗࡓ 4 ᰴࡘ࠸࡚ PCR ἲ 㡪ࡼࡾ O ⩌ูࡀ㞴ࡋ࠸ሙྜࡶ࠶ࡾ㸪PCR ἲ ࡼࡿ O ⩌ูࢆ⾜ࡗࡓࡇࢁ㸪2 ᰴ࡛ O ⩌ࡀ≉ᐃ ࡼࡿ O ⩌ูࡣ㔜せ࡞⾑Ύᆺࢆ㎿㏿᥎ᐃࡍࡿࡢ ࡛ࡁࡓ㸦⾲ 2㸧ࠋࢧࣝࣔࢿ࡛ࣛࡣᇵ㣴᮲௳➼ ᙺ❧ࡘ⪃࠼ࡽࢀࡿࠋ 3&5 ἲࡼࡿ +㸪+ ᆺู H1 ᢠཎࡘ࠸࡚ࡣ㸪3 ࡘࡢ Multiplex PCR ἲ ࡼࡾ 12 ✀㢮ࡢᆺูࡀྍ⬟࡛࠶ࡗࡓ㸦ᅗ 1B㸧ࠋ ࡓࡔࡋ㸪G-complex ࡘ࠸࡚ࡣ㸪PCR ἲࡼࡿ ளᆺࡢศ㢮ࡣᅔ㞴࡛࠶ࡗࡓࠋ H2 ᢠཎࡘ࠸࡚ࡣ㸪l , w , z6, 1-complex ཬࡧ e,n-complex ࡢ 4 ✀㢮ูࡍࡿ Multiplex PCR ἲຍ࠼㸪1-complex ࡢளᆺ 4 ✀㢮 e,n-complex ࡢளᆺ 2 ✀㢮ࡶᆺูྍ⬟࡞ Multiplex PCR ἲ ࡼࡾ 㸪ྜィ 8 ✀ 㢮ࡢ ᆺ ูࡀ ྍ⬟ ࡛࠶ ࡗࡓ 㸦ᅗ 1C,D㸧ࠋ ࢧࣝࣔࢿࣛࡣ㸪ከࡃࡢሙྜ 2 ┦ᛶࢆ♧ࡋ㸪⾑ ᅗ 2 ཬࡧ 2JURXS ࡢ㚷ู lane M1: 100 bp DNA size ladder, O9: Enteritidis, O9, Ύᆺࡢྠᐃ࠾࠸࡚ࡶ 2 ✀㢮ࡢ H ᢠཎࢆ≉ᐃࡍ 46: Hillington 㸦 ⚄ ዉ ᕝ ┴ ⾨ ⏕ ◊ ✲ ᡤ ࡼ ࡾ ศ 㸧 , ࡿ ᚲ せ ࡀ ࠶ ࡿ ࠋ 2 ࡘ ࡢ ┦ ࡢ ษ ࡾ ᭰ ࠼ ࡣ 㸪 fljAB O3,10: Anatum, O1,3,19: Senftenberg. operon ࡼࡗ࡚ไᚚࡉࢀ࡚࠾ࡾ - 23 - 11) 㸪ᇵ㣴ࡋࡓ⳦ 秋田県健康環境センター年報 第10号 2014 ࡛ࡣ㏻ᖖ㸪ࡕࡽ୍᪉ࡀⓎ⌧ࡋ࡚࠸ࡿࠋ2 ┦ 3㸧Hirose K, Itoh K, Nakajima H, Kurazono T, ┠ࢆ᳨ฟࡍࡿࡓࡵࡣ㸪1 ┦┠᳨࡛ฟࡉࢀࡓ H Yamaguchi M, Moriya K, Ezaki T, Kawamura ᢠཎᑐࡍࡿᢠ⾑Ύࢆྵࡴᇵᆅ୍ᗘ᥋✀ࡋ㸪2 Y, ┦┠ࡢ Ⓨ⌧ࢆ ㄏᑟ ࡍࡿᚲ せࡀ࠶ ࡿࠋᚑ ࡗ࡚ 㸪2 amplification of tyv (rfbE), prt (rfbS), viaB, and ࡘࡢ H ᢠཎࡢᆺࢆỴࡵࡿࡣྜィ 3 ᅇࡢᇵ㣴ࢆ fliC genes by multiplex PCR for identification of ⾜࠺ᚲせࡀ࠶ࡾ㸪ᩘ᪥ࢆせࡍࡿࠋPCR ἲ࡛ࡣ㸪 Salmonella H ᢠཎࢆⓎ⌧ࡉࡏࡿࡓࡵࡢᇵ㣴ࢆ⾜࠺ᚲせࡀ࡞ Paratyphi A, J Clin Microbiol, 40, 2002, ࠸ࡓࡵ㸪㎿㏿࡞ྠᐃࡀྍ⬟࡛࠶ࡿࠋ 633–636. Tamura K, Watanabe enterica H: serovars Selective Typhi and ⛅⏣┴࡛ࡇࢀࡲ࡛ศ㞳ࡉࢀࡓ㠀ᐃᆺⓗ࡞⾑ 4㸧Herrera-León S, Ramiro R, Arroyo M, Diez R, Ύᆺࡢ PCR ἲࡼࡿᆺู⤖ᯝࢆ⾲ 2 ♧ࡍࠋ༢ Usera MA, Echeita MA: Blind comparison of ┦ኚ␗ࡸ H ᢠཎࡢⓎ⌧ࡀ☜ㄆ࡛ࡁ࡞࠸ 7 ᰴࡢ traditional serotyping with three multiplex PCRs ෆ㸪5 ᰴ࠾࠸࡚ PCR ἲࡀ᭷ຠ࡛࠶ࡾ㸪⾑Ύᆺ for the identification of Salmonella serotypes, ࢆ᥎ᐃࡍࡿࡇࡀྍ⬟࡛࠶ࡗࡓࠋᡃࠎࡣ㸪ᖹᡂ Res Microbiol, 158, 2007, 122–127. 22 ᖺ」ᩘศ㞳ࡉࢀࡓ O7:-:1,5 ࡀ PCR ἲࡼ 5 㸧 Herrera-León S, McQuiston JR, Usera MA, ࡾ⾑Ύᆺ Thompson ྠᐃ࡛ࡁࡓࡇࢆሗ࿌ࡋ Fields PI, Garaizar J, Echeita MA. Multiplex 7) ࡚࠸ࡿ ࠋPCR ἲࡼࡿ⾑Ύᆺูࡣ㸪ᚑ᮶ἲ࡛ PCR ᆺู࡛ࡁ࡞࠸⳦ᰴࡘ࠸࡚ࡶ᭷ຠ࡛࠶ࡾ㸪ṇ☜ phase-1 flagellar antigens of Salmonella spp., J for distinguishing the most common ࡞㣗୰ẘཬࡧឤᰁࡢࢧ࣮࣋ࣛࣥࢫࡢ᭷⏝ Clin Microbiol, 42, 2004, 2581–2586. 6㸧Levy H, Diallo S, Tennant SM, Livio S, Sow ᛶࡀ♧၀ࡉࢀࡓࠋ SO, Tapia M, Fields PI, Mikoleit M, Tamboura ⾲ 㠀ᐃᆺ⾑Ύᆺࡢ 3&5 ᆺู⤖ᯝ Antigenic Formulas No. Sa O⩌ H1 H2 1440 O4 i 1558 r 1,5 1584 O13 y 1600 y 1,7 1669 O4 i 1671 O4 1684 O7 k 1924 O3,10 l,v 2165 O7 c 2605 k 1,5 2730 b e,n,x O⩌ NT NT O13 NT NT NT NT NT O4 PCRᆺู H1 H2 NT NT NT NT NT NT NT 1,2 b 1,2 NT 1,5 NT 1,6 NT NT NT NT NT B, Kotloff KL, Lagos R, Nataro JP, Galen JE, ⾑Ύᆺ Levine MM: PCR method to identify Salmonella ᫂ ᫂ ᫂ Yarrabah Typhimurium Paratyphi B Thompson London ᫂ ᫂ Abony enterica serovars Typhi, Paratyphi A, and Paratyphi B among Salmonella isolates from the blood of patients with clinical enteric fever, J Clin Microbiol, 46, 2008, 1861–1866. 7㸧㔝㈗அ: ⛅⏣┴࡛ศ㞳ࡉࢀࡓ㠀ᐃᆺⓗ࡞༢ ┦ᛶࢧࣝࣔࢿࣛ⳦ࡢᆺูἲࡢ☜❧Ⓨ⏕ືྥ ࡢゎ᫂, ᖹᡂ 24 ᖺᗘྠ⏕ཌ⏕ᴗᅋࠕᆅ 㸫 : ᳨ฟࡏࡎ㸪NT : ᳨ᰝࡏࡎ ᇦಖ⚟♴◊✲ຓᡂࠖሗ࿌㞟, 2014, 20–24. 8㸧Echeita MA, Herrera S, Garaizar J, Usera MA: ཧ⪃ᩥ⊩ Multiplex PCR-based detection and identification of the most common Salmonella 1㸧Franklin K, Lingohr EJ, Yoshida C, Anjum M, second-phase flagellar antigens, Res Microbiol, Bodrossy L, Clark CG, Kropinski AM, Karmali 153, 2002, 107–113. 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Bacteriol, 188, 2006, 950–957. - 24 - control mechanism, J 秋田県健康環境センター年報 第10号 2014 ឤᰁⓎ⏕ືྥㄪᰝᴗ ᑠඣ྾ჾᝈ⏤᮶᳨యࡽࡢཎయ᳨ฟ≧ἣ ᰘ⏣ࡕࡦࢁ బ⸨⏤⾰Ꮚ బ⸨ᐶᏊ ⛅㔝⳹Ꮚ ᩪ⸨༤அ ឤᰁⓎ⏕ືྥㄪᰝᴗࡢ୍⎔ࡋ࡚㸪ᙜࢭࣥࢱ࣮࡛ࡣཎయᐃⅬᣦᐃࡉࢀ࡚࠸ࡿ ┴ෆࡢᑠඣ⛉ ་⒪ᶵ㛵 9 タࡽᥦ౪ࡉࢀࡓ᳨యࡘ࠸࡚㸪ཎయ᳨⣴ࢆᐇࡋ࡚࠸ࡿࠋᖹᡂ 26 ᖺᗘࡣᑠඣ྾ჾ ᝈᝈ⪅ࡽ᥇ྲྀࡉࢀࡓဗ㢌ᣔ࠸ᾮ➼ࡢ྾ჾ ⏤᮶᳨య 565 ᳨య୰㸪㔜ឤᰁࢆྵࡵ 260 ᳨యࡽఱ ࡽࡢཎయࡀ᳨ฟࡉࢀࡓ㸦᳨ฟ⋡ 46.0%㸧ࠋ᳨ฟᩘࡣࣛࣀ࢘ࣝࢫ㸦83 ᳨య㸧㸪RS ࢘ࣝࢫ㸦50 ᳨య㸧㸪ࣃࣛࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫ㸦PIV㸧㸦44 ᳨య㸧ࡢ㡰ከࡗࡓࠋPIV ࡣ 7 ᭶㹼9 ᭶᳨ฟᩘ ࡢቑຍࡀㄆࡵࡽࢀ㸪ྠᮇࡣ PIV ࢆཎᅉࡋࡓ㧗㱋⪅タ࠾ࡅࡿ㞟ᅋឤᰁࡶ 2 ሗ࿌ࡉࢀࡓࠋ ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫࡣ┴ෆࡢᝈ⪅Ⓨ⏕ࡀࣆ࣮ࢡࢆྥ࠼ࡓ 1 ᭶㹼2 ᭶᥇ྲྀࡉࢀࡓ᳨యࡽࡣ ࢇ᳨ฟࡉࢀࡎ㸪་⒪⌧ሙ࠾ࡅࡿ㎿㏿デ᩿࢟ࢵࢺ⏝ࡼࡿデ᩿☜ᐃࡢ᭷↓ࡼ ࡗ࡚㸪᳨య᥇ྲྀ ೫ࡾࡀ⏕ࡌ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋRS ࢘ࣝࢫࡘ࠸࡚ࡶྠᵝ㸪ዲⓎᖺ㱋ࡉࢀࡿ 2 ṓᮍ‶ ࡢᖺ㱋⩌ࡢ㛫᳨࡛ฟ⋡ᕪࡣㄆࡵࡽࢀࡎ㸪ಖ㝤㐺⏝ࡼࡿ㎿㏿デ᩿࢟ࢵࢺ⏝ࡢᙳ㡪ࡀ ࠺ࡀ ࢃࢀࡓࠋࡇࡢࡼ࠺࡞≧ἣࢆཷࡅ㸪ᖹᡂ 28 ᖺ 4 ᭶⾜ࡉࢀࡿᨵṇឤᰁἲ࡛ࡣ㸪᳨యᩘࡸ᳨యᥦ౪㢖 ᗘ➼ࡀཝᐦつᐃࡉࢀࡿࡇ࡞ࡗࡓࠋࡇࢀࡼࡾ㸪ࡼࡾ⌧≧ࢆᫎࡋࡓཎయࢧ࣮࣋ࣛࣥࢫࡀྍ ⬟࡞ࡿࡇࡽ㸪ᚋࡶ་⒪ᶵ㛵➼ࡢ㐃ᦠࡢୗ㸪ᴗ᥎㐍ດࡵ࡚࠸ࡁࡓ࠸ࠋ ࡣࡌࡵ ⏤᮶᳨యࡢ࠺ࡕ㸪᥇ྲྀᖺ㱋ࡀ 16 ṓ௨ୖ࠶ࡿ࠸ ᅜෆ࡛Ⓨ⏕࣭ὶ⾜ࡋ࡚࠸ࡿឤᰁࡢᢕᥱࢆ┠ ࡣ྾ჾ≧ࢆ᭷ࡋ࡚࠸࡞࠸ᝈ⪅⏤᮶ࡢ᳨యࢆ ⓗ㸪 56 ᖺ 7 ᭶ࡼࡾឤᰁⓎ⏕ືྥㄪᰝࡀ 㝖እࡋࡓ 565 ᳨యࢆᑐ㇟ࡋࡓࠋ ᐇࡉࢀ࡚࠸ࡿࠋ⌧ᅾࡣࠕឤᰁࡢண㜵ཬࡧឤ ᰁࡢᝈ⪅ᑐࡍࡿ་⒪㛵ࡍࡿἲᚊࠖ㸦ᖹᡂ ᳨ᰝ 10 ᖺ 10 ᭶ᡂ❧㸪ᖹᡂ 11 ᖺ 4 ᭶⾜㸧ᇶ࡙ࡁ㸪 ᑐ㇟ཎయ ឤᰁࡢⓎ⏕ሗࢆṇ☜ᢕᥱ࣭ศᯒࡋ㸪ⓗ☜ ᳨ฟᑐ㇟ࡋࡓཎయࡣḟࡢ 10 ✀࡛࠶ࡿࠋ ሗᥦ౪ࡍࡿࡇ࡛ឤᰁࡢࡲࢇᘏࢆ㜵Ṇࡍ ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫ㸦Flu㸧㸪RS ࢘ࣝࢫ㸪 ࡿࡓࡵ㸪ᅜ࣭ᆅ᪉࣭་⒪ᶵ㛵ࡢ㐃ᦠࡢୗ㸪ᅜ ࣛࣀ࢘ࣝࢫ㸪ࣃࣛࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫ 1) ⓗ⟇ࡋ࡚ᐇࡉࢀ࡚࠸ࡿ ࠋ 㸦PIV㸧㸪ࣄࢺ࣎࢝࢘ࣝࢫ㸪ࣄࢺ࣓ࢱࢽ࣮ࣗ ࡇࡢᴗࡢ୍⎔ࡋ࡚㸪ᐇ㝿ࡢࡼ࠺࡞ ࣔ࢘ࣝࢫ㸦hMPV㸧㸪ࢹࣀ࢘ࣝࢫ㸪࢚ࣥ ཎయࡀὶ⾜㛵ࡋ࡚࠸ࡿࡢࢆ᥈ࡿࡓࡵ 㸪ᙜ ࢸࣟ࢘ࣝࢫ㸪ࣄࢺࢥࣟࢼ࢘ࣝࢫ㸦HCoV㸧㸪 ࢭࣥࢱ࣮࡛ࡣཎయᐃⅬ་⒪ᶵ㛵ᣦᐃࡉࢀ࡚ ⫵⅖࣐ࢥࣉࣛࢬ࣐ ࠸ࡿ┴ෆࡢᑠඣ⛉་⒪ᶵ㛵 9 タࡽᥦ౪ࡉࢀ ࡓ᳨యࡘ࠸࡚㸪ཎయࡢ᳨⣴ࢆᐇࡋ࡚ ࠸ࡿࠋ ᳨ᰝἲ ᥦ౪ࡉࢀࡿ᳨యࡣ྾ჾᝈࢆ⏤᮶ࡋࡓࡶࡢ MagNA Pure LC 2.0 㸦Roche Diagnostics㸧ࢆ⏝ ࡀከࡃ㸪᳨ฟࡉࢀࡿཎయࡣᏘ⠇ࡸᝈ⪅ࡢᖺ ࠸࡚᰾㓟ᢳฟࢆ⾜࠸㸪SLEK ᇵᆅ⁐ฟࡋࡓ᳨ 㱋➼ࡼࡗ࡚≉ᚩࡀ࠶ࡿࠋᅇ㸪ᖹᡂ 26 ᖺᗘࡢ య 200 μL ࡽ 50 μL ࡢ᰾㓟ᢳฟᾮࢆᚓࡓࠋ ᐇ⤖ᯝࡘ࠸࡚ࡲࡵࡓࡢ࡛ሗ࿌ࡍࡿࠋ Flu 2) 㸪RS ࢘ࣝࢫ 3) 㸪ࣛࣀ࢘ࣝࢫ 4) 㸪PIV 3) 㸪 hMPV 5) 㸪 ࢚ ࣥ ࢸ ࣟ ࢘ ࣝ ࢫ 6) 㸪 HCoV 7) ࡣ 㸪 ᪉ἲ LightCycler480 RNA Master Hydrolysis Probes ᑐ㇟᳨య 㸦Roche Diagnostics㸧ྛᩥ⊩グ㍕ࡢ࠶ࡿࣉ ᖹᡂ 26 ᖺ 4 ᭶㹼ᖹᡂ 27 ᭶ 3 ᭶ཎయᐃⅬ ་⒪ᶵ㛵ࡽᥦ౪ࡉࢀࡓဗ㢌ᣔ࠸ᾮ➼ࡢ྾ჾ ࣐࣮࣭ࣛࣉ࣮ࣟࣈࢭࢵࢺࡼࡿ real-time PCR ࢆ⾜ࡗࡓࠋྠᵝ㸪ࣄࢺ࣎࢝࢘ࣝࢫ - 25 - 8) ࣐ - 26 - - 27 - 秋田県健康環境センター年報 第10号 2014 ࡲࡵ Young French Children with Acute Bronchiolitis ᅇ㸪ᖹᡂ 26 ᖺᗘཎయᐃⅬほ ᴗࡋ by Real-Time Reverse Transcriptase PCR and ࡚ᐇࡋࡓ྾ჾឤᰁ᳨యࡽࡢཎయ᳨ฟ Phylogenetic Analysis, J. 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Trompette ་⒪㛵ࡍࡿἲᚊ⾜つ๎ࡢ୍㒊ࢆᨵṇࡍࡿ A., et. al.: Detection of Human Metapneumovirus ┬௧ࡢබᕸཬࡧ୍㒊⾜ࡘ࠸࡚㸦㏻▱㸧㸪 RNA Sequences in Nasopharyngeal Aspirates of ᖹᡂ 27 ᖺ 9 ᭶ 28 ᪥㸬 - 28 - 秋田県健康環境センター年報 第10号 2014 ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠 ᪂ᆺࣥࣇ࢚ࣝࣥࢨ➼᪂⯆࣭⯆ឤᰁ◊✲ᴗ㸦 H24㹼26 ᖺᗘ㸧 ᙉẘᆺ࠾ࡼࡧᙅẘᆺ 2ULHQWLDWVXWVXJDPXVKL ࡢ ࣐࢘ࢫ⾑୰࠾ࡅࡿ᥎⛣ బ⸨ᐶᏊ ᰘ⏣ࡕࡦࢁ ⛅㔝⳹Ꮚ బ⸨ᝋ ᩪ⸨༤அ Ᏻ㒊┿⌮Ꮚ 㰺⸨ᚿಖᏊ ࡘࡘࡀࡢཎయ Orientia tsutsugamushi㸦Ot㸧ࡢせ 6 ᆺࡢ࠺ࡕ㸪Shimokoshi ᆺࡣᙅẘᛶࡉࢀࡿ ࡀ㸪⛅⏣┴࡛ 1992 ᖺ௨㝆Ⓨ⏕ࡋࡓྠᆺࡘࡘࡀᝈ⪅ 16 ࡣ㸪࡚ࡀ㍍ࡣゝ࠼࡞ࡗࡓࠋࡑࡇ ࡛㸪16 ࡢ࠺ࡕᛴᛶᮇࡢ᳨ᰝᡤぢࡀ㧗ᗘ␗ᖖ್ࢆ♧ࡋࡓᝈ⪅⾑ᾮࡽศ㞳ࡉࢀࡓ Shimokoshi ᆺ Matsui ᰴ㸦AB742542㸧ࡘ࠸࡚㸪ICR ࣐࢘ࢫ㸦ࢡ࣮ࣟࢬࢻࢥࣟࢽ࣮㸪චṇᖖࣔࢹࣝ㸧࠾ࡼࡧ ICR ࢾ࣮ࢻ࣐ ࢘ࢫ㸦චࣔࢹࣝ㸧ᑐࡍࡿ⮴Ṛẘᛶࡢ☜ㄆඹࡑࢀࡒࢀࡢ⾑୰ Ot ࢥࣆ࣮ᩘࢆ⤒ⓗᐃ㔞ࡋ ࡓࠋẚ㍑ᑐ㇟ࡣ Kato ᆺ Kakuma-2 ᰴ㸦ᙉẘᆺᰴ㸹AB701788㸧ࢆ⏝ࡋࡓࠋᙉẘᆺ Kakuma-2 ᰴ㸦௨ୗ㸪 ᙉẘᆺᰴ㸧ࢆ᥋✀ࡋࡓ ICR ࣐࢘ࢫ࠾ࡼࡧ ICR ࢾ࣮ࢻ࣐࢘ࢫࡣࡑࢀࡒࢀ᥋✀ᚋ 10㹼11 ᪥┠Ṛஸࡋࡓࠋ Matsui ᰴ࡛ࡣ㸪ICR ࣐࢘ࢫࡣ⏕Ꮡࡋࡓࡀ㸪ICR ࢾ࣮ࢻ࣐࢘ࢫࡣ 16㹼19 ᪥┠Ṛஸࡋࡓࠋ࣐࢘ࢫ⾑୰ࡢ Ot ⃰ᗘࡣ㸪ᙉẘᆺᰴ᥋✀⣔ࡀ Matsui ᰴ᥋✀⣔ࡼࡾࡶቑຍࡀ㢧ⴭ࡛࠶ࡗࡓࠋࡉࡽ ICR ࢾ࣮ࢻ࣐࢘ࢫṚ ஸࡢ⾑୰⃰ᗘࡶᙉẘᆺᰴ᥋✀⣔ࡢ᪉ࡀ㧗࠸ഴྥ࠶ࡗࡓࠋࣄࢺᑐࡋ࡚ẚ㍑ⓗᙉ࠸ཎᛶࢆ♧ࡋࡓ Shimokoshi ᆺ Matsui ᰴࡣ ICR ࣐࢘ࢫᑐࡋ࡚ࡣᙅẘᛶ࡛࠶ࡿࡇࡀ☜ㄆࡉࢀࡓࠋ ⥴ゝ ᪉ἲ ࡘࡘࡀࡢཎయ Orientia tsutsugamushi ࣐ ࢘ ࢫ ᥋ ✀ ࡣ S hi m o ko s h i ᆺ Ma t s ui ᰴ 㸦Ot㸧ࡣ㸪࣐࢘ࢫᑐࡍࡿ⮴Ṛᛶࢆࣔࢹࣝࡋ 㸦ᙅẘᆺ㸸㔜ࡢᝈ⪅⏤᮶㸧ࢆ⏝ࡋࡓࠋᑐ ࡚ࣄࢺᑐࡍࡿཎᛶࡢᙉᙅࡀ⾲⌧ࡉࢀ࡚࠸ࡿ ㇟ࡋ࡚ Kato ᆺ Kakuma-2 ᰴ㸦ᙉẘᆺ㸸㔝㰡⏤ ࡀ㸪ࡇࢀࡣ⾑Ύᆺ࡛࠾࠾ࡼࡑࡀุูࡉࢀ࡚࠸ ᮶㸧ࢆ⏝࠸ࡓࠋ୧ᰴࡣ㸪L929 ⣽⬊࡛ᇵ㣴ᚋ DNA 1) ࡿ ࠋ᪥ᮏᅜෆ࡛Ⓨ⏕ࡍࡿࡘࡘࡀᝈ⪅ࡽศ ᢳฟࡋ㸪real-time PCR4) ࡼࡗ࡚ Ot ࢥࣆ࣮ᩘࢆᐃ 㞳ࡉࢀࡿ Ot ࡢ⾑Ύᆺࡣ 6 ✀࡛㸪ࡇࡢ࠺ࡕ 㔞ࡋࡓࠋࡑࡢᚋྛᰴࢆ PBS ࡛ 1.010 7 copies/ml Shimokoshi ᆺࡣᙅẘᛶࡉࢀ࡚࠸ࡿ 1,2) ࠋࡋࡋ㸪 㸦E7㸧࠾ࡼࡧ 1.010 5 copies/ml㸦E5㸧ᕼ㔘ㄪ ࡇࡢᆺࡢឤᰁ㛵ࡍࡿሗ࿌ࡣᑡ࡞ࡃ㸪ศ㞳 〇ࡋࡓ Ot ⁐ᾮࢆసᡂࡋࡓࠋྛᰴྛ⃰ᗘࡢ Ot ⁐ ࡉࢀࡓᰴࡣ 2014 ᖺ⌧ᅾ㸪ᅜෆ࡛ 2 ᰴࡢࡳ࡛࠶ࡿ ᾮ 100 μl ࢆ ICR ࣐࢘ࢫ ICR ࢾ࣮ࢻ࣐࢘ࢫࡑࢀ 3) ࡒࢀ 2 㢌ࡢ⭡⭍᥋✀ࡋࡓࠋࡑࡢᚋ㸪࣐࢘ࢫࡢ ࠋᮏ┴࡛ࡣࡇࡢᆺࡢᝈ⪅Ⓨ⏕ࡀ 1992 ᖺ௨㝆 3) 2013 ᖺࡲ࡛ 15 㸪2014 ᖺࡢ 1 ࢆྵࡵࡿ እぢ࣭⾜ືほᐹᑿ㟼⬦ࡽࡢ᥇⾑ࢆ 8 ᪥┠ࡲ ᅜෆ᭱ከࡢ 16 ࡀ☜ㄆࡉࢀ࡚࠸ࡿࠋࡇࡢ୰ࡣ ࡛ࡣ 1 ᪥ 1 ᅇ⾜࠸㸪9 ᪥┠௨㝆ࡣ 1㹼2 ᪥ẖ⾜ DIC ࡸ⬻⅖➼ࡢేⓎࡣ࡞࠸ࡶࡢࡢ㸪ࡀ㍍ ࡗࡓࠋ࣐࢘ࢫ⾑ᾮࡢ Ot ࢥࣆ࣮ᩘࡣ㸪ᰴࡢሙྜ ࡣゝ࠼ࡎ㸪᳨ᰝᡤぢࡽࡣ CRP ್ 10.0 mg ྠᵝ DNA ᢳฟᚋᐃ㔞ࡋ㸪ྛ⩌ࡢ⾑ᾮ 1 ml ௨ୖ㸪AST ALT ࡀ 101 IU/L ௨ୖ࡞㸪㔜 ᙜࡓࡾ⟬ࡋࡓ 2 ༉ࡢᖹᆒ್ࢆồࡵࡓࠋ ࡳ࡞ࡋᚓࡿᩘ್ࢆ♧ࡍࡶྵࡲࢀ࡚࠸ ࡓ 3) ⤖ᯝ ࠋᅜෆศ㞳ࡋ࡚ 2 ᰴ┠ࡢ Shimokoshi ᆺᰴ࡛࠶ࡿ Matsui ᰴࡣ㸪㔜ࡽศ㞳ࡉࢀࡓࠋ ,&5 ࣐࢘ࢫࡢ⾑୰ 2W ⃰ᗘ㸦ᅗ 㸧 ྠࡌ⾑Ύᆺ࡛ࡶᰴࡼࡗ࡚ࡣẘᛶᕪࡀ࠶ࡿ ICR ࣐࢘ࢫ࠾ࡅࡿᙉẘᆺᰴ᥋✀⣔࡛ึࡵ࡚ ࠸ࢃࢀ࡚࠸ࡿࡇࡽ㸪ࡇࡢࣄࢺᑐࡋ࡚ᙅẘ Ot ࡀ᳨ฟࡉࢀࡓࡢࡣ㸪E7 ࢆ᥋✀ࡋࡓ⩌㸦E7 ⩌㸧 ࡛ࡣ࡞ࡗࡓᰴࡢ࣐࢘ࢫᑐࡍࡿ⮴Ṛẘᛶࡢ☜ ࡀ 2 ᪥┠㸪E5 ࢆ᥋✀ࡋࡓ⩌㸦E5 ⩌㸧ࡀ 4 ᪥┠࡛㸪 ㄆࢆࡍࡿඹ࣐࢘ࢫࡢ⾑୰ Ot ࢥࣆ࣮ᩘࢆ⤒ ࡑࡢᚋࢥࣆ࣮ᩘࡣቑຍࡋ㸪10 ᪥┠ E7 ⩌ࡀ㸪11 ⓗᐃ㔞ࡋࡓࠋ ᪥┠ E5 ⩌ࡢ࣐࢘ࢫࡀ⾶ᙅື࡞ࡾṚஸࡋ ࡓ ࡓ ࡵ ほ ᐹ ࢆ ⤊ ࡋ ࡓ ࠋ ࡇ ࡢ Ⅼ ࡛ E7 ⩌ ࡢ - 29 - 秋田県健康環境センター年報 第10号 2014 (copies/ml) Kato ᆺ㸦Kakuma-2㸧E7 Katoᆺ㸦Kakuma-2ᰴ㸧E7 Kato ᆺ㸦Kakuma-2㸧E5 1.00E+07 Shimokoshi ᆺ㸦Matsui ᰴ㸧E7 Katoᆺ㸦Kakuma-2ᰴ㸧E5 Shimokoshi ᆺ㸦Matsui ᰴ㸧E5 1.00E+06 1.00E+05 1.00E+04 1.00E+03 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ᅗ ,&5 ࣐࢘ࢫࡢ⾑୰ 2W ⃰ᗘࡢ᥎⛣ (days) Kato ᆺ㸦Kakuma-2㸧E7 Katoᆺ㸦Kakuma-2ᰴ㸧E7 Kato ᆺ㸦Kakuma-2㸧E5 Katoᆺ㸦Kakuma-2ᰴ㸧E5 (copies/ml) Shimokoshi ᆺ㸦Matsui ᰴ㸧E7 Shimokoshiᆺ㸦Matsuiᰴ㸧E7 Shimokoshi ᆺ㸦Matsui ᰴ㸧E5 Shimokoshiᆺ㸦Matsuiᰴ㸧E5 1.00E+07 1.00E+06 1.00E+05 1.00E+04 1.00E+03 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 ᅗ ,&5 ࢾ࣮ࢻ࣐࢘ࢫࡢ⾑୰ 2W ⃰ᗘ ⾑୰ᖹᆒ Ot ࢥࣆ࣮ᩘࡣ 1. 010 5 copies/ml㸪E5 (days) እぢ⾜ືࡁ࡞ኚࡀ࡞࠸ࡲࡲ⏕Ꮡࡋࡓࠋ 5 ⩌ࡣ 1.110 copies/ml ࡛࠶ࡗࡓࠋ ୍᪉㸪Matsui ᰴ᥋✀⣔࡛ึࡵ࡚ Ot ࡀ᳨ฟࡉࢀ ,&5 ࢾ࣮ࢻ࣐࢘ࢫࡢ⾑୰ 2W ⃰ᗘ㸦ᅗ 㸧 ࡓࡢࡣ㸪E7 ⩌ࡀ᥋✀ᚋ 3 ᪥┠㸪E5 ⩌ࡀ 6 ᪥┠࡛ ICR ࢾ࣮ࢻ࣐࢘ࢫ࡛ึࡵ࡚ Ot ࡀ᳨ฟࡉࢀࡓ ࠶ࡗࡓࠋࡑࡢᚋ㸪⾑୰ࢥࣆ࣮ᩘ㢧ⴭ࡞ቑຍࡣ࡞ ࡢࡣ㸪ᙉẘᰴ࡛ࡣ E7 ⩌ࡀ 3 ᪥┠㸪E5 ⩌ࡀ 4 ᪥ ࡃ㸪E7 ⩌ࡣ 7 ᪥┠㸪E5 ⩌ࡣ 11 ᪥┠㸦ᖹᆒ 7.1 ┠࡛㸪ࡑࡢᚋࢥࣆ࣮ᩘࡣ ICR ࣐࢘ࢫࡼࡾࡶ㢧ⴭ 103 copies/ml㸧ࢆ᭱㧗್ࡋ㸪ࡑࢀࡒࢀ 12 ᪥┠௨ ࡞ቑຍࢆ♧ࡋ㸪10 ᪥┠ E7 ⩌ࡀ㸪11 ᪥┠ E5 㝆ࡣ᳨ฟ࡞ࡗࡓࠋ࣐࢘ࢫࡣࡑࡢᚋ 1 ࣨ᭶㛫㸪 ⩌ࡀṚஸࡋࡓࠋࡇࡢⅬ࡛⾑୰ᖹᆒ Ot ࢥࣆ࣮ᩘ - 30 - 秋田県健康環境センター年報 第10号 2014 ࡣࡑࢀࡒࢀ 1.010 6 copies/ml㸪2.410 6 copies/ml ࡍࡿඹ㸪ศࡾ᫆࠸༑ศ࡞ሗᥦ౪ࢆ⥅⥆ ྠᰴࢆ᥋✀ࡋࡓ ICR ࣐࢘ࢫṚஸࡢ 10 ಸ㔞 ࡋ㸪᪩ᮇཷデࢆಁࡍࡇࡀᚲせ⪃࠼ࡽࢀࡓࠋ ࢆ♧ࡋࡓࠋMatsui ᰴ࠾࠸࡚㸪Ot ࡢ᳨ฟࡀึࡵ ཧ⪃ᩥ⊩ ࡚☜ㄆࡉࢀࡓࡢࡣ୧⩌ඹ 5 ᪥┠࡛㸪ࡑࡢᚋ㸪 ࢥࣆ࣮ᩘࡣⴭࡋࡃቑຍࡋ㸪E7 ⩌ࡣ 16 ᪥┠㸪E5 ⩌ࡣ 19 ᪥┠ṚஸࡋࡓࠋࡇࡢⅬࡢ⾑୰ᖹᆒ 1 㸧 Norio OHASHI, et al.: Demonstration of 6 antigenic and genotypic variation in Orientia Ot ࢥࣆ࣮ᩘࡣࡑࢀࡒࢀ 1.910 copies/ml㸪1.9 5 10 copies/ml 㸪ྠᰴࢆ᥋✀ࡋࡓ ICR ࣐࢘ࢫ tsutsugamushi which were isolated in Japan, and ࠾ࡅࡿ᭱㧗್ࡢ 1000 ಸࢆ♧ࡋࡓࠋ their classification into type and subtype, Microbiol. Immunol., 40(9), 1996, 627-638. 㸬⪃ᐹ 2㸧Akira TAMURA, et al.: Isolation of Rickettsia tsutsugamushi antigenically different from Kato, ⮫ᗋ᳨ᰝ್ࡽẚ㍑ⓗ㔜ぢ࡞ࡋᚓࡓᝈ⪅ ⾑ᾮ⏤᮶ࡢ Shimokoshi ᆺ Matsui ᰴࡢ࣐࢘ࢫ Karp and Gilliam strains from patients, ᑐࡍࡿ⮴Ṛẘᛶࢆ☜ㄆࡋࡓࡇࢁ㸪ICR ࣐࢘ࢫ Microbiol. Immunol, 28(8) , 1984, 873-882. ࠾࠸࡚㸪Matsui ᰴࡣᙉẘᛶࡢ Kato ᆺ Kakuma-2 3㸧బ⸨ᐶᏊ㸪㸸⛅⏣┴࠾ࡅࡿ Shimokoshi ᰴࡼࡾࡶయෆ࡛ࡢቑṪࡀ㠀ᖖ⦆៏࡛࠶ࡾ㸪⮴ ᆺࡘࡘࡀࡢ䭧ཬⓗᏛㄪᰝ, ⾨⏕ື≀, vol. 65, 2014, No.4, 183-188. Ṛᛶࡣ☜ㄆࡉࢀ࡞ࡗࡓࠋࡇࢀࡼࡾ㸪ᚑ᮶ࡢ ͆Shimokoshi ᆺЍᙅẘᛶ͇࠸࠺ࢭ࣮࢜ࣜࡣࡇ 4㸧ᕝ᳃ᩥᙪ, 㸸ࣜࢣࢵࢳឤᰁࡢ㎿㏿デ᩿ ἲ㛵ࡍࡿ◊✲, 㟼ᒸ┴⎔ቃ⾨⏕⛉Ꮫ◊✲ᡤ ࡢᰴ࠾࠸࡚ࡶྠᵝ࡛࠶ࡗࡓࠋ ሗ࿌, No.5, 2011, 37㸫40. ࡇࢀࡲ࡛ࡢ࣐࢘ࢫࢆ⏝࠸ࡓ Ot ࡢẘᛶ㛵ࡍ ࡿ◊✲࠾࠸࡚㸪࣐ࢡࣟࣇ࣮ࢪෆ࡛ࡢቑṪᛶ 5㸧ከᮧ᠇㸸᜔ཎయ Orientia tsutsugamushi ࡢྍྰࡸឤᰁึᮇ TNF ⏘⏕ࡢ᭷↓ࡀ Ot ቑṪ 㸦 4㸧 㸧 , 1999, ࡢᚤ⏕≀Ꮫ, ᪥ᮏ⣽⳦Ꮫ㞧ㄅ, 54㸦 ᢚไ㛵㐃ᛶࡍࡿࡇࡀᣦࡉࢀ࡚࠸ࡿ 5, 6) 815-832. ࠋ ࡲࡓ㸪ࣄࢺࡢᛴᛶᮇ⾑ᾮࢆㄪࡿ㸪ࢧࢺ࢝ 6㸧⚟ཎṇ༤: Orientia tsutsugamushi ࡢẘຊỴᐃ ࣥࡀ㧗್ࢆ♧ࡋ࡚࠾ࡾ㸪චᛂ⟅ࡢ㐣ᛂ ᅉᏊ࠾ࡼࡧឤᰁᐟᛂ⟅㛵ࡍࡿ◊✲㸪༓ⴥ ࡀ㔜㛵㐃ࡍࡿᣦࡉࢀ࡚࠸ࡿ 7) Ꮫ, ༤ኈㄽᩥ, 58, 2006. p24-25. ࠋࡇࢀ ࡽࡢࡇࡣ㸪Ot ࡢࣄࢺᑐࡍࡿཎᛶࡣ Ot ࡑ http://mitizane.ll.chiba-u.jp/metadb/up/assist1/Y ࡢࡶࡢࡢቑṪᛶࡢᙉᙅຍ࠼࡚㸪ᐟഃࡢච 2006-16.pdf ᛂ⟅ࡀཎᛶᙳ㡪ࢆ࠼࡚࠸ࡿࡇࢆ♧ࡋ࡚ 7㸧ᒾᓮ༤㐨: ࢶࢶ࣒࢞ࢩ࠾ࡅࡿ㔜ࢧ ࠸ࡿࠋࣄࢺࡢච≧ែࡣಶࠎᵝࠎ࡛࠶ࡿࡓࡵ㸪 ࢺ࢝ࣥ⏘⏕ไᚚ: ཌ ⏕ປാ┬㸦⦅㸧: ཌ ᮏ┴࠾ࡅࡿࢃࡎ 16 ࡢ⮫ᗋ≧ࡶከ ⏕ປാ⛉Ꮫ◊✲⿵ຓ㔠㸦᪂⯆࣭⯆ឤᰁ ᵝᛶࡀぢࡽࢀࡓࡶࡢᛮࢃࢀࡿࠋᚋࡼࡾከࡃ ᴗ㸧ࣜࢣࢵࢳឤᰁࡢᅜෆᐇែㄪᰝཬࡧ᪩ ࡢࡀ㞟✚ࡉࢀࡿࡇ࡛㸪ࣄࢺᑐࡍࡿཎ ᮇデ᩿యไࡢ☜❧ࡼࡿ᪩ᮇ㆙㚝ࢩࢫࢸ࣒ࡢ ᛶࡀ᫂☜࡞ࡿᛮࢃࢀࡿࡀ㸪Shimokoshi ᆺࡣ ᵓ⠏, ཌ⏕ປാ┬, 2007, 134-144. Ⓨ⏕ࡀ⛥࡛࠶ࡿ࠸࠺㏻ㄝࡽ㸪ࢇࡢ᳨ 8㸧㇂⨾, 㸸ᒣᙧ┴࡛Ⓨ⏕ࡋࡓ Shimokoshi ᰝᶵ㛵ࡀ᳨ᰝᑐ㇟ࡋ࡚࠸࡞࠸ࠋࡑࡢࡓࡵ㸪2014 ᆺࣜࢣࢵࢳឤᰁࡼࡿࡘࡘࡀࡢ୍, ᖺ⌧ᅾ㸪ᅜෆ࡛ Shimokoshi ᆺࡢᝈ⪅Ⓨ⏕ࡀ☜ㄆ ⾨⏕ື≀㸪60, 2009, 317-321㸬 ࡉࢀ࡚࠸ࡿࡢࡣ㸪ࡇࡢᆺࢆ࣮ࣝࢳ᳨ࣥᰝຍ࠼ 9 㸧 ➉ அ ୗ ⚽ 㞝 , Shimokoshi ᆺ Orientia tsutsugamushi ឤᰁࡼࡿࢶࢶ࣒࢞ࢩࡢ 1 㸬 ࡚࠸ࡿ 5 ┴㸦᪂₲㸪ᒣᙧ㸪⚟ᓥ㸪⚟࠾ࡼࡧ⛅ ⏣┴㸧࡞ࡗ࡚࠾ࡾ 2,3,8-10) ⓶⮫ᗋ㸪55, 2013, 1181-1185. 㸪ࡇࢀࡽࡢᅜෆ ࠾ࡅࡿ᳨ᰝయไࡢᩚഛᮇᚅࡋࡓ࠸ࠋ 10 㸧 Satoshi IKEGAYA. , et al. : Tsutsugamushi disease caused by Shimokoshi-Type Orientia ࡲࡓ㸪๓㏙ࡢ࠾ࡾཎᛶࡣࣄࢺഃࡢᛂ⟅ࡶ ࡁࡃ㛵ࡍࡿࡇࡽ㸪ࡘࡘࡀࡣ᪩ᮇ tsutsugamushi㸸The first report in western Japan㸬 MINO ࡼࡿ⒪ࢆ㛤ጞࡍࡿࡇࡀ᭱ၿ⟇࡛࠶ࡿ Am㸬J㸬Trop㸬Med㸬88, 2013, 1217-1219. ࡇኚࢃࡾࡣ࡞࠸ࠋᚋࡶ┴ẸࡢၨⓎࢆᙉ - 31 - 秋田県健康環境センター年報 第10号 2014 ឤᰁⓎ⏕ືྥㄪᰝᴗ ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣࡘ࠸࡚ ⛅㔝⳹Ꮚ బ⸨⏤⾰Ꮚ ᰘ⏣ࡕࡦࢁ బ⸨ᐶᏊ ᩪ⸨༤அ ឤᰁⓎ⏕ືྥㄪᰝ࠾࠸࡚ᖹᡂ 26 ᖺᗘᑠඣ⛉ࡽ᥇ྲྀࡉࢀࡓ⣅౽᳨య 225 ᳨య࠾ࡼࡧឤᰁᛶ⫶ ⭠⅖࠸࡛౫㢗ࡀ࠶ࡗࡓ⾜ᨻ᳨ᰝ 35 㸦㣗୰ẘ 8 㸪㞟ᅋឤᰁ 27 㸧ࡘ࠸࡚㸪᳨ฟࡉࢀࡓ ࢘ࣝࢫࡢ✀㢮ࡸ᳨ฟ㢖ᗘ➼ࡢ㞟ィゎᯒࢆ⾜ࡗࡓࠋឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᳨ ฟ≧ἣࡣ㸪࢘ࣝࢫࡀ᳨ฟࡉࢀࡓ 57 ᳨యࡢ࠺ࡕ㸪ࣀࣟ࢘ࣝࢫ GII ᆺࡀ 27 㸦47.4㸣㸧᭱ࡶከࡃ㸪 ᳨ฟࡢࣆ࣮ࢡࡣ 1 ᭶࡛࠶ࡗࡓࠋḟ࠸᳨࡛ฟࡀከࡃㄆࡵࡽࢀࡓࢧ࣏࢘ࣝࢫࡣ 14 㸦24.6㸣㸧࡛㸪10 㹼12 ᭶ከⓎࡋ࡚࠸ࡓࠋ㣗୰ẘࡽࡢ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣࡣ㸪8 ࡚ࡽࣀࣟ࢘ࣝ ࢫ GII ᆺࡀ᳨ฟࡉࢀࡓࠋࡑࡢ࠺ࡕ 3 ࡣ GII ᆺ GI ᆺࡢΰྜឤᰁ࡛࠶ࡾ㸪ࡇࡢ 3 ࡣ࢝࢟ࡀឤᰁ ※☜ㄆࡉࢀࡓ࡛࠶ࡗࡓࠋ㣗୰ẘ 8 ୰ 7 ࡀ 1 ᭶㸪2 ᭶Ⓨ⏕ࡋ࡚࠸ࡓࠋ㞟ᅋឤᰁ࡛ ࡣ㸪27 ୰ 22 ࡽࣀࣟ࢘ࣝࢫ GII ᆺ㸪2 ࡽࣀࣟ࢘ࣝࢫ GI ᆺࡀ᳨ฟࡉࢀࡓࠋ᳨ฟ ᮇࡣ 4㹼6 ᭶㸪11㹼2 ᭶࡛࠶ࡗࡓࠋࢧ࣏࢘ࣝࢫࡣ 10㹼12 ᭶ࡢ㛫 3 ࡽ᳨ฟࡉࢀ㸪Ⓨ⏕ືྥㄪ ᰝࡼࡿ᳨ฟ≧ἣྠᵝࡢഴྥࡀㄆࡵࡽࢀࡓࠋࡲࡓ㸪Ⓨ⏕ືྥㄪᰝ࠾ࡼࡧ㣗୰ẘ➼㞟ᅋࡼࡾ㸪㏆ ᖺ⛅⏣┴࠾࠸᳨࡚ฟࡉࢀ࡚࠸࡞࠸㑇ఏᏊᆺ࡛࠶ࡿࣀࣟ࢘ࣝࢫ GII.17 ᆺࡀ᳨ฟࡉࢀࡓࠋᅇࡢ⤖ᯝ ࡽ㸪㣗୰ẘ➼㞟ᅋࡢⓎ⏕ࡣ㸪ᕷ୰࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ䬧≧ែࢆᫎࡋ࡚࠸ࡿࡶࡢ⪃ ࠼ࡽࢀ㸪ࡑࡢ≧ἣࢆᢕᥱࡍࡿឤᰁⓎ⏕ືྥㄪᰝࡢ⤖ᯝ┦㛵ࡍࡿࡶࡢᛮࢃࢀࡓࠋ 㸬ࡣࡌࡵ ᑐ㇟ཎయ ᡃࡀᅜ࠾ࡅࡿឤᰁࢧ࣮࣋ࣛࣥࢫࡣ㸪ឤ ឤᰁᛶ⫶⭠⅖ࡢཎᅉ࡞ࡿࣀࣟ࢘ࣝࢫ㸪ࢧ ᰁἲᇶ࡙ࡃឤᰁⓎ⏕ືྥㄪᰝᴗࡋ ࣏࢘ࣝࢫ㸪ࣟࢱ࢘ࣝࢫ㸦A ⩌㸪C ⩌㸧㸪⭠ ࡚㸪ឤᰁࡢண㜵ࡲࢇᘏ㜵Ṇࢆ┠ⓗᅜつ ⟶ࢹࣀ࢘ࣝࢫࢆᑐ㇟ࡋࡓࠋ ᶍ࡛ᐇࡉࢀ࡚࠸ࡿࠋᙜࢭࣥࢱ࣮࡛ࡣᴗࡢ୍ ⎔ࡋ࡚㸪ཎయᐃⅬ་⒪ᶵ㛵ࡼࡾᥦ౪ࡉࢀࡓ ᳨ᰝἲ ᳨యࡽࡢཎయ᳨ฟࢆ⥅⥆ⓗ⾜ࡗ࡚࠸ࡿࠋ ᳨యࡽ MagNA Pure LC2.0㸦ࣟࢩࣗ㸧ࡼࡾ ୰࡛ࡶ㸪⫶⭠⅖࢘ࣝࢫࡣ㣗୰ẘࡸ㞟ᅋឤᰁ DNA ࠾ࡼࡧ RNA ࢆᢳฟࡋࡓࠋࡑࡢᚋ㸪real-time ࡢཎᅉ࡞ࡿࡇࡽ㸪ᆅᇦ࠾ࡅࡿὶ⾜≧ PCR 1 㹼 3) ࠾ࡼ ࡧ conventional nested-PCR 4) ࡼ ࡾ ἣࡢᢕᥱࡣ㔜せ⪃࠼ࡽࢀࡿࠋᅇࡣ㸪ᖹᡂ 26 ࢘ࣝࢫࡢ᳨ฟࢆヨࡳࡓࠋ᳨ฟࡉࢀࡓ࢘ࣝࢫ ᖺᗘ᥇ྲྀࡉࢀࡓឤᰁᛶ⫶⭠⅖ᝈ⪅ࡢ⣅౽᳨య ࡘ࠸࡚ࡣ㸪ࢲࣞࢡࢺࢩ࣮ࢡ࢚ࣥࢫࡼࡾሷ ྵࡲࢀࡿ࢘ࣝࢫࡢ✀㢮ࡸ᳨ฟ㢖ᗘࡘ࠸࡚ ᇶ㓄ิࢆỴᐃࡋ㸪㑇ఏᏊᆺูࢆ⾜ࡗࡓࠋ 㞟ィゎᯒࢆ⾜ࡗࡓࡢ࡛ሗ࿌ࡍࡿࠋ 㸬⤖ᯝ⪃ᐹ 㸬᪉ἲ ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫ ᑐ㇟ ࡢ᳨ฟ≧ἣ ཎయᐃⅬ་⒪ᶵ㛵࠾࠸࡚ᖹᡂ 26 ᖺᗘ ᥦ౪ࡉࢀࡓ⣅౽᳨య 225 ᳨య୰ 57 ᳨య㸦25.3 ᑠඣ⛉ࡽ᥇ྲྀࡉࢀࡓ᳨య 1047 ᳨యࡢ࠺ࡕ⣅ 㸣㸧ࡽ࢘ࣝࢫࡀ᳨ฟࡉࢀࡓࠋឤᰁⓎ⏕ື ౽᳨య 225 ᳨య࠾ࡼࡧᖹᡂ 26 ᖺᗘ౫㢗ࡀ࠶ࡗ ྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᭶᳨ูฟ≧ἣ ࡓ⾜ᨻ᳨ᰝ 35 㸦㣗୰ẘ 8 㸸↷ 1 ࡘ࠸࡚㸪⛅⏣┴ࡢ≧ἣࢆ⾲ 1 㸪ᅜࡢ≧ἣ ྵࡴ㸪㞟ᅋឤᰁ 27 㸸↷ 1 ࢆ⾲ 2 ♧ࡍࠋ ྵࡴ㸧ࢆ㞟ィᑐ㇟ࡋࡓࠋ ᳨ฟ⫶⭠⅖࢘ࣝࢫࡢෆヂ ᳨ฟࡉࢀࡓ⫶⭠⅖࢘ࣝࢫࡢෆヂࡣ㸪ࣀࣟ࢘ - 32 - 秋田県健康環境センター年報 第10号 2014 ⾲ ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣ㸦⛅⏣┴㸸ᖹᡂ ᖺᗘ㸧 4᭶ 5᭶ 6᭶ ࣀࣟ࢘ࣝࢫ 4 2 4 ࢧ࣏࢘ࣝࢫ 2 A ⩌ࣟࢱ࢘ࣝࢫ 5 1 7᭶ 8᭶ 9᭶ 1 10 ᭶ 2 11 ᭶ 4 12 ᭶ 1᭶ 2᭶ 3᭶ ィ 1 11 4 1 27 14 4 6 13 2 1 1 C ⩌ࣟࢱ࢘ࣝࢫ ⭠⟶ࢹࣀ࢘ࣝࢫ 1 ィ 12 2 1 8 5 1 0 0 2 5 5 12 6 1 57 㸦᳨యᩘ㸧 ⾲ ឤᰁⓎ⏕ືྥㄪᰝ࠾ࡅࡿ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣ㸦ᅜ㸸ᖹᡂ ᖺᗘ㸧 4᭶ 5᭶ 6᭶ 7᭶ 8᭶ 9᭶ 10 ᭶ 11 ᭶ 12 ᭶ 1᭶ 2᭶ 3᭶ ィ ࣀࣟ࢘ࣝࢫ 303 272 104 32 37 49 32 256 497 514 464 409 2969 ࢧ࣏࢘ࣝࢫ 27 31 19 16 13 3 15 60 98 34 24 42 382 A ⩌ࣟࢱ࢘ࣝࢫ 110 108 35 6 8 5 6 2 13 40 68 155 556 C ⩌ࣟࢱ࢘ࣝࢫ 0 0 0 0 0 0 0 0 0 1 0 0 1 ⭠⟶ࢹࣀ࢘ࣝࢫ 23 19 13 8 4 7 6 5 2 7 5 9 108 ィ 463 430 171 62 62 64 59 323 610 596 561 615 4016 ฟ㸸ᅜ❧ឤᰁ◊✲ᡤཎᚤ⏕≀᳨ฟሗ㸦H27.8.21 ⌧ᅾ㸧ࢆᇶ㞟ィ 㸦᳨యᩘ㸧 ⾲ ឤᰁⓎ⏕ືྥㄪᰝ࠾࠸᳨࡚ฟࡉࢀࡓࣀࣟ࢘ࣝࢫࡢ㑇ఏᏊᆺ㸦ᖹᡂ ᖺᗘ㸧 4᭶ 5᭶ 6᭶ 7᭶ 8᭶ 9᭶ 10 ᭶ 11 ᭶ 12 ᭶ 1᭶ 2᭶ 3᭶ ィ GII.2 1 1 GII.3 1 1 GII.4 Sydney 2012 3 GII.4 2006b GII.6 1 2 9 3 15 2 1 3 2 5 GII.17 1 ィ 4 2 4 0 0 0 0 0 1 11 4 1 2 1 27 㸦ᰴᩘ㸧 ࣝࢫࡀ 27 㸦47.4㸣㸧᭱ࡶከࡃ㸪ḟ࠸࡛ࢧ࣏ ≧ࢆ࿊ࡋ࡚࠾ࡾ㸪㑇ఏᏊᆺูࡢศ㢮ࡽ⭠⟶ ࢘ࣝࢫࡀ 14 㸦24.6㸣㸧㸪A ⩌ࣟࢱ࢘ࣝࢫ ࢹࣀ࢘ࣝࢫ㸦41 ᆺ㸧☜ㄆࡉࢀࡓࠋ ࡀ 13 㸦22.8㸣㸧㸪ࢹࣀ࢘ࣝࢫࡀ 2 㸦3.5 㸣㸧㸪C ⩌ࣟࢱ࢘ࣝࢫࡀ 1 㸦1.8㸣㸧࡛࠶ࡗ ᳨ฟ⫶⭠⅖࢘ࣝࢫࡢᏘ⠇ⓗ≉ᚩ ࡓࠋࣀࣟ࢘ࣝࢫࡣࡍ࡚ GII ᆺ࡛㸪ࢧ࣏࢘ ሙὶ⾜ᮇࢆ㏄࠼ࡿࣀࣟ࢘ࣝࢫࡣ 1 ᭶ ࣝࢫࡣேឤᰁࡍࡿ GI ᆺ㸦5 㸧㸪GII ᆺ㸦5 ᭱ࡶከࡃ᳨ฟࡉࢀࡓࠋࣟࢱ࢘ࣝࢫࡣᖺᅜ 㸧㸪GIV ᆺ㸦1 㸧㸪GV ᆺ㸦3 㸧ࡢ࡚ࡢ ⓗ 2㹼5 ᭶ࣆ࣮ࢡࢆ㏄࠼ࡿࡀ ᆺࡀ᳨ฟࡉࢀࡓࠋA ⩌ࣟࢱ࢘ࣝࢫࡢ 1 ࡘ 1㹼5 ᭶᳨ฟࡀㄆࡵࡽࢀࡓࠋࢧ࣏࢘ࣝࢫࡘ ࠸࡚ࡣ࣡ࢡࢳࣥᰴ࡛࠶ࡿࡇࡀ☜ㄆࡉࢀࡓࠋ A ࠸࡚ࡣ㸪ᅜⓗࡣᖺ㛫ࢆ㏻ࡋ᳨࡚ฟࡀㄆࡵࡽ ⩌ࣟࢱ࢘ࣝࢫẚ C ⩌ࣟࢱ࢘ࣝࢫࡢ᳨ฟ ࢀ㸪ࡑࡢࣆ࣮ࢡࡣ 12 ᭶࡛࠶ࡗࡓࡀ㸪ᮏ┴࠾࠸ 5) 5㸧 㸪ᮏ┴࡛ࡶ 㢖ᗘࡣᅜⓗࡶప࠸ഴྥ࠶ࡾ 㸪ᖹᡂ 26 ᖺ ࡚ࡣ 10㹼12 ᭶ከⓎࡋ࡚࠸ࡓࠋ᪥ᮏ࠾ࡅࡿ⭠ ᗘࡣᮏ┴᳨࡛ฟࡉࢀࡓ 1 ࡀᅜ࡛ࡶ၏୍ࡢሗ ⟶ࢹࣀ࢘ࣝࢫࡢ᳨ฟࡣ㸪ࣀࣟ࢘ࣝࢫ㸪ࢧ ࿌࡛࠶ࡗࡓࠋࢹࣀ࢘ࣝࢫࡣୖୗẼ㐨⅖㸪⤖ ࣏࢘ࣝࢫ㸪ࣟࢱ࢘ࣝࢫẚ㍑ࡋ㢖ᗘࡣప࠸ ⭷⅖㸪⫶⭠⅖࡞ከᵝ࡞ᝈࡢཎᅉ࡞ࡾ࠺ࡿ ࡀ㸪ᖺ㛫ࢆ㏻ࡋ᳨࡚ฟࡀㄆࡵࡽࢀ࡚࠸ࡿࠋᮏ┴ ࢘ࣝࢫ࡛࠶ࡿࡀ㸪᳨ฟࡉࢀࡓ 2 ࡣ⫶⭠⅖ ࡛ࡣ 4 ᭶ 11 ᭶ 1 ࡎࡘࡢ᳨ฟ࡛࠶ࡗࡓࠋ - 33 - 秋田県健康環境センター年報 第10号 2014 ⾲ 㣗୰ẘ࠾ࡼࡧ㞟ᅋឤᰁࡽࡢ⫶⭠⅖࢘ࣝࢫࡢ᳨ฟ≧ἣ㸦ᖹᡂ ᖺᗘ㸧 4᭶ ࣀࣟ࢘ࣝࢫ GII ᆺ 㣗୰ẘ ឤᰁ 6᭶ 7᭶ 8᭶ 9᭶ 10᭶ 11᭶ 12᭶ 1 ᭶ 1 ࣀࣟ࢘ࣝࢫ GII ᆺ࣭G I ᆺΰྜឤᰁ ࣀࣟ࢘ࣝࢫ GII ᆺ 㞟ᅋ 5᭶ 6 5 ࣀࣟ࢘ࣝࢫ G I ᆺ 2 1 1 1 7 6 3᭶ 2 0 0 0 1 1 5 2 1 3 1 5 22 2 3 2 1 4 ィ 3 1 ࢧ࣏࢘ࣝࢫ ィ 2 2᭶ 7 7 35 0 㸦ᩘ㸧 ⾲ ⛅⏣┴࠾ࡅࡿ㣗୰ẘࡢヲ⣽㸦ᖹᡂ ᖺᗘ㸧 Ⓨ⏕ᖺ᭶ ᥎ᐃཎᅉ㣗ရ ཎᅉタ ᳨ฟ࢘ࣝࢫ ᖹᡂ 26 ᖺ 4᭶ ᫂ 㸦㣧㣗ᗑࡢ㣗㸧 㣧㣗ᗑ ࣀࣟ࢘ࣝࢫ GII ᆺ GII.4 Sydney 2012 ࢝࢟ᩱ⌮ 㣧㣗ᗑ ࣀࣟ࢘ࣝࢫ GII ᆺ GII Mix ⪁ேタ ࣀࣟ࢘ࣝࢫ GII ᆺ GII.4 Sydney 2012 ᪑㤋 ࣀࣟ࢘ࣝࢫ GII ᆺ ࣭G I ᆺ GII.4 Sydney 2012㸪 GII.17㸪GI.4 㣧㣗ᗑ ࣀࣟ࢘ࣝࢫ GII ᆺ GII.4 Sydney 2012 㓑࢞࢟ ᪑㤋 ࣀࣟ࢘ࣝࢫ GII ᆺ ࣭G I ᆺ GII.4 Sydney 2012㸪 GII.17㸪GII.13㸪GI.3 ࡋ࢞࢟ ᪑㤋 ࣀࣟ࢘ࣝࢫ GII ᆺ GII.6 㣧㣗ᗑ ࣀࣟ࢘ࣝࢫ GII ᆺ ࣭G I ᆺ GII.17㸪GII.21㸪GI.4 ᖹᡂ 27 ᖺ 1᭶ ᫂ 㸦タࡢ⤥㣗㸧 ࣏࢝࢟ࣥ㓑 ᫂ 㸦㣗ᩱ⌮㸧 ᖹᡂ 27 ᖺ 2᭶ 㓑࢞࢟ ᳨ฟࣀࣟ࢘ࣝࢫࡢ㑇ఏᏊᆺ ࣀࣟ࢘ࣝࢫ㑇ఏᏊᆺ ഛ ⪃ ↷㸸༓ⴥ┴ 㣗୰ẘࡢෆヂ 㣗୰ẘࡢヲ⣽ࡘ࠸࡚⾲ 5 ♧ࡍࠋ8 ឤᰁⓎ⏕ືྥㄪᰝ࠾࠸᳨࡚ฟࡉࢀࡓࣀࣟ ࢘ࣝࢫࡢ㑇ఏᏊᆺࡘ࠸࡚⾲ 3 ♧ࡍࠋ ࡚ࡢࡽࣀࣟ࢘ࣝࢫ GII ᆺࡀ᳨ฟࡉ 2011/12 ࢩ࣮ࢬࣥὶ⾜ࡋࡓ GII.4 Sydney 2012 ࢀ㸪ࡲࡓ㸪3 ࡽࣀࣟ࢘ࣝࢫ GI ᆺࡶ᳨ฟ ኚ␗ᰴࡀ 15 㸦55.6㸣㸧᭱ࡶከࡃ㸪ࡑࡢ᳨ฟ ࡉࢀࡓࠋΰྜឤᰁ 3 ࡚࢝࢟ࡀឤᰁ※☜ ᮇࡣὶ⾜ᮇ࡛࠶ࡿ 1 ᭶㸪2 ᭶ከࡃ࡞ࡗ࡚࠸ ㄆࡉࢀࡓ࡛࠶ࡗࡓࠋࡇࡢ 3 ࡘ࠸࡚ࡣ㸪 ࡓࠋḟ࠸࡛㸪5 㸦18.5㸣㸧࡛ㄆࡵࡽࢀࡓ GII.6 㑇ఏᏊᆺࡀ」ᩘ᳨ฟࡉࢀ࡚࠾ࡾ㸪Ⓨ⏕ືྥㄪᰝ ᆺࡣ 4㹼6 ᭶ࡢඛ᳨ฟࡀ㝈ᒁࡋ࡚࠸ࡓࠋࡲ ࠾࠸᳨࡚ฟࡀㄆࡵࡽࢀࡓ GII.17 ᆺࡶ᳨ฟࡉࢀ ࡓ㸪㏆ᖺᮏ┴࠾࠸᳨࡚ฟࡉࢀ࡚࠸࡞ࡗࡓ ࡓࠋ࢝࢟ࡸࢧࣜ➼ᯛ㈅ࡢ୰⭠⭢ࡣ」ᩘࡢ GII.17 ᆺ ࡀ 2 㸦 7.4 㸣 㸧 ࡽ ᳨ ฟ ࡉ ࢀ ࡓ ࠋ 㑇ఏᏊᆺࡢ࢘ࣝࢫࡀ✚ࡉࢀࡿࡇࡀከ࠸ࡓ ࡵ㸪ᯛ㈅ࡢႚ㣗ࡀཎᅉ࡞ࡿ㣗୰ẘ࡛ࡣΰྜ 㣗୰ẘ࠾ࡼࡧ㞟ᅋឤᰁࡽࡢ⫶⭠⅖࢘ ឤᰁ࡞ࡿྍ⬟ᛶࡀ㧗࠸⪃࠼ࡽࢀࡿ 6) ࠋ㣗୰ẘ ࣝࢫࡢ᳨ฟ≧ἣ ࡢⓎ⏕ࡣ 1 ᭶㸪2 ᭶㞟୰ࡋ࡚࠸ࡓࠋࡇࡢ 㣗୰ẘ࠾ࡼࡧ㞟ᅋឤᰁࡽࡢ⫶⭠⅖࢘ ᮇ㛫㸪࢝࢟ࡀཎᅉ㣗ရࡳࡽࢀࡓ㣗୰ẘࡀ 5 ࣝࢫࡢ᭶᳨ูฟ≧ἣࢆ⾲ 4 ♧ࡍࠋ⾜ᨻ᳨ᰝ 㸦GII ᆺឤᰁ 2 㸸ෆ 1 ࡣ↷㸪 ࡋ࡚౫㢗ࡢ࠶ࡗࡓ㞟ᅋࡢཎᅉཎయࡣࣀࣟ GII ᆺ࣭GI ᆺΰྜឤᰁ 3 㸧ᖺ࡞ࡃከ ࢘ࣝࢫࢧ࣏࢘ࣝࢫࡢ 2 ✀㢮࡛࠶ࡗࡓࠋ ࡃⓎ⏕ࡋ࡚࠾ࡾ㸪࠸ࡎࢀࡶ⏕㣗ࡲࡓࡣຍ⇕༑ - 34 - ⾲ 㞟ᅋឤᰁ࠾ࡅࡿࣀࣟ࢘ࣝࢫࡢ㑇ఏᏊᆺ㸦ᖹᡂ ᖺᗘ㸧 GII.2 4᭶ 5᭶ 1 1 6᭶ GII.3 7᭶ 8᭶ 9᭶ 10 ᭶ 11 ᭶ 12 ᭶ 1᭶ 2᭶ 3᭶ ィ 2 1 GII.4 Sydney 2012 1 1 1 2 * 1 3 8 GII.4 2006b 0 GII.6 4 4 1 9 GII.17 1 * GI.2 2 3 1 GI.3 1 1 6 ィ 6 1 2 0 0 0 0 1 3 2 5 * 12 ᭶ࡢ 2 ୰ 1 ࡼࡾ GII.4 sydney 2012 ᆺ GII.17 ᆺࡢ 2 ✀㢮ࡢ㑇ఏᏊᆺࢆ᳨ฟ 25 0 㸦ᰴᩘ㸧 ศ࡛㣗ࡉࢀ࡚࠸ࡓࠋࣀࣟ࢘ࣝࢫởᰁࡉࢀࡓ ࢝࢟ࡀὶ㏻ࡋ࡚࠸ࡓྍ⬟ᛶࡀ᥎ᐃࡉࢀࡿࠋᏘ ᯛ㈅ࢆႚ㣗ࡍࡿ㝿ࡣ༑ศ࡞ຍ⇕ฎ⌮ࡀឤ ᰁண㜵ࡘ࡞ࡀࡿ⪃࠼ࡽࢀࡿࠋ 㞟ᅋឤᰁࡢෆヂ 㞟ᅋឤᰁ 27 ୰ 22 ࡽࣀࣟ࢘ࣝࢫ GII ᆺࡀ᳨ฟࡉࢀ㸪2 ࡽࣀࣟ࢘ࣝࢫ GI ᆺࡀㄆࡵࡽࢀࡓࠋࣀࣟ࢘ࣝࢫࡢ᳨ฟᮇࡣ 4㹼6 ᭶㸪11㹼2 ᭶࡛࠶ࡗࡓࠋࢧ࣏࢘ࣝࢫࡣ 3 ࡛ㄆࡵࡽࢀ㸪ࡍ࡚ G V ᆺ࡛࠶ࡗࡓࠋ᳨ฟ ᮇࡣ 10㹼12 ᭶ࡢ㛫࡞ࡗ࡚࠾ࡾ㸪Ⓨ⏕ືྥㄪ ᅗ 㞟ᅋឤᰁ࠾ࡅࡿⓎ⏕タࡢෆヂ ᰝࡼࡿ᳨ฟ≧ἣྠᵝࡢഴྥࡀࡳࡽࢀࡓࠋࢧ 㸦ᖹᡂ ᖺᗘ㸧 ࣏࢘ࣝࢫࡣ 6 ṓ௨ୗࡢᑠඣࡢឤᰁࡀከ࠸࢘ 7) 㸪ࡇࡢ 3 ࡣಖ⫱ᅬ࠾ࡼࡧᗂ ᰝࡢ᳨ฟ≧ἣྠᵝ㸪4㹼6 ᭶ࡢඛ GII.6 ⛶ᅬ ࡛ࡢ 㞟ᅋ ឤᰁ ࡛ ࠶ࡗ ࡓࠋ 㞟ᅋ ឤᰁ 27 ᆺࡀ㞟୰ࡋ࡚ 9 㸦36.0㸣㸧ㄆࡵࡽࢀ࡚࠾ࡾ㸪11㹼2 ࠾ࡅࡿⓎ⏕タࡢෆヂࢆᅗ 1 ♧ࡍࠋಖ ᭶ࡢὶ⾜ᮇࡣ GII.4 Sydney 2012 ኚ␗ᰴࡀ 7 ⫱ᅬࡀ 14 㸦51.9㸣㸧᭱ࡶከࡃ㸪ࡘ࠸࡛ᗂ 㸦28.0㸣㸧᳨ฟࡉࢀ࡚࠸ࡓࠋࡲࡓ㸪Ⓨ⏕ືྥ ⛶ᅬࡀ 4 㸦14.8㸣㸧㸪ᑠᏛᰯࡀ 3 㸦11.1 ㄪᰝࡸ㣗୰ẘ࠾࠸࡚ࡶ᳨ฟࡉࢀࡓ 㸣㸧㸪㧗㱋⪅タࡀ 3 㸦11.1㸣㸧㸪ࡑࡢ GII.17 ᆺࡀ 3 㸦12.0㸣㸧ࡽㄆࡵࡽࢀࡓࠋ ࡀ 3 㸦11.1㸣㸧࡛࠶ࡗࡓࠋಖ⫱ᅬᗂ⛶ᅬ GII.17 ᆺࡢ᳨ฟࡘ࠸࡚ࡣ㸪ᮏ┴ྠᵝࡇࢀࡲ࡛ ࢆྜࢃࡏࡓⓎ⏕ᩘࡀ༙ᩘ௨ୖᅽಽⓗከ࠸≧ ᳨ฟࡀᑡ࡞ࡗࡓ㜰ᕷ 9) ࡸ୕㔜┴ ἣ࡛࠶ࡗࡓࠋஙᗂඣࡸ㧗㱋⪅ࡣ㸪චࡀᮍ⇍࡛ ࡚ࡶᖹᡂ 26 ᖺᗘ᳨ฟࡉࢀ࡚࠾ࡾ㸪ᘬࡁ⥆ࡁື ࠶ࡗࡓࡾ㸪పୗࡋ࡚࠸ࡓࡾࡍࡿࡇ࡛ឤᰁࡋࡸ ྥὀពࢆᡶ࠺ᚲせࡀ࠶ࡿ⪃࠼ࡿࠋ ࣝࢫ࡛࠶ࡾ 10) ࠾࠸ ࡍࡃ࡞ࡗ࡚࠾ࡾ㸪≉ஙᗂඣࡣ⾨⏕ほᛕࡀⓎ㐩 ࡋ࡚࠸࡞࠸ 8) ࠸࠺⌮⏤ࡽࡶឤᰁࡀᗈࡀࡾࡸ 㸬ࡲࡵ ࡍ࠸ࡶࡢ⪃࠼ࡽࢀࡿࠋ ᑠඣ࠾ࡅࡿឤᰁᛶ⫶⭠⅖ࡢⓎ⏕ືྥࡣ㸪ࣀ ࣟ࢘ࣝࢫ➼ࡼࡿ㞟ᅋⓎ⏕ࡢሗ࿌ᩘࡢ᥎⛣ 11) 㞟ᅋឤᰁ࠾ࡅࡿ᳨ฟࣀࣟ࢘ࣝࢫ ᴫࡡ୍⮴ࡍࡿ࠸ࢃࢀ࡚࠸ࡿ ࡢ㑇ఏᏊᆺ 26 ᖺᗘ࠾ࡅࡿ㞟ィ⤖ᯝࡽࡶ㸪㣗୰ẘ➼㞟ᅋ ࠋ⛅⏣┴ࡢᖹᡂ 㞟ᅋឤᰁ࠾࠸᳨࡚ฟࡉࢀࡓࣀࣟ࢘ࣝ ࡢⓎ⏕ࡣ㸪ឤᰁⓎ⏕ືྥㄪᰝࡼࡿ⫶⭠ ࢫࡢ㑇ఏᏊᆺࢆ⾲ 6 ♧ࡍࠋឤᰁⓎ⏕ືྥㄪ ⅖࢘ࣝࢫࡢ᳨ฟ≧ἣྠᵝࡢഴྥࡀㄆࡵࡽࢀ - 35 - 秋田県健康環境センター年報 第10号 2014 ࡓࠋ㣗୰ẘࡸ㞟ᅋឤᰁࡢⓎ⏕ண㜵࠾ࡼࡧᑐ ሗࠒࣟࢱ࢘ࣝࢫࡢᴫせ㸪ཎᚤ⏕≀᳨ฟ ⟇ࡣ㸪ᕷ୰ὶ⾜ࡋ࡚࠸ࡿཎయࡢືែࢆヲ ሗ㸪35㸪3㸪2014㸪1-4㸬 ⣽ࡘⓗ☜ᢕᥱࡍࡿࡇࡀ㔜せ࡛࠶ࡿࡇ 6㸧ᩪ⸨༤அ㸪:ࣀ࣮࢛࣮࢘ࢡᵝ࢘ࣝࢫ㸦NLV㸧 ࡽ㸪ᆅᇦࡸᝈࡢ✀㢮೫ࡾࡀ⏕ࡌ࡞࠸ᙧ࡛ࡢ ࡢ᳨ᰝ࠾ࡅࡿ୍ᮏ㙐㧗ḟᵓ㐀ከᆺ㸦SSCP㸧 ᳨య㞟ࡀᚲせ⪃࠼ࡽࢀࡿࠋᚋࡶ་⒪ᶵ㛵 ゎᯒࡢᛂ⏝㸪⮫ᗋ࢘ࣝࢫ㸪30㸪3㸪2002㸪 ࡢ㐃ᦠࢆᅗࡾ࡞ࡀࡽὶ⾜≧ἣࡢᢕᥱሗ㑏 163-171㸬 7㸧Oka T., et al.: Comprehensive review of human ඖດࡵ࡚࠸ࡁࡓ࠸ࠋ sapovirus. J. Clin. Microbiol. Rev., 28, 2015, ཧ⪃ᩥ⊩ 32-53. 8㸧ᅜ❧ឤᰁ◊✲ᡤឤᰁሗࢭࣥࢱ࣮ ࣀࣟ 1㸧Kageyama,T.,et al.:Broadly reactive and highly ࢘ࣝࢫឤᰁ ࣀࣟ࢘ࣝࢫឤᰁࡑࡢ sensitive assay for Norwalk-like viruses based ᑐᛂ࣭ண㜵㸦་⒪ᚑ⪅࣭タࢫࢱࢵࣇ⏝㸧 on real-time quantitative reverse transcription-PCR. http://idsc.nih.go.jp/disease/norovirus/taio-b.html 9㸧ධ㇂ᒎᘯ㸪:㞟ᅋ⫶⭠⅖ࡽ᳨ฟࡉࢀ J. Clin. Microbiol.,41,2003,1548-1577. 2㸧Oka, T., et al.: Detection of human sapovirus by ࡓࣀࣟ࢘ࣝࢫࡢศᏊᏛⓗゎᯒ࠾ࡼࡧᅜ real-time reverse transcription-polymerase chain ⏘ᕷ㈍⏕࢝࢟ࡢ࢘ࣝࢫởᰁㄪᰝ㸪ཌ⏕ປാ reaction. J.Med. Virol.,78,2006,1347-1353. ⛉Ꮫ◊✲㈝⿵ຓ㔠 3㸧Logan, C., et al.: Real-time reverse transcription ✲ᴗ 㣗ရࡢᏳ☜ಖ᥎㐍◊ 㣗ရ୰ࡢཎ࢘ࣝࢫࡢ᳨ฟἲ -PCR for detection of rotavirus and adenovirus 㛵ࡍࡿ◊✲ as causative agents of acute viral gastroenteritis ࿌᭩㸪2015㸪147-155㸬 in children. J. Clin. Microbiol. 44, 2006, 10㸧ᅜ❧ឤᰁ◊✲ᡤឤᰁሗࢭࣥࢱ࣮:ࠑᅜ 3189-3195. ෆሗࠒࣀࣟ࢘ࣝࢫ GII.17 ᆺࡢὶ⾜ࡑࡢ 4 㸧 Inagawa W. S., et al.: Rapid diagnosis of ≉ᚩࡘ࠸࡚㸫୕㔜┴㸪ཎᚤ⏕≀᳨ฟ adenoviral conjunctivitis by PCR and restriction fragment length polymorphism analysis. J. Clin. ᖹᡂ 26 ᖺᗘ⥲ᣓ࣭◊✲ศᢸሗ ሗ㸪36㸪5㸪2015㸪19-20㸬 11㸧㔝⏣⾨㸪:ࣀࣟ࢘ࣝࢫ㣗୰ẘ࣭ឤᰁ Microbiol.,41,1996,2113-2116. ࡽࡲࡶࡿĵ㸫ࡑࡢ▱㆑ᑐ⟇㸫㸪බ┈♫ᅋἲ ே᪥ᮏ㣗ရ⾨⏕༠ 2013㸪16-17㸬 5㸧ᅜ❧ឤᰁ◊✲ᡤឤᰁሗࢭࣥࢱ࣮:ࠑ≉ 㞟ࠒࣟࢱ࢘ࣝࢫ 2010㹼2013 ᖺࠑ≉㞟㛵㐃 - 36 - 秋田県健康環境センター年報 第10号 2014 ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠 㣗ရࡢᏳ☜ಖ᥎㐍◊✲ᴗ㸦H25㹼27 ᖺᗘ㸧 㣗ရࡽࡢ࢘ࣝࢫ᳨ฟἲࡋ࡚ࡢ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ⢭ᗘྥୖ㛵ࡍࡿ◊✲ ᩪ⸨༤அ ⛅㔝⳹Ꮚ 㔝⏣ ⾨* 1 ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡣ㸪㣗ရ᳨యྵࡲࢀࡿ࢘ࣝࢫ⢏Ꮚࢆ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ⾲㠃྾╔ࡉ ࡏ࡚ᅇࡍࡿࡇࢆᇶᮏཎ⌮ࡋ࡚࠸ࡿࠋࡑࡢᛶ㉁ୖ㸪ᢳฟࡉࢀࡓ RNA ࡣ㔞ࡢ㯤Ⰽࣈࢻ࢘⌫⳦ ⏤᮶ࡢ㑇ఏᏊࡀΰධࡍࡿࡇ࡞ࡿࡀ㸪ᴟᚤ㔞ࡢ࢘ࣝࢫ RNA ࢆᏳᐃⓗಖᣢࡍࡿ࣮࢟ࣕࣜࡋ ࡚ാࡃࡓࡵ㸪᳨ฟឤᗘᑐࡋ࡚ࡣࣉࣛࢫࡢຠᯝࡀぢ㎸ࡵࡿࠋࡑࡢ୍᪉࡛㸪㔞ࡢ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ㑇 ఏᏊࡢ୰ྵࡲࢀࡿ࢘ࣝࢫ㑇ఏᏊࢆ᳨ฟࡍࡿ࠸࠺≉␗࡞᮲௳ୗ࡛ࡢᛂ࡛࠶ࡿࡇࡽ㸪ヨ㦂ᶵ 㛵ࡼࡗ᳨࡚ฟ⢭ᗘࡀࣂࣛࡘࡃ࠸࠺ၥ㢟ࡀᣦࡉࢀࡓࠋࡑࡇ࡛㸪᳨ฟ⤖ᯝᕪࢆ⏕ࡌࡸࡍ࠸⟠ᡤ ࡘ࠸࡚ᛂ᮲௳➼ࢆ᭱㐺ࡍࡿᚲせࡀ࠶ࡗࡓࠋࡇࡢࡇࡽ㸪ḟࡢ㡯ࡘ࠸᳨࡚ウࡋࡓࠋձĮ-Amylase ⢊ᮎྵࡲࢀࡿ㈿ᙧࡀṧ␃ࡍࡿ㸪᳨ฟឤᗘᙳ㡪ࡍࡿࡇࡽ㸪ࡇࢀࢆ࠶ࡽࡌࡵ㝖ཤࡋࡓ㓝⣲ ᾮࢆㄪ〇ࡋ࡚࠾ࡃࡇ࡛ᑐᛂࡋࡓࠋᾮㄪ〇ရࡣ㸫20Υ࡛ 2 ࣨ᭶௨ୖᏳᐃಖᏑ࡛ࡁ㸪⢊ᮎࢆ┤᥋ᢞධ ࡋࡓሙྜྠ➼ࡢຠᯝࡀ࠶ࡿࡇࢆ☜ㄆࡋࡓࠋղ㯤Ⰽࣈࢻ࢘⌫⳦⏤᮶ DNA ࢆ㝖ཤࡍࡿ᪉ἲࡋ࡚㸪 RNA ᢳฟ㐣⛬࡛ࡢ࣒࢜ࣥ࢝ࣛ DNase ฎ⌮㸪RNA ᢳฟᚋࡢ᫆⇕ᛶ DNase ฎ⌮ࡢ 2 ㏻ࡾࢆᐇ࡛ࡁࡿ ࡼ࠺ࡋࡓࠋ࠸ࡎࢀࡢ᪉ἲࢆ⏝࠸࡚ࡶ㸪ࡇࢀࡲ࡛ࡢ DNase ฎ⌮࡛ၥ㢟࡞ࡗࡓࢥࣆ࣮ᩘࡢపୗࡣ㉳ࡇ ࡽ࡞ࡗࡓࠋճ㏫㌿ᛂࡣ㸪58Υ࡛ࡢ㧗 ㏫㌿ᛂࡀ㠀≉␗ᛂࡼࡿឤᗘపୗࢆᢚไࡍࡿࡢ ᭷ຠ࡛࠶ࡗࡓࠋմPCR ᛂࡣ㓝⣲〇ရࡢ㑅ᢥࡀ㔜せ࡛࠶ࡾ㸪≉࣍ࢵࢺࢫࢱ࣮ࢺᵝࡀᚲ㡲࡛࠶ࡿࡇ ࡀ☜ㄆࡉࢀࡓࠋ 㸬ࡣࡌࡵ ࡿࡓࡵࡢ⣒ཱྀࢆぢฟࡍࡇࡀ࡛ࡁࡓ 4-13) ࠋࡑࡢ ࢘ࣝࢫᛶ㣗୰ẘࡢᑐ⟇ࡋ࡚ᯛ㈅ࡢởᰁ ᚋ㸪ᖹᡂ 22~24 ᖺᗘᐇࡉࢀࡓཌ⏕ປാ⛉Ꮫ ᐇែㄪᰝࡸ㸪ㄪ⌮ᚑ⪅ࡢ⾨⏕ᩍ⫱➼ࡀ㐍ࡵ ◊✲㈝⿵ຓ㔠ࠕ㣗ရ୰ࡢཎ࢘ࣝࢫࡢࣜࢫࢡ 1,2) ࠋࡋࡋ࡞ࡀࡽ㸪ཎᅉࡋ࡚ ⟶ ⌮ 㛵 ࡍ ࡿ ◊ ✲ ࠖ㸦 H22㸫 㣗 ရ 㸫 ୍ ⯡ 㸫 013㸧 ࢃࢀࡿ㣗ရࡽࡢ࢘ࣝࢫ᳨ฟࡣ㸪ࡑࡢసᴗ ࠾࠸࡚㸪ᕷ㈍ࡢ࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࢆ⏝ ࡢᅔ㞴ࡉࡽࡇࢀࡲ࡛ࢇ᳨ウࡉࢀ࡚ࡇ࡞ ࡍࡿࡇ࡛ῧຍᢠయࡢᏳᐃ౪⤥ࡀᅗࡽࢀࡓ㸪 ࡗࡓࡓࡵ㸪ලయⓗ࡞ởᰁ࣮ࣝࢺࡢゎ᫂Ỵࡵ ᳨ฟࡋࡓ㑇ఏᏊࡢሷᇶ㓄ิゎᯒࡶྍ⬟࡞᪉ἲ ᡭࢆḞ࠸࡚࠸ࡓࠋཎᅉ≀㉁ࡋ࡚ࡣࣀࣟ࢘ࣝ ࡋ࡚Ⓨᒎࡉࡏࡿࡇࡀ࡛ࡁࡓ ࢫ㸦NoV㸧 ࡀ㒊 ศࢆ༨ ࡵ࡚࠸ࡿࡀ 㸪 ࡶࢧ 24 ᖺ ᗘ ᐇ ࡋ ࡓ ඹ ㏻ ヨ ⸆ ඹ ㏻ ᳨ య ࢆ ⏝ ࠸ ࣏ ࢘ ࣝ ࢫ 㸦 SaV㸧 ࡸ ࢹ ࣀ ࢘ ࣝ ࢫ 41 ᆺ ࡓࢥ࣭ࣛ࣎ࢫࢱࢹ࠾࠸࡚㸪ヨ㦂ᶵ㛵ࡈࡢ 㸦 AdV41㸧௦⾲ ࡉࢀࡿ ⭠⟶⣔ࢹ ࣀ࢘ࣝࢫ ⤖ᯝࡢࣂࣛࡘࡁࡀၥ㢟࡞ࡗࡓ ࡶྵࡲࢀ࡚࠸ࡿࠋࡉࡽ㸪㍺ධ㣗ရ➼ࡀཎᅉ ✲࡛ࡣ㸪⤖ᯝᕪࡀ⏕ࡌࡸࡍ࠸⟠ᡤ╔┠ࡋ㸪 ⪃࠼ࡽࢀࡿ A ᆺ⫢⅖࢘ࣝࢫ㸦HAV㸧ឤᰁ⪅ࡢ ᛂ⣔➼ࢆ᭱㐺ࡍࡿࡇࡼࡗ᳨࡚ฟ⢭ᗘࢆ ሗ࿌ࡀᛴቑࡍࡿ࡞㸪㣗ရ୰ࡢ࢘ࣝࢫࢆ᳨ฟ ྥୖࡉࡏࡿࡓࡵࡢ᳨ウࢆ⾜ࡗࡓࠋ ࡽࢀ࡚ࡁ࡚࠸ࡿ ࡍࡿ᪉ἲࡢ☜❧ࡀᛴົ࡞ࡗ࡚࠸ࡿ 3) 14-18) ࠋ୍᪉㸪ᖹᡂ 19) ࠋࡑࡇ࡛ᮏ◊ ࠋᖹᡂ 19㹼 21 ᖺ ᗘ ᐇ ࡉ ࢀ ࡓ ཌ ⏕ ປ ാ ⛉ Ꮫ ◊ ✲ ㈝ ᪉ἲ ⿵ຓ㔠ࠕ㣗ရ୰ࡢ࢘ࣝࢫࡢไᚚ㛵ࡍࡿ◊✲ࠖ ◊✲ᮦᩱ 㸦H19㸫㣗ရ㸫୍⯡㸫016㸧࠾࠸࡚㸪ᅛᙧ㸪ᾮ ᐇ㦂⏝࠸ࡿ㣗ရࡋ࡚㸪ᕷ㈍ࡉࢀ࡚࠸ࡿࡁ ≧㸪⦎ࡾ≀㸪Ἔ≀࡞ࡢ୍⯡ⓗ࡞㣗ရࡽ NoV ࡞⢊࣏ࢸࢺࢧࣛࢲࢆ⏝࠸ࡓࠋࡲࡓ㸪᳨ฟᑐ㇟ ࢆ᳨ฟࡍࡿᡭἲࡋ࡚ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉ ࡞ࡿ࢘ࣝࢫࡋ࡚㸪NoV-GII.4㸦AB293424㸧 ἲ㸦ࣃࣥࢺࣛἲ㸧ࢆ㛤Ⓨࡋ㸪ࡇࡢၥ㢟ࢆゎỴࡍ ࢆྵࡴ⣅౽ࢆ⏝࠸ࡓࠋ *1 ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ - 37 - 秋田県健康環境センター年報 第10号 2014 ࣟࢩࣗ♫〇ࠕLightCycler FastStart DNA Master ヨ⸆㢮 PLUS Hybridization Probesࠖࢆ⏝࠸ࡓࠋ 㣗ရὙᾮ Tris-HCl㸦 pH8.4㸧– 0.5 M NaCl – 0.1% Tween20 UHDOWLPH3&5 ᛂ⣔ Kageyama 21) ࡽࡢ᪉ἲᚑࡗࡓࠋ ࢆㄪ〇ࡋ࡚⏝ࡋࡓࠋ ࣐࢞ࣥࢢࣟࣈࣜࣥ〇 ⡿ᅜ Baxter ♫ࡢ 5㸣㟼ὀ⏝࣐࢞ࣥࢢࣟࣈࣜࣥ ⤖ᯝࡢᕪࡀ⏕ࡌࡸࡍ࠸⟠ᡤࡢᐃ 〇 ࠕ Gammagard ࠖ ࢆ ⏝ ࠸ ࡓ ࠋ Alfresa Pharma ヨ㦂ᶵ㛵ࡼࡗ࡚⤖ᯝࡢࣂࣛࡘࡁࡀ⏕ࡌࡸࡍ ♫ࡽ㉎ධࡋࡓࠋ ࠸㒊ศࡋ࡚㸪ḟࡢ 4 ⟠ᡤࢆᐃࡋࡓࠋࡇࡢෆ㸪 ࣃࣥࢯࣝࣅࣥ 㸦1㸧ࡣ࢘ࣝࢫ RNA ᢳฟẁ㝵㸦⊃⩏ࡢࣃࣥࢺ 㯤Ⰽࣈࢻ࢘⌫⳦ࢆ⇕ฎ⌮ࡋ࡚࣐࣍ࣝࣜࣥᅛᐃ ࣛ ἲ 㸧 ࡢ ࡶ ࡢ ࡛ ࠶ ࡾ 㸪 㸦 2㸧 ~㸦 4㸧 ࡣ ᢳ ฟ ࡋ ࡓ ࡋࡓࡶࡢࡢᠱ⃮ᾮ࡛㸪࣓ࣝࢡ♫ࡽ㉎ධࡋࡓࠋ RNA ࡢ᳨ฟẁ㝵┦ᙜࡍࡿࠋ ࣇ࢙ࣀ࣮ࣝ⣔ 51$ ᢳฟ࢟ࢵࢺ 㸦1㸧Į-Amylase ⢊ᮎࡢ⁐ᛶᡂศࡢṧ␃㛵ࡍ TRIzol-LS㸦invitrogen㸧ࢆ⏝ࡋࡓࠋ ࡿၥ㢟 㸦2㸧ᢳฟࡋࡓ RNA ⁐ᾮᑐࡍࡿ DNase ฎ⌮ࡢ ࣒࢝ࣛ᪉ᘧࡢ 51$ ᢳฟ࢟ࢵࢺ QIAamp Viral RNA Mini Kit (Qiagen)ࢆ⏝ࡋࡓࠋ ᠱ⃮ᾮ ၥ㢟 㸦3㸧㏫㌿㓝⣲ࡢ㑅ᐃᛂ᮲௳᭱㐺 2.2.5 ࡢᢳฟ࢟ࢵࢺῧࡢ AVL ᾮࢆ⏝࠸ࡓࠋ '1DVH,㸦57*UDGH㸧ཬࡧ 51DVHLQKLELWRU ࢽࢵ࣏ࣥࢪ࣮ࣥࡢ〇ရࢆ⏝ࡋࡓࠋ 㸦4㸧PCR 㓝⣲ࡢ㑅ᐃᛂ᮲௳᭱㐺 ࡞࠾㸪ࣉࣟࢺࢥࣝୖࡢ㡰 ␒ࡣ㸦1㸧ĺ㸦4㸧࡛࠶ ࡿࡀ㸪᭱㐺࠶ࡓࡗ࡚ࡣ㏲ḟ⤖ᯝࢆほᐹࡋ࡞ +HDOODELOH GRXEOH VWUDQG VSHFLILF ࡀࡽ⾜࠺ᚲせࡀ࠶ࡿࡓࡵ㸪᳨ウࡢ㡰␒ࡣ㸦4㸧ĺ '1DVH 㸦+/GV'1DVH᫆⇕ᛶ ᮏ㙐≉␗ⓗ '1DVH㸧 㸦1㸧ࡋࡓࠋࡍ࡞ࢃࡕ㸪᭱ึ㸦4㸧ࡢ᳨ウ ࣀ࢙࣮ࣝ࢘ArcticZymes ♫ࡽ㉎ධࡋࡓࠋ ᭱㐺ࢆ⾜࠸㸪ḟ㸦3㸧ࢆ᳨ウࡍࡿ㸦᭱㐺῭ ࡳࡢ㸦4㸧ࢆ⏝࠸࡚⤖ᯝࢆほᐹࡍࡿ㸧࠸࠺㡰 ࣑࣮ࣛࢮ ᯤⲡ⳦⏤᮶ Į-Amylase ⢊ᮎ㸦ග⣧⸆㸧ࢆ ࡞ࡿࠋ ⏝ࡋࡓࠋ 㣗ရฎ⌮⿄ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢయࡢᡭ㡰 ᅗ 1 ♧ࡋࡓỗ⏝ࣉࣟࢺࢥࣝᚑࡗࡓࠋᮏ◊ ࢧࢽࢫ࣌ࢵࢡࢸࢫࢺࣂࢵࢢ㸦ࢬ࣡ࣥ㸧ࢆ ⏝ࡋࡓࠋ ✲࡛ࡣ㸪ྛ✀㓝⣲ࡢ㑅ᢥᛂ᮲௳ࡢ᭱㐺ࡢ FRQYHQWLRQDO3&5 ⏝㓝⣲ ࡓࡵ㸪ࠕࣃࣥࢺࣛᢳฟ≀ࠖࡼࡿ㈇Ⲵヨ㦂ࢆ ⾲ グ㍕ࡋࡓࡶࡢࢆ⏝ࡋࡓࠋ ⾜ࡗࡓࠋࠕࣃࣥࢺࣛᢳฟ≀ࠖࡣ㸪࢘ࣝࢫࢆྵ ㏫㌿㓝⣲ ࡲ࡞࠸ᕷ㈍ࡢࡁ࡞⢊ࡽࣃࣥࢺࣛἲࡢࣉࣟࢺࢥ ⾲ グ㍕ࡋࡓࡶࡢࢆ⏝ࡋࡓࠋ ࣝࢆ⏝࠸࡚ㄪ〇ࡋࡓࠋ ᢠ 7DT ࣔࣀࢡ࣮ࣟࢼࣝᢠయ anti-Taq high㸦ᮾὒ⣳ 㸧ࢆ⏝ࡋࡓࠋ 3&5 㛵ࡍࡿ᳨ウ NoV ᳨ ᰝ ࡢ ࡓ ࡵ ᅜ ❧ ឤ ᰁ ◊ ✲ ᡤ ࡼ ࡾ 㓄 ㏫㌿ᛂ⏝࠸ࡓࣉ࣐࣮ࣛ ㏫ ㌿ ᛂ ᑓ ⏝ ࣉ ࣛ ࣐ ࣮ PANR-G2 18) ࢆ⏝ ᕸࡉࢀ࡚࠸ࡿᐃ㔞ࢥࣥࢺ࣮ࣟࣝ DNA ࢆ␃Ỉ㸪 ࠸ࡓࠋ ཬࡧࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡍࡿࡇ࡛㸪 FRQYHQWLRQDO3&5 ⏝࠸ࡓࣉ࣐࣮ࣛ 10 5 ~10 1 ࢥࣆ࣮ ȝ/ ࡢ NoV 㑇ఏᏊ㓄ิ⏤᮶ DNA COG2F 20) / G2SKR 21) 㸪 ࡲ ࡓ ࡣ G2SKF 21) / ᩿∦ࢆྵࡴ⿕᳨యࢆㄪ〇ࡋࡓࠋࡇࢀࡽࢆ㸪⾲ 1 G2SKR ࡢࣉ࣐࣮ࣛࢭࢵࢺࢆ⏝࠸ࡓࠋ ♧ࡋࡓ 32 ✀㢮ࡢ PCR ᛂ⣔࡛ቑᖜຠ⋡ࢆẚ FRQYHQWLRQDO3&5 ⨨ ㍑ࡋࡓ ࠋࣉࣛ ࣐࣮ ࢭࢵ ࢺࡣ G2SKF / G2SKR ࢫࢸࢵࢡ♫〇ࠕPC-320ࠖࢆ⏝࠸ࡓࠋ UHDOWLPH3&5 ⨨ ࣟࢩࣗ♫〇ࠕLightCycler 320Sࠖࢆ⏝࠸ࡓࠋ UHDOWLPH3&5 ⏝㓝⣲ ࢆ⏝࠸㸪ᛂᐜ㔞 25 ȝ/ ୰ୖグ⿕᳨యࡀ 5 ȝ/ ྵࡲࢀࡿࡼ࠺ࡋࡓࠋ⾲ 1 ࡛ࡣ㸪㓝⣲ࡢ✀ู ࡋ࡚ࣇ࣑࣮ࣜA㸦pol ᆺ㓝⣲㸧㸪ࣇ࣑࣮ࣜB 㸦Į ᆺ㓝⣲㸧㸪ཬࡧࡑࢀࡽࡢΰྜ㸦Blend㸧ࢆ♧ - 38 - 秋田県健康環境センター年報 第10号 2014 㣗ရ 10 g Ὑᾮ 5 mL 㸦㣗ရฎ⌮⿄୰࡛⾜࠺㸧 ㉸㡢Ἴฎ⌮ 15 ศ ࢳ࢙ ࢵࢡ ࣏ ࣥࢺ ձ Į-Amylase ⢊ᮎ 0.125 g 3,000 rpm 30 ศ ࢳ࢙ ࢵࢡ ࣏ ࣥࢺ ղ ୖΎ 5% ࣐࢞ࣥࢢࣟࣈࣜࣥ〇 ȝ/ ࣃࣥࢯࣝࣅࣥ 1.0 mL 37Υ15 ศ 㟼⨨ 3,000 rpm 20 ศ ỿ⃦ ᠱ⃮ᾮ ȝ/ TRIzol-/6 ȝ/ ࢡ࣒ࣟࣟ࣍ࣝ ȝ/ ⃭ࡋࡃ㟈┞ ࢳ࢙ ࢵࢡ ࣏ ࣥࢺ ճ 12,000 rpm 15 ศ Ỉᒙ ࢳ࢙ ࢵࢡ ࣏ ࣥࢺ մ ࢚ࢱࣀ࣮ࣝ 㸦Ỉᒙࡢ 0.8 ಸ㔞㸧 QIAamp Viral RNA Mini Kit ࡢ࣒࢝ࣛࣉࣛࡋ࡚ RNA ᢳฟ ྛ✀ࣉ࣐࣮ࣛࡼࡾ cDNA ࢆྜᡂࡋ㸪PCR ᅗ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ᧯సᡭ㡰 ͤձ 㹼մ ࡢࢳ ࢙ࢵ ࢡ࣏ ࣥ ࢺࡣ Į-Amylase ᾮ ㄪ〇 ἲ ࡛ཧ ↷ࡍ ࡿࠋ ࡋࡓࠋࡲࡓ㸪࣍ࢵࢺࢫࢱ࣮ࢺᵝࡢ᭷↓㸦”H”㸪 ᗘ㛫ࡣ㓝⣲ࡢῧㄝ᫂᭩ᚑࡗ࡚タᐃࡋࡓࠋ ཬࡧ”㸫”㸧 Mg ࡢ⤊⃰ ᗘ 㸦 mM㸧 ࡶ グ ㍕ ࡋ ࡓ ࠋ ᛂ buffer ࡣ㓝⣲ῧࡢࡶࡢࢆᇶᮏࡋ㸪ࡉࡽ ࠼ࡤ㸪ࠕA/ H /1.5 mMࠖࡢ⾲グࡣࠕࣇ࣑࣮ࣜ MgCl 2 ࢆ㏣ຍࡋ࡚ Mg 2+ ࢆቑຍࡉࡏࡓࡶࡢࡶ A ᒓࡍࡿ㓝⣲ࡀ⏝࠸ࡽࢀ㸪࣍ࢵࢺࢫࢱ࣮ࢺ ᳨ウࡋࡓࠋ ᵝ࡛㸪Mg ⃰ᗘࡣ 1.5 mM ࡛ᛂࢆ⾜࠺ࠖ࠸࠺ ࠙ࢱࢵࢳࢲ࢘ࣥ PCRࠚ ព࡞ࡿࠋࡲࡓ㸪ᑗ᮶ⓗഇ㝧ᛶᑐ⟇ࢆㅮࡎ 95Υ2 ศ ࡿᕸ▼ࡋ࡚㸪ᇶ㉁ dUTP ࢆ⏝࠸ࡓᛂ⣔ࡶ 95Υ30 ⛊㸫㸦ĺΥ㸧30 ⛊㸫72Υ30 ⛊ 5 ྠẚ㍑ࡋࡓࠋPCR ⨨ࡢ ᗘࣉࣟࢢ࣒ࣛࡣ㸪 ࢧࢡࣝ㸸ୗ⥺㒊ࡀࢱࢵࢳࢲ࢘ࣥタᐃ ḟ ♧ࡍ ࠾ࡾ ࡢࢱ ࢵ ࢳ ࢲ࢘ ࣥ PCR ࢆ ᇶ ᮏ 95Υ30 ⛊㸫50Υ30 ⛊㸫72Υ30 ⛊ 40 ࢧࢡࣝ ࡋࡓࡀ㸪denature ᗘ㛫㸪ཬࡧ extension 72Υ7 ศ 1 ࢧࢡࣝ - 39 - 1 ࢧࢡࣝ 秋田県健康環境センター年報 第10号 2014 ⾲ ẚ㍑᳨ウࡋࡓ 3&5 ⏝㓝⣲ᛂ⣔㸦ḟ㡫⥆ࡃ㸧 ࣓࣮ ࣮࢝ No. ࣓࣮ ࣮࢝ 㺪㺅㺮㺶㺎 / 㺬㺍㺢㺛㺞㺎㺢 /Mg ⃰ ᗘ No. ရྡ 㸦 ᛂ⣔ ࡢ ㏣ຍ 㸧 1 2 3 㸦 ᛂ⣔ ࡢ ㏣ຍ 㸧 greiner A /㸫/1.5 mM Taq DNA Polymerase, Standard greiner 11 Roche Diagnostics A /H/1.8 mM FastStart High Fidelity PCR System Roche Diagnostics A /H/1.5 mM 12 Taq DNA Polymerase, Hot Start A /H/2.5 mM AptaTaq Fast PCR Master 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 greiner A /㸫/2.0 mM Taq DNA Polymerase, High Yield 13 greiner 4 㺪㺅㺮㺶㺎 / 㺬㺍㺢㺛㺞㺎㺢 /Mg ⃰ ᗘ ရྡ TaKaRa TaKaRa A /㸫/1.5 mM Taq TaKaRa A /H/2.0 mM Taq DNA Polymerase, High Yield 14 TaKaRa A /H/1.5 mM Taq HS 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 TaKaRa greiner 5 A /H/4.5 mM Taq DNA Polymerase, High Yield 15 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 Blend /㸫/2.0 mM TaKaRa Ex Taq 㸦Mg ቑ 㔞 ĺP0㸧 6 7 8 9 10 greiner Blend /㸫/2.0 mM Taq DNA Polymerase, High Fidelity greiner B /㸫 /2.0 mM 17 Pfu-X DNA Polymerase TOYOBO 16 TaKaRa Blend /H /2.0 mM TaKaRa Ex Taq HS TaKaRa Blend /㸫/3.0 mM TaKaRa Z-Taq TaKaRa B /H /2.0 mM 18 KOD FX Neo Blend /H /3.0 mM TaKaRa Z-Taq 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 Agilent Technologies B /H/2.0 mM PfuTurbo Cx Hotstart DNA Polymerase 19 Roche Diagnostics A /H/2.0 mM FastStart Taq DNA Polymerase 20 TaKaRa B /H /2.0 mM Tks Gflex DNA Polymerase TaKaRa A /H/4.0 mM MightyAmp DNA Polymerse Ver.2 ࢆ☜ㄆࡋࡓࠋ ㏫㌿ᛂ㛵ࡍࡿ᳨ウ ⣅౽᳨యࡼࡾᢳฟࡋࡓ NoV ࡢ RNA ࢆ␃Ỉ㸪 ཬࡧࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ⿕᳨యࢆㄪ '1DVH ฎ⌮㛵ࡍࡿ᳨ウ 〇ࡋࡓࠋDNase ฎ⌮ࡣ⾜ࢃࡎ㸪㏫㌿ᛂᑓ ⣅౽᳨యࡼࡾᢳฟࡋࡓ NoV ࡢ RNA ⁐ᾮᑐ ⏝ࣉ࣐࣮ࣛPANR-G2 ⾲ 2 ♧ࡍ㏫㌿㓝 ࡋ࡚㸪DNase I ฎ⌮㸦37Υ 10 ศ㸧ࢆ⾜࠸㸪75 ⣲ࢆ⏝࠸࡚ cDNA ࢆྜᡂࡋࡓࠋᛂ ᗘࡣ㸪ῧ Υ 5 ศຍ⇕ࡍࡿᛂ⣔ࢆᇶᮏ㸦୍⯡ⓗ⾜ࢃࢀ ㄝ᫂᭩グ㍕ࡀ࠶ࡿ࠾ࡾ 37Υ㸪42Υ㸪55Υ㸪 ࡚࠸ࡿ᪉ἲ㸧ࡋࡓࠋࡉࡽ㸪ຍ⇕ࢆࡋ࡞࠸ 58Υ㸪ཬࡧ 60Υ࡚⾜ࡗࡓࠋᛂᐜ㔞ࡣ 20 ȝ/ ᛂ ⣔ ࡋ ࡚ 㸪 DNase I ᫆ ⇕ ᛶ 㓝 ⣲ ࡛ ࠶ ࡿ ࡛㸪ୖグ⿕᳨యࡀ 10 ȝ/ ྵࡲࢀࡿࡼ࠺ࡋࡓࠋ HL-dsDNase ࢆ ⏝ ࠸ ࡓ ሙ ྜ ࢆ ẚ ㍑ ࡋ ࡓ ࠋ ࡲ ࡓ 㸪 30 ศࡢ㏫㌿ᛂᚋ㸪␃Ỉࢆ 20 ȝ/ ຍ࠼ࡓࡶ ࣃࣥࢺࣛἲ࡛ RNA ᢳฟ ࢆ⾜࠺㏵୰࡛ DNase I ࡢ㸦2 ಸᕼ㔘㸧ࢆ 5 ȝ/ ྲྀ ࡾ㸪real-time PCR ඛ ࢆస⏝ࡉࡏࡿࠕ࣒࢜ࣥ࢝ࣛ DNase I ฎ⌮ࠖࡘ ᭱㐺ࢆ῭ࡲࡏࡓ conventional PCR 㸦ࣉࣛ ࠸࡚ࡶ᳨ウࡋࡓ㸦ᅗ 7㸧ࠋࡇࢀࡽࡢ᪉ἲ࡛ DNase ࣐࣮ࢭࢵࢺࡣ COG2F / G2SKR㸧ࡢᛂ⣔ࡼࡾ ฎ⌮ࢆ⾜ࡗࡓᚋ㸪ඛ᭱㐺ࢆ῭ࡲࡏࡓ㏫㌿ ቑᖜຠ⋡ࢆẚ㍑ࡋࡓࠋconventional PCR ࡛ቑᖜ ᛂ⣔ࡼࡾ cDNA ࢆྜᡂࡋ㸪real-time PCR ࡛ ࣂࣥࢻࡀほᐹࡉࢀࡓሙྜࡣษࡾฟࡋ࡚ሷᇶ㓄ิ ࢥࣆ࣮ᩘࢆ ᐃࡋࡓࠋ - 40 - 秋田県健康環境センター年報 第10号 2014 ⾲ 㸦๓㡫ࡽࡢ⥆ࡁ㸧 ࣓࣮ ࣮࢝ No. 21 ࣓࣮ ࣮࢝ 㺪㺅㺮㺶㺎 / 㺬㺍㺢㺛㺞㺎㺢 /Mg ⃰ ᗘ No. ရྡ 㺪㺅㺮㺶㺎 / 㺬㺍㺢㺛㺞㺎㺢 /Mg ⃰ ᗘ ရྡ 㸦 ᛂ⣔ ࡢ ㏣ຍ 㸧 㸦 ᛂ⣔ ࡢ ㏣ຍ 㸧 New England BioLabs KAPA BIOSYSTEMS A /㸫/2.0 mM Taq DNA Polymerase with ThermoPol Buffer A /H/4.5 mM KAPA2G Robust HotStart PCR Kit, buffer A 27 㸦Mg ቑ 㔞 ĺ mM㸧 New England BioLabs 22 KAPA BIOSYSTEMS A /H/2.0 mM Taq DNA Polymerase with ThermoPol Buffer A /H/1.5 mM KAPA2G Fast HotStart PCR Kit 28 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 New England BioLabs 23 KAPA BIOSYSTEMS A /H/4.5 mM Taq DNA Polymerase with ThermoPol Buffer A /H/4.5 mM KAPA2G Fast HotStart PCR Kit 29 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 㸦Mg ቑ 㔞 ĺ mM㸧 㸦Mg ቑ 㔞 ĺ mM㸧 24 Nippongene A /H/1.5 mM THUNDER Taq Gold DNA Polymerase LD BIOLINE 30 Nippongene 25 26 BIOLINE A /H/4.5 mM THUNDER Taq Gold DNA Polymerase LD A /㸫/2.0 mM HybriPol DNA Polymerase 31 A /H/2.0 mM HybriPol DNA Polymerase 㸦Mg ቑ 㔞 ĺ mM㸧 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 KAPA BIOSYSTEMS BIOLINE A /H/1.5 mM KAPA2G Robust HotStart PCR Kit, buffer A A /H/4.5 mM HybriPol DNA Polymerase 32 㸦ᢠ Taq ࣔ ࣀࢡ ࣮ࣟ ࢼࣝ ᢠ యῧ ຍ㸧 㸦Mg ቑ 㔞 ĺ mM㸧 㸦2㸧ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ Į-Amyalse Ș$P\ODVH ฎ⌮㛵ࡍࡿ᳨ウ ᅗ 10 ♧ࡋࡓ࠾ࡾࡢᡭ㡰࡛ Į-Amylase ⢊ᮎ ⢊ᮎ 0.125 g ࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ࠾ ࢆᾮㄪ〇ࡋࡓࠋ㸫20Υ࡚ 2 ࣨ᭶ಖᏑࡋࡓᚋ㸪 ࠸࡚㸪ୖΎࢆࢹ࢝ࣥࢺࡼࡾูࢳ࣮ࣗࣈ⛣ࡋ ຠᯝࡢ☜ㄆࢆ⾜ࡗࡓࠋⅣỈ≀㝖ཤຠᯝ☜ㄆࡢ ࡓࡶࡢ ࡓࡵ㸪0.1%㤿㕥䭏ࢹࣥࣉࣥᾮ㸦␃Ỉ࡚ຍ 㸦3㸧ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ Į-Amyalse ⢶㸧࣏ࢸࢺࢧࣛࢲ 10 g ࢆ 50 mL ࡢ␃Ỉ ⢊ᮎ 0.125 g ࢆຍ࠼㸪ࡑࡢᚋࡢ㐲ᚰࢆ⾜ࢃ࡞ ᠱ⃮ࡋ࡚㣗ရฎ⌮⿄ࡢࣇࣝࢱ࣮ࢆ㏻ࡋࡓℐ ࡗࡓࡶࡢ ᾮࢆ⏝࠸ࡓࠋࡇࢀࡽࡢ⿕᳨య 50 mL Į-Amylase 㸦4㸧ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ Į-Amyalse ⢊ᮎ 0.125 g㸪ࡲࡓࡣᾮㄪ〇ရ 1 mL ࢆῧຍࡋ ᾮㄪ〇ရ 1 mL ࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ ࡚㸪37Υ 15 ศ㟼⨨ࡋ㸪3,000 rpm 30 ศ㐲ᚰࡋࡓ ࠾࠸࡚㸪ୖΎࢆࢹ࢝ࣥࢺࡼࡾูࢳ࣮ࣗࣈ ୖΎࢆ 5 mL ศྲྀࡋ㸪ࡑࡇ᪥ᮏ⸆ᒁ᪉ࠕ࣮ࣚ ⛣ࡋࡓࡶࡢ ࢻࢳࣥ࢟ࠖ50 ȝ/ ࢆຍ࠼࡚ࣚ࢘⣲ࢹࣥࣉࣥᛂ ᢳฟࡋࡓ RNA ࢆ㗪ᆺࡋ࡚ඛ᭱㐺ࢆ῭ࡲ ࢆほᐹࡋࡓࠋ ࡏࡓ㏫㌿ᛂ⣔࡛ cDNA ࢆྜᡂࡋ㸪real-time ḟࣃࣥࢺࣛἲࢆᐇࡍࡿୖ࡛ࡢᙳ㡪ࢆ᳨ウ PCR ᳨࡛ฟࢆヨࡳࡓࠋ ࡍࡿࡓࡵ㸪㣗ရὙᾮ NoV ࢆῧຍࡋࡓࡶࡢ 㸦㣗ရࡣධࢀ࡞࠸㸧ᑐࡋ࡚㸪ḟࡢ᧯సࢆ⾜࠸㸪 ⤖ᯝ RNA ᢳฟᾮࢆᚓࡓࠋ 3&5 ⏝࠸ࡿ㓝⣲ࡢ㑅ᐃᛂ᮲௳᭱㐺 㸦1㸧ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ Į-Amyalse NoV 㑇ఏᏊ㓄ิ⏤᮶ DNA ࢆ␃Ỉ࡛ẁ㝵ᕼ ⢊ᮎ 0.125 g ࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ࠾ 㔘ࡋࡓ⿕᳨యࢆ㗪ᆺࡋ㸪㏻ᖖࡢࢹ࢜࢟ࢩࢾࢡ ࠸࡚㸪ࢫ࣏ࢺࢆ⏝࠸࡚ୖΎࢆูࢳ࣮ࣗࣈ ࣞ࢜ࢳࢻ㸦dATP㸪dGTP㸪dCTP㸪dTTP㸧ࢆᇶ㉁ ᑀศྲྀࡋࡓࡶࡢ ࡋ ࡚ PCR ࢆ ⾜ࡗ ࡓሙ ྜࡣ 㓝⣲ 〇ရ 㛫ࡢ ᕪ - 41 - dNTP dUTP M 1 2 3 4 5 M 1 2 3 4 5 No.21 M: φX174/HaeϪ New England Biolabs 1: 10 5 ࢥࣆ࣮ Taq with PolBuffer 2: 10 4 ࢥࣆ࣮ 䠄14.84 /25 μl reaction䠅 3: 10 3 ࢥࣆ࣮ 4: 10 2 ࢥࣆ࣮ Family A 5: 10 1 ࢥࣆ࣮ No.7 greiner Pfu-X 䠄50 /25 μl reaction䠅 Family B No.15 TaKaRa TaKaRa Ex-Taq 䠄67.5 /25 μl reaction䠅 Blend ᅗ $ ␃Ỉ࡛ẁ㝵ᕼ㔘ࡋࡓ 1R9VW3&5 ⏘≀ᑐࡍࡿ QG3&5 “dNTP”⾲ グ ࡋ࡚ ࠶ࡿ ᛂ ࡣ dATP㸪dGTP㸪 dCTP㸪dTTP ࢆྛ 200 μM ࡚ ⏝ “dUTP”⾲ グ ࡋ࡚ ࠶ࡿ ᛂ ࡣ dATP㸪dGTP㸪dCTP㸪ࢆ ྛ 200 μM㸪dUTP ࢆ 600 μM ࡚ ⏝ ࡣ ࢇ ㄆ ࡵ ࡽ ࢀ ࡞ ࡗ ࡓ ࠋ ୍ ᪉ 㸪 dTTP ࢆ ࡀ࡛ࡁ࡞ࡗࡓࠋ dUTP ⨨ࡋࡓሙྜࡣ㸪ࣇ࣑࣮ࣜ A ᒓ ୍᪉࡛㸪NoV 㑇ఏᏊ㓄ิ⏤᮶ DNA ࢆࣃࣥࢺ ࡍࡿ㓝⣲〇ရࡢࡳࡀ᭷ຠ࡛㸪ࣇ࣑࣮ࣜ B ᒓ ࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ⿕᳨యࢆ㗪ᆺࡋࡓሙ ࡍࡿ㓝⣲〇ရ㸪ཬࡧࣇ࣑࣮ࣜ A ࣇ࣑࣮ࣜ ྜࡣ㓝⣲〇ရ㛫࡛ࡁ࡞ᕪࡀ⏕ࡌࡓࠋ࠼ࡤ㸪 B ࡢΰྜ〇ရ࡛ࡣቑᖜࡀ㉳ࡇࡽ࡞ࡗࡓ㸦ᅗ 2A ⾲ 1 ࡢ No.21 ࡣ␃Ỉࡼࡿᕼ㔘㸦ᅗ 2A㸧࡛ࡣ ୍㒊ࢆ♧ࡍ㸧ࠋእࡋ࡚㸪⾲ 1 ࡢ No.9 ࡣࣇ Ⰻዲ࡞㑇ఏᏊቑᖜࡀᚓࡽࢀࡓࡀ㸪ᅗ 2B ᕥ㸦ࣃ ࣑࣮ࣜB ᒓࡍࡿࡀ㸪 dUTP ࢆྲྀࡾ㎸ࡵࡿࡼ ࣥࢺࣛᢳฟ≀ࡼࡿᕼ㔘㸧࡛ࡣឤᗘࡀⴭࡋࡃప ࠺㓝⣲⺮ⓑࡀᨵኚࡉࢀ࡚࠸ࡿࠋࡲࡓ㸪⾲ 1 ࡢ ୗࡋ࡚࠸ࡓࠋྠࡌ㓝⣲〇ရᢠ Taq ࣔࣀࢡ࣮ࣟ No.20 ࡣࣇ࣑࣮ࣜ A ࡛ ࠶ࡿࡇࡽ㸪⌮ㄽⓗ ࢼࣝᢠయࢆຍ ࠼࡚ ”࣍ࢵ ࢺࢫࢱ࣮ࢺ ” ࡍࡿ㸪 dUTP ࢆ⏝࠸ࡿࡇࡀྍ⬟ࡔࡀ㸪〇ရᵓᡂ ᅗ 2B ࡢ No.22 ♧ࡍ࠾ࡾ␃Ỉ࡛ᕼ㔘ࡋࡓ ࡋ࡚ buffer ࢹ࢜࢟ࢩࢾࢡࣞ࢜ࢳࢻࡀࣉ࣑ࣞࢵ ሙྜྠ➼ឤᗘࡀᅇࡋࡓࠋ୍᪉㸪ᇶ㉁ࡢ୰ ࢡࢫࡉࢀ࡚࠸ࡿࡇࡽ dUTP ⨨ࡍࡿࡇ ࡢ dTTP ࢆ dUTP ⨨ࡋࡓᛂ⣔࡛ࡣឤᗘࡀ - 42 - dNTP dUTP M 1 2 3 4 5 M 1 2 3 4 5 No.21 New England Biolabs M: φX174/HaeϪ Taq with PolBuffer 1: 10 5 ࢥࣆ࣮ ق14.84 /25 μl reactionك 2: 10 4 ࢥࣆ࣮ Family A 4: 10 2 ࢥࣆ࣮ 3: 10 3 ࢥࣆ࣮ 5: 10 1 ࢥࣆ࣮ No.22 New England Biolabs ᢠ Taq ᢠ య ࡼࡿ Taq with PolBuffer ࣍ࢵ ࢺࢫ ࢱ࣮ ࢺ + anti-Taq high 䠄34.84 /25 μl reaction䠅 Family A No.23 New England Biolabs ᢠ Taq ᢠ య ࡼࡿ Taq with PolBuffer ࣍ࢵ ࢺࢫ ࢱ࣮ ࢺ + anti-Taq high + MgCl 2 MgCl 2 ࡢቑ 㔞㸦 4.5 mM㸧 䠄34.84 /25 μl reaction䠅 Family A ᅗ % ࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ 1R9VW3&5 ⏘≀ᑐࡍࡿ QG3&5 “dNTP”⾲ グࡋ ࡚࠶ ࡿ ᛂ ࡣ dATP㸪 dGTP㸪dCTP㸪dTTP ࢆྛ 200 μM ࡚ ⏝ “dUTP”⾲ グࡋ ࡚࠶ ࡿ ᛂ ࡣ dATP㸪 dGTP㸪dCTP㸪ࢆ ྛ 200 μM㸪 dUTP ࢆ 600 μ㹋 ࡚ ⏝ ࡸࡸప࠸ࡶࡢࡢ㸪 No.23 ࡢࡼ࠺ Mg ⃰ᗘࢆ ࡀ ON / OFF ࡉࢀࡿ⤌ࡳ㸦ࣉࢱ࣐࣮㸧ࡀ 4.5mM ࡲ ࡛ ቑ 㔞 ࡍ ࡿ ࡇ ࡛ ࠶ ࡿ ⛬ ᗘ ࡢ ᨵ ၿ ᑟධࡉࢀ࡚࠸࡚ᛂ≉␗ᛶࡢྥୖᐤࡋ࡚ ࡀぢࡽࢀࡓࠋࡑࡇ࡛㸪࣍ࢵࢺࢫࢱ࣮ࢺᵝ࡛ ࠸ࡿࡀ㸪PCR ࡢ➨୍ࢧࢡࣝࡔࡅࡣ㸪ࡼࡾ㧗 ࠶ࡿ ࡇ 㸪Mg ࢆ ቑ 㔞 ࡍࡿ ࡇ ╔║ ࡋ 㸪 ࡢ࣍ࢵࢺࢫࢱ࣮ࢺࡀᮃࡲࡋ࠸ࡓࡵᢠ Taq ࣔ ᇶ㉁ࡋ࡚ dUTP ࢆ⏝࠸࡚ࡶ࡞࠾ᅗ 2A ࡢୖ ࣀࢡ࣮ࣟࢼࣝᢠయࢆῧຍࡍࡿࡇ࡛⿵ࡋࡓࠋ ẁྑྠ➼ࡢឤᗘࡀᚓࡽࢀࡿᛂ⣔ࢆ᳨ウࡋ ࡓࡇࢁ㸪ᅗ 2C ♧ࡍ 3 ࡘ㸦⾲ 1 ࡢ No.12㸪 ㏫㌿㓝⣲ࡢ㑅ᐃᛂ᮲௳᭱㐺 No.27㸪No.32㸧ࡀ㑅ᢥࡉࢀࡓࠋࡇࡢෆ㸪No.12 NoV RNA ࢆ␃Ỉ࡛ẁ 㝵ᕼ㔘ࡋࡓ⿕᳨య 㸦ᅗ 2C ୗẁ㸧ࡘ࠸࡚ࡣ᭱ึࡽ dUTP ࡀ ࢆ㗪ᆺࡋࡓሙྜࡣ⾲ 2 ♧ࡋࡓ㓝⣲〇ရ ⤌ࡳ㎸ࡲࢀࡓࣉ࣑ࣞࢵࢡࢫ〇ရ࡛࠶ࡾ㸪ᇶ㉁ 㛫ࡢᕪࡣࢇㄆࡵࡽࢀ࡞ࡗࡓࠋࡋࡋ㸪 ࢆධࢀ᭰࠼ࡿᚲせࡀ࡞࠸ࡇࡽ➨୍㑅ᢥ ࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ⿕᳨య࡛ࡣ〇 ࡋࡓࠋࡇࡢ㓝⣲〇ရࡣ㸪 55Υࢆቃ㓝⣲άᛶ ရ㛫࡛ᕪࡀ⏕ࡌ㸪RNA ࡀ ప⃰ᗘ࡞ࡿࡘࢀ - 43 - dNTP dUTP M 1 2 3 4 5 M 1 2 3 4 5 No.27 KAPA BIOSYSTEMS KAPA2G Robust MgCl 2 ࡢቑ 㔞㸦4.5 mM㸧 HotStart DNA Polymerase with buffer A and enhancer1 + MgCl 2 㸦99.8 /25 μl reaction㸧 Family A No.32 BIOLINE HybriPol DNA Polymerase ᢠ Taq ᢠ య ࡼࡿ +anti-Taq high ࣍ࢵ ࢺࢫ ࢱ࣮ ࢺ + MgCl 2 MgCl 2 ࡢቑ 㔞㸦4.5 mM㸧 㸦75.5 /25 μl reaction㸧 Family A Fusion No.12 dUTP ⤌ ㎸ ῭ Roche Diagnostics ᢠ Taq ᢠ య ࡼࡿ AptaTaq Fast PCR Master ࣍ࢵ ࢺࢫ ࢱ࣮ ࢺ +anti-Taq high 䠄132 /20 μl reaction䠅 Family A ᅗ & ࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ 1R9 VW3&5 ⏘≀ᑐࡍࡿ QG3&5 “dNTP”⾲グࡋ࡚࠶ࡿᛂࡣ dATP㸪dGTP㸪dCTP㸪dTTP ࢆྛ 200 μM ࡚⏝ࠋ “dUTP”⾲グࡋ࡚࠶ࡿᛂࡣ dATP㸪dGTP㸪dCTP㸪ࢆྛ 200 μM㸪dUTP ࢆ 600 μM ࡚⏝ ⾲ ẚ㍑᳨ウࡋࡓ㏫㌿㓝⣲ᛂ ᗘ No. ࣓࣮࣮࢝ ရྡ ᛂ ᗘ㸦Υ㸧 1 New England BioLabs M-MLV Reverse Transcriptase 37 2 TOYOBO ReverTra Ace 42 3 TaKaRa PrimeScript 42 4 Roche Diagnostics Transcriptor 55㹼65 5 invitrogen SuperScript II 42 6 invitrogen Thermoscript 55㹼65 ࡑࡢᕪࡣ㢧ⴭ࡞ࡗࡓࠋᅗ 3A ࡣ 10 5 ࢥࣆ࣮ 2 ࡢ No.1㸧࡛ࡣ᫂ࡽቑᖜຠ⋡ࡀⴠࡕ࡚࠸ /μL ࡢ RNA ࢆ⿕᳨యࡋ ࡓሙྜࡢቑᖜ᭤⥺ࢆ ࡓࠋ10 3 ࢥࣆ࣮/μL㸦ᅗ 3B㸧㸪10 2 ࢥࣆ࣮/μL㸦ᅗ ♧ࡋࡓࡀ㸪37Υ࡛ᛂࢆ⾜࠺㏫㌿㓝⣲㸦⾲ 3C㸧⃰ᗘࡀప࠸⿕᳨య࡞ࡿ 42Υ࡛ᛂ - 44 - 10 5 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ Fluorescence (F1/F2) No.4 Transcriptor (55Υ ) No.2 ReverTraAce (42Υ ) No.5 SuperScript II (42Υ ) No.1 M-MLV RT (37Υ ) DW Cycle Number ᅗ $ ㏫㌿㓝⣲ࡢẚ㍑㸦㸧 No.4 Transcriptor (55Υ ) Fluorescence (F1/F2) 10 3 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ No.5 SuperScript II (42Υ ) No.2 ReverTraAce (42Υ ) No.1 M-MLV RT (37Υ ) DW Cycle Number ᅗ % ㏫㌿㓝⣲ࡢẚ㍑ 㸦㸧 10 2 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ Fluorescence (F1/F2) No.4 Transcriptor (55Υ ) No.5 SuperScript II (42Υ ) No.2 ReverTraAce (42Υ ) No.1 M-MLV RT (37Υ ) DW Cycle Number ᅗ & ㏫㌿㓝⣲ࡢẚ㍑㸦㸧 - 45 - 10 3 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ Fluorescence (F1/F2) 60Υ 55Υ DW Cycle Number ᅗ $ 7UDQVFULSWRU ࡢᛂ ᗘẚ㍑㸦 㸧 60Υ 10 3 Fluorescence (F1/F2) 58Υ ࢥࣆ ࣮/μL 58Υ 10 2 58Υ 60Υ 10 1 60Υ Cycle Number ᅗ % 7UDQVFULSWRU ࡢᛂ ᗘẚ㍑㸦 㸧 ࢆ⾜࠺㏫㌿㓝⣲㸦⾲ 2 ࡢ No.2㸪No.5㸧࠾࠸ ⣲㸦⾲ 2 ࡢ No.4㸧ࢆ⏝࠸࡚㸪55Υ 60Υ࡛ᛂ ࡚ ࡶ ቑ ᖜ ຠ ⋡ ࡢ ప ୗ ࡀ ほ ᐹ ࡉ ࢀ ࡓ ࠋ ᅗ 3A㹼 ᅗ ࡉࡏࡓሙྜ㸪ᚋ⪅ࡢቑᖜຠ⋡ࡀࡼࡾ㧗ࡗࡓ㸦ᅗ 3C ࡢቑᖜ᭤⥺࡛ࡣ㸪ᶓ㍈ࡽࡢ❧ࡕୖ ࡀࡾࢧ 4A㸧ࠋḟᛂ ᗘࢆ 58Υ 60Υ࡛ẚ㍑ࡋࡓ ࢡࣝࢆぢࡿ㝈ࡾ〇ရ㛫ࡢᕪࡣࢇ࡞ࡃ㸪⿕᳨ ࡇࢁ㸪ప⃰ᗘࡢ⿕᳨య࡞ࡿ๓⪅ࡢቑᖜ యྵࡲࢀࡿ RNA ࡢࢥࣆ࣮ᩘࡶࡰྠࡌ⟬ᐃ ຠ⋡ࡀ㧗ࡗࡓ㸦ᅗ 4B㸧ࠋᛂ ᗘࡀ 58Υ࡛ ࡉࢀࡓࡀ㸪⦪㍈㸦ቑᖜຠ⋡㸧࠾࠸࡚ᕪࢆ⏕ࡌ࡚ ᭱㐺ࡉࢀࡓࡇࡽ㸪㧗 ᛂᑐᛂࡋࡓ㏫ ࠾ࡾ㸪᳨ฟ㝈⏺ᙳ㡪ࡀཬࢇ࡛࠸ࡓ㸦37Υ࡛ᛂ ㌿㓝⣲㛫࡛ࡢẚ㍑᳨ウࢆ⾜ࡗࡓ㸦⾲ 2 ࡢ No.4 ࢆ⾜࠺㓝⣲࡛ࡣᅗ 3B ᅗ 3C ࠾࠸࡚ࡣ㸪ࡶࡣ No.6㸧ࠋᅗ 5A ᅗ 5B ࡢ࠾ࡾ㸪୧⪅ࡢẚ ࡸቑᖜࡀぢࡽࢀ࡞࠸㸧ࠋ௨ ୖ ࡢ ࡇ ࡽ 㸪 ㏫ ㌿ ㍑࡛ࡣ⾲ 2 ࡢ No.4 ♧ࡋࡓ㏫㌿㓝⣲ࡢቑᖜຠ ᛂ࠾࠸࡚ࡣᛂ ᗘࡀ㔜せ࡛࠶ࡿࡇࡀ ⋡ࡀ㧗ࡗࡓࠋẚ㍑ᑐ↷ࡋ࡚㸪 42Υ࡛ᛂࢆ ࢃࡗࡓࡓࡵ㸪 ᗘࢆࡉࡽୖ᪼ࡉࡏࡓሙྜ ⾜࠺㏫㌿㓝⣲㸦⾲ 2 ࡢ No.3㸧ࡶྠ⏝࠸ࡓ ࡘ࠸᳨࡚ウࡋࡓࠋ㧗 ᛂᑐᛂࡋࡓ㏫㌿㓝 ࡀ㸪๓㏙ࡢ࠾ࡾ 58Υࡢ㧗 ㏫㌿㓝⣲ࡼࡾࡶ - 46 - 10 3 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ Fluorescence (F1/F2) No.4 Transcriptor (58Υ) No.6 ThermoScript (58Υ) No.3 PrimeScript (42Υ) DW Cycle Number ᅗ $ 㧗 ㏫㌿㓝⣲ࡢẚ㍑㸦㸧 10 2 ࢥࣆ࣮RNA / μL ࣃࣥࢺࣛᢳฟ≀ Fluorescence (F1/F2) No.4 Transcriptor (58Υ) No.6 ThermoScript (58Υ) No.3 PrimeScript (42Υ) DW Cycle Number ᅗ % 㧗 ㏫㌿㓝⣲ࡢẚ㍑㸦㸧 ቑᖜຠ⋡ࡣపࡗࡓࠋ ᢳฟࡋࡓ 51$ ⁐ᾮᑐࡍࡿ '1DVH ฎ⌮ἲࡢ᳨ウ ḟᅗ 3㹼5 ࠾࠸࡚ real-time PCR ᳨࡛ウࡋ 㧗 ㏫㌿ᛂࡼࡗ࡚㸪㯤Ⰽࣈࢻ࢘⌫⳦⏤ ࡓ⿕᳨యࢆ㸪ඛ᭱㐺ࢆ῭ࡲࡏࡓ conventional ᮶ࡢ㑇ఏᏊ➼㉳ᅉࡍࡿ㠀≉␗ᛂࢆᢚไࡍࡿ PCR ࡛ቑᖜࡋ࡚㸪㠀≉␗ᛂࡢ᭷↓ࢆ☜ㄆࡋࡓ ࡇࡀྍ⬟࡞ࡗࡓࡀ㸪㝜ᛶ᳨యࢆ semi-nested 㸦ᅗ 6㸧ࠋ37Υ࡛ᛂࢆ⾜࠺㓝⣲࡛ࡣࣂࣥࢻࡀ PCR ࡛ቑᖜࡋࡓሙྜ㸪NoV ࡣ↓㛵ಀࡢࣂ ほᐹࡉࢀࡎ㸦ᅗ 6 ᕥ㸧㸪42Υ࡛ᛂࢆ⾜࠺㓝⣲ ࣥࢻࡀฟ⌧ࡍࡿࡇࡀ࠶ࡗࡓࠋࡇ࠺ࡋࡓ㠀≉␗ ࡛ࡣప⃰ᗘࡢ⿕᳨య࡞ࡿ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ ࣂࣥࢻࡣ㸪real-time PCR㸦ࣁࣈࣜヨ㦂┦ᙜ㸧 ࣂࣥࢻ㸦16s ࣜ࣎ࢯ࣮࣒㸧ࡀฟ⌧ࡋࡓ㸦ᅗ 6 ୰ ࡛ࡣᛂࡋ࡞࠸ࡇࡽㄗุᐃࡣ⮳ࡽ࡞࠸ࡶ ิ㸧ࠋ㧗 ㏫㌿㓝⣲ࢆ⏝࠸ࡓሙྜ㸦ᅗ 6 ྑิ㸧㸪 ࡢࡢ㸪ቑᖜࢧࢬࡼࡗ࡚ࡣ୍ⓗ࡞ΰࢆࡁ 55Υࡢᛂ࡛ࡣ㯤Ⰽࣈࢻ࢘⌫⳦ࡢࣂࣥࢻࡀⱝᖸ ࡓࡍ࠾ࡑࢀࡀ࠶ࡿࡓࡵ㸪㏫㌿ᛂࡢ๓ ㄆࡵࡽࢀࡓࡶࡢࡢ㸪58Υ࡛ࡣ NoV ࡢࣂࣥࢻࡢࡳ DNase ฎ⌮ࢆ⾜࠺ࡇࡀ᥎ዡࡉࢀ࡚࠸ࡿࠋ୍᪉ ࡀほᐹࡉࢀࡓࠋ ࡛㸪୍⯡ᗈࡃ⏝࠸ࡽࢀ࡚࠸ࡿ DNase I ࢆ⏝࠸ - 47 - 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 10 5 → ẁ㝵ᕼ㔘 ᅗ ቑᖜࣂࣥࢻ࠾ࡅࡿ㏫㌿㓝⣲ᛂ ᗘࡢᙳ㡪 5 ࣭10 ࢥࣆ࣮/μL ࡢ NoV RNA ࢆࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓࡶࡢࢆ㸪ྛ✀㓝⣲࡛㏫㌿ࡋ㸪COG2F/G2SKR ࡢ ࣉ࣐࣮ࣛࢭࢵࢺ AptaTaq㸦ᢠ Taq ᢠయῧຍ㸧ࡼࡾ PCR ࢆ⾜ࡗࡓࠋྛ No.ࡣ⾲ 2 グ㍕ࡋࡓ㓝⣲ࢆ♧ࡍࠋቑ ᖜࡋࡓ DNA ᩿∦ࢆࢩ࣮ࢡ࢚ࣥࢫࡍࡿࡇ࡛ NoV ࡢࣂࣥࢻ࡛࠶ࡿྰࢆ☜ㄆࡋࡓࠋ ࡓࣉࣟࢺࢥ࡛ࣝࡣ㸪ᛂᚋせ࡞ࡗࡓ Ꮫⓗ࡛࠶ࡿࡇࡽ RNase inhibitor ࡢῧຍ DNase I ࢆኻάࡉࡏࡿࡓࡵ 75Υ 5 ศࡢຍ⇕ฎ ࡛ࡣ㜵ࡄࡇࡀ࡛ࡁ࡞࠸ࠋᚑࡗ࡚㸪ゎỴ⟇ࡋ ⌮ࡀᚲせ࡛࠶ࡾ㸪ࡑࡢࡇࡀ࢘ࣝࢫ RNA ࡢ ࡚ຍ⇕ࢆ⾜ࢃ࡞࠸᪉ἲࢆ⪃ࡍࡿᚲせࡀ࠶ࡗࡓࠋ ศゎࢆࡶࡓࡽࡍ࠸࠺ࢪ࣐ࣞࣥࡀᣦࡉࢀ࡚࠸ ᪉ἲࡢ 1 ࡘࡋ࡚㸪ᴟᾏ⏕ᜥࡍࡿ࢚ࣅ⏤᮶ ࡿࠋຍ⇕ࡼࡿ RNA ศゎࡣ㸪࣮ࣜ࣎ࢫࡢ OH ࡢ᫆⇕ᛶ DNase㸦HL-dsDNase㸧ࡢ᭷⏝ᛶ╔┠ ᇶࡢồ᰾⨨ᛂ⏤᮶ࡍࡿࡶࡢ࡛࠶ࡾ㸪⣧⢋ ࡋࡓࠋࡇࡢ㓝⣲ࡣ 37Υ࡛ 2 ᮏ㙐 DNA ࢆ≉␗ⓗ - 48 - 秋田県健康環境センター年報 第10号 2014 TRIzol ᢳฟᚋࡢỈᒙ 0.8 ಸࡢ EtOH ΰྜ࣭ࢫࣆࣥࢲ࢘ࣥ ༙㔞ࢆ࣒࢝ࣛࣉࣛ 8,000 rpm, 1 ศ ࢥࣞࢡࢩࣙࣥࢳ࣮ࣗࣈ ṧࡾ༙㔞 ࢆ࢝ࣛ ࣒ ࣉ ࣛ 8,000 rpm, 1 ศ ࢥࣞࢡࢩࣙࣥࢳ࣮ࣗࣈ AW1, 500 ȝO 8,000 rpm, 1 ศ ࢥࣞࢡࢩࣙࣥࢳ࣮ࣗࣈ ࢽࢵ࣏ࣥࢪ࣮ࣥ '1DVH, ⏝ 40 ȝO ࢆ࣒࢝ࣛ୰ᚰࡢࣇࣝࢱ࣮ 㸦ูࡢࢳ࣮ࣗࣈ࡛ΰྜ࣭1 ᳨యᙜࡓࡾ㸧 DW 16 ȝO ᐊ ࡛ 5 ศ㛫 10×ῧࣂࢵࣇ࣮ 4 ȝO 㟼⨨ '1DVH,XQLWȝO ȝO AW2,500 ȝO 8,000 rpm, 1 ศ ࢥࣞࢡࢩࣙࣥࢳ࣮ࣗࣈ AW2, 500 ȝO 15,000 rpm, 3 ศ ᚤ㔞㐲ᚰࢳ࣮ࣗࣈ AVE 60 ȝO ࡚⁐ฟ᧯స ᅗ 4,$DPS9LUDO51$0LQL.LW ࡢ࣒࢜ࣥ࢝ࣛ '1DVH, ฎ⌮ࡢ⤌㎸ࡳ ศゎࡋ㸪50Υ࡛άᛶࢆ♧ࡉ࡞ࡃ࡞ࡾ㸪55Υ࡛ ࡀ࠶ࡿࠋࡑࡇ࡛㸪୍⯡ⓗὶ㏻ࡋ࡚࠸ࡿ DNase I ྍ㏫ⓗኻάࡍࡿᛶ㉁ࢆ᭷ࡋ࡚࠸ࡿࠋᚑࡗ࡚㸪 ࢆ⏝࠸ࡓୖ࡛㸪ຍ⇕ࢆ⾜ࢃ࡞࠸᪉ἲࡋ࡚࢜ࣥ ඛ ᭱㐺 ࢆ ῭ࡲ ࡏࡓ 58Υ࡛ࡢ 㧗 ㏫ ㌿ ࣒࢝ࣛฎ⌮ࢆ⪃ࡋࡓࠋู࣓࣮࣮࢝ࡢ RNA ᢳ ᛂࢆ⤌ࡳྜࢃࡏࡿ࡞ࡽࡤ㸪DNase ฎ⌮ᚋ 75Υ ฟ࢟ࢵࢺࡢ୰ࡣ㸪࣒࢜ࣥ࢝ࣛ DNase I ฎ⌮ᕤ ࡢຍ⇕ࢆ⾜࠺ᚲせࡣ࡞ࡃ࡞ࡿࠋ࡞࠾㸪ࡇࡢ㓝⣲ ⛬ ࡀ ⤌ ㎸ ࡲ ࢀ ࡚ ࠸ ࡿ ࡶ ࡢ ࡶ ࠶ ࡿ ࡀ 㸪ࠕ QIAamp ࡣࡲࡔᡃࡀᅜ࡛ࡣὶ㏻ࡋ࡚࠸࡞࠸ࡓࡵ㸪ࣀࣝ࢘ Viral RNA Mini Kitࠖࡣࡑࡢࣉࣟࢭࢫࡀ࡞࠸ࡓ ࢙࣮ࡢ ArcticZymes ♫ࡽ┤᥋ྲྀࡾᐤࡏࡿᚲせ ࡵ㸪ᨵࡵ࡚⤌㎸ࢇࡔࣉࣟࢺࢥࣝࢆᅗ 7 ♧ࡋࡓࠋ - 49 - 1.0E+04 1.0E+03 1.0E+02 1.0E+01 1.0E+00 ᅗ $ ␃Ỉ୰ࡢ 1R951$ 㔞ཬࡰࡍ '1DVH ฎ⌮ࡢᙳ㡪 1.0E+03 1.0E+02 1.0E+01 1.0E+00 ᅗ % ࣃࣥࢺࣛᢳฟ≀୰ࡢ 1R951$ 㔞ཬࡰࡍ '1DVH ฎ⌮ࡢᙳ㡪 NoV RNA ࢆྵࢇࡔ␃Ỉ㸪ࡲࡓࡣࣃࣥࢺࣛ ࡓሙྜ࡛ࡶῶᑡࡀぢࡽࢀࡓࠋ୍᪉㸪HL-dsDNase ᢳฟ≀ᑐࡋ࡚ୖグࡢ DNase ฎ⌮ࢆ⾜࠸㸪ඛ ฎ⌮࣒࢜ࣥ࢝ࣛ DNase I ฎ⌮࡛ࡣ࠸ࡎࢀࡶࢥ ᭱㐺ࢆ῭ࡲࡏࡓ㧗 ㏫㌿ᛂ㸦⾲ 2 ࡢ No.4㸧 ࣆ࣮ᩘࡢῶᑡࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ ࡼࡗ࡚ cDNA ࢆྜᡂࡋࡓᚋ㸪real-time PCR ࡛ ࢥࣆ࣮ᩘࢆ ᐃࡋࡓ⤖ᯝࢆᅗ 8A ᅗ 8B ♧ࡋ Ș$P\ODVH ⢊ᮎࡢ⁐ᛶᡂศࡢṧ␃㛵ࡍ ࡓࠋDNase ฎ⌮ࢆ⾜ࢃ࡞ࡗࡓሙྜẚ㍑ࡋ࡚㸪 ࡿ᳨ウ 75Υ 5 ศࡢຍ⇕ࢆక࠺ DNase I ฎ⌮࡛ࡣࢥࣆ࣮ 㣗ရ୰ከࡃྵࡲࢀࡿⅣỈ≀ࡣ≀⌮Ꮫⓗ ᩘࡀ 1/5㹼1/10 ῶᑡࡋࡓࠋ༢⣧ຍ⇕ࢆ┬࠸ ᣲືࡀ᰾㓟㸦࣏࣮ࣜࣜ࣎ࢫ㸧㢮ఝࡋ࡚࠸ࡿࡇ - 50 - ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺղ ᅗ Ș$P\ODVH ⢊ᮎࡢ⁐ᛶᡂศ α-Amylase ⢊ᮎ㸦ග⣧⸆㸧 5g PBS(-) 20 ml ⁐ゎ㸦ᠱ⃮㸧 1.5 ml ࣐ࢡࣟࢳ࣮ࣗࣈ 20㹼24 ᮏศὀ 15,000 rpm 5 ศ ୖΎ 50 ml 㐲ỿ⟶ࡲࡵࡿ 0.22 μm ⁛⳦ࣇࣝࢱ࣮࡛ℐ㐣 㔞 ᐃ㸦㐲ỿ⟶┠┒࡛ྍ㸧 ➼㔞ࡢࢢࣜࢭ࣮ࣟࣝ㸦ΰྜ㸧 㸫20ΥಖᏑ㸪1 ᳨యᙜࡓࡾ 1 ml ⏝ ᅗ Ș$P\ODVH ⢊ᮎࡢᾮㄪ〇ᡭ㡰 ࡽ㸪PCR ࡢ㜼ᐖ≀㉁ࡉࢀ࡚࠸ࡿࠋ࢝࢟ࡢ ᅗ 9 ♧ࡋࡓ࠾ࡾ㐲ᚰࡼࡗ࡚ỿ⃦ࡋ࡚ࡃࡿࠋ ᳨ᰝἲ࠾࠸࡚ࡶ㸪ΰධࡋ࡚࠸ࡿࢢࣜࢥ࣮ࢤࣥ ࣓࣮࣮࢝ၥ࠸ྜࢃࡏࡓࡇࢁ㸪⁐ᛶᡂศࡣ ࢆศゎࡍࡿࡓࡵ α-Amylase ⢊ᮎࡀ⏝࠸ࡽࢀ࡚ 㘄࡞ࡢ〇㐀࠺㈿ᙧ࠸࠺ࡇ࡛࠶ࡗ ࠸ࡿࠋࣃࣥࢺࣛἲ࡛ࡣ㸪ᅗ 1 ♧ࡍ࠾ࡾ ࡓࠋලయⓗ࡞ᡂศࡣ㠀බ㛤࡛࠶ࡗࡓࡀ㸪୍⯡ⓗ α-Amylase ⢊ᮎࢆῧຍࡍࡿࣉࣟࢭࢫࡀ࠶ࡿࠋࡋ ࣓ࢳࣝࢭ࣮ࣝࣟࢫࡸ↓Ỉࢣ㓟࡞ࡀ⏝࠸ࡽ ࡋ㸪ࡇࡢ⢊ᮎࡣ⁐ᛶᡂศࡀྵࡲࢀ࡚࠾ࡾ㸪 ࢀࡿࡇࡀከ࠸ࠋ㐲ᚰᚋୖΎࢆᑀูࢳ - 51 - ᮍฎ⌮ ᮍฎ⌮ ⢊ᮎ ᾮㄪ〇ရ ␃Ỉ ᪥ᮏ⸆ᒁ᪉ࠕ࣮ࣚࢻࢳࣥ࢟ࠖῧຍ ᅗ $ Ș$P\ODVH ᾮㄪ〇ရࡢຠᯝࡢ☜ㄆ 㸦ࣚ࢘⣲ࢹࣥࣉࣥᛂ࣭ࢹࣥࣉࣥᾮ㸧 ᮍฎ⌮ ᮍฎ⌮ ⢊ᮎ ᾮㄪ〇ရ ␃Ỉ ᪥ᮏ⸆ᒁ᪉ࠕ࣮ࣚࢻࢳࣥ࢟ࠖῧຍ ᅗ % Ș$P\ODVH ᾮㄪ〇ရࡢຠᯝࡢ☜ㄆ 㸦ࣚ࢘⣲ࢹࣥࣉࣥᛂ࣭࣏ࢸࢺࢧࣛࢲ㸧 ࣮ࣗࣈ⛣ࡋ㸪⁐ᛶᡂศࢆྲྀࡾ㝖ࡃࡼ ࡎࢀ࠾࠸࡚ࡶ㸪ᾮㄪ〇ရࢆῧຍࡍࡿࡇ ࠺ࡍࢀࡤၥ㢟ࡣ㉳ࡇࡽ࡞࠸ࡀ㸪ᡭ㛫ࡀࡿ ࡼࡗ࡚ࣚ࢘⣲ࢹࣥࣉࣥᛂࡀᾘኻࡋࡓࠋ ࡇࡽࢹ࢝ࣥࢺ࡛῭ࡲࡏࡿࢣ࣮ࢫࡶᐃࡉࢀ ḟࣃࣥࢺࣛἲࡢࣉࣟࢺࢥࣝ࠾࠸࡚㸪⁐ ࡿࡇࡢሙྜ㸪α-Amylase ࡢ⁐ᛶᡂศࡀ௨ᚋࡢ ᛶᡂศࡀṧ␃ࡋࡓሙྜ㸪࠶ࡽࡌࡵ⁐ᛶᡂ ᕤ⛬ṧ␃ࡍࡿࡇ࡞ࡾ㸪᭱⤊ⓗ࡞᳨ฟ⢭ᗘ ศࢆྲྀࡾ㝖࠸ࡓᾮㄪ〇ရࢆ⏝࠸ࡓሙྜ᳨࡛ ᙳ㡪ࢆཬࡰࡍࡇࡀࢃࡗࡓࡓࡵ㸪ࡑࡢゎỴ ฟ⤖ᯝࡢᙳ㡪ࢆẚ㍑ࡋࡓࠋᅗ 12A ࡢ No.2 ⟇ࢆ᳨ウࡋࡓࠋα-Amylase ࢆࢢࣜࢭ࣮ࣟࣝ⁐ᾮ No.3 ࡣ⁐ᛶᡂศࡀㄆࡵࡽࢀࡓࠋTRIzol ᢳฟᚋ ࡋࡓᾮయ㓝⣲〇ရࡶ࠶ࡿࡀ㸪20 μL ⛬ᗘࡢ ࡢỈᒙ 㸦ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺճ㸧ࡣ↓Ⰽ㏱ ᛂ⣔ࢆᐃࡋ࡚࠸ࡿࡓࡵ㸪50 mL ࡢࣃࣥࢺࣛἲ ࡛᫂࠶ࡗࡓࡀ㸪࣒࢝ࣛࣉࣛࡍࡿ┤๓࢚ ࡢࣉࣟࢺࢥ࡛ࣝ⏝࠸ࡿࡣࢥࢫࢺ㠃࠾࠸࡚⌧ ࢱࣀ࣮ࣝࢆῧຍࡋࡓࡇࢁ㸦ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ ᐇⓗ࡛ࡣ࡞࠸ࠋࡑࡇ࡛㸪α-Amylase ⢊ᮎࢆᾮ ࣥࢺմ㸧㸪⁐ᛶᡂศ⏤᮶ࡍࡿⓑ⃮ࡀ⏕ࡌ ㄪ 〇 ࡍ ࡿ ᪉ ἲ ࢆ ⪃ ࡋ ࡓ 㸦 ᅗ 10 㸧 ࠋ ࡇ ࡢ ࡓ㸦ᅗ 12A㸧ࠋ୍᪉㸪α-Amylase ᾮㄪ〇ရࢆῧ α-Amylase ᾮㄪ〇ရࢆ㸫20Υ 2 ࣨ᭶ಖᏑࡋ ຍࡋࡓሙྜࡣࡇࡢࡼ࠺࡞ⓑ⃮ࡣ⏕ࡌࡎ㸪 ࡓᚋ㸪ࢹࣥࣉࣥࡢศゎຠᯝࢆほᐹࡋࡓࠋࢹࣥࣉ RNA ᢳฟᕤ⛬ࢆ⤊࠼ࡿࡇࡀ࡛ࡁࡓࠋᢳฟࡋ ࣥᾮ㸦ᅗ 11A㸧࣏ࢸࢺࢧࣛࢲ㸦ᅗ 11B㸧ࡢ࠸ ࡓ RNA ࢆ㗪ᆺࡋ࡚㸪ඛ᭱㐺ࢆ῭ࡲࡏࡓ㧗 - 52 - 1㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 2㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 ᅗ $ 3㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 4㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 5 Ș$P\ODVH ᾮㄪ〇ရࡢຠᯝࡢ☜ㄆ 㸦51$ ⢭〇࣒࢝ࣛࡢࣉࣛ㸧 1㸸ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ α-Amylase ⢊ᮎࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ࠾࠸࡚㸪 ࢫ࣏ࢺࢆ⏝࠸࡚ୖΎࢆูࢳ࣮ࣗࣈᑀศྲྀࡋࡓࡶࡢ 2㸸ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ α-Amylase ⢊ᮎࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ࠾࠸࡚㸪 ୖΎࢆࢹ࢝ࣥࢺࡼࡾูࢳ࣮ࣗࣈ⛣ࡋࡓࡶࡢ 3㸸ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ α-Amylase ⢊ᮎࢆຍ࠼㸪ࡑࡢᚋࡢ㐲ᚰࢆ⾜ࢃ࡞ࡗࡓࡶࡢ 4㸸ᅗ 1 ࡢࢳ࢙ࢵࢡ࣏ࣥࢺձ࡛ α-Amylase ᾮㄪ〇ရࢆຍ࠼㸪ࢳ࢙ࢵࢡ࣏ࣥࢺղ ࠾࠸࡚㸪ୖΎࢆࢹ࢝ࣥࢺࡼࡾูࢳ࣮ࣗࣈ⛣ࡋࡓࡶࡢ 5㸸␃Ỉ ㏫㌿ᛂࡼࡾ cDNA ࢆྜᡂࡋ㸪real-time ᯝࡣྠࡌ࡛࠶ࡗࡓࠋࡑࡇ࡛ᮏ◊✲࡛ࡣ㸪⤖ᯝᙳ PCR ᳨࡛ฟࢆヨࡳࡓ⤖ᯝࢆᅗ 12B ♧ࡋࡓࠋ 㡪ࢆཬࡰࡍ⪃࠼ࡽࢀࡿ⟠ᡤࢆḟࡢ 4 Ⅼ⤠ࡾ㸪 ⁐ᛶᡂศࡀṧ␃ࡋࡓ≧ែ࡛ࡣ㸪ᅗ 12A ࡛ⓑ⃮ࢆ ࡑࢀࡒࢀ࠾࠸࡚᭱㐺ࢆᅗࡗࡓࠋ ⏕ࡌࡓࡔࡅ࡛ࡣ࡞ࡃ㸪PCR ࡶ㜼ᐖࡉࢀࡓࠋ⁐ 㸦1㸧α-Amylase ⢊ᮎࡢ⁐ᛶᡂศࡢΰධ ᛶᡂศࢆ࠶ࡽࡌࡵ㝖ཤࡋࡓᾮㄪ〇ရࢆ⏝࠸ 㸦2㸧DNase ฎ⌮ࡶ࡞࠺ RNA ศゎ ࡿࡇ࡛㸪ࡇ࠺ࡋࡓ㒔ྜࡀ࡞ࡃ࡞ࡾ㸪᳨ฟ⤖ 㸦3㸧㏫㌿ᛂ⣔ࡢ᭱㐺 ᯝࢆṇᖖࡍࡿࡇࡀ࡛ࡁࡓࠋ 㸦4㸧PCR ᛂ⣔ࡢ᭱㐺 ᭱㐺ࡣ⤖ᯝࢆ㏲ḟほᐹࡋ࡞ࡀࡽ⾜࠺ᚲせࡀ࠶ 㸬⪃ᐹ ࡿࡓࡵࠋ᳨ウࡢ㡰␒ࡣࣉࣟࢺࢥࣝࡣ㏫㸦4㸧 ⤖ᯝࡢࣂࣛࡘࡁࡀ㉳ࡇࡾࡸࡍ࠸⟠ᡤ →㸦1㸧ࡋࡓ㸦㸦1㸧→㸦4㸧ࡢ᳨ウ࡛ࡣ㸪᭱㐺 ࡉࢀ࡚࠸࡞࠸ࣉࣟࢭࢫࢆ⤒ࡓ⤖ᯝࢆぢࡿࡇ ᮏἲࡣࡍ࡛」ᩘࡢᐇ࠾࠸࡚㣗ရࡽ ࡢ NoV ࡢ᳨ฟᡂຌࡋ࡚࠸ࡿࡇࡽ㸪ࣉࣟࢺ ࡞ࡾṇ☜㸧ࠋ ࢥࣝࡋ࡚ࡣᡂࡢᇦ࠶ࡿࡶࡢ⪃࠼ࡽࢀࡿࠋ ࡋࡋ㸪ᐇࡍࡿᶵ㛵ࡀቑ࠼ࡿࡘࢀ࡚㸪᳨ฟ⢭ 3&5 ᛂ⣔ࡢ᳨ウ ᗘࣂࣛࡘࡁࢆ⏕ࡌࡿࡇ࡞ࡿࡓࡵ㸪࠶ࡽࡌ Real-time PCR ⏝࠸ࡿヨ⸆ࡢẚ㍑᳨ウࡣᖹ ࡵᑐ⟇ࢆㅮࡌ࡚࠾ࡃࡇࡀᚲせ࡛࠶ࡿࠋᙜึ㸪⤖ ᡂ 24 ᖺᗘ⾜ࡗ࡚࠸ࡿࡇࡽ㸪ᮏ◊✲࡛ࡣ ᯝࡢࣂࣛࡘࡁࢆ⏕ࡌࡸࡍ࠸⟠ᡤࡋ࡚㸪ࣃࣥࢯࣝ conventional PCR ࡢヨ⸆㑅ᐃᛂ᮲௳ࡢ᭱㐺 ࣅࣥࡢ㍺㏦୰ࡢ⤖ࡸ㸪RNA ⢭〇࣒࢝ࣛࣉ ࡘ࠸᳨࡚ウࡋࡓࠋ᳨ウ࠶ࡓࡗ࡚ࡣ㸪PCR ࣛࡍࡿ㝿ῧຍࡍࡿ࢚ࢱࣀ࣮ࣝࡢ㔞ࡘ࠸࡚ ࢆ⾜࠺┤๓ࡢẁ㝵࡛࠶ࡿ cDNA ྜᡂࡲ࡛ࡣၥ㢟 ࡶᐃࡋ࡚࠸ࡓࠋ๓⪅ࡘ࠸࡚ࡣ㸪ពᅗⓗ⤖ ࡀ࡞࠸ࡇࢆ๓ᥦࡋ࡞ࡅࢀࡤ࡞ࡽ࡞ࡗࡓࠋ ⼥ゎࢆ⧞ࡾ㏉ࡋࡓࣃࣥࢯࣝࣅࣥࢆ⏝࠸࡚ࡶ⤖ᯝ ࡑࡇ࡛㸪⤖ᯝᙳ㡪ࡀཬࡪせᅉࢆ㝖ࡍࡿࡓࡵ ᙳ㡪ࡋ࡞࠸ࡇࡀ☜ㄆࡉࢀࡓࠋᚋ⪅ࡘ࠸࡚ࡣ㸪 㸪࢘ࣝࢫࢆྵࡲ࡞࠸㣗ရࡽㄪ〇ࡋࡓࣃࣥ ῧຍࡍࡿ࢚ࢱࣀ࣮ࣝࡢ㔞ࢆẁ㝵ⓗኚ࠼࡚ࡶ⤖ ࢺࣛᢳฟ≀ࢆ࣮࣋ࢫࡋ㸪ࡑࡇ NoV ࡢ㑇ఏᏊ - 53 - Fluorescence (F1/F2) ⣅౽࢜ࣜࢪࢼࣝ 1 4 2 3 5 Cycle Number ᅗ % Ș$P\ODVH ᾮㄪ〇ရࡢຠᯝࡢ☜ㄆ 㸦ቑᖜ᭤⥺㸧 ͤࢧࣥࣉࣝ 1㹼5 ࡢෆᐜࡣᅗ 12A ࢆཧ↷ 㓄ิ⏤᮶ࡢ DNA ᩿∦ࢆຍ࠼ࡓ⿕᳨యࢆ⏝࠸ࡓࠋ ♧ࡋࡓ࠾ࡾ㸪࣍ࢵࢺࢫࢱ࣮ࢺᵝࡀᚲ㡲࡛ ᳨ฟឤᗘࡢ᳨ウࡢࡓࡵẁ㝵ᕼ㔘ヨ㦂ࡀ୍⯡ⓗ ࠶ࡗࡓࠋࡇࢀࡣ㸪㔞Ꮡᅾࡍࡿ㯤Ⰽࣈࢻ࢘⌫ ⾜ࢃࢀ࡚࠸ࡿࡀ㸪ࡇࡇ࡛ࡣ␃Ỉ࡛ᕼ㔘ࡍࡿ ⳦ࡢ㠀≉␗ᛂࢆᢚ࠼ࡿࡇࡀ㔜せ࡛࠶ࡿࡇ ௦ࢃࡾࣃࣥࢺࣛᢳฟ≀ࢆ࠺ࡇ࡛㸪㔞ࡢ ࢆ♧ࡋ࡚࠸ࡿࠋࡲࡓ㸪ᇶ㉁ࡋ࡚ dUTP ࢆ 㯤Ⰽࣈࢻ࢘⌫⳦㑇ఏᏊࡢᏑᅾୗ࠸࠺㈇Ⲵⓗ᮲ ⏝ࡍࡿሙྜ ࡣ Mg2+ ⃰ ᗘࢆቑࡸࡍ ᚲせࡀ࠶ ࡿ ௳ࢆタᐃࡋ࡚࠸ࡿࠋࡲࡓ㸪ᑗ᮶ⓗഇ㝧ᛶᑐ⟇ ࡇ ࡶ ♧ ࡉ ࢀ ࡓ ࠋ dUTP ࡣ ᮏ ᮶ ࡢ ሷ ᇶ ࡛ ࠶ ࡿ ࡶど㔝ධࢀࡿᚲせࡀ࠶ࡾ㸪ഇ㝧ᛶࡢ᭱ࡢࣜ dTTP ࡼࡾࡶྲྀࡾ㎸ࡳ⋡ࡀⴠࡕࡿࡇࡽ㸪 ࢫࢡせᅉ࡛࠶ࡿ㑇ఏᏊゎᯒసᴗ㸦㟁ẼὋືࡸࢤ ᛂ⣔ 3 ಸ㔞ࢆຍ࠼ࡿࡇ࡛ᑐᛂࡋ࡚࠸ࡿ ࣝࡽࡢษࡾฟࡋ⢭〇㸧୧❧ࡉࡏࡿࡓࡵ㸪 㸦dTTP:200 μM㸪dUTP:600 μM㸧ࠋࢾࢡࣞ࢜ࢳࢻ ᇶ㉁ࡋ࡚ dUTP ࢆ࠼ࡿࡼ࠺᳨ウࢆ㐍ࡵࡓࠋ ࡣᙅ࠸࡞ࡀࡽࡶ࣮࢟ࣞࢺຠᯝࡀ࠶ࡿࡓࡵ㸪 PCR ࡛⏝࠸ࡿ㓝⣲㸦DNA polymerase㸧ࡣ㸪⏤᮶ Mg 2+ ࡢ ᐇ ຠ ⃰ ᗘ ࡀ ప ୗ ࡍ ࡿ ศ ࡔ ࡅ ቑ 㔞 ࡍ ࡿ ᚲ ࡼࡗ࡚ࣇ࣑࣮ࣜA ࣇ࣑࣮ࣜB ูࡉ せࡀ࠶ࡿࡶࡢ⪃࠼ࡽࢀࡿࠋdUTP ࢆ⏝࠸ࡓୖ ࢀࡿࠋᚋ⪅ࡣ㸪ᛂ୰㛫㐪ࡗࡓሷᇶࡀྲྀࡾ ࡛㸪␃Ỉࡼࡿẁ㝵ᕼ㔘⤖ᯝྠ➼ࡢឤᗘࢆ ㎸ࡲࢀࡓሙྜࡸࡾ┤ࡍ࠸࠺ᰯṇάᛶࡀ࠶ࡿ ᭷ࡍࡿᛂ⣔ࡋ࡚㸪᭱⤊ⓗᅗ 2C ♧ࡍ 3 ࡀ㸪๓⪅ࡣ࡞࠸ࠋࡋࡋ㸪ྠࣇ࣑࣮ࣜ ࡘࡀ㑅ᢥࡉࢀࡓࠋࡉࡽ౽ᛶࢆ⪃៖ࡋ㸪⾲ 1 B ᒓࡍࡿ㓝⣲ࡣ dUTP ࢆྲྀࡾ㎸ࡵ࡞࠸࠸ ࡢ No.12 ࢆ➨୍㑅ᢥࡋࡓࠋdUTP ࢆྲྀࡾ㎸ࢇ ࠺ᛶ㉁ࡶ࠶ࡿࠋᅗ 2A ♧ࡋࡓ࠾ࡾ㸪dUTP ࢆ ࡔቑᖜ᩿∦ࡣ㸪Uracil-N-Glycosylase ࢆస⏝ࡉࡏ ᇶ㉁ࡋ࡚⏝࠸ࡿࡇࡀ࡛ࡁࡿ㓝⣲ࡣࣇ࣑ ࡿࡇ࡛㑅ᢥⓗศゎ㝖ཤࡀྍ⬟࡛࠶ࡿࡓࡵ㸪 ࣮ࣜA ᒓࡍࡿࡶࡢࡔࡅ࡛࠶ࡗࡓࠋࣇ࣑࣮ࣜ ഇ㝧ᛶࡢ㜵Ṇ᭷ຠ࡛࠶ࡿࡶࡢ⪃࠼ࡽࢀࡿࠋ A ࡣᰯṇάᛶࡀ࡞࠸ࡓࡵ㸪ሷᇶࡢㄞࡳ㛫㐪࠸ ࡀⓎ⏕㸦ᩘሷᇶ 1 ࡘ㸧ࡋ㸪㑇ఏᏊゎᯒࡢṇ ㏫㌿ᛂ⣔ࡢ᳨ウ ☜ࡉࡀᦆ࡞ࢃࢀࡿᠱᛕࡶ࠶ࡿࠋࡋࡋ㸪㏻ᖖ⾜ PCR ᛂ⣔ࡢ᭱㐺ࡢሙྜྠᵝ㸪㏫㌿ ࢃࢀ࡚࠸ࡿ PCR ⏘≀ࡢࢲࣞࢡࢺࢩ࣮ࢡ࢚ࣥ ᛂ┤๓ࡢẁ㝵ࡲ࡛ࡣၥ㢟ࡀ࡞࠸ࡇࢆ๓ᥦ ࢫ࡛ࡣ㸪ࡇ࠺ࡋࡓ࢚࣮ࣛࡣࣂࢵࢡࢢࣛ࢘ࣥࢻࣀ ࡋ᳨࡚ウࢆ⾜࠺ᚲせࡀ࠶ࡿࡇࡽ㸪NoV RNA ࢬໟྵࡉࢀ࡚㢧ᅾࡋ࡞࠸ࡓࡵၥ㢟ࡣ࡞࠸ ࢆࣃࣥࢺࣛᢳฟ≀࡛ẁ㝵ᕼ㔘ࡋࡓ⿕᳨యࢆ⏝࠸ ⪃࠼ࡽࢀࡿࠋ⾲ 1 ࡢᛂ⣔ࡽ dUTP ࢆྲྀࡾ ࡓࠋ⾲ 2 ♧ࡋࡓ㓝⣲ࢆ⏝࠸࡚㏫㌿ᛂࢆ⾜ ㎸ࡵࡓࡶࡢࢆᢤࡁฟࡋ㸪ࡉࡽࣃࣥࢺࣛᢳฟ≀ ࡗࡓᚋࡣ㸪᭱㐺ࡢ῭ࢇࡔ real-time PCR㸪ࡲࡓ ࡛㈇Ⲵࢆࡅࡓ᮲௳᳨࡛ウࡋࡓࡇࢁ㸪ᅗ 2B ࡣ conventional PCR ࡛⤖ᯝࢆẚ㍑᳨ウࡋࡓࠋ⾲ - 54 - 秋田県健康環境センター年報 第10号 2014 2 ♧ࡋࡓ㏫㌿㓝⣲ࡣ㸪⮳㐺ᛂ ᗘࡼࡗ ࡞ࡿࠋࡇࡢࡼ࠺࡞㯤Ⰽࣈࢻ࢘⌫⳦ᑐࡍࡿ㠀≉ ࡚ 3 ࡘศࡅࡽࢀࡿࠋM-MLV㸦ࣔࣟࢽ࣮࣭࣐࢘ ␗ᛂࡣ 55Υ࡛ࡣⱝᖸṧࡿࡶࡢࡢ㸪58Υ࡛ࡣ ࢫ࣭ⓑ⾑࢘ࣝࢫ㸧⏤᮶ࡢ㏫㌿㓝⣲ࡣ㸪37 ࡞ࡃ࡞ࡾ㸪NoV 㑇ఏᏊࡔࡅࡀຠ⋡ⓗቑᖜ Υ࡛ᛂࡀ⾜ࢃࢀࡿ㸦⾲ 2 ࡢ No.1㸧ࡀ㸪ࡇࢀࢆ ࡛ࡁ࡚࠸ࡿࠋࡍ࡞ࢃࡕ㸪real-time PCR ࠾࠸࡚ ᨵኚࡋ࡚㸪ᛂ ᗘࢆ 42Υ㧗ࡵࡓࡶࡢࡀ No.2㸪 ⺯ගᙉᗘࡀ㧗ࡗࡓ⤖ᯝࢆࡅ࡚࠸ࡿࡶࡢ No.3㸪No.5 ࡛࠶ࡿࠋ୍᪉㸪ࡶࡶ⮳㐺ᛂ ⪃࠼ࡽࢀࡿࠋ ᗘࡀ㧗࠸ AMV㸦ࢺࣜ㦵㧊ⱆ⌫࢘ࣝࢫ㸧⏤ '1DVH ฎ⌮ἲࡢ᳨ウ ᮶ࡢ㏫㌿㓝⣲ࢆᨵኚࡋ࡚㸪ࡉࡽ㧗 ᛂࢆ ⾜࠼ࡿࡼ࠺ࡋࡓࡶࡢࡀ No.4 No.6 ࡛࠶ࡿࠋ ࡇࢀࡲ࡛࢝࢟➼ࡢ࢘ࣝࢫ᳨ᰝ࠾࠸࡚㸪 ᅗ 3A㸪ᅗ 3B㸪ᅗ 3C ࡛ࡣ㸪RNA ࡀప⃰ᗘ࡛࠶ ࢝࢟ᮏయ⏤᮶ࡢ DNA ࢆ㝖ཤࡍࡿࡓࡵ㏫㌿ ࡿ㸪㧗 ㏫㌿ᛂࡀ᭷ຠ࡛࠶ࡿࡇࡀ♧ ᛂࡢ๓ DNase ฎ⌮ࢆ⾜࠺ࡇࡀ᥎ዡࡉࢀ࡚ ࡉࢀ࡚࠸ࡿࠋᶓ㍈ࡽࡢ❧ࡕୖࡀࡾࢧࢡࣝ ࡁࡓࠋࣃࣥࢺࣛἲ࡛ᢳฟࡉࢀࡓ RNA ࡶ㔞 ࡁ࡞㐪࠸ࡣ࡞࠸ࡶࡢࡢ㸪⦪㍈ࡢ⺯ගᙉᗘᕪ ࡢ㯤Ⰽࣈࢻ࢘⌫⳦⏤᮶ࡢ DNA ࡀΰධࡍࡿࡇ ࡀぢࡽࢀࡿࡇࡽ㸪㧗 ࡞ࡿቑᖜຠ⋡ ࡽ㸪㠀≉␗ᛂࢆᢚไࡍࡿࡓࡵࡶ DNase ฎ ࡀ㧗ࡃ࡞ࡗ࡚࠸ࡿ࠸࠼ࡿࠋᅗ 3A ࡢࡼ࠺㧗 ⌮ࡣ᭷ຠ࡞ᡭἲ⪃࠼ࡽࢀࡿࠋࡋࡋ୍᪉࡛㸪 ⃰ᗘࡢ RNA ࢆቑᖜࡋ᳨࡚య୰ࡢึᮇࢥࣆ࣮ᩘ DNase ฎ ⌮ ࡛ ୍ ⯡ ⓗ ⏝ ࠸ ࡽ ࢀ ࡿ 㓝 ⣲ ࡛ ࠶ ࡿ 㸦ᶓ㍈౫Ꮡ㸧ࢆ⟬ᐃࡋ࡚ࡶ㐪࠸ࡀ⌧ࢀ࡞࠸ࡀ㸪 DNase I ࡣ㸪ᛂᚋࡢኻά 75Υ 5 ศࡢຍ⇕ࢆ ⺯ගᙉᗘࡀప࠸㸦ቑᖜຠ⋡ࡀప࠸㸧ᛂ⣔࡛ࡣ ᚲせࡍࡿࡇࡽ㸪⫢ᚰࡢ࢘ࣝࢫ RNA ࡀ ᳨ฟ㝈⏺࠾࠸࡚࡞ࡿ㸦ᅗ 3B㸪ᅗ 3C㸧ࠋ ศゎ࣭ῶᑡࡋ࡚ࡋࡲ࠺࠸࠺㞴Ⅼࡀ࠶ࡗࡓ㸦ᅗ ḟ㧗 ㏫㌿ᛂࡢ᭱㐺ࢆ⾜ࡗࡓࡇࢁ 8A㸪ᅗ 8B㸧ࠋᴟᚤ㔞ࡢ࢘ࣝࢫ RNA ࢆᢅ࠺㣗 58Υࡼࡿᛂࡀ᭱ࡶቑᖜຠ⋡ࡀ㧗ࡗࡓ㸦ᅗ ရ᳨ᰝ࠾࠸࡚㸪ࡇࡢၥ㢟ࡣ┳㐣ࡋᚓ࡞࠸ࡶࡢ 4A㸪ᅗ 4B㸧ࠋ㧗 ࡞ࡿ㠀≉␗ᛂࡣῶࡿ ⪃࠼ࡽࢀࡿࠋࡲࡓ㸪༢⣧ຍ⇕ࢆ┬࠸ࡓࡔࡅ ࡶࡢࡢ㸪ࣉ࣐࣮ࣛࡢࢽ࣮ࣜࣥࢢຠ⋡ࡶୗࡀ ࡛ࡣ㸪ṧᏑࡍࡿ DNase I ࡼࡗ࡚㸪ྜᡂࡉࢀࡓ ࡿࡓࡵ㸪⥲ྜⓗ࡞ቑᖜຠ⋡ࡣ୧⪅ࡢࣂࣛࣥࢫ࡛ cDNA ࡶᦆയࢆཷࡅࡿࡓࡵ᰿ᮏⓗ࡞ゎỴࡣ࡞ Ỵࡲࡿࠋ58Υ࠸࠺ ᗘ᮲௳ࡣ㸪ࡑࡢࣂࣛࣥࢫ ࡾᚓ࡞࠸ࠋࡑࡇ࡛㸪50Υ࡛άᛶࡀ࡞ࡃ࡞ࡾ㸪55 ࡢୖᡂࡾ❧ࡗ࡚࠸ࡿࡶࡢ⪃࠼ࡽࢀࡿࡀ㸪㧗 Υ ࡛ ྍ ㏫ ⓗ ኻ ά ࡍ ࡿ ᫆ ⇕ ᛶ DNase ㏫㌿ᛂࡀྍ⬟࡞㓝⣲ྠኈ㸦⾲ 2 ࡢ No.4 㸦HL-dsDNase㸧ࢆ⏝࠸ࡿࡇ࡛㸪75Υ 5 ศࡢ No.6㸧ࢆẚ㍑ࡋࡓࡇࢁ No.4 ࡢቑᖜຠ⋡ࡀ ຍ⇕ࢆ⾜ࢃࡎ㏫㌿ᛂ⛣⾜ࡍࡿࡇࡀ࡛ 㧗 ࡗ ࡓ 㸦 ᅗ 5A㸪 ᅗ 5B㸧ࠋ No.4 ࡢ 㓝 ⣲ ࡣ ࡁࡓࠋࡇࡢࣉࣟࢭࢫࢆᐇ⾜ࡍࡿࡓࡵࡣ㸪ඛ RNaseH άᛶࡀṧᏑࡋ࡚࠾ࡾ㸪No.6 ࡢ㓝⣲࡛ࡣ ᭱㐺 ࢆ ῭ࡲ ࡏࡓ 58Υ ࡢ㧗 ㏫ ㌿ ᛂ ⤌ 㝖ཤࡉࢀ࡚࠸ࡿ࠸࠺㐪࠸ࡀ࠶ࡿࠋcDNA ྜᡂ ࡳྜࢃࡏࡿࡇࡀᚲ㡲࡛࠶ࡾ㸪42Υࡢ㏫㌿ ᚋࡣ㸪㗪ᆺ࡞ࡗࡓ RNA 㙐ࡣせ࡞ࡾ㸪ࡴ ᛂࡣ㸪άᛶࢆⓎ⌧ࡉࡏࡓ DNase ࡼࡗ࡚ cDNA ࡋࢁ PCR ࡢຠ⋡ࢆᝏࡃࡍࡿࡇࡽ RNaseH ά ࡀᦆയࢆཷࡅࡿࡓࡵ㐺ᙜ࡛࠶ࡿࠋ࡞࠾㸪ࡇࡢ㓝 ᛶࡀṧᏑࡋ࡚࠸ࡿ᪉ࡀ᭷࡛࠶ࡿࡶࡢ⪃࠼ࡽ ⣲ࡣᡃࡀᅜ࡛ὶ㏻ࡋ࡚࠸࡞࠸ࡓࡵ㸪ࣀ࢙࣮ࣝ࢘ ࢀࡿࠋ ࡽ┤᥋ྲྀࡾᐤࡏࡿᚲせࡀ࠶ࡿࠋࡑࡇ୍࡛⯡ⓗ ⥆࠸࡚㸪real-time PCR ࢆ⏝࠸࡚⾜ࡗࡓ᳨ウෆ ⏝࠸ࡽࢀ࡚࠸ࡿ DNase I ࢆ⏝࠸ࡓ᪉ἲࡋ࡚㸪 ᐜࢆ conventional PCR ࡚⾜࠸㸪㠀≉␗ᛂࡢ ᅗ 7 ♧ࡋࡓ࣒࢜ࣥ࢝ࣛฎ⌮ἲࢆ⪃ࡋࡓࠋࣃ ⛬ᗘࢆẚ㍑ࡋࡓ㸦ᅗ 6㸧ࠋ37Υࡼࡿ㏫㌿ᛂ ࣥࢺࣛἲࡢ RNA ᢳฟᕤ⛬⤌ࡳධࢀࡿࡇࡀ ⣔࡛ࡣቑᖜࣂࣥࢻࡀㄆࡵࡽࢀ࡞࠸ࡀ㸪ࡇࢀࡣᅗ ࡛ࡁࡿࡓࡵ㸪᭱ࡶ౽ᛶࡀ㧗࠸ࡀ㸪DNA ࢘ࣝ 3A~ᅗ 3C ࠾࠸࡚⺯ගᙉᗘࡀపࡗࡓ⤖ᯝ ࢫ࡛࠶ࡿࢹࣀ࢘ࣝࢫࡀᐃࡉࢀࡿࢣ࣮ࢫ࡛ ୍ ⮴ ࡋ ࡚ ࠸ ࡿ ࠋ 42 Υ ࡢ ᛂ ⣔ ࡛ ࡣ 㸪 㧗 ⃰ ᗘ ࡢ ࡣ⏝࡛ࡁ࡞࠸ࠋ᫆⇕ᛶ DNase ฎ⌮࢜ࣥ࢝ࣛ NoV RNA ᑐࡋ࡚ࡣṇࡋࡃቑᖜ࡛ࡁࡿࡶࡢࡢ㸪 ࣒ DNase I ฎ⌮ ࡢ࠸ ࡎ ࢀ ࠾࠸ ࡚ࡶ ࢘ ࣝࢫ ప⃰ᗘ࡛ࡣ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ㠀≉␗ቑᖜࡀඃ RNA ࡢῶᑡࡣㄆࡵࡽࢀ࡞ࡗࡓ㸦ᅗ 8A㸪ᅗ 8B㸧 ࡞ࡗ࡚࠸ࡿࠋࡇࡢࡇࡣ㸪ᅗ 3B㸪ᅗ 3C㸪ᅗ ࡇࡽ㸪ྛᶵ㛵ࡢᐇ≧ྜࢃࡏ࡚㑅ᢥࡍࡿࡇ 5A㸪ᅗ 5B ࠾࠸࡚⺯ගᙉᗘࡀపࡗࡓ⌮⏤ ࡀྍ⬟࡛࠶ࡿࠋ - 55 - 秋田県健康環境センター年報 第10号 2014 Ș$P\ODVH ࡢ⁐ᛶᡂศࡢᙳ㡪㛵ࡍࡿ᳨ウ ჾලࡸヨ⸆ࢆศู⏝ࡍࡿ࡞㸪ᐇ㦂᧯సࢆៅ ࣃࣥࢺࣛἲ࡛ࡣ㣗ရ୰ࡢⅣỈ≀ࢆศゎ㝖ཤ 㔜⾜࠺࠸࠺ࡇ௨እ✚ᴟⓗ࡞ഇ㝧ᛶᑐ⟇ ࡍࡿࡓࡵ Į-Amylase ⢊ᮎࢆῧຍࡋ࡚࠸ࡿࡀ㸪 ࡣྲྀࡽࢀ࡚ࡇ࡞ࡗࡓࠋࡋࡋ㸪㣗ရࡢ࢘ࣝ ࡇࡢ⢊ᮎࡣ㈿ᙧࡀྵࡲࢀ࡚࠸ࡿࡓࡵ ࢫ᳨ᰝࡀ⌧ᐇⓗ࡞ࡶࡢ࡞ࡗࡓẁ㝵࡛ࡣ㸪ࡑࡢ ࡣ⁐ゎࡋ࡞࠸㸦ᅗ 9㸧ࠋࡑࡢᚋࡢᕤ⛬࠾࠸࡚㸪 ♫ⓗ࡞ᙳ㡪㚷ࡳ࡚㸪୍ᒙࡢഇ㝧ᛶᑐ⟇ࢆㅮ 3,000 rpm 30 ศࡢ㐲ᚰࡼࡗ࡚㣗ရṧ´ࢆỿ⃦ ࡌࡿᚲせࡀ࠶ࡿࡶࡢ⪃࠼ࡽࢀࡿࠋᮏ◊✲ࡼ 㝖ཤࡍࡿ㝿㸪ࡇࡢ⁐ᛶᡂศࡶ୍⥴ྲྀࡾ㝖 ࡾ᭱⤊ⓗᚓࡽࢀࡿ DNA ᩿∦࢘ࣛࢩࣝࡀྵ ࡃࡇࡀྍ⬟࡛࠶ࡿࡀ㸪ࡑࡢࡓࡵࡣࢫ࣏ࢺ ࡲࢀࡿࡼ࠺ࢹࢨࣥࡍࡿࡇࡀ࡛ࡁࡓࡓࡵ㸪 ࡛ୖΎࢆᑀูࢳ࣮ࣗࣈศ㞳ࡍࡿᚲせࡀ࠶ Uracil-N-Glycosylase ฎ⌮ࡼࡿഇ㝧ᛶ㜵Ṇ⟇ࢆ ࡿࠋ᳨యᩘࡀከ࠸ሙྜࡣ㸪ࡇࡢ᧯సࡣ࡞ࡾ ྲྀࡿࡇࡀྍ⬟࡛࠶ࡿࠋࡑࡢ᪉ἲࡢホ౯ࡣᚋ ࡢ㈇ᢸ࡞ࡾᚓࡿࠋࡋࡋ㸪ࢹ࢝ࣥࢺࡼࡗ࡚ ࡢㄢ㢟ࡍࡿࠋ ⡆౽ୖΎࢆศ㞳ࡋࡓሙྜࡣ㸪⁐ᛶᡂศࡢ ࡉࡽ㸪ᮏἲࡀ᭷ຠά⏝ࡉࢀࡿࡓࡵࡣ㸪 ṧ␃ࡣ㑊ࡅࡽࢀࡎ㸪⢭〇࣒࢝ࣛࣉࣛࡍࡿ 㐺ษ࡞㣗ရࢧࣥࣉࣝࡢ☜ಖࡀ㔜せ࡛࠶ࡿࠋලయ ┤๓࢚ࢱࣀ࣮ࣝࢆῧຍࡋࡓ㝿ⓑ⃮ࢆ⏕ࡌ ⓗࡣ㸪ᐇ㝿㣗༟౪ࡏࡽࢀࡿẁ㝵ࡢ᳨㣗㸦ㄪ 㸦ᅗ 12A ࡢ No.2㸪No.3㸧㸪᳨ฟ⤖ᯝࡶᙳ㡪ࡍ ⌮ࡽ┒ࡾࡅࡢࣉࣟࢭࢫࢆ⤒ࡓࡶࡢ㸧ࢆಖᏑ ࡿ㸦ᅗ 12B ࡢ No.2㸪No.3㸧ࠋࡑࡇ࡛㸪ᮏ◊✲࡛ ࡍࡿ࠸࠺ཎ๎ࢆ㸪ᴗ⪅࿘▱ࡍࡿᚲせࡀ࠶ ࡣ Į-Amylase ⢊ᮎࡽ⁐ᛶᡂศࢆ㝖ཤࡋ࡚ᾮ ࡿࠋࡲࡓ㸪࢘ࣝࢫࡣ㣗ရ୰࡛ࡣቑ࠼ࡎ╔ࡍ ㄪ〇㸦ᅗ 10㸧ࡍࡿࡇ࡛ၥ㢟ゎỴࢆᅗࡗࡓࠋ ࡿࡢࡳ࡛࠶ࡿࡇࡽ㸪ศྲྀࡋࡓ㣗ရࢧࣥࣉࣝ ᾮㄪ〇ࡋࡓ Į-Amylase ࡣ㸪⢊ᮎྠᵝⅣỈ ╔ࡋ࡚࠸࡞ࡅࢀࡤ㝜ᛶ࡞ࡗ࡚ࡋࡲ࠺ࠋࡑ ≀ࢆศゎ࡛ࡁࡿࡇࡀ☜ㄆࡉࢀࡓ㸦ᅗ 11A㸪 ࡢࡓࡵ㸪ࢧࣥࣉࣜࣥࢢࣉࣛࣥࡸ㸪ࢫࢣ࣮ࣝࢵ ᅗ 11B㸧ࠋࡲࡓ㸪⢭〇࣒࢝ࣛࣉࣛࡍࡿᕤ⛬ ࣉࡢ᪉ἲࡘ࠸࡚ࡶ᳨ウࡍࡿవᆅࡀṧࡉࢀ࡚࠸ࡿࠋ ࠾࠸࡚ⓑ⃮ࡏࡎ㸦ᅗ 12A㸧㸪᳨ฟ⤖ᯝࡶࡇࢀࡲ ࡛ࡢ⢊ᮎࢆῧຍࡍࡿ᪉ἲྠ➼࡛࠶ࡗࡓ㸦ᅗ ࡲࡵ 12B㸧ࠋࡇࢀࡽࡢ⤖ᯝࡣ㸪㸫20Υ࡛ 2 ࣨ᭶㛫ಖᏑ ࡋࡓᚋࡢᾮㄪ〇ရࢆ⏝࠸࡚ᚓࡽࢀࡓࡶࡢ࡛࠶ ᮏ◊✲࡛ᚓࡽࢀࡓ▱ぢࢆ⥲ᣓࡍࡿḟࡢ 4 Ⅼ 㞟⣙ࡉࢀࡿࠋ ࡾ㸪࠶ࡽࡌࡵ༑ศ㔞ࢆㄪ〇ࡋ࡚࠾࠸࡚ࡶ㒔 ➨୍㸪ࣃࣥࢺࣛἲῧຍࡍࡿ Į-Amylase ࡣ ྜࡣ࡞࠸ࡇࢆ♧ࡋ࡚࠸ࡿࠋ࢝࢟ࡢ࢘ࣝࢫ᳨ ࠶ࡽࡌࡵᾮㄪ〇ࡋࡓࡶࡢࢆ⏝࠸ࡿ᪉ࡀ⡆౽ ᰝࡢᕤ⛬ࡶ Į-Amylase ⢊ᮎࡀ⏝࠸ࡽࢀ࡚࠸ࡿ ࡛㸪ࡑࡢᚋࡢࢺࣛࣈࣝࢆ㜵Ṇ࡛ࡁࡿࠋ ➨㸪㯤Ⰽࣈࢻ࢘⌫⳦ࡢ DNA ࢆ㝖ཤࡍࡿ ࡀ㸪ᾮㄪ〇ရ⨨ࡁ࠼ࡿࡇ࡛⡆౽ࡀᅗ ࡓࡵࡣ RNA ᢳฟ࣒࢜ࣥ࢝ࣛ DNase I ฎ⌮ ࡽࢀࡿࡶࡢ⪃࠼ࡽࢀࡿࠋ ࢆ⾜࠺㸪ᢳฟᚋ᫆⇕ᛶ DNase ฎ⌮ࢆ⾜࠺ࡇ ᚋࡢㄢ㢟 ࡀ᭷ຠ࡛࠶ࡿࠋ࠸ࡎࢀࡢ᪉ἲࡶ㠀ຍ⇕࡛࠶ࡿ ᮏἲࡣᐇ㝿ࡢ㣗୰ẘ㐺⏝ࡉࢀ㸪㣗ရ ࡽࡢ࢘ࣝࢫ᳨ฟᡂຌࡋ࡚࠾ࡾ ാ┬㏻▱ 26) ࡸ㣗ရ⾨⏕᳨ᰝᣦ㔪 22-25) 27) ࡓࡵ㸪RNA ࡢศゎ࣭ῶᑡࡣ㉳ࡇࡽ࡞࠸ࠋ ➨୕㸪58Υࡢ㧗 ㏫㌿ᛂࢆ⾜࠺ࡇ࡛㸪 㸪ཌ⏕ປ グ㍕ࡉࢀ ࡓࡇࡽ㸪ᚋࡢᬑཬࡀぢ㎸ࡲࢀࡿࠋࡑࡢ୍ 㯤Ⰽࣈࢻ࢘⌫⳦⏤᮶ࡢ RNA㸦16s ࣜ࣎ࢯ࣮࣒㸧 ᑐࡍࡿ㠀≉␗ᛂࢆᢚไ࡛ࡁࡿࠋ ᪉࡛㸪ᐇ㦂ᐊෆởᰁࡼࡿഇ㝧ᛶࡢၥ㢟ࡀᾋୖ ➨ᅄ㸪PCR ࡢᛂ⣔ࢆ㐺ษ㑅ᢥ㸦㓝⣲ࡢ ࡋ࡚ࡃࡿࡇࡀᐃࡉࢀࡿࠋᮏ◊✲ࡼࡾຠ⋡ ✀㢮㸪࣍ࢵࢺࢫࢱ࣮ࢺᵝ㸪Mg ⃰ᗘ㸧ࡍࡿࡇ ⓗ㑇ఏᏊゎᯒྍ⬟࡞ DNA ᩿∦ࢆᚓࡿࡇࡀ ࡛㸪ቑᖜຠ⋡ࢆ᭱㐺࡛ࡁ㸪ᑗ᮶ⓗ࡞ഇ㝧ᛶ ࡛ࡁࡿࡼ࠺࡞ࡗࡓࡀ㸪ࡑࢀࡣ୧ลࡢ࡛࠶ࡿ ᑐ⟇ࡢᕸ▼ࡋ࡚ dUTP ࢆ⏝࠸ࡿࡇࡀ࡛ࡁࡿࠋ ࡇ␃ពࡍࡿᚲせࡀ࠶ࡿࠋࡍ࡞ࢃࡕ㸪㑇ఏᏊ ཧ⪃ᩥ⊩ ゎᯒࢆ✚ᴟⓗᐇࡍࢀࡤ㸪ࡑࢀࡔࡅసᴗ⎔ቃ ୰ࡢ PCR ቑᖜ⏘≀ởᰁࡀ✚ࡉࢀ࡚㸪᪂つࡢ㣗 ရ᳨ᰝ࠾࠸࡚ᝏᙳ㡪ࢆཬࡰࡍ࠸࠺ࢪ࣐ࣞࣥ 1㸧ᅜ❧ឤᰁ◊✲ᡤឤᰁሗࢭࣥࢱ࣮㸪ᅜ❧ ࡛࠶ࡿࠋࡇࢀࡲ࡛ࡣ㸪సᴗࡍࡿᐇ㦂ᐊࢆศࡅ࡚㸪 ឤᰁ◊✲ᡤ ࢘ࣝࢫ➨ 㒊:ࣀࣟ࢘ ࣝࢫ - 56 - 秋田県健康環境センター年報 第10号 2014 㞟ᅋⓎ⏕ᑐࡋ࡚ឤᰁ࠾ࡼࡧ㣗ရ㒊 ࡼࡿ㣗ရ᳨ᰝἲࡢᵓ⠏㸦᳨ウ 1㸧㸪ཌ⏕ປ ᒁࡀඹྠ࡛ᐇࡍࡿึᮇᐇᆅᏛㄪᰝ࠾ࡼ ാ⛉Ꮫ◊✲㈝⿵ຓ㔠 ࡧᚤ⏕≀Ꮫⓗ᳨ᰝࡢ࣏ࣥࢺ㸦➨ 1 ∧㸸ᖹᡂ ᥎㐍◊✲ᴗ 19 ᖺ 11 ᭶ 18 ᪥ࡅ㸧㸪2007, 16-17. 㛵ࡍࡿ◊✲ 2㸧ᒣົ㸦┘ಟ㸧 : ᨵゞ 㣗ရ୰ࡢ࢘ࣝࢫࡢไᚚ ᖹᡂ 21 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ ࿌᭩, 2010, 45-60. ࣀࣟ࢘ࣝࢫ⌧ሙ 12㸧ᮾ᪉⨾ಖ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ᑐ⟇, 2007, 35-36. 3㸧⸆࣭㣗ရ⾨⏕ᑂ㆟ ࡼࡿ㣗ရ᳨ᰝἲࡢᵓ⠏㸦᳨ウ 2㸧㸪ཌ⏕ປ 㣗ရ⾨⏕ศ⛉㣗୰ ẘ㒊: ࣀࣟ࢘ࣝࢫ㣗୰ẘᑐ⟇ࡘ࠸࡚ ാ⛉Ꮫ◊✲㈝⿵ຓ㔠 㸦ᥦゝ㸧㸪2007, 1-2. ᥎㐍◊✲ᴗ 4㸧ᩪ⸨༤அ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ 㛵ࡍࡿ◊✲ ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠 ☜ಖ᥎㐍◊✲ᴗ 㣗ရࡢᏳᚰ࣭Ᏻ☜ಖ 㣗ရ୰ࡢ࢘ࣝࢫࡢไᚚ ᖹᡂ 21 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ ࿌᭩,2010, 187-197. ࡼࡿ㣗ရ᳨యࡽࡢࣀࣟ࢘ࣝࢫࡢᅇ㸪ཌ ᚚ㛵ࡍࡿ◊✲ 㣗ရࡢᏳᚰ࣭Ᏻ☜ಖ 13㸧ᩪ⸨༤அ㸪㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ 㣗ရࡢᏳᚰ࣭Ᏻ ࡼࡿ㣗ရ୰ࡢࣀࣟ࢘ࣝࢫ᳨ᰝἲࡢᵓ⠏ 㸪 㣗ရ୰ࡢ࢘ࣝࢫࡢไ ⛅⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ㸪6㸪2010㸪59-69. ᖹᡂ 19 ᖺᗘ ⥲ᣓ࣭ศᢸ◊ 14㸧ᩪ⸨༤அ㸪㸸㣗ရ᳨యࡢཎ࢘ࣝࢫ᳨ ✲ሗ࿌᭩, 2008, 103-111. 5㸧ᮾ᪉⨾ಖ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ฟࢆྍ⬟ࡋࡓỗ⏝ᆺࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵ ࡼࡿ㣗ရ᳨యࡽࡢࣀࣟ࢘ࣝࢫࡢᅇ㸦᳨ ࣉἲࡢ㛤Ⓨ㸪⛅⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ㸪 ウ 2㸧㸪ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠 7㸪2011㸪43-53. 㣗ရ୰ࡢ࢘ 15㸧ᩪ⸨༤அ, : ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ᖹᡂ 19 ᖺᗘ ⥲ ࡼࡿ㣗ရࡽࡢ࢘ࣝࢫ᳨ฟἲ, ཎᚤ Ᏻᚰ࣭Ᏻ☜ಖ᥎㐍◊✲ᴗ ࣝࢫࡢไᚚ㛵ࡍࡿ◊✲ 㣗ရࡢ ⏕≀᳨ฟሗ, 32,12, 2011, 4-5. ᣓ࣭ศᢸ◊✲ሗ࿌᭩, 2008, 125-133. 16㸧ᩪ⸨༤அ, 㸸㣗ရ୰ࡢ࢘ࣝࢫ᳨ᰝྥ 6㸧ᩪ⸨༤அ㸪㸸㣗ရ᳨యࡢࣀࣟ࢘ࣝࢫ᳨ᰝ ྥࡅࡓࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ㛤Ⓨ㸪 ࡅ࡚ࡢࣃࣥࢯࣝࣅࣥࢺࣛࢵࣉἲࡢỗ⏝㸪ཌ ⛅⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ㸪4㸪2008㸪75-81. ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ㔠 7㸧ᮾ᪉⨾ಖ㸪㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ᥎㐍◊✲ᴗ 㣗ရࡢᏳ☜ಖ 㣗ရ୰ࡢཎ࢘ࣝࢫࡢࣜ ᖹᡂ 22 ᖺᗘ ⥲ᣓ ࡼࡿ㣗ရ᳨యࡽࡢࣀࣟ࢘ࣝࢫࡢᅇ᳨ ࢫࢡ⟶⌮㛵ࡍࡿ◊✲ ウ(➨ 1 ሗ㸧㸪⚟┴⾨⏕⎔ቃ◊✲ࢭࣥࢱ࣮ᖺ ࣭ศᢸ◊✲ሗ࿌᭩, 2011, 45-57. 17㸧ᩪ⸨༤அ: 㣗ရࡢࣀࣟ࢘ࣝࢫ᳨ᰝࡢỗ⏝ ሗ ,7, 2008, 69-72. 8㸧ᩪ⸨༤அ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ ࢆ┠ᣦࡋࡓࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ㛤 ᐇ⏝ྥࡅࡓᨵⰋ㸦᳨ウ 1㸧㸪ཌ⏕ປാ⛉ Ⓨ , ᪥ ᮏ 㣗 ရ ᚤ ⏕ ≀ Ꮫ 㞧 ㄅ , 29, 1, 2012, Ꮫ◊✲㈝⿵ຓ㔠 32-37. 㣗ရ୰ࡢ࢘ࣝࢫࡢไᚚ㛵ࡍ 18㸧ᩪ⸨༤அ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ᖹᡂ 20 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ࿌᭩, ࡼࡗ࡚㣗ရ᳨యࡽ᳨ฟࡉࢀࡓࣀࣟ࢘ ◊✲ᴗ ࡿ◊✲ 㣗ရࡢᏳᚰ࣭Ᏻ☜ಖ᥎㐍 2009, 27-38. ࣝࢫࡢ㑇ఏᏊゎᯒἲࡢ㛤Ⓨ㸪ཌ⏕ປാ⛉Ꮫ◊ 9㸧ᮾ᪉⨾ಖ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ ✲㈝⿵ຓ㔠 㣗ရࡢᏳ☜ಖ᥎㐍◊✲ᴗ ᐇ⏝ྥࡅࡓᨵⰋ㸦᳨ ウ 2㸧㸪ཌ⏕ປാ⛉ 㣗ရ୰ࡢཎ࢘ࣝࢫࡢࣜࢫࢡ⟶⌮㛵ࡍ Ꮫ◊✲㈝⿵ຓ㔠 ࡿ◊✲ ◊✲ᴗ ࡿ◊✲ 㣗ရࡢᏳᚰ࣭Ᏻ☜ಖ᥎㐍 2013, 35-48. 㣗ရ୰ࡢ࢘ࣝࢫࡢไᚚ㛵ࡍ ᖹᡂ 20 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ࿌ ᖹᡂ 24 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ࿌᭩, 19㸧㔝⏣⾨㸪㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢ ᭩,2009, 181-190. ከᶵ㛵ホ౯ヨ㦂⤖ᯝ㸪ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ 10㸧ᩪ⸨༤அ㸪 㸸 㣗ရ᳨యࡢࣀࣟ࢘ࣝࢫ ຓ㔠 ᳨ᰝྥࡅࡓࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡢᐇ 㣗ရࡢᏳ☜ಖ᥎㐍◊✲ᴗ ୰ࡢཎ࢘ࣝࢫࡢࣜࢫࢡ⟶⌮㛵ࡍࡿ◊ ᖹ ᡂ 24 ᖺ ᗘ ⥲ ᣓ ࣭ ศ ᢸ ◊ ✲ ሗ ࿌ ᭩ , ⏝ᛶྥୖ㛵ࡍࡿ◊✲㸪⛅⏣┴ᗣ⎔ቃࢭࣥ ✲ ࢱ࣮ᖺሗ㸪5㸪2009㸪54-62. 2013, 77-110. 11㸧ᩪ⸨༤அ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ 㣗ရ 20㸧 Kojima S., et. al.: Genogroup-specific PCR - 57 - 秋田県健康環境センター年報 第10号 2014 primers for detection of Norwalk-like viruses. J. ࡢཎ࢘ࣝࢫࡢࣜࢫࢡ⟶⌮㛵ࡍࡿ◊✲ Virol. Method, 100, 2002, 107-114. ᖹᡂ 24 ᖺᗘ ⥲ᣓ࣭ศᢸ◊✲ሗ࿌᭩, 2013, 175-180. 21 㸧 Kageyama T., et. al.: Broady reactive and highly sensitive assay for Norwalk-lile viruses based on real-time quantitative 24㸧ྂ⏣ᩄᙪ, : ᯇᕷෆ࠾ࡅࡿࣀࣟ࢘ reverse ࣝࢫ㞟ᅋ㣗୰ẘ, ཎᚤ⏕≀᳨ฟሗ, 35, 7, 2014, 4-5. transcription-PCR. J. Clin. Microbiol., 41, 2003, 1548-1577. 25㸧ᅵᒇ♸ྖ, : ࣃࣥཎᅉࡋࡓࣀࣟ࢘ ࣝࢫ㞟ᅋ㣗୰ẘ, ᪥ᮏ㣗ရᚤ⏕≀Ꮫ 22㸧୕ዲ㱟ஓ, : 㣗ရ୰ࡽࣀࣟ࢘ࣝࢫ㑇 㞧ㄅ, 32, 3, 2015, 153-158. ఏᏊࡀ᳨ฟࡉࢀࡓ㣗୰ẘ, ཎᚤ⏕≀ 26㸧ཌ⏕ປാ┬㏻▱: ࠕࣀࣟ࢘ࣝࢫࡢ᳨ฟἲ ᳨ฟሗ, 32, 12, 2011, 13-14. ࡘ࠸࡚ࠖࡢ୍㒊ᨵṇࡘ࠸࡚, 㣗Ᏻ┘Ⓨ 23㸧㣤ሯ⠇Ꮚ, 㸸ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ 1022 ➨ 1 ྕ, ᖹᡂ 25 ᖺ 10 ᭶ 22 ᪥. ࡼࡿ㣗ရࡽࡢࣀࣟ࢘ࣝࢫ㑇ఏᏊࡢ᳨ ฟ㸫ᘚᙜᒇࢆཎᅉタࡋࡓࣀࣟ࢘ࣝࢫ 27㸧ᩪ⸨༤அ, : 㣗ရ࣭⮫ᗋᮦᩱ࣭ᣔࡁྲྀࡾ 㞟ᅋ㣗୰ẘ㸫㸪ཌ⏕ປാ⛉Ꮫ◊✲㈝⿵ຓ ࡢ๓ฎ⌮ἲ, 㣗ရ⾨⏕᳨ᰝᣦ㔪 2015㸦ᚤ⏕≀ 㔠 ⦅㸧, 2015, 607-617. 㣗ရࡢᏳ☜ಖ᥎㐍◊✲ᴗ 㣗ရ୰ - 58 - 秋田県健康環境センター年報 第10号 2014 㣗ရ୰ࡢṧ␃㎰⸆ཬࡧṧ␃ື≀⏝་⸆ရࡢ୍ᩧศᯒἲ㛵ࡍࡿ◊✲㸦ᖹᡂ 24㹼26 ᖺᗘ㸧 ␆Ỉ⏘≀୰ࡢື≀⏝་⸆ရ୍ᩧศᯒἲࡢ᳨ウཬࡧጇᙜᛶホ౯ࡘ࠸࡚ ᯇῲளᕼᏊ 㔝⚘ᮁ ⌋⏣ᑦಇ* 1 ື≀⏝་⸆ရ 115 ᡂศࢆᑐ㇟㸪ࡼࡾຠ⋡ⓗ࡞୍ᩧศᯒἲࢆᵓ⠏ࡍࡿࡓࡵ㸪ᢳฟ᧯సࡸᅛ┦࣒࢝ࣛ ࡼࡿ⢭〇➼ࡢ᳨ウࢆ⾜ࡗࡓࠋᢳฟ᧯సࡘ࠸࡚ࡣ⡆␎ࡢࡓࡵ㸪࢞ࣛࢫࣅ࣮ࢬᢳฟ⁐፹ࢆຍ࠼ࡓ ࠸ᤞ࡚㐲ᚰ⟶ෆ࡛ࡢ࠺ᢳฟἲࢆ᥇⏝ࡋࡓࠋࡲࡓ㸪᳨ウࡢ⤖ᯝ㸪⢭〇⏝࠸ࡿᅛ┦࣒࢝ࣛࡣ㸪࢜ ࢡࢱࢹࢩࣝࢩࣜࣝࢩࣜ࢝ࢤࣝࡀ᭱㐺࡛࠶ࡾ㸪⁐ฟᾮࡣ㸪㓟ᛶሷᇶᛶࡢ 2 ✀㢮ࡢ⁐፹ࢆ⏝ࡍࡿ ᚲせࡀ࠶ࡗࡓࠋ ᳨ウࡋࡓศᯒἲࡘ࠸࡚㸪ཌ⏕ປാ┬ࡢ࢞ࢻࣛࣥᇶ࡙ࡁ㸪7 㣗ရ㸦㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ㸪 㭜༸㸪∵ங㸪ࡣࡕࡳࡘ㸧ࡢጇᙜᛶホ౯ヨ㦂ࢆᐇࡋࡓ㸦ῧຍ⃰ᗘ 0.01 ཬࡧ 0.04 ȝJJ㸪ศᯒ⪅ 2 ࡲࡓ ࡣ 3 ྡࡀ 2 ⃰ᗘῧຍヨᩱࢆ 1 ᪥ 2 ヨ⾜㸪3 ࡲࡓࡣ 2 ᪥㛫ศᯒ㸧ࠋ ࢞ࢻࣛࣥࡢ┠ᶆ್ࢆ‶ࡓࡋࡓᡂศࡢྜࡣ㸪㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ㸪㭜༸࡛⣙ 8 㸪∵ஙཬ ࡧࡣࡕࡳࡘ࡛⣙ 9 ࡞ࡾ㸪ᴫࡡⰋዲ࡞⤖ᯝࡀᚓࡽࢀࡓࠋᮏἲࡣ㧗⢭ᗘ࡞㎿㏿୍ᩧศᯒἲࡋ࡚᭷⏝ ⪃࠼ࡽࢀࡓࠋ 㸬ࡣࡌࡵ ᢠ⏕≀㉁ࡸ㥑⸆➼ࡢື≀⏝་⸆ရࡣ㸪␆Ỉ 㸬᪉ἲ ⏘≀ࡢẼࡢ⒪ࡸண㜵ࡢࡓࡵ⏝ࡉࢀ㸪⏕ ヨᩱ ⏘ᛶࡢྥୖᐤࡋ࡚࠸ࡿࠋ୍᪉㸪㐺ษᢞ ࢞ࢻࣛࣥࡢ♧ࢆཧ⪃㸪㭜⫗㸪㇜⫗㸪 ࡉࢀࡎ␆Ỉ⏘㣗ရṧ␃ࡋࡓሙྜ㸪ࣄࢺࡢ ∵⫗㸪ࡉࡅ㸪㭜༸㸪∵ங㸪ࡣࡕࡳࡘࡢ 7 ✀㢮ࢆ ᗣ⿕ᐖࡸ⪏ᛶ⳦ࡢⓎ⌧ࢆᘬࡁ㉳ࡇࡍྍ⬟ᛶࡀ ヨᩱࡋࡓࠋ ࠶ࡿࠋ 㭜⫗㸪㇜⫗㸪∵⫗ࡣヨᩱ㔞ᑐࡋ࡚༙㔞ࡢỈ ື≀⏝་⸆ရࡢつไࡣ㸪ᖹᡂ 18 ᖺࡽ࣏ࢪࢸ ࢆຍ࠼㸪ࣇ࣮ࢻࣉࣟࢭࢵࢧ࣮࡛࣮࣌ࢫࢺ≧࡞ ࣈࣜࢫࢺไ࡞ࡾ㸪つไᑐ㇟ရ┠ࡀᖜቑ ࡿࡲ࡛☻○ᆒ୍ࡋࡓࠋࡉࡅ㸪㭜༸ࡣỈࢆຍ࠼ ຍࡋࡓࠋࡇࡢࡓࡵ㸪ከᩘࡢື≀⏝་⸆ရࢆ㎿㏿ ࡎ㸪ࡑࡢࡲࡲ☻○ᆒ୍ࡋࡓࠋ∵ங㸪ࡣࡕࡳࡘ ࡘ⢭ᗘࡼࡃ᳨ฟࡍࡿ୍ᩧศᯒἲࡀồࡵࡽࢀ࡚ ࡣ᭱ึࡽᆒ㉁࡞≧ែ࡛࠶ࡿࡓࡵ㸪ࡑࡢࡲࡲ⏝ ࠸ࡿࠋࡉࡽ㣗ရ⾨⏕ἲࡢつ᱁ᇶ‽ࡢ㐺ྰࡢ ࠸ࡓࠋ ☻○ᆒ୍ࡋࡓᚋ㸪50 mL ࣏ࣜࣉࣟࣆࣞࣥ〇 ุ᩿⏝࠸ࡿศᯒἲࡘ࠸࡚ࡣ㸪ཌ⏕ປാ┬ࡢ ࢞ࢻࣛࣥ 1,2) ‽ࡌ㸪ጇᙜᛶホ౯ヨ㦂ࢆ⾜࠺ ࡼ࠺㔞ࡾ㸪㸫30Υ࡛⤖ಖᏑࡋࡓࠋ⏝ࡍࡿ ࡇࡀᚲ㡲࡞ࡗࡓࠋ ᙜ ࢭ ࣥ ࢱ࣮ ࡢ ື≀ ⏝ ་⸆ရ ࡢ ᳨ ᰝࡣ 㸪 ㏻ ▱ ἲ 3) ࢆᨵⰋࡋࡓ⊂⮬ἲ 㐲ᚰ⟶ 5.0 g ┦ᙜ㔞㸦ࡣࡕࡳࡘ 2.0 g㸧࡞ࡿ 4,5) ᪥ࡢ๓᪥ࡲࡓࡣᙜ᪥ゎࡋࡓࠋ ࡛ᑐᛂࡋ࡚ࡁࡓࡀ㸪୍ 㒊᧯సࡀ↹㞧࡞ࡇࡸ⢭〇ࡀ༑ศ࡞ࡇ➼ࡢ ᑐ㇟ᡂศ ⾲ 3 ♧ࡋࡓ㸪ື≀⏝་⸆ရ 115 ᡂศࢆᑐ㇟ ၥ㢟ࡀ࠶ࡗࡓࠋࡑࡇ࡛㸪୍ᩧศᯒἲࡢᑟධ࠶ ࡓࡾ᧯సࢆຠ⋡ࡋ㸪࡞࠾ࡘ⢭〇ຠᯝࢆ㧗ࡵ ࡋࡓࠋ ࡿࡓࡵ㸪ᢳฟ᪉ἲࡸᅛ┦࣒࢝ࣛࡼࡿ⢭〇➼ ࡘ࠸᳨࡚ウࡋࡓࠋ᳨ウࡢ⤖ᯝ㸪ࡼࡾ㎿㏿࡛㧗⢭ ᶆ‽ᾮཬࡧヨ⸆➼ ᶆ‽ᾮࡣ㸪ᕷ㈍ࡢΰྜᶆ‽ᾮಶูᶆ‽ရࢆ ᗘ࡞᪂つ୍ᩧศᯒἲࢆᵓ⠏ࡍࡿࡇࡀ࡛ࡁࡓࠋ ేࡏ࡚ጇᙜᛶホ౯ヨ㦂ࢆᐇࡋ㸪ጇᙜᛶࢆ☜ㄆ ⤌ࡳྜࢃࡏ࡚ㄪ〇ࡋࡓࠋ ΰྜᶆ‽ᾮࡣᯘ⣧⸆ᰴᘧ♫㸦௨ୗ㸪ᯘ⣧⸆㸧 ࡋࡓࡢ࡛ሗ࿌ࡍࡿࠋ 〇ࡢ PL ື≀⸆ LC/MS Mix1㸦ࢧࣝࣇ㸪ⴥ㓟 *1 ⛅⏣┴㎰ᯘỈ⏘㒊Ỉ⏘⯆ࢭࣥࢱ࣮㈨※㒊 - 59 - 秋田県健康環境センター年報 第10号 2014 25%ࣥࣔࢽỈ㸦≉⣭㸧ࢆ⏝࠸ࡓࠋ ྜᡂᣕᢠ㸧㸪PL ື≀⸆ LC/MS Mix2㸦࢟ࣀࣟ ࣥ㸧ྛ ȝJP/㸪ග⣧⸆ᰴᘧ♫㸦௨ୗ㸪 ⢭〇⏝࣒࢝ࣛࡣ㸪GL ࢧ࢚ࣥࢫᰴᘧ♫〇 ග⣧⸆㸧〇ࡢື≀⏝་⸆ရΰྜᶆ‽ᾮ㸦࣐ࢡ ࡢ࢜ࢡࢱࢹࢩࣝࢩࣜࣝࢩࣜ࢝ࢤ࣒ࣝ࢝ࣛ ࣟࣛࢻ㸧㸪ື≀⏝་⸆ရΰྜᶆ‽ᾮ㸦࣍ࣝࣔ 㸦InertSep C18㸹500 mg/6mL㸪1000 mg/6mL㸪௨ ࣥ㸧ྛ ȝJP/ ࢆ⏝ࡋࡓࠋ ୗ C18 ࣒࢝ࣛ㸧㸪࣑ࣀࣉࣟࣆࣝࢩࣜࣝࢩࣜ ಶูᶆ‽ရࡣ㸪ග⣧⸆〇ࡢ࣋ࣥࢪࣝ࣌ࢽࢩ ࢝ࢤ࣒ࣝ࢝ࣛ㸦InertSep NH2㸹500 mg/6 mL㸪௨ ࣜࣥ㸪ࣥࣆࢩࣜࣥ㸪࢜࢟ࢧࢩࣜࣥ㸪ࢪࢡࣟ࢟ ୗ NH2 ࣒࢝ࣛ㸧㸪࢚ࢳࣞࣥࢪ࣑ࣥ-N-ࣉࣟࣆ ࢧ ࢩ ࣜ ࣥ㸪 ࢭ ࣇ ࣆ ࣜࣥ㸪 ࢭ ࣇ ࢰ ࣜ ࣥ㸪 ࢭ ࣝࢩࣜࣝࢩࣜ࢝ࢤ࣒ࣝ࢝ࣛ㸦InertSep ࣇ࢛࣌ࣛࢰࣥ㸪࣑ࣟࢧ࣐ࢩࣥ㸪ࢪࣙࢧ࣐ࢩ 500 mg/6mL㸪௨ୗ PSA ࣒࢝ࣛ㸧ࢆ⏝࠸ࡓࠋ㝖⢏ ࣥ㸪ࣇ࢙࣋ࣥࢲࢰ࣮ࣝ㸪ࣇ࢙ࣂࣥࢸࣝ㸪࢜ࢡࢫ Ꮚ⏝࣓ࣥࣈࣛࣥࣇࣝࢱ࣮ࡣ ADVANTEC ♫〇 ࣇ࢙ࣥࢲࢰ࣮ࣝ㸪࢜ࢡࢫࣇ࢙ࣥࢲࢰ࣮ࣝࢫࣝ࣍ DISMIC㸦Ꮝᚄ ȝP㸪ぶỈᛶ PTFE㸧ࢆ⏝࠸ࡓࠋ ࣥ㸪ࣉࣛࢪࢡࣥࢸࣝ㸪ࣔࣛࣥࢸࣝ㸪ࣇࣝ࣋ࣥ ࢞ ࣛ ࢫ ࣅ ࣮ ࢬ ࡣ ࢬ ࣡ ࣥ ♫ 〇 ࡢ ┤ ᚄ 2.5 㹼 3.5 ࢲࢰ࣮ࣝ㸪࢜࢟ࢩ࣋ࣥࢲࢰ࣮ࣝ㸪ࢺࣜࢡࣛ࣋ࣥ mm ࡢࡶࡢࢆ⏝࠸ࡓࠋ PSA㸹 ࢲࢰ࣮ࣝ㸪ࢺࣜࢡࣛ࣋ࣥࢲࢰ࣮ࣝ࢜࢟ࢯࣥ㸪ࢳ ჾලཬࡧ⨨ ࣋ࣥࢲࢰ࣮ࣝ㸪5-ࣄࢻࣟ࢟ࢩࢳ࣋ࣥࢲࢰ࣮ ࣝ㸪࣓࣋ࣥࢲࢰ࣮ࣝ㸪ࣞࣂ࣑ࢰ࣮ࣝ㸪ࢡࣟࣆࢻ ࣇ࣮ࢻࣉࣟࢭࢵࢧ࣮ࡣ᪥❧〇సᡤ〇᪥❧ࢡࢵ ࣮ࣝ㸪࣋ࣥࢰ࢝ࣥ㸪ࢨ࣌ࣟࣥ㸪ࣈࣟࢳࢰࣛ ࢟ࣥࢢ࢝ࢵࢱ࣮FV-F3㸪 㧗㏿࣍ࣔࢪࢼࢨ࣮ࡣ ࣒㸪࢟ࢩࣛࢪࣥ㸪ࢺࣝࣇ࢙ࢼ࣒㓟㸪࣓ࣟ࢟ࢩ࢝ DREMEL ♫〇 MH-1000 ࢆ⏝࠸ࡓࠋ㐲ᚰศ㞳ᶵ ࣒㸪ࢣࢺࣉࣟࣇ࢙ࣥ㸪࢝ࣝࣉࣟࣇ࢙ࣥ㸪ࣇࣝࢽ ࡣ᪥❧ᕤᶵ〇 himacCR20GϪ㸪Sigma ♫〇 Sigma ࢟ࢩࣥ㸪ࣄࢻࣟࢥࣝࢳࢰࣥ㸪࣋ࢱ࣓ࢱࢰࣥ㸪ࢡ 1-15K ࢆ ⏝ ࠸ ࡓ ࠋ ࣝ ࣑ ࣈ ࣟ ࢵ ࢡ ᜏ ᵴ ࡣ 㸪 ࣐ࣟࣝࢪࣀࣥ㸪㓑㓟࣓ࣞࣥࢤࢫࢸ࣮ࣟࣝ㸪ࣀ࣎ EYELA 〇 MG-2100 ࢆ⏝࠸ࡓࠋ ࣅ࢜ࢩࣥ㸪ࣜࣥࢥ࣐ࢩࣥ㸪ࣜࣇ࢟ࢩ࣑ࣥ㸪 LC-MS/MS ࡣ Agilent Technologies ♫〇 Agilent ࣓ࣥࣈࢺࣥ㸪ࢺࣜ࣌ࣞࢼ࣑ࣥ㸪ࣇ࣒ࣇ࣮ࣝ㸪 1100㸦㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ㸧ཬࡧ Sciex ♫ ࢚ࢺࣂ࣮࣌ࢺ㸪ࣟ࢟ࢧࣝࢯࣥ㸪ࢭࣇࣟࢽ࣒࢘㸪 〇 API4000㸦㉁㔞ศᯒ⨨㸧ࢆ⏝࠸ࡓࠋ ࢭࣇ࢟ࣀ࣒㸪ࣇ࢙ࣀ࢟ࢩ࣓ࢳࣝ࣌ࢽࢩࣜࣥ㸪ࢡ ᐃ᮲௳ ࣟ࢟ࢧࢩࣜࣥ㸪ࢭࣇ࢟ࣀ࣒㸪ࢡ࣒ࣟࣛࣇ࢙ࢽࢥ ࣮ࣝ㸪ࣇࣟࣝࣇ࢙ࢽࢥ࣮ࣝ㸪ࢳ࣒ࣇ࢙ࢽࢥ࣮ ศᯒ࣒࢝ࣛࡣ㸪Ꮫ≀㉁ホ౯◊✲ᶵᵓ ࣝ㸪࢜࢟ࢩࢡࣟࢨࢽࢻ㸪ࢡࣟࣝࢫࣟࣥ㸪ࢪࢽࢺ 〇ࡢ L-column 2㸦2.1 mm i.d150 mm㸪⢏Ꮚᚄ ࣑ࣝࢻ㸪ࢪࢡࣛࢬࣜࣝ㸪ᯘ⣧⸆〇ࡢ࣐࣍ࣉࣛࢪ 3 ȝm㸧ࢆ⏝ࡋࡓࠋ ࣥ㸪㛵ᮾᏛᰴᘧ♫〇ࡢ࣓ࢩࣜࢼ࣒㸪ࢼࣇࢩ ⛣ື┦ࡣ㸪A ᾮ 0.1%ࡂ㓟Ỉ⁐ᾮ㸪B ᾮ ࣜࣥ㸪࢜ࣞࣥࢻ࣐ࢩࣥ㸪ࢼ࢝ࣝࣂࢪࣥ㸪 MeCN ࢆ⏝ࡋࡓࠋ࣒࢝ࣛ ᗘࡣ 40Υ㸪ὶ㏿ࡣ ␆Ỉ⏘ရṧ␃Ᏻ༠㆟〇ࡢࣝࢺࣞࣀࢤࢫ 0.2 mL/min㸪ὀධ㔞ࡣ 10 ȝL ࡋࡓࠋ ࢜ ࣥ ἲ ࡣ ESI ࡼ ࡿ ࣏ ࢪ ࢸ ࣈ 㸦 Posi- ࢺ㸪ࣂ࣮ࢪࢽ࣐ࢩࣥ㸪ࣉࣜࣇࢽ࣒࢘ࢆ ⏝ࡋࡓࠋࡇࢀࡽࡢಶูᶆ‽ရࡣࢭࢺࢽࢺࣜࣝ㸪 tive㸧ཬࡧࢿ࢞ࢸࣈ࣮ࣔࢻ㸦Negative㸧࡛⾜࠸㸪 N,N-ࢪ ࣓ ࢳ ࣝ ࣍ ࣝ ࣒ ࣑ ࢻ ➼ ࢆ ⏝ ࠸ ࡚ 200 㹼 ศ ᯒ ࣔ ࣮ ࢻ ࡣ Scheduled Multiple Reaction 500 ȝJP/ ㄪ〇ࡋࡓࠋ Monitoring㸦Scheduled MRM㸧ࡋࡓࠋ ࡚ࡢΰྜᶆ‽ᾮಶูᶆ‽ᾮྜࢃࡏ㸪 ࢜ࣥࢫࣉ࣮ࣞ㟁ᅽࡣ 5 kV㸦Positive㸧 㸪-4 kV 1 ȝJP/ ࡞ ࡿ ࡼ ࠺ ࢭ ࢺ ࢽ ࢺ ࣜ ࣝ ࡛ ᕼ 㔘 㸦 Negative 㸧 ࡋ 㸪 ࢜ ࣥ ※ ᗘ ࡣ 500 Υ ࡋ㸪ΰྜᶆ‽ᾮࡋࡓࠋ 㸦Positive㸧㸪400Υ㸦Negative㸧ࡋࡓࠋ HPLC ࡢࢢࣛࢪ࢚ࣥࢺ᮲௳ࡣ㸪Positive 㸸0 min ⁐፹ࡣ㸪ග⣧⸆〇ࡢࢭࢺࢽࢺࣜࣝ㸦ṧ␃ ㎰⸆ศᯒ⏝ཬࡧ LC/MS ⏝㸪௨ୗ MeCN㸧㸪࣓ࢱ 㸦B ᾮ 5%㸧Ѝ1 min㸦25%㸧Ѝ3 min㸦25%㸧Ѝ ࣀ ࣮ ࣝ 㸦 ṧ ␃ ㎰ ⸆ ศ ᯒ ⏝ 㸪 ௨ ୗ MeOH㸧 㸪 Ỉ 12 min㸦65%㸧Ѝ14 min㸦65%㸧Ѝ18 min㸦95%㸧 㸦 LC/MS ⏝ 㸧 㸪 N,N- ࢪ ࣓ ࢳ ࣝ ࣍ ࣝ ࣒ ࣑ ࢻ Ѝ21 min㸦95%㸧Ѝ21.5 min㸦5%㸧Ѝ30 min㸦5%㸧㸪 㸦Infinity Pure㸧ࢆ⏝࠸ࡓࠋヨ⸆ࡣග⣧⸆〇ࡢ Negative 㸸0 min㸦B ᾮ 5%㸧Ѝ0.5 min㸦5%㸧Ѝ 㓑㓟ࣥࣔࢽ࣒࢘㸦≉⣭㸧㸪ࡂ㓟㸦LC/MS ⏝㸧㸪 1 min㸦40%㸧Ѝ3 min㸦40%㸧Ѝ10 min㸦95%㸧 - 60 - 秋田県健康環境センター年報 第10号 2014 Ѝ15 min㸦95%㸧Ѝ15.1 min㸦5%㸧Ѝ25 min㸦5%㸧 Ỉ࡛ 2 mL ᐃᐜࡋࡓࠋࡇࢀࢆࣇࣝࢱ࣮ ࡋࡓࠋ ㏻ࡋ㸪ヨ㦂⁐ᾮࡋࡓࠋᐃᐜᚋ㸪ヨ㦂⁐ᾮࡀⓑ MRM ᮲௳ࡣ 50㹼100 ng/mL ࡢ⠊ᅖ࡛ㄪ〇ࡋ ࡃ⃮ࡿሙྜࡣ㸪㐲ᚰศ㞳㸦10,000g㸪10 ศ㛫㸪 ࡓྛᑐ㇟ᡂศࡢᶆ‽⁐ᾮࢆࢩࣜࣥࢪ࣏ࣥࣉࢆ⏝ 0Υ㸧ࢆ⾜࠸㸪ᚓࡽࢀࡓ㏱࡞ᾮࢆࣇࣝࢱ࣮ ࠸࡚┤᥋ MS/MS ὀධࡋ㸪⮬ື᭱㐺ࡼࡾ ㏻ࡋ㸪ヨ㦂⁐ᾮࡋࡓࠋ Ỵᐃࡋࡓࠋ⮬ື᭱㐺ࡢ⤖ᯝ㸪ឤᗘࡀ᭱ࡶⰋዲ ࡛࠶ࡗࡓ࢜ࣥࢆ㑅ᢥࡋ㸪ᐃ㔞⏝࢜ࣥタᐃ ヨᩱ 5.0 g ┦ᙜ ࡋࡓ㸦⾲ 3㸧ࠋ 㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ M eCNځ0.1%ࡂ㓟Ỉ⁐ᾮ㸦9:1㸧ΰᾮ 20 mL 㸦ࡂ㓟⤊⃰ᗘ0.01%㸧 㭜༸ M eCNځ0.1%ࡂ㓟Ỉ⁐ᾮ㸦7:3㸧ΰᾮ 15 mL 㸦ࡂ㓟⤊⃰ᗘ0.03%㸧 ∵ங M eCN 15 mL ヨ㦂⁐ᾮࡢㄪ〇 ヨ㦂⁐ᾮࡢㄪ〇ἲࢆᅗ 1-1㸪1-2 ♧ࡍࠋ 㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ㸪㭜༸㸪∵ஙࡢ ࢞ࣛࢫࣅ࣮ࢬ ㄪ〇 1ศ㛫 ࠺ ヨᩱ࢞ࣛࢫࣅ࣮ࢬ⣙ 3 g㸪MeCN0.1%ࡂ㓟 㐲ᚰศ㞳 Ỉ⁐ᾮ㸦9:1㸧ΰᾮ 20 mL 㸦㭜༸ࡣ MeCN0.1% ࡂ㓟Ỉ⁐ᾮ㸦7:3㸧ΰᾮ 15 mL㸪∵ஙࡣ MeCN 15 1300×g 㸪5ศ㛫㸪10Υ ṧ␃≀ mL㸧ࢆຍ࠼㸪ᡭ࡛ 1 ศ㛫⃭ࡋࡃ࠺ࡋࡓࠋ ࠺࣭㐲ᚰศ㞳 ࠺ᚋ㸪㐲ᚰศ㞳㸦1300g㸪5 ศ㛫㸪10Υ㸧ࢆ ⾜࠸㸪ୖΎࢆ 50 mL ᐜ࣓ࢫࣇࣛࢫࢥ㸦㭜༸㸪∵ ୖࠉΎ ஙࡣ 25 mL ᐜ㸧⛣ࡋࡓࠋṧ␃≀ୖグࡢΰᾮ M eCN 15 mL㸦㭜༸㸪∵ஙࡣྛᾮ 7 mL㸧ࢆຍ࠼㸪 ᐃࠉᐜ 50 mL ࠉ㸦㭜༸㸪∵ங 25 mL㸧 ࠺ཬࡧ㐲ᚰศ㞳ࢆ⾜࠸㸪ୖΎࢆྜࢃࡏࡓࠋࡇࢀ ศࠉྲྀ 6 mL 㸦㭜༸㸪∵ங 4 mL㸧 pH ㄪᩚࡢࡓࡵࡢ 1M 㓑㓟ࣥࣔࢽ࣒࢘Ỉ⁐ 㐲ᚰศ㞳 ᾮ 200 ȝL ࢆຍ࠼㸪MeCN ࡛ 50 mL㸦㭜༸㸪∵ங 㸸25 mL㸧ᐃᐜࡋࡓࠋ 㭜༸䠈∵ங ୖࠉΎ ୖࠉΎ 㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅࡘ࠸࡚ࡣᐃᐜࡋࡓ ศࠉྲྀ 5 mL ᢳฟᾮࡣ⃮ࡾࡀࡳࡽࢀࡓࡓࡵ㸪⣙ 6 mL ࢆู ⃰ࠉ⦰ ᥇ࡾ㸪㐲ᚰศ㞳㸦1300g㸪5 ศ㛫㸪10Υ㸧ࢆ⾜ M eCN ࠸㸪㏱࡞ᾮࢆᚓࡓࠋḟ㏱࡞ᾮ 5 mL ࢆ 1 mL ᐃࠉᐜ 1 mL ࣓ࢫࡁ࢞ࣛࢫヨ㦂⟶⛣ࡋ㸪❅⣲Ẽὶୗ࡛⃰ ⦰ࡋ㸪MeCN ࡛ 1mL ᐃᐜࡋࡓࠋ ศࠉྲྀ 0.8 mL 㸦㭜༸㸪∵ங 2 mL㸧 㭜༸㸪∵ஙࡣ㸪⣙ 4 mL ࢆู᥇ࡾ㸪㐲ᚰศ C18 㸦1000 mg㸧 ࠉ㺘㺻㺡㺼㺆㺚㺌㺤㺻㺖㺼 ࠉM eCN 5 mLЍM eCNځỈ㸦4:1㸧ΰᾮ 5 mL ⁐ࠉฟ 㞳ࢆ⾜࠸㏱࡞ᾮࢆᚓࡓࠋ C18 ࣒࢝ࣛ㸦1000 mg㸧㸪MeCN 5mL㸪MeCN Ỉ㸦4:1㸧ΰᾮ 5 mL ࢆ㡰ὀධࡋ㸪ࢥࣥࢹ ࢩࣙࢽࣥࢢࢆ⾜ࡗࡓࠋࡇࢀᐃᐜࡋࡓୖΎࡢ࠺ ࡕ㸪ヨᩱ 0.4 g ┦ᙜࡢ 0.8 mL㸦㭜༸㸪∵ங㸸2 mL㸧 ࠉ⁐ฟᾮ ࠉձ0.2%ࡂ㓟ྵ᭷ M eCNځỈ㸦4:1㸧ΰᾮ 7 mL ࠉղ0.5%㺏㺻㺱㺤㺏ྵ᭷ M eCNځỈ㸦4:1㸧 ΰᾮ 3 mL ⃰ࠉ⦰ ࢆ㈇Ⲵࡋࡓࠋ0.2%ࡂ㓟ྵ᭷ MeCNỈ㸦4:1㸧ΰ ᾮ 7 mL㸪0.5%ࣥࣔࢽྵ᭷ MeCNỈ㸦4:1㸧 ΰᾮ 3 mL ࢆ㡰ὀධࡋ㸪࡚ࡢࢁᾮࢆ 2 mL ࣓ 0 H&1ȝ/㸪Ỉ ᐃࠉᐜ 2 mL 㺪㺆㺷㺞㺎ࢁ㐣 ࢫࡁ࢞ࣛࢫヨ㦂⟶ᅇࡋࡓࠋᅇࡋࡓࢁᾮ 1 M 㓑㓟ࣥࣔࢽ࣒࢘Ỉ⁐ᾮ 100 ȝL ࢆຍ࠼㸪 ヨ㦂⁐ᾮ 5ಸᕼ㔘 ❅⣲Ẽὶୗ㸪1 mL ௨ୗࡲ࡛⃰⦰ࡋࡓࠋ ᅗ ヨᩱ⁐ᾮࡢㄪ〇ࣇ࣮ࣟ ⃰⦰ࡋࡓࢁᾮ㸪MeCN 600 ȝL ࢆຍ࠼㸪ࡉࡽ - 61 - 秋田県健康環境センター年報 第10号 2014 ᾮ㸦௨ୗ࣐ࢺࣜࢵࢡࢫ⁐ᾮ㸧ࢆ⏝࠸࡚ᐃᐜࡋ 20 ࡣࡕࡳࡘࡢㄪ〇 ng/mL ᾮࡋࡓࠋࡇࡢ 20 ng/mL ᾮࢆ࣐ࢺࣜࢵࢡ ヨᩱ 30 mM 㓑㓟ࣥࣔࢽ࣒࢘Ỉ⁐ᾮ 8 mL ࢆຍ࠼㸪⁐ゎࡍࡿࡲ࡛ᡭ࡛⃭ࡋࡃ࠺ ࢫ⁐ᾮ࡛ 10㸪5㸪1 ng/mL ࡞ࡿࡼ࠺ᕼ㔘ࡋࡓࠋ ࡋ㸪ୖグࡢỈ⁐ᾮ࡛ 10 mL ᐃᐜࡋࡓࠋࡇࢀࢆ ࡉࡽ 1 ng/mL ᾮࢆ 0.5㸪0.1 ng/mL ࡞ࡿࡼ࠺ 㔞ู᥇ࡾ㸪㐲ᚰศ㞳㸦1300g㸪5 ศ㛫㸪10 ᕼ㔘ࡋࡓࠋ0 ng/mL ࡣ࣐ࢺࣜࢵࢡࢫ⁐ᾮࢆ⏝ Υ㸧ࢆ⾜࠸㸪㏱࡞ᾮࢆᚓࡓࠋ ࠸ࡓࠋ࡚ࡢᡂศ࡛Ⰻዲ࡞┤⥺ᛶ㸦r = 0.99 ௨ୖ㸧 C18 ࣒࢝ࣛ㸦500 mg㸧㸪MeCN 3 mL㸪Ỉ 6 mL ࡀᚓࡽࢀࡓࠋ ࢆ㡰ὀධࡋ㸪ࢥࣥࢹࢩࣙࢽࣥࢢࢆ⾜ࡗࡓࠋ ␗ᛶయΰྜ≀ࡢࢳ࣑ࣝࢥࢩࣥࣔࣛࣥࢸࣝ㸪ྛ ࡇࢀୖグࡢᾮ 2.0 mL㸦ヨᩱ 0.4 g ┦ᙜ㸧ࢆ㈇ ␗ᛶయࡀ༢⊂ࡢ Į-ࢺࣞࣥ࣎ࣟࣥ ȕ-ࢺࣞࣥ࣎ࣟ Ⲵࡋ㸪⁐ฟᾮࢆᤞ࡚ࡓࠋࡉࡽỈ 3 mL ࢆὀධ ࣥࡘ࠸࡚ࡣ㸪ᅇࡢࢢࣛࢪ࢚ࣥࢺ᮲௳࡛ࡣ㸪 ࡋ⁐ฟᾮࢆᤞ࡚ࡓࠋ ྛࣆ࣮ࢡࡀ㏆᥋ࡋ࡚࠸ࡿࡓࡵ㸪ࡲࡵ࡚ 1 ࣆ࣮ 0.2%ࡂ㓟ྵ᭷ MeCNỈ㸦4:1㸧ΰᾮ 5 mL㸪0.5% ࢡࡋ࡚ᐃ㔞ࡋࡓࠋ ࣥࣔࢽྵ᭷ MeCNỈ㸦4:1㸧ΰᾮ 2 mL ࢆ㡰 ὀධࡋ㸪࡚ࡢࢁᾮࢆ 2 mL ࣓ࢫࡁ࢞ࣛࢫ ጇᙜᛶホ౯ヨ㦂 ヨ㦂⟶ᅇࡋࡓࠋᅇࡋࡓࢁᾮ 1 M 㓑㓟 ጇᙜᛶホ౯ヨ㦂ࡣ㸪࢞ࢻࣛࣥཬࡧᙜࢭࣥ ࣥࣔࢽ࣒࢘Ỉ⁐ᾮ 100 ȝL ࢆຍ࠼㸪❅⣲Ẽὶୗ㸪 ࢱ࣮ࡢᴗົ⟶⌮つᐃ‽ᣐࡋࡓࠋ 1 mL ௨ୗࡲ࡛⃰⦰ࡋࡓࠋ⃰⦰ࡋࡓࢁᾮ㸪MeCN ῧຍ⃰ᗘࡣ 2 ⃰ᗘࡋ㸪ΰྜᶆ‽ᾮࢆヨᩱ୰ 600 ȝL ࢆຍ࠼㸪ࡉࡽỈ࡛ 2 mL ᐃᐜࡋࡓࠋ ⃰ᗘࡀ 0.01 ཬࡧ 0.04 ȝg/g ࡞ࡿࡼ࠺ྛヨᩱ ࡇࢀࢆࣇࣝࢱ࣮㏻ࡋ㸪ヨ㦂⁐ᾮࡋࡓࠋ ῧຍࡋ㸪30 ศ௨ୖ⤒㐣ࡋ࡚ࡽヨᩱࡢ๓ฎ⌮ ࢆ⾜ࡗࡓࠋ ጇᙜᛶホ౯ヨ㦂ࡣ㸪ศᯒ⪅ 3 ྡࡀ 2 ヨ⾜ 2 ᪥ ヨᩱ 2.0 g 30 mM 㓑㓟㺏㺻㺱㺤㺑㺯Ỉ⁐ᾮ 8 mL ࠺ 㛫㸦㭜⫗㸪㇜⫗㸪㭜༸㸪∵ங㸪ࡣࡕࡳࡘ㸧ཬࡧ 1ศ㛫 ศᯒ⪅ 2 ྡࡀ 2 ヨ⾜ 3 ᪥㛫㸦∵⫗㸪ࡉࡅ㸧ศᯒ ᐃࠉᐜ 10 mL 㐲ᚰศ㞳 ࡍࡿィ⏬ࡼࡾᐇࡋࡓࠋᚓࡽࢀࡓ⤖ᯝࡣ㸪 Microsoft Excel ࢆ⏝࠸୍࡚ඖ㓄⨨ࡢศᩓศᯒ 1300×g 㸪5ศ㛫㸪10Υ ࡼࡿゎᯒࢆ⾜࠸㸪┿ᗘ㸪ే⾜⢭ᗘ㸪ᐊෆ⢭ᗘࢆ ୖࠉΎ ồࡵࡓࠋ ศࠉྲྀ 2.0 mL C18䠄500 mg䠅 Ὑࠉί 㑅ᢥᛶࡘ࠸࡚ࡣ㸪㣗ရẖࡢࣈࣛࣥࢡヨᩱࡢ 㺘㺻㺡㺼㺆㺚㺌㺤㺻㺖㺼 M eCN 3 mLЍỈ6 mL ࣆ࣮ࢡ㠃✚ࡀ㸪ῧຍ⃰ᗘ 0.01 ȝg/g ᑐᛂࡍࡿ⃰ Ỉ 3 mL ᗘࡢᶆ‽ᾮ㸦2 ng/mL㸧ࡽᚓࡽࢀࡿࣆ࣮ࢡ㠃✚ ࡢ 1/3 ᮍ‶࡛࠶ࡿࡇࢆ᮲௳ࡋࡓࠋ ⁐ࠉฟ ࠉ⁐ฟᾮ ࠉձ0.2%ࡂ㓟ྵ᭷ M eCNځỈ㸦4:1㸧ΰᾮ 5 mL ࠉղ0.5%㺏㺻㺱㺤㺏ྵ᭷ M eCNځỈ㸦4:1㸧ΰᾮ 2 mL ⃰ࠉ⦰ 0 H&1ȝ/㸪Ỉ ᐃ㔞㝈⏺ࡣ S/NӍ10 ࡞ࡿ᳨㔞⥺㸦㣗ရẖࡢ ࣐ࢺࣜࢵࢡࢫῧຍ᳨㔞⥺㸧ࡢ᭱ᑠ⃰ᗘࡋ㸪ῧ ຍ⃰ᗘ 0.01 ȝg/g ௨ୗ࡛࠶ࡿࡇࢆ᮲௳ࡋࡓࠋ ᐃࠉᐜ 2 mL 㸬⤖ᯝཬࡧ⪃ᐹ 㺪㺆㺷㺞㺎ࢁ㐣 ヨ㦂⁐ᾮ ᢳฟ⁐፹ࡢ᳨ウ 5ಸᕼ㔘 ື≀⏝་⸆ရ ࡢᢳฟ⁐ ፹ࡣ㸪MeCN ࡸ MeOH㸪Ỉࡢΰᾮ࡛⏝ࡉࢀࡿࡇࡀከ ᅗ ヨᩱ⁐ᾮࡢㄪ〇ࣇ࣮ࣟ㸦ࡣࡕࡳࡘ㸧 ࠸ ᐃ㔞ἲ 3,6 㹼 9) ࠋᅇࡢᑐ㇟ᡂศࡢ࠺ࡕ ȕ㸫ࣛࢡࢱ࣒ཬ ࡧࢭࣇ࢙࣒⣔ᢠ⏕≀㉁࠾࠸࡚㸪MeOH ୰࡛ศ ᳨㔞⥺ࡣ࣐ࢺࣜࢵࢡࢫῧຍ᳨㔞⥺ࡋ㸪⃰ᗘ ゎࡀࡳࡽࢀࡓࡓࡵ㸪MeCN ࢆ⏝࠸ࡿࡇࡋࡓࠋ ⠊ᅖࡣ 0㸪0.1㸪0.5㸪1㸪5㸪10㸪20 ng/mL ࡋࡓࠋ ࡓࡔࡋ㸪MeCN ࡢࡳ࡛ࡣࢱࣥࣃࢡ㉁ࡀจᅛࡋ㸪 ΰྜᶆ‽⁐ᾮ 20 ȝ/ ࢆ 1 mL ࣓ࢫࣇࣛࢫࢥ᥇ ᢳฟ᧯సᨭ㞀ࡀ⏕ࡌࡓࡓࡵ㸪Ỉࢆΰࡐࡿࡇ ࡾ㸪ᑐ㇟ᡂศࢆῧຍࡋ࡚࠸࡞࠸ྛ㣗ရࡢヨ㦂⁐ ࡋࡓࠋ - 62 - ࡲࡓ㸪㝖ࢱࣥࣃࢡຠᯝࢆୖࡆࡿࡓࡵ㸪Ⓨ ᛶࡢ࣓ࢱࣜࣥ㓟ࡢ௦ࢃࡾࡂ㓟ࢆ⏝࠸ࡓ 10) C18 ࠋ㓟 ࡢ⃰ᗘࡣ 0.1㹼1% ๓ᚋ࡛ㄪ〇ࡉࢀ ࡿ ࡀከ ࠸ 11,12) ࡀ 㸪㓟ᛶୗ࡛Ᏻᐃ࡞ᡂศࡀ࠶ࡿࡓࡵ 13,14) ప⃰ᗘࡢ᪉ࡢ 0.1%ࡋࡓࠋ ḟ㸪⤌ᡂࡢ␗࡞ࡿ MeCN 0.1%ࡂ㓟Ỉ⁐ ᾮࡢΰᾮࢆ࠸ࡃࡘㄪ〇ࡋ㸪ࡑࢀࡽࢆ㭜⫗㭜 ༸ຍ࠼㸪㧗㏿࣍ࣔࢪࢼࢨ࣮ࡼࡿ᧠ᢾࢆヨ ࡳࡓࠋࡑࡢ⤖ᯝ㸪㭜⫗ 9:1㸪㭜༸ 7:3 ࡢࡁ㸪༑ ศ࡞᧠ᢾࡀ࡛ࡁࡓࠋ∵ஙࡣỈศࡀከ ࡃ㸪MeCN ࡢࡳ࡛༑ศ࡞᧠ᢾࡀ࡛ࡁࡓࠋ NH2 ࡣࡕࡳࡘࡘ࠸࡚ࡣ㸪MeCN㸫Ỉΰᾮࢆຍ࠼ࡿ 㸪ᡂศࡢ⢾㢮ࡀᅛࡍࡿࡓࡵ 15)㸪30 mM 㓑㓟 ࣥࣔࢽ࣒࢘Ỉ⁐ᾮ㸦pH6㸧ࢆ⏝࠸ࡿࡇࡋࡓࠋ ྛ⁐፹࡛ࡢᢳฟຠ⋡ࢆ☜ㄆࡍࡿࡓࡵ㸪㭜⫗㸪 㭜༸㸪∵ஙࡢྛ 5 g ΰྜᶆ‽ᾮࢆ 0.2 μg/g ࡞ ࡿࡼ࠺ῧຍࡋ㸪ᅇヨ㦂ࢆᐇࡋࡓ㸦ྛ n=1㸧ࠋ ᨩᢾᢳฟࢆ 2 ᅇ⾜࠸㸦㭜⫗㸸20 mL2㸪㭜༸㸪 ∵ங㸸15㸪7 mL㸧㸪Ỉ࡛ 40 ಸᕼ㔘ࡋࡓヨ㦂⁐ ᾮࢆ ᐃࡋࡓࠋ⤖ᯝࢆᅗ 2 ♧ࡍࠋ3 ヨᩱࡶ 8 PSA ௨ୖࡢᡂศ࡛ᅇ⋡ 70㹼120%ෆ࡛࠶ࡗࡓࠋ 㭜 ⫗ 97 11 7 MeCN / 0.1% ࡂ㓟Ỉ⁐ᾮ㸦9:1㸧 㭜 96 ༸ 12 7 MeCN / 0.1% ࡂ㓟Ỉ⁐ᾮ㸦7:3㸧 ∵ 95 ங 9 11 MeCN 115 ᡂศ ᅇ⋡㸦%㸧 70㹼120 ڧ㸺70 115 ᡂศ 㸼120 ᅇ⋡㸦%㸧 ⢭〇ࡢ᳨ウ ␆Ỉ⏘≀ከࡃྵࡲࢀࡿ⬡㉁ࢆຠ⋡ࡼࡃ㝖ཤ ࡍࡿࡓࡵ㸪ᅛ┦࣒࡛࢝ࣛࡢ⢭〇᪉ἲࢆ᳨ウࡋࡓࠋ ᅛ┦࣒࢝ࣛࡣ C18㸪PSA㸪NH2 ࣒࢝ࣛ㸦ྛ 500 3,16) 㸪┠ⓗᡂศࢆຠ⋡ࡼࡃᅇࡍ 㸼120 ᅗ ྛᅛ┦࣒࢝ࣛࡢ⁐ฟヨ㦂⤖ᯝ㸦Q 㸧 ࡿࡓࡵ㸪⤌ᡂࡢ␗࡞ࡿ 8 ✀ࡢ⁐ฟᾮࢆㄪ〇ࡋ㸪 3 ✀ ࡢ ࢝ ࣛ ࣒ ࡢ࠺ ࡕ 㸪 C18 ࢝ ࣛ ࣒ ࡛ ⁐ ฟ ᾮ ⁐ฟヨ㦂ࢆ⾜ࡗࡓ㸦ྛ n=1㸧ࠋ 20 ng/mL ᕼ㔘ࡋࡓΰྜᶆ‽ᾮ 1 mL ࢆྛ࢝ 50㹼70 ڧ70㹼120 a MeCN b MeCN / Ỉ㸦4:1㸧 ΰᾮ c 0.2% ࡂ㓟ྵ᭷ eCN d 0.5% 㺏㺻㺱㺤㺏ྵ᭷ MeCN e 0.2% ࡂ㓟ྵ᭷ MeCN / Ỉ㸦4:1㸧ΰᾮ f 0.5% 㺏㺻㺱㺤㺏ྵ᭷ MeCN / Ỉ㸦4:1㸧ΰᾮ g 0.02% ࡂ㓟ྵ᭷ MeCN / Ỉ㸦4:1㸧ΰᾮ h 2% 㺏㺻㺱㺤㺏ྵ᭷ MeCN/ Ỉ㸦4:1㸧ΰᾮ ᅗ ྛᢳฟ⁐፹ࡼࡿᅇヨ㦂⤖ᯝ㸦Q 㸧 mg㸧ࢆ㑅ᢥࡋ <50 MeCN㸫Ỉΰᾮࡢሙྜ 㸪᭱ࡶከࡃࡢ┠ⓗᡂศ ࣒ࣛ㈇Ⲵࡋ㸪ྛ⁐ฟᾮ 5 mL ࢆὀධࡋࡓࠋḟ ࢆ⁐ฟ࡛ࡁࡓࠋᙅ㝜࢜ࣥ┦ࡢ NH2 ཬࡧ 㸪⁐ฟࡋࡓࢁᾮࢆ⃰⦰ࡋ㸪Ỉ࡛ 2 mL ᐃᐜ PSA ࣒࡛࢝ࣛࡣ྾╔ࡀ㉳ࡁ㸪⁐ฟࡉࢀ࡞࠸ᡂศ ࡋࡓࡶࡢࢆ ᐃࡋࡓࠋ⤖ᯝࢆᅗ 3 ♧ࡍࠋ ࡀከࡃ࠶ࡗࡓࠋ - 63 - RSD㸦%㸧 C18 ࣒࢝ࣛ࠾࠸࡚ࡣ㸪0.2%ࡂ㓟ྵ᭷ࢭࢺ ∵⫗ ࢽࢺࣜࣝỈ㸦4:1㸧ΰᾮ㸦e ᾮ㸧ࡼࡿ⁐ฟࡀ᭱ ࡶⰋዲ࡛࠶ࡗࡓࠋ୍᪉㸪࢚ࣜࢫ࣐ࣟࢩࣥ➼ࡢ ᩘಶࡢᡂศ࡛ࡣ㸪ሷᇶᛶࡢࣥࣔࢽྵ᭷ MeCNỈ㸦4:1㸧ΰᾮ㸦f㸪h ᾮ㸧ࡼࡿ⁐ฟࡢ ᪉ࡀⰋዲ࡛࠶ࡗࡓࠋ ࡋࡓࡀࡗ࡚㸪ᅛ┦ࡣ C18 ࣒࢝ࣛࢆ⏝࠸㸪e ཬ ࡧ f ᾮࡢ 2 ᾮ࡛⁐ฟࡍࡿࡇࡋࡓࠋࡲࡓ⢭〇 ຠᯝࢆୖࡆࡿࡓࡵ㸪ሸ㔞 1000 mg ࢆ⏝࠸ࡿࡇ ࡋࡓࠋ ᖹᆒᅇ⋡㸦%㸧 ࡣࡕࡳࡘࡢሙྜࡣ㸪⬡㉁ࢆྵࡲࡎ⢾ศࡀከ࠸ ࡓࡵ 17) RSD㸦%㸧 㸪C18㸦500 mg㸧୍᪦┠ⓗᡂศࢆಖᣢ ࡉࡅ ࡉࡏ㸪Ỉ 3 mL ࡛⢾ศࢆὙίࡋ࡚ࡽ㸪e ཬࡧ f ᾮ࡛⁐ฟࡍࡿࡇࡋࡓࠋὙίỈ 3 mL ࡛ࡣ㸪 ⁐ฟࡍࡿ┠ⓗᡂศࡀ↓࠸ࡇࢆ☜ㄆࡋࡓࠋ ᢳฟ᪉ἲࡢ┬ຊࡢ᳨ウ 㧗㏿࣍ࣔࢪࢼࢨ࣮ࡼࡿᨩᢾᢳฟ㸦௨ୗ㸪 ᨩᢾᢳฟ㸧࡛ࡣ㸪ヨᩱẖࡢลࡢὙίປຊࢆせ ࡍࡿࠋࡲࡓὙίࡀ༑ศ࡛࠶ࡿࢥࣥࢱ࣑ࢿ࣮ ࢩࣙࣥࡸ ᐃ್ࡢࡤࡽࡘࡁࡢᜍࢀࡀ࠶ࡿࠋࡑࡇ ࡛㸪ᢳฟᕤ⛬ࡢ┬ຊࡢࡓࡵ㸪࠸ᤞ࡚ࡢ㐲ᚰ ᖹᆒᅇ⋡㸦%㸧 ⟶ヨᩱࢆ᥇ࡾ㸪⁐፹࢞ࣛࢫࣅ࣮ࢬࢆຍ࠼㸪 㧗㏿࣍ࣔࢪࢼࢨ࣮ࡼࡿ᧠ᢾᢳฟ㸦n=3㸧 ࢞ࣛࢫࣅ࣮ࢬࡼࡿ࠺ᢳฟ㸦n=3㸧 ᡭ࡛⃭ࡋࡃ࠺ࡍࡿ᪉ἲ㸦௨ୗ㸪࠺ᢳฟ㸧 ࢆ᳨ウࡋ㸪ྛ᪉ἲࡢᢳฟຠ⋡ࢆẚ㍑ࡋࡓࠋ ᅗ ᧠ᢾཬࡧ࠺ᢳฟࡼࡿྛᡂศࡢศᕸ ∵⫗㸪ࡉࡅΰྜᶆ‽ᾮࢆ 0.01 μg/g ࡞ࡿࡼ ࠺ ῧ ຍࡋ 㸪 MeCN0.1%ࡂ 㓟 Ỉ ⁐ᾮ 㸦 9:1㸧 ΰ 2 ⃰ᗘ࡛ᅇ⋡ 70㹼120%㸪RSD% < 25㸦0.01 ᾮࢆຍ࠼㸪ྛᢳฟ᪉ἲࢆ 2 ᅇ⧞ࡾ㏉ࡋࡓࠋḟ μg/g㸧ཬࡧ㸺15㸦0.04 μg/g㸧࡛࠶ࡗࡓࡢࡣ㸪࣐ ྛᢳฟᾮࢆᐇ㦂᪉ἲ 2.6.1 ᚑࡗ࡚ㄪ〇ࡋ㸪ᅇ ࢺࣜࢵࢡࢫῧຍ᳨㔞⥺ࡀ 104 ᡂศ㸪⤯ᑐ᳨㔞⥺ ヨ㦂ࢆ⾜ࡗࡓ㸦ྛヨᩱཬࡧ᪉ἲ n=3㸧ࠋ⤖ᯝ ࡀ 93 ᡂศ࡛࠶ࡗࡓࠋࡇࡢࡇࡽ㸪ዃ㞧ᡂศࡢ ࢆᅗ 4 ♧ࡍࠋ ᙳ㡪ࢆ⿵ṇࡍࡿᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡓࡓࡵ㸪 ∵⫗㸪ࡉࡅࡶ࠺ᢳฟࡢ᪉ࡀ㸪ᖹᆒᅇ ࣐ࢺࣜࢵࢡࢫῧຍ᳨㔞⥺ࢆ⏝࠸ࡿࡇࡋࡓࠋ ⋡ 70㹼120%㸪┦ᑐᶆ‽೫ᕪ㸦RSD%㸧25%ᮍ ‶ࢆ ‶ࡓ ࡋࡓ ᡂศ ᩘࡀ ከ ࡗ ࡓࠋ ᅇ⋡ 50㹼 ጇᙜᛶホ౯ヨ㦂 70%ࡢᡂศࢆྜࢃࡏࡿ 9 ௨ୖ࡛ RSD25%ᮍ 7 㣗ရࢆᑐ㇟㸪᳨ウࡋࡓศᯒἲࡢጇᙜᛶホ ‶࡛࠶ࡗࡓࠋࡋࡓࡀࡗ࡚㸪ࡼࡾ᧯సࡀ⡆౽࡞ ౯ヨ㦂ࢆᐇࡋࡓࠋ⤖ᯝࢆ⾲ 1 ࡢ࠾ࡾ A㹼E ࠺ᢳฟࢆ⾜࠺ࡇࡋࡓࠋ ศ㢮ࡋࡓࠋ㣗ရูࡢ⤖ᯝࢆ⾲ 2 㸪ᡂศูࡢ ⤖ᯝࢆ⾲ 3 ♧ࡍࠋ ᳨㔞⥺ࡢ᳨ウ ศ㢮ࡣ㸪A㸦2 ⃰ᗘ࡛┿ᗘ㸪⢭ᗘࡢ┠ᶆ್ࢆ‶ ㄪ〇ࡋࡓヨ㦂⁐ᾮṧ␃ࡍࡿዃ㞧ᡂศࡢᙳ㡪 ࡓࡋ㐺ྜࡍࡿሙྜ㸧㸪B㸦㧗⃰ᗘῧຍ࡛┿ᗘ㸪 ࢆ☜ㄆࡍࡿࡓࡵ㸪⤯ᑐ᳨㔞⥺࣐ࢺࣜࢵࢡࢫῧ ⢭ᗘࡢ┠ᶆ್ࢆ‶ࡓࡍ㸧㸪B’㸦ప⃰ᗘῧຍ࡛┿ ຍ᳨㔞⥺ࡼࡿᐃ㔞⤖ᯝࢆẚ㍑ࡋࡓࠋ ᗘ㸪⢭ᗘࡢ┠ᶆ್ࢆ‶ࡓࡍ㸧㸪C㸦2 ⃰ᗘ࡛┿ᗘ ∵ங 5 g 㸪ΰྜᶆ‽ᾮࢆ 0.01 ཬࡧ 0.04 μg/g ࡞ࡿࡼ࠺ῧຍࡋ㸪ᐇ㦂᪉ἲ 2.6.1 ᚑࡗ࡚ 50%௨ୖ 70%ᮍ‶㸪⢭ᗘࡀ┠ᶆ್ࢆ‶ࡓࡍ㸧㸪 D㸦A㸪B㸪B'㸪C ௨እ㸧㸪E㸦 ᐃ⬟㸧ࡋࡓࠋ ㄪ〇ࡋࡓヨ㦂⁐ᾮࢆ ᐃࡋࡓ㸦ྛ⃰ᗘ n=3㸧ࠋ ホ౯ A ࡢྜࡣ㸪㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ㸪 - 64 - 秋田県健康環境センター年報 第10号 2014 ⾲ ጇᙜᛶホ౯ヨ㦂⤖ᯝ㸦㣗ရู㸧 ⾲ ホ౯ศ㢮 ホ౯ ᮲ ௳ A 2 ⃰ᗘ࡛┠ᶆ್ࢆ‶ࡓࡋ㐺ྜࡍࡿሙྜ A ┿ᗘ㸦%㸧 㸸70 ӌ 㹼 ӌ120 ⢭ᗘ㸦%㸧 㸸 < 25㸪< 30 㸦ȝJJ㸧 ే⾜㸪ᐊෆ < 15㸪< 20 㸦ȝJJ㸧 ȝJJ ࡢࡳ┠ᶆ್ࢆ‶ࡓࡍ B ┿ᗘ㸦%㸧 ⢭ᗘ㸦%㸧 ే⾜㸪ᐊෆ 㸸70 ӌ 㹼 ӌ120 㸸< 15㸪< 20 ȝJJ ࡢࡳ┠ᶆ್ࢆ‶ࡓࡍ B' ┿ᗘ㸦%㸧 ⢭ᗘ㸦%㸧 ే⾜㸪ᐊෆ 㸸70 ӌ 㹼 ӌ120 㸸< 25㸪< 30 2 ⃰ᗘࡶ௨ୗࡢ᮲௳ࢆ‶ࡓࡍ C D E ┿ᗘ㸦%㸧 㸸50 ӌ 㹼 㸺70 ⢭ᗘ㸦%㸧 㸸 < 25㸪< 30 㸦ȝJJ㸧 ే⾜㸪ᐊෆ < 15㸪< 20 㸦ȝJJ㸧 㸸< 50㸪120 < ⢭ᗘ㸦%㸧 㸸25ӌ㸪30ӌ 㸦ȝJJ㸧 ే⾜㸪ᐊෆ 15ӌ㸪20ӌ 㸦ȝJJ㸧 ᐃ⬟ ⫗ ㇜ ⫗ ∵ ⫗ ࡉ ࡅ 㭜 ༸ ∵ ங 98* (85)** 92 (80) 97 (84) 98 (85) 90 (78) 108 (94) 110 (96) ࡣࡕࡳࡘ 0 1 (1) 1 (1) 2 (2) 2 (2) 0 1 (1) B' 1 (1) 1 (1) 5 (4) 5 (4) 12 (10) 1 (1) 2 (2) C 0 7 (6) 3 (3) 0 1 (1) 1 (1) 2 (1) D 16 (14) 13 (11) 9 (8) 10 (9) 10 (9) 5 (4) 0 E Total 0 115 1 (1) 115 0 115 0 115 0 115 0 115 0 115 *㸸ྛホ౯ヱᙜࡍࡿᡂศᩘ **㸸ᑐ㇟ᡂศᑐࡍࡿྜ㸦%㸧 A㸪B㸪B'㸪C ௨እ 2 ⃰ᗘࡲࡓࡣࡕࡽࡢ⃰ᗘ࡛㸪ḟࡢ ࠸ࡎࢀ 1 ࡘ࡛ࡶヱᙜ ┿ᗘ㸦%㸧 㭜 B ࢺࣜࢡࣛ࣋ࣥࢲࢰ࣮ࣝ㸪ࣟ࢟ࢧࣝࢯࣥ㸪࢜࢟ࢩ ࢡࣟࢨࢽࢻࡀ㢧ⴭ࡛࠶ࡗࡓࠋࡇࢀࡽࡢᡂศࡘ ࠸࡚ࡣ㸪㐺⏝ྍ⬟࡞⃰ᗘ⠊ᅖࡸ㏣ຍ⢭〇➼ࡢ᳨ ウࡀᚲせ⪃࠼ࡽࢀࡓࠋ ᳨࣭㔞⥺ࡢ┤⥺ᛶࡀ↓࠸ ࣭ࣈࣛࣥࢡࣆ࣮ࢡࡀࡁࡃ 㑅ᢥᛶࡀప࠸ ࣭ᐃ㔞㝈⏺ࡀチᐜ⠊ᅖእ ࡲࡵ ␆Ỉ⏘≀ࢆᑐ㇟㸪ື≀⏝་⸆ရ 115 ᡂศࡢ ୍ᩧศᯒἲࢆ᳨ウࡋࡓࠋᢳฟ⁐፹㸪ᅛ┦࣒࢝ࣛ ͤ A㹼D ࡣ㑅ᢥᛶ㸪ᐃ㔞㝈⏺ࡢ᮲௳ࢆ‶ࡓࡍ ࡼࡿ⢭〇➼ࡢ᳨ウࢆ⾜࠸㸪ࡼࡾ㎿㏿࡛㧗⢭ᗘ 㭜༸࡛⣙ 8 㸪∵ஙཬࡧࡣࡕࡳࡘ࡛⣙ 9 ࡞୍ᩧศᯒἲࢆᵓ⠏ࡋࡓࠋ ࡞ࡾ㸪ᴫࡡⰋዲ࡞⤖ᯝࡀᚓࡽࢀࡓࠋ7 㣗ရࢃ ᵓ⠏ࡋࡓศᯒἲࡘ࠸࡚㸪7 㣗ရ㸦㭜⫗㸪㇜ ࡓࡾホ౯ A ࡞ࡗࡓࡢࡣ㸪115 ᡂศ୰ 66 ᡂศ࡛ ⫗㸪∵⫗㸪ࡉࡅ㸪㭜༸㸪∵ங㸪ࡣࡕࡳࡘ㸧ࢆᑐ ࠶ࡗࡓࠋ ㇟㸪ጇᙜᛶホ౯ヨ㦂ࢆᐇࡋࡓࠋ㐺ྜࡋࡓᡂ యⓗホ౯ A ࡢḟホ౯ D ࡢྜࡀ㧗ࡃ㸪 ศࡢྜࡣ㸪㭜⫗㸪㇜⫗㸪∵⫗㸪ࡉࡅ㸪㭜༸࡛ ホ౯ B㸪C ࡣప࠸ഴྥ࡛࠶ࡗࡓࠋE ヱᙜࡋࡓ ⣙ 8 㸪∵ஙཬࡧࡣࡕࡳࡘ࡛⣙ 9 ࡞ࡾ㸪ᴫ ࡢࡣ㸪㇜⫗ࡢࢭࣇࣆࣜࣥࡢࡳ࡛࠶ࡾ㸪ࣈࣛࣥ ࡡⰋዲ࡞⤖ᯝࡀᚓࡽࢀࡓࠋᮏἲࡣ㧗⢭ᗘ࡞㎿㏿ ࢡࣆ࣮ࢡࡀࡁࡃ㑅ᢥᛶࡸᐃ㔞ୗ㝈ၥ㢟ࡀ ୍ᩧศᯒἲࡋ࡚᭷⏝࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋ ᚋࡣ㸪㐺ྜ࡞ࡗࡓᡂศࡢ᳨ウࡸ㸪 ࠶ࡗࡓࠋ ࡣࡕࡳࡘࡣ㸪ホ౯ D㸪E ࡢᡂศࡀ࡞ࡃ㸪≉ Ⰻዲ࡞⤖ᯝ࡛࠶ࡗࡓࠋࡇࢀࡣ LC-MS/MS ᐃࡢ ᅇ ᐃᑐ㇟ࡋ࡞ࡗࡓᡂศࡘ࠸࡚ᮏἲࡢ㐺 ⏝ࢆᣑࡍࡿணᐃ࡛࠶ࡿࠋ ጉᐖ࡞ࡿ⬡⫫ࢆྵࢇ࡛࠸࡞࠸ࡓࡵ⪃࠼ࡽࢀ ཧ⪃ᩥ⊩ ࡓࠋ 㭜༸࡛ࡣ㸪ホ౯ B’ࡢྜࡀホ౯ A ḟ࠸࡛㧗 ࡗࡓࠋヱᙜࡍࡿᡂศࢆࡃࢃࡋࡃࡳࡿ㸪ࡍ 1㸧ཌ⏕ປാ┬་⸆㣗ရᒁ㣗ရᏳ㒊㛗㏻▱㸸㣗 ࡚㧗⃰ᗘῧຍࡢᐊෆ⢭ᗘࡀእࢀ࡚࠸ࡓࠋࡢ ရ୰ṧ␃ࡍࡿ㎰⸆➼㛵ࡍࡿヨ㦂ἲࡢጇ ࢇࡢᡂศࡣ㸪ే⾜࣭ᐊෆ⢭ᗘࢆ‶ࡓࡋ࡚࠾ ᙜᛶホ౯࢞ࢻࣛࣥࡘ࠸࡚㸪ᖹᡂ 19 ᖺ ࡾ㸪ᅇࡢᐊෆ⢭ᗘࡢ㐓⬺ࡣഅ↛㉳ࡁࡓ⪃ 11 ᭶ 15 ᪥㸪㣗ᏳⓎ➨ 115001 ྕ㸬 ࠼ࡽࢀࡿࠋ 2㸧ཌ⏕ປാ┬་⸆㣗ရᒁ㣗ရᏳ㒊㛗㏻▱㸸㣗 ホ౯ C㸪D ヱᙜࡍࡿᡂศࡣ㸪」ᩘࡢ㣗ရ ရ୰ṧ␃ࡍࡿ㎰⸆➼㛵ࡍࡿヨ㦂ἲࡢጇ ࢃࡓࡗ࡚࠸ࡿሙྜࡀከࡃ㸪≉࣑ࣟ࢟ࢧࢩࣥ㸪 ᙜᛶホ౯࢞ࢻࣛࣥࡢ୍㒊ᨵṇࡘ࠸࡚㸪 - 65 - 秋田県健康環境センター年報 第10号 2014 ᖹᡂ 22 ᖺ 12 ᭶ 24 ᪥㸪㣗ᏳⓎ➨ 1124 ➨ 1 ྕ㸬 ື≀⏝་⸆ရ୍ᩧศᯒἲ㸪㣗ရ⾨⏕Ꮫ㞧ㄅ㸪 54㸪4㸪2013㸪290-297㸬 3㸧ཌ⏕ປാ┬་⸆㣗ရᒁ㣗ရᏳ㒊㛗㏻▱㸸㣗 10㸧ྜྷ⏣⤮⨾Ꮚ㸸ຍᕤ㣗ရ୰ࡢື≀⏝་⸆ရ ရṧ␃ࡍࡿ㎰⸆㸪㣫ᩱῧຍ≀ཪࡣື≀⏝་ ⸆ရ ࡢ ᡂศ ࡛ ࠶ࡿ ≀ ㉁ࡢ ヨ㦂 ἲ ࡘ ࠸ ࡚ 㸪 ㎿㏿୍ᩧヨ㦂ἲࡢ᳨ウ㸪㣗ရ⾨⏕Ꮫ㞧ㄅ㸪52㸪 ࠕ HPLC ࡼ ࡿ ື ≀ ⏝ ་ ⸆ ရ ➼ࡢ ୍ ᩧ ヨ 㦂 1㸪2011㸪59-65㸬 11㸧ᇼỤṇ୍㸸LC-MS/MS ࡼࡿ␆Ỉ⏘㣗ရ ἲϨ㹼Ϫ㸦␆Ỉ⏘≀㸧ࠖ㸪ᖹᡂ 17 ᖺ 1 ᭶ 24 ᪥㸪㣗ᏳⓎ➨ 0124001 ྕ㸬 ୰ࡢ ࢽ ࢺࣟ ࣇ ࣛࢰ ࣥ ࡢᐃ 㔞㸪 㣗 ရ⾨ ⏕ Ꮫ㞧 4㸧ᯇῲளᕼᏊ㸸LC-MS/MS ࢆ⏝࠸ࡓ␆Ỉ⏘≀ ㄅ㸪49㸪3㸪2008㸪204-210㸬 ୰ࡢ ື ≀⏝ ་ ⸆ရ ୍ ᩧศ ᯒ ࠾ ࡅࡿ ᅛ ┦ᢳ 12㸧ᯘὒ㸸HPLC ࡼࡿ␆⏘㣗ရ୰ࡢࢩ࣐ࣟ ฟཬࡧࣜࣥ⬡㉁ࢡ࣮ࣜࣥࢵࣉἲࡢ᳨ウ㸪⛅ ࢪࣥศᯒἲ㸪ᮾி㒔ᗣᏳ◊✲ࢭࣥࢱ࣮ᖺ ⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ㸪7㸪2011㸪70-82㸬 ሗ㸪57㸪2006㸪187-191㸬 13㸧⏿㔝ᘯ㸸LC/MS/MS ࡼࡿஙཬࡧங〇ရ 5㸧ᯇῲளᕼᏊ㸸LC-MS/MS ࢆ⏝࠸ࡓࣁࢳ࣑ࢶ ୰ࡢື≀⏝་⸆ရ୍ᩧศᯒἲࡢ᳨ウ, ⛅⏣ ୰ࡢ࣌ࢽࢩࣜࣥ⣔ᢠ⏕≀㉁ࡢ㎿㏿ྠᐃ㔞㸪 ┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ㸪7㸪2011㸪83-90㸬 ⚟ᒸᕷಖ⎔ቃ◊✲ᡤሗ㸪28㸪2002㸪79-83㸬 6㸧ஂ⏣ಇ⿱㸸ᾮయࢡ࣐ࣟࢺࢢࣛࣇࢱࣥࢹ 14㸧➉ୖᬕ⨾㸸㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ࣮ ࣒ᆺ㉁㔞ศᯒィ㸦LC/MS/MS㸧ࢆ⏝࠸ࡓ␆Ỉ ㉁㔞 ศ ᯒἲ ࡼࡿ ங ୰ࡢ ࣐ࢡ ࣟ ࣛ ࢻ ⣔ᢠ ⏘㣗ရ୰ࡢື≀⏝་⸆ရ㎿㏿ศᯒἲࡢ᳨ウ ⏕ ≀ ㉁ ࡢ ᐃ 㔞 㸪 ศ ᯒ Ꮫ 㸪 55 㸪 9 㸪 2006 㸪 651-659㸬 㸦➨ 1 ሗ㸧㸪⇃ᮏ┴ಖ⎔ቃ⛉Ꮫ◊✲ᡤሗ㸪 15㸧すᮧಟ୍㸸LC-MS/MS ࡼࡿࣁࢳ࣑ࢶ୰ ➨ 39 ྕ㸪2009㸪39-44㸬 7㸧ᇼỤṇ୍㸸ᚤ⏕≀Ꮫⓗヨ㦂ἲࡼࡿ␆Ỉ⏘ ࡢṧ␃ື≀⏝་⸆ရ➼ࡢ୍ᩧศᯒ㸪㮵ඣᓥ┴ ⎔ቃಖࢭࣥࢱ࣮ᡤሗ㸪8㸪2007㸪125-130㸬 ≀୰ṧ␃ࡍࡿᢠ⳦ᛶ≀㉁ࡢ㧗ឤᗘ ᐃἲ㸪 16㸧㏆⸨㈗ⱥ㸸ศᩓᅛ┦ᢳฟ࠾ࡼࡧከᶵ⬟࢝ ศᯒᏛ㸪56㸪12㸪2007㸪1097-1103㸬 ࣒ࣛࢆ⏝࠸ࡓ GC-MS/MS ࡼࡿ␆⏘≀୰ࡢ 8㸧㛗༡㝯ኵ㸸HPLC ࡼࡿ␆Ỉ⏘㣗ရ୰ࡢ࢟ ࣀࣟࣥࡢ୍ᩧศᯒἲ㸪㣗ရ⾨⏕Ꮫ㞧ㄅ㸪49㸪 ṧ␃㎰⸆୍ᩧศᯒἲ㸪㣗ရ⾨⏕Ꮫ㞧ㄅ㸪53㸪 3㸪2008㸪244-248 2㸪2012㸪75-84㸬 9㸧ᒣཱྀ㈗ᘯ㸸ศᩓᅛ┦࠾ࡼࡧᅛ┦࣮࢝ࢺࣜࢵ 17㸧ゞቑ⿵᪥ᮏ㣗ရᶆ‽ᡂศ⾲ ࢪ࣒࢝ࣛࢆ⏝࠸ࡓ LC-MS/MS ࡼࡿ㣗⫗୰ࡢ - 66 - 秋田県健康環境センター年報 第10号 2014 ⾲ ጇᙜᛶホ౯ヨ㦂⤖ᯝ㸦ᡂศู㸧 No. ᑐ㇟ᡂศྡ RT 1 ) (min) 㺱㺤㺞㺎㺐㺓㺻 (m/z ) 㭜⫗ ㇜⫗ ∵⫗ ࡉࡅ 㭜༸ ∵ங ྛホ౯ࡢศᕸ 2 ) ࡣࡕ ࡳࡘ A B B' C D E 1 Positive 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 Cephapirin 5.8 424.2 > 292.2 D E D D A A A 3 0 0 0 3 5-Hydroxythiabendazole 5.8 218.0 > 191.0 A A D B' C A A 4 0 1 1 1 0 Sulfisomidine Lincomycin Lebamisole Cefquinome Ampicillin Diaveridine Thiabendazole Norfloxacin Ofloxacin Ciprofloxacin M arbofloxacin Trimethoprim Clopidol Danofloxacin Ormetoprim Neospiramycin䊠 Enrofloxacin Sulfadiazine Cephalonium Sulfathiazole M ecillinam Orbifloxacin Azaperone Sulfapyridine Spiramycin䊠 Sarafloxacin Xylazine Difloxacin Sulfamerazine M orantel Cefazolin Tripelennamine Sulfadimidine Sulfamonomethoxine Sulfisozole Pyrimethamine M afoprazine Tilmicosin (cis,trans) 5.9 279.1 > 124.0 A A B' B' A A A 5 0 2 0 0 0 5.9 407.2 > 126.0 A A D D A D A 4 0 0 0 3 0 6.0 205.1 > 178.0 A A A A A A A 7 0 0 0 0 0 6.0 529.2 > 134.0 A A D D D D A 3 0 0 0 4 0 6.1 350.1 > 106.3 A A D D A A A 5 0 0 0 2 0 6.1 261.1 > 245.1 A A B' A A A A 6 0 1 0 0 0 6.1 202.1 > 175.1 A A A A A A A 7 0 0 0 0 0 6.2 320.1 > 276.1 A A B' B' A A A 5 0 2 0 0 0 6.2 362.1 > 318.3 A A A A A A A 7 0 0 0 0 0 6.2 332.1 > 288.2 A A A A A A A 7 0 0 0 0 0 6.2 363.1 > 72.1 A A B' B' A A A 5 0 2 0 0 0 6.2 291.1 > 230.1 A A A A A A A 7 0 0 0 0 0 6.2 192.0 > 101.0 A A A A A A A 7 0 0 0 0 0 6.3 358.2 > 340.2 A A A A B' A A 6 0 1 0 0 0 6.4 275.1 > 259.1 A A A A A A A 7 0 0 0 0 0 6.5 350.3 > 174.3 A C C A A A A 5 0 0 2 0 0 6.5 360.2 > 316.3 A A A A A A A 7 0 0 0 0 0 6.5 251.1 > 155.9 A A A A A A A 7 0 0 0 0 0 6.5 459.1 > 337.1 A A A D D D A 4 0 0 0 3 0 6.6 256.1 > 155.9 A A A A A A A 7 0 0 0 0 0 6.7 326.2 > 167.2 A A A D A A A 6 0 0 0 1 0 6.7 396.1 > 352.2 A A A A A A A 7 0 0 0 0 0 6.7 328.1 > 165.0 A A A A A A A 7 0 0 0 0 0 6.8 250.1 > 156.1 A A A A A A A 7 0 0 0 0 0 6.9 422.4 > 174.1 A A A A A A A 7 0 0 0 0 0 7.0 386.1 > 342.2 A A A A A A A 7 0 0 0 0 0 7.0 221.1 > 90.1 A A A A A A A 7 0 0 0 0 0 7.1 400.1 > 356.1 A A A A A A A 7 0 0 0 0 0 7.1 265.1 > 155.9 A A A A A A A 7 0 0 0 0 0 7.3 221.1 > 123.0 A A A A A A A 7 0 0 0 0 0 7.5 455.1 > 323.2 A A D D B' A A 4 0 1 0 2 0 7.7 256.2 > 211.2 A A A A A A A 7 0 0 0 0 0 7.8 279.1 > 186.0 A A A A A A A 7 0 0 0 0 0 7.9 281.2 > 156.0 A A A A A A A 7 0 0 0 0 0 8.3 240.0 > 156.0 A A A A A A A 7 0 0 0 0 0 8.5 249.1 > 233.0 A A A A A A A 7 0 0 0 0 0 8.6 402.2 > 193.1 A A A A A A A 7 0 0 0 0 0 8.7 869.7 > 174.0 A A A A B' A A 6 0 1 0 0 0 Sulfamethoxypyridazine 8.7 281.2 > 155.9 A A A A A A A 7 0 0 0 0 0 Cefoperazone Oxibendazole Sulfachloropyridazine Oxfendazole M irosamicin Oleandomycin Sulfadoxine Sulfamethoxazole 8.9 646.1 > 530.3 A B' A A D A A 5 0 1 0 1 0 8.9 250.1 > 218.1 A C A A A A A 6 0 0 1 0 0 9.2 285.1 > 155.9 A A A A A A A 7 0 0 0 0 0 9.3 316.1 > 159.0 A A A A A A A 7 0 0 0 0 0 9.6 728.4 > 158.1 A A A A A A A 7 0 0 0 0 0 9.7 688.4 > 158.2 A A A A A A A 7 0 0 0 0 0 9.7 311.2 > 155.9 A A A A A A A 7 0 0 0 0 0 9.8 254.1 > 156.0 A A A A A A A 7 0 0 0 0 0 Sulfaethoxypyridazine 9.8 295.0 > 156.0 A A A A A A A 7 0 0 0 0 0 - 67 - 秋田県健康環境センター年報 第10号 2014 ⾲ ⥆ࡁ No. ᑐ㇟ᡂศྡ RT 1 ) (min) 㺱㺤㺞㺎㺐㺓㺻 (m/z ) 㭜⫗ ㇜⫗ ∵⫗ ࡉࡅ 㭜༸ ∵ங ྛホ౯ࡢศᕸ 2) ࡣࡕ ࡳࡘ A B B' C D E 51 Sulfatroxazole 9.9 268.1 > 156.0 B' B A A A A A 5 1 1 0 0 0 52 Erythromycin 10.1 734.5 > 158.2 A A A A A A A 7 0 0 0 0 0 53 Sulfisoxazole 10.2 268.1 > 156.0 A A A A B A A 6 1 0 0 0 0 54 Roxarson 10.2 263.9 > 245.8 D D D A D D B' 1 0 1 0 5 0 55 M iloxacin 10.3 263.9 > 215.2 D D D A D D B' 1 0 1 0 5 0 56 Oxolinic acid 10.4 262.2 > 216.1 A A A A A A A 7 0 0 0 0 0 57 Tylosin 10.5 916.6 > 174.1 A A A A A A A 7 0 0 0 0 0 0 58 Sulfabenzamide 10.9 277.1 > 156.0 A A A A A A A 7 0 0 0 0 59 Prednisolone 10.9 361.3 > 147.1 A A A A B' A A 6 0 1 0 0 0 60 Ethopabate 10.9 238.1 > 206.0 A A A A A A A 7 0 0 0 0 0 61 Kitasamycin 11.0 772.4 > 109.3 A A A A A A A 7 0 0 0 0 0 62 Tiamulin Fumarate 11.0 494.4 > 192.1 A A A A A A A 7 0 0 0 0 0 63 Sulfadimethoxine 11.1 311.2 > 156.1 A A A A A A A 7 0 0 0 0 0 64 Sulfaquinoxaline 11.1 301.2 > 156.0 A A A A A A A 7 0 0 0 0 0 65 Hydrocortisone 11.1 363.3 > 327.3 A A B B' A A A 5 1 1 0 0 0 0 66 Prifinium 11.1 306.3 > 86.1 A A A A A A A 7 0 0 0 0 67 Oxfendazole sulfone 11.4 332.1 > 300.1 A A A A A A A 7 0 0 0 0 0 68 M ebendazole 11.6 296.1 > 264.1 A C A A A A A 6 0 0 1 0 0 69 Josamycin 11.9 828.6 > 174.0 A A A A A A A 7 0 0 0 0 0 70 Benzocaine 11.9 166.0 > 138.0 A A A A A A A 7 0 0 0 0 0 71 Benzylpenicillin 12.0 335.1 > 160.1 A A A A B' A A 6 0 1 0 0 0 72 Methylprednisolone 12.0 375.1 > 356.9 A A A A A A A 7 0 0 0 0 0 73 Flubendazole 12.2 314.1 > 282.1 D C A A A A A 5 0 0 1 1 0 74 Betamethasone 12.2 393.2 > 373.1 A A A A B' A B 5 1 1 0 0 0 0 75 Nalidixic acid 12.4 233.2 > 215.2 A A A A A A A 7 0 0 0 0 76 Flumequine 12.7 262.2 > 202.2 A A A A A A A 7 0 0 0 0 0 77 Sulfabromomethazine 12.9 357.0 > A A A A A A A 7 0 0 0 0 0 0 92.1 78 7UHQERORQHĮȕ 13.0 271.2 > 253.2 A A A A A A A 7 0 0 0 0 79 Oxacillin 13.4 402.2 > 160.1 A A A A B' A A 6 0 1 0 0 0 80 Fenbendazole 13.4 300.1 > 268.1 D D A D A A A 4 0 0 0 3 0 81 Virginiamycin 13.4 526.3 > 337.1 A A A A B' A A 6 0 1 0 0 0 82 Brotizolam 13.6 392.9 > 314.0 A A A A A A A 7 0 0 0 0 0 83 Piromidic acid 13.8 289.2 > 243.2 A A A A A A A 7 0 0 0 0 0 84 M enbutone 14.0 259.3 > 159.1 A A A A B' A A 6 0 1 0 0 0 85 Nafcillin 14.4 415.3 > 199.2 A A A A B' A A 6 0 1 0 0 0 86 Ketoprofen 14.4 255.2 > 209.1 A A A A A A A 7 0 0 0 0 0 87 Praziquantel 14.5 313.2 > 203.1 A A A A A A A 7 0 0 0 0 0 88 M eloxicam 14.8 352.0 > 115.0 A A C A B A A 5 1 0 1 0 0 89 Dicloxacillin 15.1 470.1 > 160.1 A A A A B' A A 6 0 1 0 0 0 90 Flunixin 15.3 297.1 > 279.0 A A A A A A A 7 0 0 0 0 0 91 Triclabendazole Oxon 15.5 329.0 > 167.9 D D A A B' A A 4 0 1 0 2 0 0 92 Famphur 15.6 326.0 > 93.0 A A A A A A A 7 0 0 0 0 93 Rifaximin 15.7 786.4 > 754.4 A C B' A D A A 4 0 1 1 1 0 94 Altrenogest 15.8 311.4 > 227.1 D D C A A B' A 3 0 1 1 2 0 95 Carprofen 16.0 274.1 > 228.0 D D A A D A A 4 0 0 0 3 0 96 Clostebol 16.2 323.0 > 143.0 A C A A A A A 6 0 0 1 0 0 97 Febantel 17.1 447.1 > 415.1 A A A A A A A 7 0 0 0 0 0 98 Novobiocin 17.8 613.3 > 189.0 D D A D D A A 3 0 0 0 4 0 0 99 Triclabendazole 18.4 358.9 > 344.0 D D A D D A C 2 0 0 1 4 100 Chlormadinone 18.4 405.2 > 345.1 D D A A A A A 5 0 0 0 2 0 101 M elengestrol acetate 18.4 397.2 > 337.3 D C A A A A A 5 0 0 1 1 0 102 Tolfenamic Acid 19.2 262.1 > 209.1 D D A A A A A 5 0 0 0 2 0 - 68 - 秋田県健康環境センター年報 第10号 2014 ⾲ ⥆ࡁ No. ᑐ㇟ᡂศྡ RT 1 ) (min) 㺱㺤㺞㺎㺐㺓㺻 (m/z ) 㭜⫗ ㇜⫗ ∵⫗ ࡉࡅ 㭜༸ ∵ங ྛホ౯ࡢศᕸ 2 ) ࡣࡕ ࡳࡘ A B B' C D E Negative 103 Thiamphenicol 6.2 353.9 > 185.1 A A D B A A A 5 1 0 0 1 0 104 Cefuroxime 6.7 423.0 > 206.9 A A A A A A A 7 0 0 0 0 0 105 Florfenicol 7.4 355.8 > 184.9 A A A A A A A 7 0 0 0 0 0 106 Dinitolmide 7.4 224.0 > 180.9 A A A A A A A 7 0 0 0 0 0 107 Chloramphenicol 7.6 321.0 > 152.0 A A A A A A A 7 0 0 0 0 0 108 Clorsulon 7.8 379.8 > 343.9 A A A A A A A 7 0 0 0 0 0 109 Sulfanitran 8.9 334.1 > 136.1 A A A B A A A 6 1 0 0 0 0 110 P enoxymethylpenicillin 9.0 348.9 > 208.0 A A A A A A A 7 0 0 0 0 0 111 Zeranol 9.7 321.1 > 277.1 A A A A A A A 7 0 0 0 0 0 112 Cloxacillin 9.8 433.9 > 292.8 A A A A A A A 7 0 0 0 0 0 113 Nicarbazin 10.8 301.0 > 136.9 D D A A A A A 5 0 0 0 2 0 114 Diclazuril 11.5 404.9 > 333.8 D D A A A A A 5 0 0 0 2 0 115 Oxyclozanide 12.2 397.8 > 175.9 D D A A D C C 2 0 0 2 3 0 1㸧RT㸸Retention time㸸ಖᣢ㛫 2㸧ᡂศࡈࡢ 7 㣗ရࡢྛホ౯㸦A㹼E㸧ࡢᩘ - 69 - ! ! ! ! !ؠཅྱࣂ౹! ! 秋田県健康環境センター年報 第10号 2014 㸬ᏛⓎ⾲ ᰴ㸦BLNAR㸧ࡀ209ᰴ㸪TEMᆺࡢβ-lactamaseࢆ ಖ᭷ࡋ࡚࠸ࡓᰴࡣ59ᰴ࡛㸪ࡑࡢ࠺ࡕftsIࡢኚ␗ࢆ ࢪ࢜࢟ࢧࣥศゎ⬟ࡀపୗࡋࡓάᛶởἾ ࡢฎ⌮⬟ຊྥୖࡢ᳨ウ ేࡏᣢࡗ࡚࠸ࡓᰴ㸦BLPACR㸧ࡣ28ᰴ࡛࠶ࡗࡓࠋ ࡲࡓ㸪ࡇࢀࡲ࡛⛥ࡉࢀ࡚ࡁࡓ ROBᆺ β-lactamaseࢆಖ᭷ࡋ࡚࠸ࡓᰴࡀ44ᰴ☜ㄆࡉࢀ㸪 Ⳣཎ ๛㸪ᑠᯘ㈗ྖ㸪ᑠᕝ༓㸪ඵᰗ ࡑ ࡢ ࢇ ࡣ ftsIࡢ ኚ ␗ ࢆ ే ࡏ ᣢࡗ ࡚ ࠸ ࡓ ࠋ ₶ ROBᆺ β-lactamaseಖ᭷ ᰴ ࡣ 6᭶ࢆࣆ ࣮ࢡ ศ ➨ 23 ᅇ⎔ቃᏛウㄽ 㞳ࡀࡳ ࡽࢀ 㸪ᩘ ᰴࢆ 㝖 ࡁ 2⣔⤫ ࡢPFGE 2014 ᖺ 5 ᭶㸪ி㒔ᗓ ࣃࢱ࣮ࣥࡀᏑᅾࡋࡓࠋ ࡲࡓ㸪TEMᆺẚ Ampicillin㸪 Cefaclor ᙉ ࠸ ⪏ ᛶ ࢆ ♧ ࡍ ഴ ྥ ࡀ 㞴ศゎᛶ࡛࠶ࡿ 1,4-ࢪ࢜࢟ࢧࣥࡣ୍⯡ⓗ࡞ฎ ࠶ࡗࡓࠋ ⌮ᕤ⛬࡛ࡣฎ⌮ࡀ㞴ࡋ࠸ࡉࢀ࡚࠸ࡓࡀ㸪┴ෆ ROB ᆺ β-lactamase ࢆಖ᭷ࡍࡿࣥࣇ࢚ࣝࣥࢨ ࡢᗫỈฎ⌮タ࡛ࡣ⏕≀ฎ⌮ࡼࡾ≉␗ⓗຠ ⳦ࡣ㸪ftsI ࡢኚ␗ࢆేࡏᣢࡘഴྥࡀ࠶ࡗࡓࡇ ⋡ࡼࡃศゎฎ⌮ࡉࢀ࡚࠸ࡿࠋᖹᡂ 24 ᖺᗘࡢ ࡽ㸪ࡑࡢቑຍࡣᙉຊ࡞⪏ᛶ⳦࡛࠶ࡿ BLPACR ඛ㸪ࡇࡢᗫỈฎ⌮タ࠾࠸࡚㝖ཤ⋡ࡀపୗࡋ㸪 ࡢቑຍࡘ࡞ࡀࡿྍ⬟ᛶࡀ࠶ࡾ㸪ࡑࡢືྥࡘ ฎ⌮Ỉ୰ 1,4-ࢪ࢜࢟ࢧࣥ⃰ᗘࡀୖ᪼ࡍࡿࡀ ࠸࡚ὀពࡀᚲせ⪃࠼ࡽࢀࡿࠋ Ⓨ⏕ࡋࡓࠋฎ⌮⬟ຊࢆྥୖࡍࡿࡓࡵ㸪ᗫỈὶධ ࢆṆࡋ㸪1,4-ࢪ࢜࢟ࢧࣥ࠾ࡼࡧࣜࣥ㓟ࡢῧຍ ࢆ⾜ࡗࡓ⤖ᯝ㸪⣙ 3 㐌㛫ᚋᅇࡍࡿࡇࡀ࡛ ࡁࡓࠋฎ⌮⬟ຊపୗࡢཎᅉࡣ࡛᫂࠶ࡿࡀ㸪ࣈ ࡘࡘࡀࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡢ ᅾࡾ᪉ ࣮ࣟ࣡ࡢὶ㔞ࡸỈ㉁ࡢᏘ⠇ኚືࡀ㛵ࡋ࡚࠸ࡿ బ⸨ᐶᏊ ⪃࠼ࡽࢀࡓࠋ ⾨⏕ᚤ⏕≀ᢏ⾡༠㆟➨35ᅇ◊✲ ⛅⏣┴࠾ࡅࡿࣥࣇ࢚ࣝࣥࢨ⳦ࡢ⸆⪏ ᛶ㛵㐃㑇ఏᏊࡢ᳨ฟ≧ἣ 2014 ᖺ 6 ᭶㸪ᮾி㒔 㣗ရࡢᏳᛶࡸ⎔ቃၥ㢟࠾࠸࡚ᗈࡃά⏝ࡉ 㔝㈗அ ࢀ࡚࠸ࡿࠕࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥ㸻ᗣ༴ ᶵ㸦ࣜࢫࢡ㸧ࡘ࠸࡚ࡢሗពぢࢆ࡚ ➨ 88 ᅇ᪥ᮏឤᰁᏛᏛ⾡ㅮ₇ ࡢ㛵ಀ⪅㛫࡛⾜࠸㸪ࡼࡾࡼ࠸ᨻ⟇Ỵᐃࢆ┠ᣦࡍ 2014 ᖺ 6 ᭶㸪⚟ᒸᕷ 㐣⛬ࠖࡣ㸪ឤᰁᑐ⟇ࡶ✚ᴟⓗྲྀࡾධࢀࡿ ࡇࡀồࡵࡽࢀ࡚࠸ࡿࠋᅇ㸪⛅⏣┴࡛Ⓨ⏕ࡋ ࣥࣇ࢚ࣝࣥࢨ⳦㸦Haemophilus influenzae㸧 ࡓࡘࡘࡀ㛵ࡍࡿᗣ༴ᶵⓎ⏕㝿ࡋ⾜ ࡢ⸆⪏ᛶࡢᶵᵓࡋ࡚ࡣ β-lactamaseࡢ⏘⏕ ࢃࢀࡓࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡢᐇࢆ2 ࡼࡾ⪏ᛶࡍࡿሙྜ㸪࣌ࢽࢩࣜࣥ⤖ྜࢱࣥ ⤂ࡍࡿࠋ ࣃࢡ㉁ࡢ㑇ఏᏊ㸦ftsI㸧ኚ␗ࡀ⏕ࡌ࡚⪏ᛶࡍ 1㸧2008ᖺ8᭶㸪ᮏ┴࠾࠸࡚15ᖺࡪࡾ࢝ࢶ ࡿሙྜࡀ࠶ࡿࠋ≉㏆ᖺ㸪β-lactamaseࡼࡽ࡞ ࢶ࣒࢞ࢩ፹ᛶࡢࡘࡘࡀࡀⓎ⏕ࡋࡓࠋࡇࢀ ࠸⪏ᛶ⳦ࡢቑຍࡀ㢧ⴭ࡞ࡗ࡚࠸ࡿࠋ ࢆཷࡅ㸪ᙜࢭࣥࢱ࣮ࡣཌປ⛉◊ㄪᰝࢳ࣮࣒ࡢ༠ ᅇ㸪ᖹᡂ25ᖺ1᭶ࡽ12᭶┴ෆ་⒪ᶵ㛵 ຊࡼࡾ㸪ᝈ⪅ឤᰁ᥎ᐃᆅ࡛ࡢ࢝ࢶࢶ࣒࢞ࢩ ࡽ⸆⪏ᛶ㑇ఏᏊࡢ☜ㄆࡢࡓࡵ㏦ࡉࢀࡓࣥ ⏕ᜥ☜ㄆᤕ⋓ࡋࡓ㔝㰡ࡽࡢཎయศ㞳ࢆᡂ ࣇ ࣝ ࢚ ࣥ ࢨ ⳦ 338 ᰴ ࡘ ࠸ ࡚ 㸪 β-lactamase ࡋᚓࡓࠋࡉࡽ㸪࢝ࢶࢶ࣒࢞ࢩࡢ⏕ᜥᇦࡀⴭ 㸦bla TEM , bla ROB 㸧ཬࡧftsIࡢኚ␗ࡢ᳨ฟࢆPCR ྡほගᆅཬࢇ࡛࠸ࡓࡇࡽ㸪┴ෆࡢࡳ࡞ࡽ ࡼࡾ⾜ࡗࡓࠋ ࡎ┴እྥࡅࡓၨⓎ➼ࡀᚲせ࡞ࡗࡓࠋ ࠸ࡎࢀࡢ⪏ᛶ㑇ఏᏊࡶ᳨ฟࡉࢀ࡞ࡗࡓᰴ ࡑࡢࡓࡵ㸪ᑓ㛛་་ᖌ࠾ࡼࡧᆅඖ⮬య 㸦BLNAS㸧ࡣ26ᰴ㸪ftsIࡢኚ␗ࡢࡳ᳨ฟࡉࢀࡓ ࡢ༠ຊࡼࡾ㸪᪂⪺㸪ほගࣃࣥࣇࣞࢵࢺ࡛ὀព - 70 - 秋田県健康環境センター年報 第10号 2014 ႏ㉳ࢆ⾜࠺࡞㸪ࠕኟᏘࡢࡘࡘࡀࠖᑐ⟇ࡀ Food samples were rinsed by a homogenizing ㅮࡌࡽࢀࡿࡼ࠺࡞ࡗࡓࠋ buffer and centrifuged to obtain food emersion. 2㸧2013ᖺ5᭶㸪ࡘࡘࡀࡼࡿṚஸ⪅ࡀᮏ┴ Anti NoV GII.4 serum was added into the food ࡛ࡣ15ᖺࡪࡾⓎ⏕ࡋࡓࡀ㸪ᙜึࡣሗබ㛤ࡢ emersion to make NoV-IgG complexes, then つᐃࡼࡾබ⾲ࡉࢀ࡞ࡗࡓࠋࡋࡋ㸪ᝈ⪅Ⓨ PANSORBIN ® was added into the samples and ⏕ᆅᇦࡢಖᡤᙜࢭࣥࢱ࣮ࡽࡢሗᥦ౪㸪 re-centrifuged ᑓ㛛་࠾ࡼࡧ་ᖌࡢാࡁࡅࡼࡾ㸪༶ adsorbed on the surface of PANSORBIN ® . NoV つᐃࡀぢ┤ࡋࡉࢀᙜヱṚஸࡀබ⾲࡞ࡗࡓࠋ RNA was extracted from the pellet using a ࡉࡽ㸪ᝈ⪅Ⓨ⏕ࡢሗබ㛤ࡢࢱ࣑ࣥࢢࡶ commercial extraction kit, then RT real-time PCR ぢ┤ࡋࡉࢀࡓࠋ process was performed. The recovery rate from to collect NoV-IgG complexes ᗣ༴ᶵ࣭⿕ᐖⓎ⏕ࢆᮍ↛㜵ࡄ࠶ࡿ࠸ࡣ⿕ potato salad was 78.3±10.8% in contrast to ᐖపῶࡢࡓࡵࡢࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥ࠾ 0.33±0.08% when no anti NoV GII.4 serum was ࠸࡚㸪ᆅ᪉⾨⏕◊✲ᡤࡀᯝࡓࡍࡁᙺࡣࡁ added. Similar results were obtained from the stir ࠸⪃࠼ࡽࢀࡿࠋṇ☜࡞ࣜࢫࢡㄆ㆑࣭ศᯒే fried noodle, 81.5±10.2% vs 1.15±0.25%. We also applied a commercially available human ࡏ࡚㸪ᗈࡃ୍⯡☜ᐇ࿘▱࡛ࡁࡿሗබ㛤᪉ gamma globulin agent to the PANtrap method to ἲࡀᚋࡢㄢ㢟ࡋ࡚ᣲࡆࡽࢀࡿࠋ detect other viruses and succeeded to recover another genotypes of NoV as well as sapoviruses, 1RURYLUXVHV 51$ GHWHFWLRQ LQ FRQWDPLQDWHG IRRGV E\ D 3$1WUDS PHWKRG adenovirus type 41 and hepatitis A virus. These results sho w the potentiality of the PANtrap method for the detection of food bone viruses in contaminated food which bring many Hiroyuki Saito , Tomoyuki Tanaka Miho Toho *2 Tomoichiro Oka *4 , Mamoru Noda *1 advantages to prevent food-borne gastroenteritis. *3 , Kazuhiko Katayama *4 nd The 2 . AFSA Conference on Food Safety and *1 㸸ሜᕷ⾨⏕◊✲ᡤ *2 㸸⚟┴⾨⏕⎔ቃ◊✲ࢭࣥࢱ࣮ *3 㸸ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ *4 㸸ᅜ❧ឤᰁ◊✲ᡤ Security 2014ᖺ8᭶, Bien Hoa City, Vietnam Noroviruses (NoVs) are one of a major cause of ⛅⏣┴ࡢࡘࡘࡀᑐ⟇࠾ࡅࡿㄢ㢟 ᒎᮃ gastroenteritis mostly related not only to food poisoning but a dominant cause of food-related బ⸨ᐶᏊ㸪ᰘ⏣ࡕࡦࢁ㸪⛅㔝⳹Ꮚ outbreaks. బ⸨⏤⾰Ꮚ㸪ᩪ⸨༤அ㸪Ᏻ㒊┿⌮Ꮚ Foodborne gastroenteritis is being 㡲⸨ᜏஂ *1 increasingly recognized as a public health problem, however, identification of the etiological agents responsible has been restricted due to the difficulty ➨ 60 ᅇ᪥ᮏᐤ⏕Ꮫ to detect the agents in food samples. A PANtrap ᪥ᮏ⾨⏕ື≀Ꮫ᪥ᮏᨭ㒊ྜྠ ® ((PANSORBIN (Staphylococcus aureus fixed with 2014 ᖺ 10 ᭶㸪┒ᒸᕷ formalin) trap) method we developed for the detection and quantification of NoVs from 㣗ရࡢᏳᛶࡸ⎔ቃၥ㢟࠾࠸࡚ᗈࡃά⏝ࡉ different food samples included solid, liquid, paste ࢀ࡚࠸ࡿࠕࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥ㸻ᗣ༴ and oily states. We spiked NoV GII.4 stool ᶵ㸦ࣜࢫࢡ㸧ࡘ࠸࡚ࡢሗពぢࢆ࡚ suspension that includes 10 5 copies of NoV RNA ࡢ㛵ಀ⪅㛫࡛⾜࠸㸪ࡼࡾࡼ࠸ᨻ⟇Ỵᐃࢆ┠ᣦࡍ into 10 grams of potato salad and stir fried noodle . 㐣⛬ࠖࡣ㸪ឤᰁᑐ⟇ࡶ✚ᴟⓗྲྀࡾධࢀࡿ - 71 - 秋田県健康環境センター年報 第10号 2014 ࡇࡀồࡵࡽࢀ࡚࠸ࡿࠋᅇ㸪⛅⏣┴࡛Ⓨ⏕ࡋ ࣟࣈࣜࣥ〇ࢆᑟධࡍࡿࡇ࡛㸪ᮏἲࢆỗ⏝ⓗ ࡓࡘࡘࡀ㛵ࡍࡿᗣ༴ᶵⓎ⏕㝿ࡋ⾜ ⏝࠸ࡿࡇࡀྍ⬟࡛࠶ࡿࠋࡇࢀࡲ࡛㸪࣐࢞ࣥ ࢃࢀࡓࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡢᐇࢆ⤂ ࢢࣟࣈࣜࣥ〇ࡋ࡚㸪་⒪⏝ࡢࡶࡢࢆ⏝࠸࡚ ࡍࡿࠋ ࠸ࡓࡀ㸪᳨ᰝἲࡢᬑཬᙜࡓࡗ࡚ࡣ㸪ヨ㦂◊✲ 2013ᖺ5᭶㸪ࡘࡘࡀࡼࡿṚஸ⪅ࡀᮏ┴࡛ ⏝➼ࡢ࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࡀᮃࡲࡋ࠸ࠋᮏ◊ ࡣ15ᖺࡪࡾⓎ⏕ࡋࡓࡀ㸪ᙜึࡣሗබ㛤ࡢつ ✲࡛ࡣ㸪ࡇࡢၥ㢟ࢆゎỴࡋ㸪㣗ရࡢ࢘ࣝࢫ᳨ ᐃࡼࡾබ⾲ࡉࢀ࡞ࡗࡓࠋࡋࡋ㸪ᝈ⪅Ⓨ⏕ ᰝࡢ࡞ᬑཬ⧅ࡆࡿࡓࡵ㸪་⸆ရ௨እ࡛ ᆅᇦࡢಖᡤᙜࢭࣥࢱ࣮ࡽࡢሗᥦ౪㸪ᑓ Ᏻᐃⓗ⏝࡛ࡁࡿᤕᤊᢠయ౪⤥※ࢆ᳨ウࡋ 㛛་࠾ࡼࡧ་ᖌࡢാࡁࡅࡼࡾ㸪༶つ ࡓࠋ ᐃࡀぢ┤ࡋࡉࢀᙜヱṚஸࡀබ⾲࡞ࡗࡓࠋࡇ NoV-GII.4 ࢆྵࡴ 50 mL ࡢ㣗ရὙᾮࡢᕤ ࡢࡇࡼࡾ㸪┴Ẹࡢὀពႏ㉳ࡀಁࡉࢀ㸪⤖ ᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥῧຍ㔞ࡢ᳨ウ࡛ࡣ㸪5㸣⁐ ᯝࡋ࡚ᩆ࡛ࡁࡓࡶ࠶ࡗࡓࠋࡑࡢᚋ㸪ᝈ ᾮࢆ 150 μL ຍ࠼ࡓ᮲௳ࡀ᭱ࡶ㧗࠸ᅇ⋡ࢆ♧ ⪅Ⓨ⏕ࡢሗබ㛤ࡢࢱ࣑ࣥࢢࡘ࠸࡚ぢ┤ ࡋࡓࠋࡇࡇ࡛᭱㐺ࡉࢀࡓῧຍ㔞ࢆ⏝࠸࡚㸪ୖ ࡋࡀᅗࡽࢀࡓࠋᖺ࡞ࡗ࡚㸪⛅⏣┴࡛ࡣ4ᖺࡪ グࡢ 5 ✀㢮ࡢ NoV ࡘ࠸࡚㣗ရὙᾮࡽࡢᅇ ࡾ࢝ࢶࢶ࣒࢞ࢩ፹ᛶࡢKatoᆺࡘࡘࡀ ࢆヨࡳࡓࡇࢁ㸪㣗୰ẘࡢཎᅉ࢘ࣝࢫࡋ ᝈ⪅ࡀⓎ⏕ࡋ㸪┴Ẹࡢሗᥦ౪ࡀ㏿ࡸ⾜ ࡚᭱ከࡢ NoV-GII.4 ᑐࡋ࡚ᕤᴗ⏝࣐࢞ࣥࢢࣟ ࢃࢀࡓࠋ ࣈࣜࣥࢆ⏝࠸ࡓሙྜࡢᅇ⋡ࡣ 52.0㸣࡛㸪་⒪ ᗣ༴ᶵ࣭⿕ᐖⓎ⏕ࢆᮍ↛㜵ࡄ࠶ࡿ࠸ࡣ⿕ ⏝࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࢆ⏝࠸ࡓሙྜࡣ 46.8㸣 ᐖపῶࡢࡓࡵࡢࣜࢫࢡࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥ࠾ ࡛࠶ࡗࡓࠋ௨ୗ㸪NoV-GII.2 ᑐࡋ࡚ࡣ 38.6㸣 ࠸࡚ࡣ㸪ヨ㦂◊✲ᶵ㛵㸪⾜ᨻᶵ㛵࠾ࡼࡧ་⒪ᶵ /38.2㸣㸦ᕤᴗ⏝/་⒪⏝㸧㸪NoV-GII.6 ᑐࡋ࡚ 㛵ࡀ୍య࡞ࡗ࡚ṇ☜࡞ࣜࢫࢡㄆ㆑࣭ศᯒࢆࡍ ࡣ 15.5㸣/13.3㸣㸪NoV-GI.4 ᑐࡋ࡚ࡣ 35.7㸣 ࡿᚲせࡀ࠶ࡿࠋేࡏ࡚㸪ᗈࡃ୍⯡☜ᐇ࿘▱ /37.6㸣㸪NoV-GI.6 ᑐࡋ࡚ࡣ 12.1㸣/6.1㸣࡛࠶ ࡛ࡁࡿሗබ㛤ࡢయไࢆ⥔ᣢࡍࡿࡇࡀồࡵࡽ ࡾ㸪ẚ㍑ࡋࡓ࡚࠾࠸࡚୧⪅ࡣྠ➼࡛࠶ࡗࡓࠋ ࢀࡿࠋ ࡲࡓ㸪ᐇ㝿ࡢởᰁ㣗ရࢆࣔࢹࣝࡋࡓẚ㍑ヨ㦂 *1 ࠾࠸࡚ࡶ㸪ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࢆ⏝࠸ࡓ 㸸⛅⏣Ꮫ་Ꮫ㒊ྡᩍᤵ ⣔ࡣ࣏ࢸࢺࢧࣛࢲ࡛ 40.6㸣㸪↝ࡁࡑࡤ࡛ 33.5㸣 㸪་⒪⏝ࡢࡑࢀ㸦࣏ࢸࢺࢧࣛࢲ࡛ 34.7㸣㸪↝ 㣗ရ᳨యࡢࣀࣟ࢘ࣝࢫ᳨ᰝࣃࣥࢯࣝࣅ ࣭ࣥࢺࣛࢵࣉἲࢆ⏝࠸ࡿ㝿ࡢᤕᤊᢠయ౪⤥ ※㛵ࡍࡿ᳨ウ ࡁࡑࡤ࡛ 32.4㸣㸧ྠ➼௨ୖࡢᅇ⋡ࢆ♧ࡋࡓࠋ ࡇࢀࡽࡢࡇࡽ㸪⏝࠾࠸࡚≉ẁࡢไ⣙ࡢ ↓࠸ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࢆヨ⸆ࡋ࡚ࣃࣥ ࢺࣛἲᑟධࡍࡿࡇࡣኚྜ⌮ⓗ࡛࠶ࡾ㸪ỗ ᩪ⸨༤அ㸪⛅㔝⳹Ꮚ㸪⏣୰ᬛஅ 㔝⏣ ⾨ *1 ⏝ᛶࢆᢸಖࡍࡿព࡛ࡶ✚ᴟⓗ࡞ά⏝ὶ㏻ *2 ࣮ࣝࢺࡢᩚഛࡀᮃࡲࢀࡿࡶࡢ⪃࠼ࡽࢀࡿࠋ ➨ 35 ᅇ᪥ᮏ㣗ရᚤ⏕≀ᏛᏛ⾡⥲ *1 㸸ሜᕷ⾨⏕◊✲ᡤ *2 㸸ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ 2014 ᖺ 10 ᭶㸪ሜᕷ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ㸦ࣃࣥࢺࣛἲ: ᩪ ⸨༤அ㸪᪥ᮏ㣗ရᚤ⏕≀Ꮫ㞧ㄅ㸪Vol.29㸪No.1㸪 32-37㸪 2012㸧 ࡣ 㸪 㣗 ရ ᳨ య ࡽ ࣀ ࣟ ࢘ ࣝ ࢫ 㣗ရ᳨యࡽࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ ࡼ ࡾ ࣀ ࣟ ࢘ ࣝ ࢫ RNA ࢆ ᢳ ฟ ࡍ ࡿ 㝿 ࡢ α-Amylase ฎ⌮㛵ࡍࡿ᳨ウ 㸦NoV㸧ࢆ᳨ฟࡍࡿࡓࡵࡢᐇ㊶ⓗ࡞ᡭἲ࡛࠶ࡿࠋ ⛅㔝⳹Ꮚ㸪ᩪ⸨༤அ㸪⏣୰ᬛஅ *1 ᮏἲࡢᇶᮏཎ⌮ࡣ㸪㯤Ⰽࣈࢻ࢘⌫⳦ࡢ⾲㠃㸪 㔝⏣ ᤕᤊᢠయࢆࡋ࡚࢘ࣝࢫ⢏Ꮚࢆ྾╔ࡉࡏ࡚ᅇ ⾨ *2 ᳨࣭ฟࡍࡿࡇ࡛࠶ࡿࠋᤕᤊᢠయࡋ࡚㸪ከ ➨ 35 ᅇ᪥ᮏ㣗ရᚤ⏕≀ᏛᏛ⾡⥲ ᵝ࡞࢘ࣝࢫᑐࡍࡿᢠయࢆྵࡴࡇ࣐࢞ࣥࢢ - 72 - 秋田県健康環境センター年報 第10号 2014 2014 ᖺ 10 ᭶㸪ሜᕷ బ⸨ᐶᏊ㸪ᰘ⏣ࡕࡦࢁ㸪బ⸨⏤⾰Ꮚ ⛅㔝⳹Ꮚ㸪ᩪ⸨༤அ㸪Ᏻ㒊┿⌮Ꮚ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ㸦ࣃࣥࢺࣛἲ: ᩪ 㰺⸨ᚿಖᏊ㸪ཌ⏕ປാ⛉Ꮫ◊✲Ᏻ⸨⌜ ⸨༤அ㸪᪥ᮏ㣗ရᚤ⏕≀Ꮫ㞧ㄅ㸪Vol.29㸪No.1㸪 ➨ 12 ᅇ⛅⏣┴බ⾗⾨⏕ᏛᏛ⾡ 32-37㸪2012㸧ࡣ㸪ࣀࣟ࢘ࣝࢫ㸦NoV㸧௦⾲ 2014 ᖺ 11 ᭶㸪⛅⏣ᕷ ࡉࢀࡿ㣗୰ẘ㉳ᅉ࢘ࣝࢫࢆ㣗ရ᳨యࡽ᳨ฟ ࡍࡿࡓࡵ㛤Ⓨࡉࢀࡓࠋ㣗ရ୰ࡣ PCR ࢆ㜼ᐖ ࡍࡿᵝࠎ࡞≀㉁ࡀྵࡲࢀ࡚࠸ࡿࡀ㸪≉ⅣỈ 2008ᖺ㸪⛅⏣┴࠾࠸࡚15ᖺࡪࡾ࢝ࢶࢶ ≀ࡣ≀⌮Ꮫⓗᣲືࡀ᰾㓟㸦࣏࣮ࣜࣜ࣎ࢫ㸧 ࣒࢞ࢩ፹ᛶࡢKatoᆺࡘࡘࡀᝈ⪅ࡀⓎ⏕ࡋ 㢮ఝࡋ࡚࠸ࡿࡇࡽ㸪㐺ษ࡞ศゎ㝖ཤฎ⌮ࡀ ࡓࡇࢆዎᶵ㸪2009ᖺࡽ࢝ࢶࢶ࣒࢞ࢩࡢ ᚲせ࡛࠶ࡿࠋ࢝࢟ࡢ᳨ᰝἲ࠾࠸࡚ࡣ㸪ΰᅾࡋ ⏕ᜥᇦㄪᰝࢆ㛤ጞࡋࡓࠋㄪᰝࡣ㞝≀ᕝୖὶ㒊ࡢ ࡚࠸ࡿࢢࣜࢥ࣮ࢤࣥࢆศゎࡍࡿࡓࡵ ἑᕷࡽ୰ὶ㒊ࡢᕷ⮳ࡿἙᕝᩜࢆᑐ㇟ α-Amylase ⢊ᮎࡀ⏝࠸ࡽࢀ࡚࠸ࡿࡀ㸪ࣃࣥࢺࣛ ᆅᇦࡋ㸪㯮ᕸぢྲྀࡾἲࡼࡾࢶࢶ࣒࢞ࢩࡢ᥇ ἲ࠾࠸࡚ࡶᕤ⛬ྠᵝࡢฎ⌮ࡀ⤌ࡳ㎸ࡲࢀ࡚ 㞟ࢆ⾜ࡗࡓࠋࡲࡓ㸪2009ᖺ㹼2014ᖺ10᭶ࡲ࡛ࡢ ࠸ࡿࠋ୍᪉࡛㸪⌧ᅾධᡭྍ⬟࡞ α-Amylase ⢊ᮎ ኟᏘࡢ ࡘࡘࡀ ᝈ⪅ 8ࡘ ࠸࡚ឤ ᰁ᥎ ᐃᆅ ࡣ㘄࡞ࡢ〇㐀࠺㈿ᙧ┦ᙜࡍࡿ ࠾ࡼࡧసᴗෆᐜ⮫ᗋᡤぢࢆ㞟ィࡋࡓࠋ ⁐ᛶᡂศࡀྵࡲࢀ࡚࠾ࡾ㸪᭱⤊ⓗ࡞᳨ฟ⢭ᗘ ㄪᰝᆅᇦࡢ᭱ୖὶ㒊࠶ࡓࡿἑᕷ㑹እ࡛ࡣ㸪 ᙳ㡪ࢆཬࡰࡍࡇࡀࢃࡗࡓࠋศᏊ⏕≀Ꮫ⏝ 39ᖺࡢグ㘓࡛ࡣᝈ⪅Ⓨ⏕ᆅࡉࢀ࡚࠸ࡓࡀ㸪 ࢢࣜࢭ࣮ࣟࣝ⁐ᾮࡋ࡚౪⤥ࡉࢀ࡚࠸ࡿ ࢝ࢶࢶ࣒࢞ࢩࡢ⏕ᜥࡣ☜ㄆࡉࢀࡎ㸪ࡑࡇࡽ α-Amylase ࡢ⏝ࡣ㸪ࢥࢫࢺ㠃࡛⌧ᐇⓗ࡛ࡣ࡞ ⣙3 kmୗὶ⮳ࡗ࡚ࡽ☜ㄆࡉࢀࡓࠋࡉࡽୗ ࠸ࠋࡑࡇ࡛㸪ᮏ◊✲࡛ࡣ α-Amylase ⢊ᮎࢆᾮ ὶࡢᶓᡭᕷࢆὶࢀࡿᨭὶࡢྜὶⅬ㏆࡙ࡃ ㄪ〇ࡍࡿ᪉ἲࢆ⪃ࡋ㸪ⅣỈ≀ࡢศゎ㝖ཤຠ ࡘࢀ㸪ከࡃࡢ࢝ࢶࢶ࣒࢞ࢩࡀ☜ㄆࡉࢀࡿࡼ࠺ ᯝࡘ࠸᳨࡚ウࡋࡓࠋࡑࡢ᳨ウ⤖ᯝࢆᑟධࡍࡿ ࡞ࡗࡓࠋ୍᪉࡛ㄪᰝ⠊ᅖࡢ᭱ୗὶ㒊⨨ࡍ ࡇࡼࡾࣃࣥࢺࣛἲࡼࡿ㣗ရ᳨ᰝࡢ⢭ᗘྥ ࡿᕷ㑹እ㒊ࡢグ㘓ୖࡢᝈ⪅Ⓨ⏕ᆅ࡛ࡶ ୖ⧅ࡆࡿࡇࢆ┠ⓗࡋ࡚࠸ࡿࠋ ࢝ࢶࢶ࣒࢞ࢩࡢ⏕ᜥ☜ㄆࡀฟ᮶࡞ࡗࡓࡀ㸪ࡑ ࣚ࢘⣲ࢹࣥࣉࣥᛂࢆほᐹࡍࡿࡇ࡛㸪ᾮ ࡇࡽ18 kmୖὶᆅⅬ௨༡࡛ࡣ☜ㄆࡉࢀࡿࡼ࠺ ㄪ〇ࡋࡓ α-Amylase ࡣ㸪α-Amylase ⢊ᮎࢆ┤᥋ ࡞ࡾ㸪㞝≀ᕝὶᇦ࠾ࡅࡿ᭱ࡶ㧗ᐦᗘ࡞⏕ᜥ ᢞධࡋࡓሙྜྠ➼ࡢⅣỈ≀㝖ཤຠᯝࢆ☜ㄆ ᇦࡣⰼⅆሙ࿘㎶࠶ࡿࡇࡀ᥎ᐹࡉࢀࡓࠋ ࡛ࡁࡓࠋࡇࡢຠᯝࡣ㸪ᾮㄪ〇ᚋ -20Υ࡛ 6 ࣨ ࡲࡓ㸪࢝ࢶࢶ࣒࢞ࢩࡢ⏕ᜥ⎔ቃࡣᗈ࠸Ἑᕝᩜ ᭶ಖᏑࡋࡓᚋ࡛ࡶᦆ࡞ࢃࢀ࡞ࡗࡓࠋࡲࡓ㸪 ࡢ୰࡛ࡶỈ㝿ࡢ◁ᆅ㸪≉୰ᕞ࡛ࡢ⏕ᜥᩘࡀ α-Amylase ⢊ᮎࡢ⁐ᛶᡂศࡀṧ␃ࡋࡓ≧ែ࡛ ከ࠸ഴྥ࠶ࡗࡓࡀ㸪Ἑᕝἢ࠸ࡢබᅬࡸ㔮ࡾሙ ࣃࣥࢺࣛἲࢆ⾜ࡗࡓሙྜࡣ㸪㏵୰࡛ RNA ⢭ ࠸ࡗࡓቑỈỈἐࡍࡿ⟠ᡤࡶ☜ㄆࡉࢀࡓࠋ 〇࣒࢝ࣛࡀ┠ワࡲࡾࢆ㉳ࡇࡍ࡞ࡢ᧯సୖࡢࢺ 2009ᖺ௨㝆 ࡢኟ Ꮨࡢ ࡘࡘࡀ 8 ࡢ࠺ ࡕ࢝ ࣛࣈࣝࡀ㉳ࡇࡾࡸࡍࡃ࡞ࡾ㸪real-time PCR ࡛ࡶ ࢶࢶ࢞ ࣒ࢩ፹ ᛶ ࡛࠶ࡿ ࡇࡀ ࢃࢀ ࡓࡢ ࡣ 3 ഇ㝜ᛶ࡞ࡗࡓࠋࡇ࠺ࡋࡓၥ㢟ࢆᅇ㑊ࡍࡿࡓࡵ ࡛㸪ឤᰁ᥎ᐃ᭶ࡣ8᭶ࡀ2㸪9᭶ࡀ1࡛࠶ࡗ 㸪ࡇࢀࡲ࡛ࡣ⁐ᛶᡂศࢆࢫ࣏ࢺ࡛ᑀ ࡓࠋ࠸ࡎࢀࡢᝈ⪅ࡶỈ㝿࡛ࡢసᴗᚋⓎࡋ࡚ ྲྀࡾ㝖ࡃᚲせࡀ࠶ࡗࡓࡀ㸪ᾮㄪ〇ရࢆ⏝࠸ࡿ ࠾ࡾ㸪ࡘࡘࡀ㛵ࡍࡿ▱㆑ࡀ࡞ࡗࡓ㸪 ࡇ࡛㸪ࡇ࠺ࡋࡓ㒔ྜࡀ࡞ࡃ࡞ࡾ㸪᧯సࡢ⡆ ࠶ࡿ࠸ࡣ▱ࡗ࡚࠸࡚ࡶᙜࡣὀពࢆᡶࡗ࡚࠸࡞ ౽᳨ฟ⢭ᗘࡢྥୖࡀᅗࡽࢀࡓࠋ ࡗࡓࡶࡢ᥎ᐹࡉࢀࡿࠋ *1 㸸ሜᕷ⾨⏕◊✲ᡤ *2 㸸ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ ࡘࡘࡀࡢᝈ⪅Ⓨ⏕ᆅࢶࢶ࣒࢞ࢩࡢ⏕ែ ࠾ࡼࡧ⏕ᜥᇦࡣᐦ᥋࡞㛵㐃ᛶࡀ࠶ࡿࡇࡽ㸪 ᚋࡶຠᯝⓗ࡞ࡘࡘࡀᑐ⟇࣭ၨⓎࡢࡓࡵࡢ ሗࢆⓎಙࡋ࡚࠸ࡁࡓ࠸⪃࠼࡚࠸ࡿࠋ ኟᏘࡢࡘࡘࡀⓎ⏕≧ἣ࢝ࢶࢶ࣒࢞ ࢩ⏕ᜥㄪᰝ - 73 - 秋田県健康環境センター年報 第10号 2014 ⛅⏣┴࠾ࡅࡿṚᅉࡢᴫせ㸫㒔㐨ᗓ┴ูᖺ 㱋ㄪᩚṚஸ⋡ᇶ࡙ࡃ᳨ウ㸫 ࢆぢࡍ࠼ࡘࡘᘬࡁ⥆ࡁᑐ⟇ࢆᙉࡋ࡚࠸ࡃᚲせ ⏣୰㈗Ꮚ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ࠾ࡅࡿᤕᤊᢠ యࡋ࡚ࡢᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࡢ᭷⏝ ᛶࡢ᳨ド ࡀ࠶ࡿᛮࢃࢀࡿࠋ ➨ 12 ᅇ⛅⏣┴බ⾗⾨⏕ᏛᏛ⾡ 2014 ᖺ 11 ᭶㸪⛅⏣ᕷ ᩪ⸨༤அ㸪⛅㔝⳹Ꮚ㸪⏣୰ᬛஅ *1 ⛅⏣┴ࡢண㜵ᑐ⟇㛵ࡍࡿ་⒪ཬࡧ⾜ᨻ 㔝⏣ ⾨ *2 ⟇❧ࢆᨭࡍࡿࡓࡵ㸪ᖺ㱋ㄪᩚṚஸ⋡ࡢ ➨ 62 ᅇ᪥ᮏ࢘ࣝࢫᏛᏛ⾡㞟 ࢹ࣮ࢱࢆ⏝࠸࡚ᇶ♏ⓗ࡞ࢹ࣮ࢱࢆ㞟ゎᯒࡋ㸪 2014 ᖺ 11 ᭶㸪ᶓᕷ ⛅⏣┴ࡢഴྥࢆ᫂ࡽࡍࡿࠋ2010 ᖺ㒔㐨 ᗓ┴ูᖺ㱋ㄪᩚṚஸ⋡㸫ேཱྀືែ⤫ィ≉Ṧሗ࿌ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ㸦ࣃࣥࢺࣛἲ㸧ࡣ㸪 㸫㸦ཌ⏕ປാ┬⮧ᐁᡣ⤫ィሗ㒊㸧ࡽ㸪⛅ ⏣┴ࡢᖺ㱋ㄪᩚṚஸ⋡㸦ேཱྀ 10 ᑐ㸧ࢆᢳฟࡋ 㣗ရ᳨యࡽࣀࣟ࢘ࣝࢫ㸦NoV㸧ࢆ᳨ฟࡍࡿ ࡓࠋ2010 ᖺࡢࢹ࣮ࢱࡘ࠸࡚ᮏ┴ᅜࡢ㧗 ࡓࡵࡢᐇ㊶ⓗ࡞ᡭἲ࡛࠶ࡿࠋᮏἲࡣ࢘ࣝࢫ⢏ పཬࡧ㸪ᮏ┴ࡢㄪᰝ㛤ጞᖺ 2010 ᖺࡢቑῶࢆẚ Ꮚࡢᅇ㯤Ⰽࣈࢻ࢘⌫⳦ࢆ⏝࠸㸪୧⪅ࢆ⤖ࡧ ㍑ࡋ 4 㡿ᇦ㸦A㸪B㸪C㸪D㸧ศ㢮ࡋࡓࠋᑐ㇟ ࡘࡅࡿࡓࡵࡢᤕᤊᢠయࢆᚲせࡍࡿࠋ㛤Ⓨẁ㝵 Ṛᅉࡣᝏᛶ᪂⏕≀㸦௨ୗ㸪ࡀࢇ㸧㸪ᚰᝈ㸪⬻ ࡛⏝࠸࡚ࡁࡓ࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࡣ㸪ከࡃࡢ ⾑⟶ᝈ㸪⫵⅖㸪⮬ẅ㸪៖ࡢᨾ㸪⫢ᝈ㸪 ࢘ࣝࢫᑐࡍࡿᢠయࢆྵࢇ࡛࠸ࡿࡇࡽ㸪 ⭈㸪⢾ᒀ㸪៏ᛶ㛢ሰᛶ⫵ᝈ㸪⪁⾶➼ࡢ ᤕᤊᢠయࡋ࡚ࡢỗ⏝ᛶࡣ㧗࠸ࠋࡇࢀࡲ࡛㸪࢞ ィ 31 Ṛᅉ࡛࠶ࡗࡓࠋ ࣐ࣥࢢࣟࣈࣜࣥ〇ࡣ㸪་⒪⏝ࡢࡶࡢࢆ⏝ࡋ ᅜࡼࡾపࡃㄪᰝ㛤ጞᖺࡽῶᑡࡋ࡚࠸ࡿ㹂 ࡚ࡁࡓࡀ㸪ᅇᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࢆ 㡿ᇦࡢṚᅉࡣ㸪⏨࡛ࡣ 7 Ṛᅉ㸪ዪ࡛ࡣ 9 Ṛᅉ࡛ ධᡭࡍࡿࡇࡀ࡛ࡁࡓࠋࡑࡇ࡛㸪ᕤᴗ⏝㏵⏕ ࠶ࡾ㸪⾑ᛶᚰᝈ㸪ᛴᛶᚰ➽᱾ሰ㸪⫢ࡀࢇ㸪 ⏘ࡉࢀ࡚࠸ࡿ࣐࢞ࣥࢢࣟࣈࣜࣥࢆࣃࣥࢺࣛἲ ⫢ᝈ㸪៏ᛶ㛢ሰᛶ⫵ᝈ㸪⪁⾶ࡣ⏨ዪඹ㏻࡛ ᑟධࡋ㸪ࡉࡽ࡞ࡿỗ⏝ᛶࢆ㏣ồࡍࡿࡇࢆ┠ⓗ ࠶ࡗࡓࠋᅜࡼࡾప࠸ࡀㄪᰝ㛤ጞᖺࡽቑຍࡋ ࡋ᳨࡚ウࢆ⾜ࡗࡓࠋ ࡚࠸ࡿ C 㡿ᇦࡣ⏨࡛ࡣ 3 Ṛᅉ㸪ዪ࡛ࡣ 4 Ṛᅉ࡛ NoV-GII.4 ࢆྵࡴ 50 mL ࡢ㣗ရὙᾮࡢᕤ ࠶ࡗࡓࠋẼ⟶࣭⫵ࡀࢇࡣ⏨ዪඹ㏻࡛࠶ࡗࡓࠋㄪ ᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥῧຍ㔞ࡢ᳨ウ࡛ࡣ㸪5㸣⁐ ᰝ㛤ጞᖺࡽࡳࡿῶᑡࡋ࡚࠸ࡿࡶࡢࡢᅜࡼ ᾮࢆ 150 μL ຍ࠼ࡓ᮲௳ࡀ᭱ࡶ㧗࠸ᅇ⋡ࢆ♧ ࡾ㧗࠸㹀㡿ᇦࡣ㸪⏨࡛ࡣ 10 Ṛᅉ㸪ዪ࡛ࡣ 13 Ṛ ࡋࡓࠋࡇࡇ࡛᭱㐺ࡉࢀࡓῧຍ㔞ࢆ⏝࠸࡚㸪ୖ ᅉ࡛࠶ࡾ㸪⏨ዪඹ㏻ࡣᝏᛶ᪂⏕≀㸪⬻⾑⟶ᝈ㸪 グࡢ 5 ✀㢮ࡢ NoV ࡘ࠸࡚㣗ရὙᾮࡽࡢᅇ ⫵⅖㸪⫶ࡀࢇ㸪⬻᱾ሰ㸪៖ࡢᨾ㸪⬻ෆฟ⾑㸪 ࢆヨࡳࡓࡇࢁ㸪࡚࠾࠸࡚་⒪⏝࣐࢞ࣥ 㣗㐨ࡀࢇ㸪ࡃࡶ⭷ୗฟ⾑࡛࠶ࡗࡓࠋᅜࡼࡾ㧗 ࢢࣟࣈࣜࣥ〇ྠ➼௨ୖࡢᅇ⋡ࡀᚓࡽࢀ ࡃㄪᰝ㛤ጞᖺࡽቑຍࡋ࡚࠸ࡿ㸿㡿ᇦࡣ㸪⏨࡛ ࡓࠋࡲࡓ㸪ᐇ㝿ࡢởᰁ㣗ရࢆࣔࢹࣝࡋࡓẚ㍑ ࡣ⮬ẅ㸪⭠ࡀࢇ㸪⮅ࡀࢇ㸪ᩚ⬦࣭ఏᑟ㞀ᐖ㸪 ヨ㦂࠾࠸࡚ࡶ㸪ᕤᴗ⏝࣐࢞ࣥࢢࣟࣈࣜࣥࢆ⏝ ⭈㸪⫹ࡢ࠺ࡀࢇ㸪๓❧⭢ࡀࢇ㸪⢾ᒀࡢ 8 ࠸ ࡓ ⣔ ࡣ ࣏ ࢸ ࢺ ࢧ ࣛ ࢲ ࡛ 40.6㸣 㸪 ↝ ࡁ ࡑ ࡤ ࡛ Ṛᅉ㸪ዪ࡛ࡣஙࡀࢇ㸪⭈㸪༸ᕢࡀࢇ㸪ᝏᛶ 33.5㸣㸪་⒪⏝ࡢࡑࢀ㸦࣏ࢸࢺࢧࣛࢲ࡛ 34.7 ࣜࣥࣃ⭘ࡢ 4 Ṛᅉ࡛࠶ࡗࡓࠋ⏨ዪඹ㏻ࡣ⭈ 㸣㸪↝ࡁࡑࡤ࡛ 32.4㸣㸧ྠ➼௨ୖࡢᅇ⋡ࢆ ࡛࠶ࡗࡓࠋᖺ㱋ㄪᩚṚஸ⋡ࡀῶᑡࡋ࡚࠸ࡿ㹀ཬ ♧ࡋࡓࠋࡇࢀࡽࡢࡇࡽ㸪ᕤᴗ⏝࣐࢞ࣥࢢࣟ ࡧ㹂㡿ᇦࡣ㸪⏨࡛ࡣ 17 Ṛᅉ㸪ዪ࡛ࡣ 22 Ṛᅉ࡛ ࣈࣜࣥࢆヨ⸆ࡋ࡚ࣃࣥࢺࣛἲᑟධࡍࡿࡇ ࠶ࡗࡓࠋᮏ┴࡛ࡣከࡃࡢṚᅉࡀῶᑡഴྥ࠶ࡾ㸪 ࡣኚྜ⌮ⓗ࡛࠶ࡾ㸪ὶ㏻࣮ࣝࢺࡀᩚഛࡉࢀࢀ ࡇࢀࡲ࡛ࡢ╔ᐇ࡞ಖ་⒪ᑐ⟇ࡢຠᯝࡀఛࢃࢀ ࡤỗ⏝ᛶࡀࡉࡽ㧗ࡲࡿࡶࡢ⪃࠼ࡽࢀࡿࠋ ࡿࠋࡋࡋ㸪⭈ࡢࡼ࠺ᅜ࡛ࡢ㡰ࡀ *1 㸸⚟┴⾨⏕⎔ቃ◊✲ࢭࣥࢱ࣮ ࡁࡃୖࡀࡗࡓṚᅉࡶ࠶ࡿࡇࡽ㸪ᚋࡢືྥ *2 㸸ᅜ❧ឤᰁ◊✲ᡤ - 74 - 秋田県健康環境センター年報 第10号 2014 ⛅⏣┴ෆࡢ་⒪ᶵ㛵࠾ࡅࡿ $PS& ᆺșࣛ ࢡࢱ࣐࣮ࢮ⏘⏕⳦ࡢศ㞳≧ἣ⸆ឤཷᛶ ࡢゎᯒ⤖ᯝ ⚟ᓥཎⓎᨾࡼࡿ⛅⏣┴ෆࡢ㣗ရࡢᨺ ᑕ⬟ᙳ㡪ࡘ࠸࡚ ᑠᕝ༓㸪Ⳣཎ బ▱Ꮚ *2 㸪ᑠᯘ㈗ྖ㸪ᒾ㇂㔠ோ *3 ᶔᑿᣅᏊ㸪⇃㇂ඃᏊ㸪㔝㈗அ 㧘ᶫᚿಖ㸪⏣ᜨ⌮Ꮚ㸪ඵᰗ ๛ *1 㸪㧗ᶫ▱Ꮚ ₶ ➨ 38 ᅇ⛅⏣┴་Ꮫ᳨ᰝᏛ ➨ 38 ᅇ⛅⏣┴་Ꮫ᳨ᰝᏛ 2014 ᖺ 11 ᭶㸪⛅⏣ᕷ 2014 ᖺ 11 ᭶㸪⛅⏣ᕷ ᖹᡂ 23 ᖺ 3 ᭶ࡢᮾ᪥ᮏ㟈⅏క࠸Ⓨ⏕ࡋࡓ AmpC ᆺ β ࣛࢡࢱ࣐࣮ࢮ⏘⏕⳦ࡢ࠺ࡕ AmpC ⚟ᓥ➨୍ཎᏊຊⓎ㟁ᡤࡢᨾ㸦⚟ᓥཎⓎᨾ㸧 㐣⏘⏕ᰴ㸪ࣉࣛࢫ࣑ࢻᛶ AmpC ⏘⏕ᰴࡣ ESBL ࡼࡾேᕤᨺᑕᛶ≀㉁ࡀ⎔ቃ୰ᣑᩓࡋࡓࡇ ⏘⏕⳦ྠᵝ➨ 3 ୡ௦ࢭࣇ࢙࣒⣔ᢠ⏕≀㉁➼ ࢆཷࡅ㸪⛅⏣┴ࡣ㣗ရࡢᏳᛶࢆ☜ಖࡍࡿࡓࡵ㸪 ⪏ᛶࢆ♧ࡍࡇࡽ㸪㝔ෆឤᰁཎᅉ⳦ࡋ࡚ ┴ෆࡢ㣗ရྵࡲࢀࡿᨺᑕᛶ≀㉁ࡢㄪᰝࢆ㛤ጞ 㔜せどࡉࢀ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽ㸪་⒪ᶵ㛵࡛ ࡋࡓࠋᅇࡣ㸪ᖹᡂ 23 ᖺ 7 ᭶ࡢ᳨ᰝ㛤ጞࡽᖹ ᐇྍ⬟࡞ AmpC ⏘⏕ᰴࡢ⡆౽࡞᳨ᰝἲࡀࡇࢀ ᡂ 26 ᖺ 7 ᭶ࡲ࡛ᙜࢭࣥࢱ࣮࡛ᐇࡋࡓ㣗ရࡢ ࡲ࡛ᬑཬࡋ࡚࠸࡞ࡗࡓࡇࡽ㸪⛅⏣┴ෆࡢ ᨺᑕᛶ≀㉁᳨ᰝ⤖ᯝ㸪⚟ᓥཎⓎᨾࡼࡿ┴ ་⒪ᶵ㛵࠾ࡅࡿ AmpC ⏘⏕⳦ࡢศ㞳≧ἣࡣ ෆࡢ㣗ရࡢᨺᑕ⬟ᙳ㡪ࡘ࠸࡚ሗ࿌ࡍࡿࠋ ࡛᫂࠶ࡗࡓࠋ㏆ᖺ㸪AmpC ⏘⏕ᰴࢆ⡆౽᳨ฟ ┴ෆ⏘㣗ရཬࡧ┴ෆ࡛ὶ㏻ࡋ࡚࠸ࡿ┴እ⏘㣗 ྍ⬟࡞ AmpC/ESBL 㚷ูࢹࢫࢡࡀᕷ㈍ࡉࢀࡓ ရ 1,262 ᳨యࡢᨺᑕᛶ≀㉁⃰ᗘࢆ㸪බᐃἲᚑ ࡇࡽ㸪ᅇࡇࢀࢆ⏝ࡋ࡚⛅⏣┴ෆࡢ ࠸ࢤ࣐ࣝࢽ࣒༙࢘ᑟయ᳨ฟჾࡼࡾ ᐃࡋࡓࠋ AmpC ⏘⏕ᰴࡢศ㞳≧ἣ⸆ឤཷᛶࡘ࠸࡚ ࡑࡢ⤖ᯝ㸪ᨺᑕᛶࣚ࢘⣲㸦I-131㸧ࡣ࡚ࡢ㣗ရ ㄪᰝࡋࡓࠋࡑࡢ⤖ᯝ㸪ᖹᡂ 25 ᖺ 4 ᭶ࡽᖹᡂ ࡽ᳨ฟࡉࢀ࡞ࡗࡓࠋ୍᪉㸪ᨺᑕᛶࢭࢩ࣒࢘ 26 ᖺ 2 ᭶ࡲ࡛㸪┴ෆࡢ་⒪ᶵ㛵ࡽ ESBL ⏘⏕ 㸦Cs-134 ཬࡧ Cs-137㸧ࡣ㸪␆⏘≀ࡸᒣ࣭⳯ࡁࡢ ࠸ᰴࡋ࡚㏦ࡉࢀࡓ 636 ᰴࡢ࠺ࡕ㸪52 ᰴ ࡇ㢮࡞ 248 ᳨యࡽ᳨ฟࡉࢀ㸪4 ᳨యࡀᇶ‽ 㸦8.2%㸧ࡀ AmpC ⏘⏕ᰴ࡛࠶ࡿࡇࡀ᫂ࡽ ್ࢆ㉸࠼ࡓࠋࡑࡢ࠺ࡕ㸪∵⫗ࡘ࠸࡚ࡣ㸪ᨾ ࡞ ࡗ ࡓ ࠋ AmpC ⏘ ⏕ ᰴ 52 ᰴ ࡢ ࠺ ࡕ E.coli 㸪 ┤ᚋࡢᖹᡂ 23 ᖺ 7 ᭶ 1 ᳨యࡀ 520 Bq/kg ᬻ Enterobacter ᒓ⳦㸪Citrobacter ᒓ⳦ࡢィ 22 ᰴࡀ ᐃつไ್㸦500 Bq/kg㸧ࢆ㉸࠼ࡓࡀ㸪⩣ᖺࡣ᳨ ➨ 3 ୡ௦ࢭࣇ࢙࣒⣔⸆ࡢ CTX CAZ ࡢ୧᪉ ฟ್᭱ࡀ 4 Bq/kg ࡲ࡛ῶᑡࡋ࡚࠾ࡾ㸪୍ⓗ ⪏ᛶࢆ♧ࡋࡓࠋ ࡞ࡶࡢ࡛࠶ࡗࡓࠋࡡࡲࡀࡾࡓࡅ㸦㔝⏕ᒣ⳯㸧 E.coli 16 ᰴࡘ࠸࡚㸪᪤ሗࢆࡶࣉࣛࢫ࣑ ࡘ࠸࡚ࡣᖹᡂ 25 ᖺ 6 ᭶ᖹᡂ 26 ᖺ 6 ᭶ 3 ᳨ ࢻᛶ ampC ࡢ᥈⣴ࢆ⾜ࡗࡓࡇࢁ㸪4 ᰴࡀ bla CMY-2 యࡀ 110㹼120 Bq/kg ᇶ‽್㸦100 Bq/kg㸧ࢆ㉸ ࢆಖ᭷ࡍࡿᰴ࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡾ㸪⛅⏣ ࠼ࡓࡀ㸪࠸ࡎࢀࡶᐑᇛ┴ࡢ┴ቃ㏆ࡽ᥇ྲྀ ┴ෆࡢ་⒪ᶵ㛵ࣉࣛࢫ࣑ࢻᛶ ampC ಖ᭷ᰴࡀ ࡉࢀࡓࡶࡢ࡛࠶ࡗࡓࠋ Ꮡᅾࡍࡿࡇࡀึࡵ࡚☜ㄆࡉࢀࡓࠋ ௨ୖࡽ㸪ᨺᑕᛶ≀㉁⃰ᗘࡀᇶ‽್ࢆ㉸࠼ࡓ ࡲࡓ㸪ᅇ☜ㄆࡉࢀࡓ AmpC ⏘⏕ᰴࡢ 38.5%ࡀ 㣗ရࡣ㸪୍ⓗ࡞ࡶࡢ㸪ࡶࡋࡃࡣ⏕⏘ᆅࡀࡈࡃ ᒀ⏤᮶࡛࠶ࡗࡓࠋᒀࡣ㝔ෆឤᰁࡢ㔜せ࡞ឤᰁ※ 㝈ࡽࢀࡓᆅᇦࡢࡶࡢ࡛࠶ࡿࡓࡵ㸪⚟ᓥཎⓎᨾ ࡛࠶ࡿࡇࡽ㸪AmpC ⏘⏕ᰴࡘ࠸࡚ࡶᒀࢆ ࡼࡿ┴ෆࡢ㣗ရࡢᨺᑕ⬟ᙳ㡪ࡣ㸪ᴟࡵ࡚ᑡ ࡋ࡚㝔ෆឤᰁࢆᘬࡁ㉳ࡇࡍྍ⬟ᛶ␃ពࡍࡿ ࡞ࡃ㝈ᐃⓗ࡞ࡶࡢ࡛࠶ࡗࡓ⪃࠼ࡽࢀࡓࠋ ᚲせࡀ࠶ࡿࠋ⪏ᛶ⳦ࡢᘏ㜵Ṇ㐺ษ࡞ᢠ⳦⸆ *1 㸸ᖹ㮵ᆅᇦ⯆ᒁ⚟♴⎔ቃ㒊 ⒪ࡢࡓࡵ㸪ᚋࡶ AmpC/ESBL 㚷ูࢹࢫ *2 㸸ඖᗣ⎔ቃࢭࣥࢱ࣮ ࢡࣉࣛࢫ࣑ࢻᛶ ampC ᳨ฟ⏝ PCR ἲࡼࡾ⛅ *3 㸸⛅⏣ᆅᇦ⯆ᒁ㤋⚟♴⎔ቃ㒊 ⏣┴࠾ࡅࡿ AmpC ⏘⏕ᰴࡢ᳨ฟືྥὀពࡍ ࡿࡇࡀᚲせ࡛࠶ࡿࠋ - 75 - 秋田県健康環境センター年報 第10号 2014 ㎰స≀࠾ࡅࡿṧ␃㎰⸆୍ᩧศᯒἲࡢጇᙜ ᛶホ౯ 㔝⏣ ⌋⏣ᑦಇ㸪ᯇῲளᕼᏊ㸪ኳ᫂ࡉ࠾ࡾ *1 ⾨ *2 ➨ 108 ᅇ᪥ᮏ㣗ရ⾨⏕ᏛᏛ⾡ㅮ₇ 2014 ᖺ 12 ᭶㸪㔠ἑᕷ ➨ 51 ᅇᅜ⾨⏕Ꮫᢏ⾡༠㆟ᖺ 2014 ᖺ 11 ᭶㸪ูᗓᕷ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲ㸦ࣃࣥࢺࣛἲ㸧ࡣ㸪 㣗ရ᳨యࡽࣀࣟ࢘ࣝࢫ㸦NoV㸧ࢆ᳨ฟࡍࡿ ṧ␃㎰⸆ከᡂศศᯒࡢ㎿㏿ཬࡧ⢭ᗘྥୖࡢ ࡓࡵ㛤Ⓨࡉࢀࡓᐇ㊶ⓗ࡞ᡭἲ࡛࠶ࡿࠋࡲࡓ㸪 ࡓࡵ㸪⡆᫆㎿㏿ἲ࡛࠶ࡿࢡ࢚ࢵࢳ࣮ࣕࢬἲࢆᇶ ᤕᤊᢠయࡋ࡚࣐࢞ࣥࢢࣟࣈࣜࣥࢆ⏝ࡍࡿࡇ 㸪᪂ࡓ࡞๓ฎ⌮ἲࢆ᳨ウࡋࡓࠋ ࡛㸪NoV ࡢࡳ࡞ࡽࡎࢧ࣏࢘ࣝࢫ㸦SaV㸧 ᢳฟᕤ⛬࡛ࡣ㸪ᚑ᮶㸪⏝࠸ࡿ ᐃᶵჾẖู ࡶ㐺⏝࡛ࡁࡿࡇࡀࢃࡗ࡚࠸ࡿࠋࣃࣥࢺࣛἲ ࡢᢳฟἲ࡛࠶ࡗࡓࡀ㸦GC-MS/MS㸸㉸⮫⏺ὶయ ࡣ࢘ࣝࢫ⢏Ꮚࡢᅇ㯤Ⰽࣈࢻ࢘⌫⳦㸦௨ ᢳฟ㸪 LC-MS/MS㸸࣓ࢱࣀ࣮ࣝᢳฟ㸧㸪సᴗ ୗ㸪ࣈ⳦㸧ࡀ⏝࠸ࡽࢀ࡚࠸ࡿࡇࡽ㸪ᚓࡽࢀ 㛫ࡢ▷࠸ࢡ࢚ࢵࢳ࣮ࣕࢬᢳฟ⤫୍ࡋ㸪᧯సࡢ ࡓ RNA ࢧࣥࣉࣝࡣࣈ⳦⏤᮶ࡢ㑇ఏᏊࢆ㔞 ㎿㏿ࢆᅗࡗࡓࠋ ྵࢇ࡛࠾ࡾ㸪RT-PCR ⏝࠸ࡿヨᩱࡋ࡚ࡣ≉ ⢭〇ᕤ⛬࡛ࡣ㸪ᅛ┦࣒࢝ࣛࢆ⏝࠸ࡿࡇࡋ㸪 Ṧ࡞ࡶࡢ࠸࠼ࡿࠋࡇࢀࡲ࡛㸪NoV ᑐࡍࡿ GC ཬࡧ LC-MS/MS ⏝ู࡛ࠎ⢭〇ࢆ⾜࠺ࡇ RT-PCR ࡢᛂ⣔㸦ࣉ࣐࣮ࣛࡸᛂ ᗘ࡞㸧 ࡛⢭〇ຠᯝࢆࡼࡾ㧗ࡵࡿࡇࡋࡓ㸦௨ୗྛἲ ࡘ࠸࡚ࡣ᭱㐺ࡀ࡞ࡉࢀ࡚࠸ࡿ㸦➨ 106 ᅇᏛ ࢆ GC ἲ㸪LC ἲࡍࡿ㸧ࠋᨵⰋࡼࡾ㸪୍ᚊᇶ ⾡ㅮ₇㸧ࠋ୍᪉㸪SaV 㛵ࡋ࡚ࡣ㸪⣅౽᳨య ‽࡛ࣞ࣋ࣝ GC ἲ 275 㡯┠㸪LC ἲ 161 㡯┠ࡢ ᑐࡍࡿ RT-PCR ࡢᛂ⣔ࡣሗ࿌ࡉࢀ࡚࠸ࡿࡶ ᐃࡀྍ⬟࡞ࡗࡓࠋ ࡢࡢ㸪ࡇࢀࡽࢆࣃࣥࢺࣛἲ⏝࠸ࡿࡓࡵࡣ㸪 GC ἲཬࡧ LC ἲࡘ࠸࡚㸪7 㣗ရ㸦ࡾࢇࡈ㸪 ᨵࡵ࡚᭱㐺ࢆ⾜࠺ᚲせࡀ࠶ࡿࠋᮏ◊✲ࡣ㸪ࣃ ࢇࢀࢇࡑ࠺㸪⋞⡿㸪ࡡࡂ㸪ࡤࢀ࠸ࡋࡻ㸪࢟ࣕ ࣥࢺࣛἲࡼࡗ࡚ᢳฟࡉࢀࡓ SaV-RNA ࡢ᳨ฟ ࣋ࢶ㸪ࣞࣔࣥ㸧ࢆᑐ㇟㸪ཌ⏕ປാ┬㏻▱ࡢ࢞ ⣔ࢆ᭱㐺ࡋ㸪ᐇ㝿ࡢ㣗୰ẘᑐᛂ࡛ࡁࡿ ࢻࣛࣥᚑ࠸ጇᙜᛶホ౯ヨ㦂ࢆᐇࡋࡓ ࡶࡢࡍࡿࡇࢆ┠ⓗࡋ࡚࠸ࡿࠋ 㸦ῧຍ⃰ᗘ㸸0.01 ཬࡧ 0.05 μg/g㸪ศᯒ⪅ 3 ྡࡀ cDNA ࡢẁ㝵ᕼ㔘⣔ิ࠾࠸࡚㸪PCR ࡛ࡢ᳨ ྛ 2 ⃰ᗘῧຍヨᩱࢆ 1 ᪥ 2 ヨ⾜㸪2 ᪥㛫ศᯒ㸧ࠋ ฟ㝈⏺ࢆẚ㍑ࡋࡓࡇࢁ㸪␃Ỉࡼࡿᕼ㔘࡛ GC ἲ࡛ࡣ㸪࢞ࢻࣛࣥࡢ┠ᶆ್ࢆ‶ࡓࡋ ࡣ RT-PCR ཎἲࡼࡾ 10 -4 ᕼ㔘ࡲ࡛ 800 bps ࡢ ࡓ㎰⸆ࡣྛ㣗ရ࡛⣙ 8 ࡞ࡾ㸪ᴫࡡⰋዲ࡞⤖ ቑᖜࣂࣥࢻࡀ☜ㄆ࡛ࡁࡓࡀ㸪ࣃࣥࢺࣛἲᢳฟ≀ ᯝ࡛࠶ࡗࡓࠋ ࢆྵࡴᕼ㔘⣔ิ࡛ࡣࣈ⳦⏤᮶ࡢ㠀≉␗ࣂࣥࢻࡀ LC ἲ࡛ࡣ㸪ࡡࡂ௨እࡢ 6 㣗ရ࡛⣙ 8 ࡞ ከࡃ㸪SaV ⏤᮶ࡢቑᖜࣂࣥࢻࡣ 10 -2 ᕼ㔘ࡲ࡛ࡋ ࡾ㸪ࡡࡂࡣ⣙ 6 ࡛࠶ࡗࡓࠋࡡࡂࡣ┿ᗘࡀ㐺ྜ ᳨ฟ࡛ࡁ࡞ࡗࡓࠋࢽ࣮ࣜࣥࢢ ᗘࢆ 60Υ ࡋ࡞࠸ࡶࡢࡀẚ㍑ⓗከࡗࡓࠋ タᐃࡋࡓሙྜࡣ㸪ࣛࣥࢲ࣒ࣉ࣐࣮ࣛࢆ⏝࠸ ᚋ㸪LC ἲࡢࡡࡂࡢᅇ⋡ࡢᨵၿ㸪ᅇ ࡚㏫㌿ᛂࢆ⾜ࡗࡓሙྜࡣ㸪ࣈ⳦⏤᮶ࡢ㠀 ጇᙜ ᛶホ ౯ヨ 㦂ࢆ ᐇ ࡋ ࡓ 7 㣗ရ௨ እࡢ 㣗ရ ≉␗ࣂࣥࢻࡣ┠❧ࡓ࡞ࡃ࡞ࡗࡓࡶࡢࡢ㸪SaV ⏤ 㸦㇋㢮➼㸧ࡘ࠸࡚ጇᙜᛶホ౯ࢆ㐍ࡵ࡚࠸ࡃண ᮶ࡢቑᖜࣂࣥࢻࡣ 10 -1 ᕼ㔘ࡲ࡛ࡋㄆࡵࡽࢀ࡞ ᐃ࡛࠶ࡿࠋ ࡗࡓࠋLNA ಟ㣭ࢆᑟධࡋࡓࣉ࣐࣮ࣛࢆ⏝࠸ *1 㸸ᒣᮏᆅᇦ⯆ᒁ⚟♴⎔ቃ㒊 ࡚㸪 ࢽ ࣮ࣜ ࣥࢢ ᗘ ࢆ 60Υタ ᐃࡋ ࡓࡇ ࢁ㸪ࣃࣥࢺࣛἲᢳฟ≀Ꮡᅾୗ࡛ࡶ㸪10 -4 ᕼ㔘ࡲ ࡛ SaV ⏤᮶ࣂࣥࢻࡀほᐹ࡛ࡁࡓࠋḟ㸪 RNA 㣗୰ẘ࠾ࡅࡿ㣗ရࡢࢧ࣏࢘ࣝࢫ᳨ ᰝࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࢆ⏝࠸ࡿ㝿 ࡢ RNA ᳨ฟ⣔ࡢ᭱㐺 ࢆ␃Ỉ࡛ẁ㝵ᕼ㔘ࡋࡓࡶࡢࢆࣛࣥࢲ࣒ࣉࣛ ࣐ ࣮ ࡛ ㏫ ㌿ ࢆ ⾜ ࡗ ࡓ ሙ ྜ ࡣ 10 -5 ᕼ 㔘 ࡲ ࡛ SaV ࡢࣂࣥࢻࡀㄆࡵࡽࢀࡓࡀ㸪ࣃࣥࢺࣛἲᢳฟ ≀Ꮡᅾୗ࡛ࡣ 10 -3 ᕼ㔘ࡲ࡛ࡋ᳨ฟ࡛ࡁ࡞ࡗ ᩪ⸨༤அ㸪⛅㔝⳹Ꮚ㸪⏣୰ᬛஅ *1 㸪 ࡓࠋ㏫㌿ᛂᑓ⏝ࣉ࣐࣮ࣛ㸦PANR-SV㸧ࢆ - 76 - 秋田県健康環境センター年報 第10号 2014 ⏝࠸ࡿࡇ࡛ 10 -5 ᕼ㔘ࡲ᳨࡛ฟ࡛ࡁࡿࡼ࠺࡞ ᵝ⾜ࡗࡓࠋ ࡗࡓࠋࡇࡇ࡛ᚓࡽࢀࡓᡂ⦼ࡣ㸪㣗ရ᳨యࡢ SaV ᙉẘᆺᰴࡢ᥋✀⣔࠾ࡅࡿ Ot ᳨ฟ㛤ጞࡣ㸪A ᳨ᰝࣃࣥࢺࣛἲࢆᑟධࡍࡿ㝿᭷⏝⪃࠼ࡽ ⩌ࡀ 1 ᪥┠㸪B ⩌ࡀ 2 ᪥┠࡛㸪ࡑࡢᚋࢥࣆ࣮ᩘ ࢀࡓࠋ ࡣቑຍࡋ㸪9 ᪥┠ A ⩌ࡀ㸪10 ᪥┠ B ⩌ࡢ࣐ *1 㸸⚟┴⾨⏕⎔ቃ◊✲ࢭࣥࢱ࣮ ࢘ࢫࡀ⾶ᙅື࡞ࡾẅฎศࡋࡓࠋࡇࡢⅬ *2 㸸ᅜ❧ឤᰁ◊✲ᡤ ࡛ A ⩌ࡣ 1.0E5㸪B ⩌ࡣ 1.1E5 ࡛࠶ࡗࡓࠋ୍᪉㸪 ᙅẘᆺᰴࡢ᥋✀⣔࡛ࡢ Ot ᳨ฟ㛤ጞࡣ㸪A ⩌ࡀ᥋ ✀ᚋ 2 ᪥┠㸪B ⩌ࡀ 5 ᪥┠࡛࠶ࡗࡓࠋࡑࡢᚋ㸪 ᝈ ⪅ ࠾ ࡼ ࡧ ࣐ ࢘ ࢫ ࡢ ⾑ ୰ Orientia tsutsugamushi ࡢᐃ㔞㔜㛵ࡍࡿ⪃ᐹ ࢥࣆ࣮ᩘࡣ㢧ⴭ࡞ቑຍࡀ࡞ࡃ㸪A ⩌ࡣ 2 ᪥┠㸪 B ⩌ࡣ 10 ᪥┠㸦7.1E3㸧ࡀ᭱㧗್࡛ 11 ᪥┠௨ 㝆ࡣ᳨ฟ࡞ࡾ㸪࣐࢘ࢫࡣ 1 ࣨ᭶㛫⏕Ꮡࡋࡓࠋ *1 బ⸨ᐶᏊ㸪ᕝ᳃ᩥᙪ 㸪⸨⏣༤ᕫ *3 Ᏻ⸨Ꮚ 㸪㛛㤿┤ኴ 2* C ⩌ D ⩌ࡶྠᵝᙉẘ࣭ᙅẘᰴࢆྛࠎ᥋✀ࡋ *4 ࡓࡇࢁ㸪୧⩌ඹṚஸࡋࡓࡀ㸪Ṛஸࡢ Ot ࢥ ࣆ࣮ᩘࡣ㸪A ⩌ B ⩌ẚ㍑ࡋ࡚ᙉẘᆺᰴࡀ⣙ 10 ➨ 24 ᅇ⛅⏣ᛂ⏝⏕⛉Ꮫ◊✲ ಸ㸪ᙅẘᆺᰴࡀ 100㹼1000 ಸ࡛࠶ࡗࡓࠋᙉẘᆺ 2014 ᖺ 12 ᭶㸪⛅⏣ᕷ ᰴࡣ㸪ᙅẘᆺᰴࡼࡾࡶ࣐࢘ࢫయෆ࡛ᛴ⃭ቑṪ ࡍࡿࡓࡵ㸪චᛂ⟅ࡀ㛫ྜࢃࡎ㸪࣐࢘ࢫࡣṚ ࡘ ࡘ ࡀ ࡢ ཎ య Orientia tsutsugamushi ⮳ࡿࡶࡢ᥎ᐹࡋࡓࠋᝈ⪅⾑ᾮ୰ࡢ Ot ࡢࢥ 㸦Ot㸧ࡣ⾑Ύᆺࡼࡾ࣐࢘ࢫᑐࡍࡿẘᛶࡀ␗ ࣆ࣮ᩘࡣ㸪DIC 㠀ేⓎ/⒪๓㸦ᖹᆒ᪥ 4.8 ࡞ࡾ㸪ࣄࢺࡶࡑࢀ‽ࡎࡿࡉࢀࡿࡀ㸪ᮏ┴ ᪥㸧 DIC ేⓎ/⒪ᚋ㸦ྠ 5.5 ᪥㸧ࡢᖹᆒ್ ࠾࠸࡚Ⓨ⏕ࡍࡿࡘࡘࡀᝈ⪅ࡢ⮫ᗋ≧ࡣᚲ ࡣඹ 1.1E4 ࡛࠶ࡗࡓࠋ୍᪉㸪DIC ేⓎ/⒪๓ ࡎࡋࡶࡇࢀ୍⮴ࡋ࡞࠸ࠋࡑࡇ࡛㸪ᝈ⪅⾑ᾮ࠶ 㸦ྠ 4.0 ᪥㸧ࡣ㸪ᖹᆒ 2.3E5 DIC ేⓎ/⒪ ࡿ࠸ࡣ㔝⏕ື≀ࡽศ㞳ࡉࢀࡓ Ot ᰴࡢ࠺ࡕ㸪ẘ ᚋ࠾ࡼࡧ㠀ేⓎ/⒪๓ࡢ⣙ 20 ಸ࡛࠶ࡗࡓࠋ ᛶࡀ␗࡞ࡿࡉࢀࡿ⾑Ύᆺࡢ 2 ᰴࡘ࠸࡚㸪࣐ ⾑Ύᢠయ౯ࡽ 26 ࡣ࡚ Karp ᆺឤᰁ ࢘ࢫᑐࡍࡿ⮴Ṛẘᛶࡢ☜ㄆඹ⾑୰ Ot ࢥ ุ᩿ࡉࢀࡓࡀ㸪ࡇࡢ࠺ࡕ DIC ేⓎ/⒪๓ࡢ 5/8 ࣆ࣮ᩘࢆ⤒ⓗᐃ㔞ࡋࡓࠋేࡏ࡚⛅⏣┴࡛ 㸪DIC 㠀ేⓎࡢ 3/14 ࡀ࣐࢘ࢫᑐࡋ࡚ᙅẘ 2011 ᖺ㹼2013 ᖺⓎ⏕ࡋࡓࡘࡘࡀᝈ⪅ 26 ᛶࡢ JP-1 ᆺឤᰁ࡛࠶ࡾ㸪ྠᆺ࡛ࡶ⮫ᗋ⤒㐣⾑ 㸦DIC ేⓎ/⒪๓ 8 DIC ేⓎ/⒪ᚋ 4 ୰ Ot ᐃ㔞್ᕪࡀㄆࡵࡽࢀࡓࠋ26 ࡢᇶ♏ ࡞ࡽࡧ DIC 㠀ేⓎ/⒪๓ 14 㸧ࡘ࠸࡚ ᝈࡸ⅖࣐࣮࣮࢝ࡢᐃ㔞್➼ࡣᮍ᳨ウ࡛࠶ࡿ ࡶᐃ㔞ࡋ㸪⾑୰ࢥࣆ࣮ᩘࡢẚ㍑ࢆ⾜ࡗࡓࡢ࡛ሗ ࡀ㸪DIC ేⓎ㸪㠀ేⓎࡢ᭷ຠ⒪ࡲ࡛ࡢ᪥ᩘ ࿌ࡍࡿࠋ ᕪࡣࢃࡎ 1 ᪥࡛࠶ࡗࡓࠋᅇࡢ᳨ウ࡛㸪㔜 ࣐࢘ࢫ᥋✀ࡣ Kato ᆺ Kakuma-2 ᰴ㸦ᙉẘᆺ 㜵Ṇࡣ㸪᪩ᮇࡢ᭷ຠ⒪ࡼࡾయෆ࡛ࡢ Ot 㸸㔝㰡⏤᮶㸧࠾ࡼࡧ Shimokoshi ᆺ Matsui ᰴ㸦ᙅ ቑṪࢆ᪩ᛴᢚไࡍࡿࡇࡢᚲせᛶࡀ⾑୰ࢥ ẘᆺ㸸ᝈ⪅⏤᮶㸧ࢆ⏝ࡋࡓࠋ୧ᰴࡣ㸪L929 ⣽ ࣆ࣮ᩘࡢᐃ㔞ࡼࡿᩘ್ࡽㄞࡳྲྀࢀࡓࠋ ⬊࡛ᇵ㣴ᚋ DNA ᢳฟᚋ㸪ࢥࣆ࣮ᩘࡢᐃ㔞ࡼ 1* 㸸㟼ᒸ┴⎔ቃ⾨⏕⛉Ꮫ◊✲ᡤ ࡾ ࣜ ࢣ ࢵ ࢳ ⁐ ᾮ ࢆ 1.0E7 ࠾ ࡼ ࡧ 1.0E5 2* 㸸㤿ཎ࢝ࣜ་Ꮫ◊✲ᡤ copies/ml ㄪ〇ࡋ㸪ྛᰴྛ⃰ᗘ 100 μl ࢆ ICR 3* 㸸㮵ඣᓥᏛඹྠ⋇་Ꮫ㒊 ࣐࢘ࢫ 2 㢌㸦1.0E7㸸A ⩌㸪1.0E5㸸B ⩌㸧㸪ICR 4* 㸸⚟ᓥ┴⾨⏕◊✲ᡤ Nude ࣐࢘ࢫ 2 㢌㸦1.0E7㸸C ⩌㸪1.0E5㸸D ⩌㸧 ࡢ⭡⭍᥋✀ࡋࡓࠋࡑࡢᚋ㸪࣐࢘ࢫࡢእぢ࣭⾜ ືほᐹᑿ㟼⬦ࡽࡢ᥇⾑ࢆ 1 ᪥ 1 ᅇ 8 ᪥㛫⾜ ࠸㸪9 ᪥┠௨㝆ࡣ 1㹼2 ᪥ẖ⾜ࡗࡓࠋ࣐࢘ࢫ⾑ ⛅⏣┴࠾ࡅ ࡿ Rickettsia helvetica ಖ᭷ ࣐ࢲࢽ่ညึ☜ㄆឤᰁ᥎ᐃᆅࡢㄪᰝ ᾮࡣ㸪Ot ᰴࡢሙྜྠ ᵝ DNA ᢳฟᚋ㸪Ot ࢥࣆ࣮ᩘࢆᐃ㔞ࡋ㸪ྛ⩌ࡢ 1 ml ᙜࡓࡾ⟬ࡋ బ⸨ᐶᏊ㸪ᮧ༤ᐉ *1 㸪ᰘ⏣ࡕࡦࢁ ࡓᖹᆒ್ࢆồࡵࡓࠋᝈ⪅⾑ᾮ୰ࡢ Ot ᐃ㔞ࡶྠ ⛅㔝⳹Ꮚ㸪㰺⸨ᚿಖᏊ㸪⸨⏣༤ᕫ *2 - 77 - 秋田県健康環境センター年報 第10号 2014 Ᏻ⸨Ꮚ *3 ඵ㑻†࠾ࡼࡧ࿘㎶ᆅୗỈὶᇦ࠾ࡅࡿࣜࣥ ⃰ᗘࡢኚ㑄 ➨ 21 ᅇࣜࢣࢵࢳ◊✲ྜྠ◊✲Ⓨ⾲ 2014 ᖺ 12 ᭶㸪ᮾி㒔 ⋢⏣ᑗᩥ㸪బ⸨Ύ㝯㸪ῲᚿఙ ⛅⏣┴ෆ࠾࠸࡚㸪࣐ࢲࢽ่ညࡋ࡚ཷデ ࡋ ࡓ ᝈ ⪅ ࡽ ฟ ࡉ ࢀ ࡓ ࣐ ࢲ ࢽ ࡽ Rickettsia ➨ 62 ᅇ᪥ᮏ⏕ែᏛ helveticaࡀ᳨ฟࡉࢀࡓࡓࡵ㸪ࡑࡢᚋࡢㄪᰝే 2015 ᖺ 3 ᭶㸪㮵ඣᓥᕷ ࡏ࡚ሗ࿌ࡍࡿࠋ ඵ㑻†ࡣ 1977㸦S52㸧ᖺࡢᅜႠඵ㑻₲ᖸᣅ ᝈ⪅ࡣ69ᡯዪᛶࠋ9᭶28᪥㸪⛅⏣㥖ࣨᓅࢆⓏᒣࠋ ⩣᪥㸪ᚋ㢌㒊ࡢ╔≀Ẽࡃࠋ9᭶30᪥㸪㏆་ ᴗᚋ㸪ᐩᰤ㣴క࠺Ỉ㉁ở⃮ࡀ㐍⾜ࡋ㸪 ࢆཷデ㸪ྠ᪥Ἠ⓶⛉ࢡࣜࢽࢵࢡࢆ⤂ཷデ㸪 2007㸦H19㸧ᖺࡣ†Ỉ㉁ಖ≉ูᥐ⨨ἲ ᚋ㢌㒊ࡽ࣐ࢲࢽ1༉ࡀฟࡉࢀ㸪࣑ࣀ࣐ࢩࣥ ᇶ࡙ࡃỈ㉁⎔ቃᇶ‽್ࡢ☜ಖࡀ⥭せ࡞ᣦᐃ† 㸦200 mg/day㸧ࢆ10᪥㛫᭹⏝㸪⓶⅖ࡸⓎ⇕ ᣦᐃࡉࢀࡓࠋ⛅⏣┴࡛ࡣ㸪ඵ㑻†ࡢỈ㉁ᨵၿ ࡞ࡢ≧ࡀ࡞࠸ࡲࡲ⤒㐣ࠋ ࡢࡓࡵඵ㑻†⎔ቃᑐ⟇ᐊࢆタ⨨ࡋ㸪ࠕඵ㑻† ฟࡋࡓ࣐ࢲࢽ✀ࡣࣄࢺࢶࢺࢤ࣐ࢲࢽࡢ㞤ᡂ ࡛࠶ࡗࡓࠋࡇࡢయࡽDNAࢆᢳฟࡋ17 kDa ಀࡿ†Ỉ㉁ಖィ⏬㸦➨ 1 ᮇ㸧ࠖࢆ⟇ᐃࡋ㸪 Ỉ㉁ಖ⟇ࢆ⥲ྜⓗㅮࡌ࡚ࡁࡓࠋࡋࡋ㸪 ࠾ࡼࡧgltA㡿ᇦ࠾ࡅࡿPCR࠾ࡼࡧࢲࣞࢡࢺ ᐩᰤ㣴ࡢཎᅉ≀㉁ࡢ㸯ࡘ࡛࠶ࡿࣜࣥ㸦T-P㸧ࡣ㸪 ࢩ࣮ࢣࣥࢫࡼࡾ⣚ᩬ⇕⩌ࣜࢣࢵࢳ㑇ఏᏊࡢ ᖺ㛫ࢆ㏻ࡋ࡚ࡰỈ㉁⎔ቃᇶ‽್ࢆ㉸㐣ࡋ࡚࠾ ᳨ฟศᏊ⣔⤫ᶞゎᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝ㸪᳨ ࡾ㸪2012㸦H24㸧ᖺࡣ࢜ࢥࡀⓎ⏕ࡋ 㸪ඵ ฟ ࡉ ࢀ ࡓ 㑇 ఏ Ꮚ ࡀ R.helvetica ┦ ྠ ᛶ ࡀ 㧗 ࠸ ࡇ 㑻†ࡢỈ㉁ࡣ㐣ཤ᭱ᝏࡢᩘ್࡞ࡗࡓࠋ ࡀ♧ࡉࢀࡓࠋࡲࡓ㸪ᝈ⪅⾑ᾮࢆ࣐ࢲࢽ่ည☜ ୍᪉㸪⛅⏣┴ࡢ᪤ ㄪᰝࡼࡾ㸪ඵ㑻†ᖸᣅ ㄆࡽ10᪥┠㸦10/9㸧21᪥┠㸦10/20㸧᥇ྲྀ ᆅ༡㒊࠾࠸࡚㸪T-P ⃰ᗘࡀᴟࡵ࡚㧗࠸Ỉ㊰ ࡋ㸪⣚ᩬ⇕⩌㸪SFTS࠾ࡼࡧ㔝ඡࡢᢠయ౯ ᐃ ࡀⓎぢࡉࢀࡓࠋࡇࡢỈ㊰ࡽඵ㑻†ὶධࡍ ࢆ⾜ࡗࡓࡀᢠయ౯ୖ᪼ࡣ☜ㄆࡉࢀ࡞ࡗࡓࠋ ࡿ T-P ᥎ᐃᖺ㛫㈇Ⲵ㔞ࡣ㸪㈇Ⲵ㔞ࡢ 26.5㸣 10/26 ឤ ᰁ ᥎ ᐃ ᆅ ࡉ ࢀ ࡿ ⛅ ⏣ 㥖 ࣨ ᓅ ࡢ ୰ ヨ⟬ࡉࢀ࡚࠸ࡿࠋ ࡇࡢཎᅉࡋ࡚㸪ྠᖸᣅᆅ༡ ⏕ಖෆⓏࡾཱྀ㏆࡛ࣇࣛࣥࢿࣝᕸࡼࡿࡎࡾ 㒊ࡢ᪉ୖᆅ༊࿘㎶࠾ࡅࡿ㧗⃰ᗘ T-P ฟỈࡢ ἲ࡛⾜ࡗࡓࠋ࣐ࣖࢺ࣐ࢲࢽ♂㸪࢟ࢳ࣐ࢲࢽ♀ࡢᡂ Ꮡᅾࡀᣦࡉࢀ࡚࠾ࡾ㸪ඵ㑻†ࡢỈ㉁ở⃮ࡢ୍ ྛ1ಶయࡀᤕ㞟ࡉࢀࡓࡀ㸪᪉ࡽ⣚ᩬ⇕⩌ࣜ せᅉࡋ࡚┳㐣࡛ࡁ࡞࠸ࡇࡽ㸪ࡇࢀࡲ࡛⛅ ࢣࢵࢳࡣ᳨ฟࡉࢀ࡞ࡗࡓࠋᮏࡣ࣐ࢲࢽ ⏣┴ࡣ㸪㧗⃰ᗘ T-P ฟỈ࣭ᆅୗỈ࣭ᆅ㉁➼ ่ညᚋࡢ᪩ᮇฟᢠ⏕ᢞ㛤ጞࡀዌຌࡋ㸪 㛵ࡍࡿྛ✀ㄪᰝࢆᐇࡋ࡚ࡁࡓࠋࡑࡢ⤖ᯝ㸪㧗 Ⓨࢆᮍ↛㜵ࡄࡇࡀ࡛ࡁࡓࡶࡢ⪃࠼ࡽࢀ ⃰ᗘ T-P ฟỈࡢ㉳※ࡋ࡚㸪ᆅ୰ሁ✚≀୰ ࡿࠋ⛅⏣┴ࡣ㸪ࡘࡘࡀࡢၨⓎ᳨ᰝయไࢆ ྵ᭷ࡉࢀࡓࣜࣥࡢ⁐⬺㸪ṇ㠃ሐ㜵ୗ㒊ᆅୗỈࡢ ᩚ࠼་⒪ᶵ㛵┴Ẹࡢព㆑ࢆ㧗ࡵࡿດຊࢆ⥆ࡅ ᙳ㡪࠾ࡼࡧᖸᣅᆅෆࡢỈ⏣➼ᅵተ⾲ᒙࡽࡢὶ ࡚ࡁࡓࡀ㸪ᅇࡢࡼࡾ⣚ᩬ⇕⩌ࣜࢣࢵࢳ ධྍ⬟ᛶ➼ࡀᣦࡉࢀ࡚࠸ࡿࠋ ࡘ࠸࡚ࡶ᪩ᛴయไࢆ☜❧ࡋ㸪ၨⓎࢆ᥎ ࡋࡋ㸪ᐃ㔞ⓗ࡞ฟᶵᵓ㛵ࡋ࡚᫂࡞Ⅼ 㐍ࡍࡿඹ┴ෆࡢ࣐ࢲࢽ⏕ᜥ≧ἣࢆᢕᥱࡍࡿ ࡀከࡃ㸪1993㸦H5㸧ᖺ௨㝆㸪㠃ⓗㄪᰝࡀᐇࡉ ᚲせᛶࢆㄆ㆑ࡉࡏࡽࢀࡓࠋࡲࡓ㸪⌧ᅾ㸪ឤᰁ ࢀ࡚࠾ࡽࡎ㸪⌧≧ࡀ࡛᫂࠶ࡿࠋ ἲ ࠾ ࠸ ࡚ R.japonica௨ እ ࡢ ⣚ ᩬ ⇕ ⩌ ࣜ ࢣ ࢵ ࢳ ࡑࡇ࡛ᮏ◊✲࡛ࡣ㸪ྠᖸᣅᆅ༡㒊࠾ࡅࡿ㎰ ឤᰁࡣᒆฟᑐ㇟࡞ࡗ࡚࠸࡞࠸ࡀ㸪㏆ᖺࡢ ᴗ⏝Ỉ㊰⾲ᒙỈ࠾ࡼࡧほ ᡞỈ୰㧗⃰ᗘ ᪥ᮏ࠾ࡅࡿᝈ⪅Ⓨ⏕ࢆ㋃ࡲ࠼㸪ࡑࡢᐇែᢕ T-P ࡢ⌧≧ࢆᢕᥱࡍࡿࡓࡵ㸪ࣜࣥ⃰ᗘㄪᰝࢆ ᥱཎᛶࡢゎ᫂ࡀᮃࡲࢀࡿࠋ ᐇࡋࡓࠋ *1 㸸Ἠ⓶⛉ࢡࣜࢽࢵࢡ *2 㸸㤿ཎ࢝ࣜ་Ꮫ◊✲ᡤ ⃰ᗘࡣ㸪0.063㹼2.5 mg/L, ᖹᆒ 0.72 mg/L ࡛࠶ *3 㸸㮵ඣᓥᏛඹྠ⋇་Ꮫ㒊 ࡾ㸪᪤ሗࡼࡾࡸࡸప࠸್࡛࠶ࡗࡓࡀ㸪࿘㎶㎰ᆅ ࡑࡢ⤖ᯝ㸪㎰ᴗ⏝Ỉ㊰⾲ᒙỈ࠾ࡅࡿࣜࣥ ࡽࡢỈࡼࡿᙳ㡪➼ࡢ㸪Ꮨ⠇ኚືࡶண - 78 - 秋田県健康環境センター年報 第10号 2014 ࡉࢀࡿࡓࡵ㸪ࡑࢀࡽࡢせᅉࢆ⪃៖ࡋࡓ⥅ ⥆ⓗ ࡞ㄪᰝࡀᚲせ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋࡲࡓほ ᡞỈ୰ T-P ⃰ᗘࡣ㸪3.7㹼26.9 mg/L ࢆ♧ࡋࡓࠋ ࡇࡢㄪᰝᡞ࡛ࡣ㸪ᖺ㛫ࢆ㏻ࡌ࡚⥅⥆ⓗ࡞ T-P ⃰ᗘࡢࣔࢽࢱࣜࣥࢢࡀ࡞ࡉࢀ࡚࠾ࡾ㸪⌧ᅾ࡛ࡶ 30 mg/L ๓ ᚋ ࡢ ⃰ ᗘ ࡀ ᳨ ฟ ࡉ ࢀ ࡚ ࠸ ࡿ ࡇ ࡽ㸪ࡰྠࣞ࣋ࣝࡢ⃰ᗘ࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋ ࡉࡽ㸪ࡑࡢほ ᡞ 6 ⟠ᡤ࠾࠸࡚ࡶ㸪T-P ᑐࡍࡿ PO4 3- ⃰ᗘྜࡀ⣙ 70%௨ୖ࡛࠶ࡾ㸪 ᪤ ྠᵝࡢ⤖ᯝࡀᚓࡽࢀ㸪⌧ᅾ࡛ࡶ㧗⃰ᗘ T-P ᆅୗỈࡢฟࡼࡿᙳ㡪ࡀ㎰ᴗ⏝Ỉ㊰⾲ ᒙỈཬࢇ࡛࠸ࡿࡇࡀ♧၀ࡉࢀࡓࠋ - 79 - 秋田県健康環境センター年報 第10号 2014 㸬ㄅᥖ㍕ Shioko Saito , Yoshito Iwade*1 , Eisuke Tokuoka *2 , Tomohiro Nishio *3 , ᮾ᪥ᮏ㟈⅏ࡼࡿ⅏ᐖᗫᲠ≀ཷධࢀక ࠺⛅⏣┴ෆࡢ⎔ቃᨺᑕ⬟ㄪᰝ⤖ᯝࡘ࠸࡚ Yoshimitsu Otomo *4 , Emiko Araki *5 , Hirotaka Konuma *5 , Hiroshi Nakagawa *6 , ⏣ᮧ㧗ᚿ *1 㸪⋢⏣ᑗᩥ㸪Ⳣཎ 㧗ᶫⱥஅ *2 㸪㧗ᔱ ྖ㸪㧗ᶫ Hiroyuki Tanaka *7 , Kanji Sugiyama *3 , ๛ Akio Hasegawa *8 , ᾈ *2 *9 Yoshiko Sugita-Konishi *8,9 and Yukiko Hara-Kudo *8,9 ᅜ⎔ቃ◊ㄅ㸪39㸪2㸪2014㸪10-15㸬 FOODBORNE PATHOGENS AND DISEASE㸪 12㸪2㸪2015㸪131-138㸬 ᮾ᪥ᮏ㟈⅏ࡢ⯆ᨭࡋ࡚㸪⛅⏣ᕷ➼ࡢ ┴ෆ 6 ࢝ᡤࡢᗫᲠ≀ฎ⌮タ࠾࠸࡚ᐇࡋࡓ ᒾᡭ┴ᐑྂᕷ࠾ࡼࡧ㔝⏣ᮧࡢ⅏ᐖᗫᲠ≀ᗈᇦฎ Vibrio parahaemolyticus carrying the tdh gene, ⌮࠾࠸࡚㸪ᗈᇦฎ⌮ᐇ๓ᚋ࠾ࡅࡿ㛵㐃⎔ encoding the thermostable direct hemolysin (TDH), ቃヨᩱࡢᨺᑕ⬟➼ࡢ ᐃࢆᐇࡋࡓࠋ or the trh gene, encoding the TDH-related ࡑࡢ⤖ᯝ㸪ᆅୗỈ࣭ᨺὶỈ㸪ἙᕝỈ㸪ởἾ㸪 hemolysin (TRH), are both considered virulent ᗏ㉁ヨᩱࡽ㸪ᨺᑕᛶࢭࢩ࣒࢘ࡣ๓࣭ᚋㄪ strains. There are, however,disproportionally fewer ᰝ࠸ࡎࢀࡽࡶࢇ᳨ฟࡉࢀ࡞ࡗࡓࠋᅵ reports ተヨᩱࡽࡣᨺᑕᛶࢭࢩ࣒࢘ࡀ᳨ฟࡉࢀࡓࡀ㸪 contaminated with trh-positive strains than by ࢭࢩ࣒࢘ 137 ࡀ᳨ฟࡉࢀࡓྜࡣ๓ㄪᰝ࡛ࡣ seafood contaminated with tdh-positive strains. 67 ᳨య୰ 9 ᳨య㸦13㸣㸧㸪ᚋㄪᰝ࡛ࡣ 135 ᳨ Bivalves such as clams and oysters are the major య୰ 36 ᳨య㸦27㸣㸧࡛࠶ࡗࡓࠋࡇࡢ࠺ࡕࢭࢩ࢘ seafood varieties associated with the infections. In ࣒ 134 ࡶ᳨ฟࡉࢀࡓࡢࡣᚋㄪᰝ࠾ࡅࡿ 2 ᳨ this study, the prevalence of strains possessing the యࡢࡳ࡛࠶ࡗࡓࠋ tdh and trh genes was investigated in Japan in 74 of infections caused by seafood samples collected in 2007–2008 and in 177 ࢭࢩ࣒࢘ 137 ࡢࡳࡀ᳨ฟࡉࢀࡓ᳨యࡘ࠸࡚ ࡣࢳ࢙ࣝࣀࣈࣜཎⓎᨾࡢᙳ㡪ࡔᛮࢃࢀ samples ࡿࠋࢭࢩ࣒࢘ 134㸪ࢭࢩ࣒࢘ 137 ࡢ୧᪉ࡀ᳨ฟ bivalves,bloody clams, ࡉࢀࡓ 2 ᳨యࡘ࠸࡚ࡣ⚟ᓥ➨୍ཎⓎᨾࡢᙳ clams, and rock oysters. The tdh-positive and 㡪ࡶᫎࡉࢀࡓᛮࢃࢀࡿࡀ㸪ࢭࢩ࣒࢘ 134 ࡢ trh-negative, tdh-negative and trh-positive, and ᳨ฟ⃰ᗘࡣ 6.2 Bq/kg㸪6.4 Bq/kg㸪ࢭࢩ࣒࢘ 137 tdh-positive and trh-positive samples represented ࡢ᳨ฟ⃰ᗘࡣ 12 Bq/kg㸪6.8 Bq/kg ప࠸ᩘ್࡛ 5.4%, 12.2%, and 4.1% of all samples collected in ࠶ࡗࡓࠋ 2007–2008, and 5.1%, 18.6%, and 5.6% of all samples ࡇࢀࡽࡢࡇࡽ㸪⅏ᐖᗫᲠ≀ཷධࡼࡿᨺ collected collected in in 2010 hen 2010, of domestic clams, short-neck respectively. As ᑕ⬟ࡢᙳ㡪ࡣᑡ࡞ࡃ㸪ᗈᇦฎ⌮ࢆ⾜ࡗࡓタࡢ determined by polymerase chain reaction, the እ㒊⎔ቃࡢᨺᑕᛶࢭࢩ࣒࢘ࡢ㣕ᩓࢆᠱᛕࡍࡿ prevalence of tdh negative and trh positive in all ᚲせࡣ࡞࠸᥎ᐹࡉࢀࡓࠋ samples was two to four times higher than that of *1 㸸⏤ᆅᇦ⯆ᒁ⚟♴⎔ቃ㒊 tdh positive and trh negative. In the samples *2 㸸ඖᗣ⎔ቃࢭࣥࢱ࣮ collected in 2010, the tdh-negative and trh-positive V. parahaemolyticus (20 samples) was more often isolated than tdh-positive V.parahaemolyticus (SLGHPLRORJLFDO(YLGHQFHRI/HVVHU5ROH RI 7KHUPRVWDEOH 'LUHFW +HPRO\VLQ 7'+̽ 5HODWHG+HPRO\VLQ75+ 7KDQ7'+ RQ 9LEULRSDUDKDHPRO\WLFXV3DWKRJHQLFLW\ (7 and trh-negative samples). The most common serotype of tdh-positive isolates (22 of 24 strains) was pandemic O3:K6. The trh-positive isolates (61 strains) were various serotypes including OUT:KUT. In 330 V. parahaemolyticus - 80 - 秋田県健康環境センター年報 第10号 2014 outbreaks and sporadic infections in Japan, most ࡘࡘࡀࡣ㸪୕ᚩࡤࢀࡿࠕⓎ⇕ࠖ outbreaks and sporadic infections were caused by యᖿ㒊୰ᚰࡢࠕⓎ⑈ࠖ㸪ࠕ่ࡋཱྀࠖࡤࢀ tdh-positive and trh-negative strains (89.4%). The ࡿࢶࢶ࣒࢞ࢩࡢ྾╔㊧࡛࠶ࡿ⓶ኚࡀᣲࡆࡽ frequencies of infections caused by tdh-negative ࢀࡿࡀ㸪ࡇࡢ୍⯡ⓗ࡞⏕Ꮫⓗ⾑ᾮ᳨ᰝ࡛ࡣ㸪 and trh-positive, and both tdh- and trh-positive ᝈࡢ㚷ู☜ᐃデ᩿┤⤖ࡍࡿሗࢆᚓ strains were 1.2% and 3.0%, respectively. This ࡿࡇࡣᅔ㞴࡛࠶ࡿࠋ☜ᐃデ᩿ࡣ㸪㑇ఏᏊデ finding suggests that the virulence of trh might be ᩿ἲ࠶ࡿ࠸ࡣ⾑ΎᏛⓗデ᩿ἲࡀⓗ☜࡛࠶ࡿࡀ㸪 less trhpositive ࠸ࡎࢀࡶࡘࡘࡀࢆᐃࡍࡿࡇࡀ๓ᥦ࡞ contaminated ࡿࡓࡵ㸪ᝈ⪅ࡢヲ⣽࡞ၥデࡀ᪩ᮇデ᩿⒪ than that of V.parahaemolyticus tdh, although frequently bivalves. *1 :Mie ࡢ㘽࡞ࡿࠋࡋࡋ㸪ࡇࡢ☜ᐃ᳨ᰝデ᩿ࡣᑐᛂ Prefecture Health and Environment ࡛ࡁࡿ᳨ᰝ◊✲ᶵ㛵ࡀከࡃ࡞࠸ࡇຍ࠼㸪ᑐ Research Institute, Yokkaichi, Japan. *2 :Kumamoto Prefectural Institute of ᛂ࡛ࡁࡿᢠཎᆺࡀ㝈ᐃࡉࢀ࡚࠸ࡿᶵ㛵ࡶ࠶ࡿࡇ Public ࡽ㸪⒪ࡣࡉࢀ࡚ࡶᒆฟ⤖ࡧࡘ࡞࠸ Health and Environmental Science, Udo, Japan. ࡀᑡ࡞ࡃ࡞࠸ᛮࢃࢀࡿࠋࡑࡢࡓࡵ㸪ᒆฟᩘ *3 :Shizuoka *4 :Hirosaki University, Hirosaki, Japan. ᳨ᰝయไࡢᅜⓗ࡞ᩚഛࢧ࣮࣋ࣛࣥࢫࡢᙉ *5 :Department of Fisheries, School of Marine 㸪ᆅᇦࡢᐃᮇⓗ࡞ሗⓎಙࡢ⥅⥆ࡀᮃࡲࢀ Institute of Environment and ┿ࡢᝈ⪅ᩘࡀᫎࡉࢀ࡚࠸࡞࠸ࡶ⪃࠼ࡽࢀࡿ Hygiene, Shizuoka, Japan. ࡇࡽ㸪་⒪ᶵ㛵බ⾗⾨⏕⾜ᨻࡢ㐃ᦠࡢඖ㸪 Science and Technology, Tokai University, ࡿࠋ Shizuoka, Japan. *6 :BML Food Science Solutions, Tokyo, Japan. *7 :Japan Food Research Laboratories, Tokyo, Japan. *8 :The University of Tokyo, Tokyo, Japan. *9 :Division of Microbiology, National Institute ⛅⏣┴࠾ࡅࡿ6KLPRNRVKLᆺࡘࡘࡀࡢ 嚳ཬⓗᏛㄪᰝ బ⸨ᐶᏊ㸪ᰘ⏣ࡕࡦࢁ㸪ᩪ⸨༤அ㸪㡲⸨ᜏஂ *1 of Health Sciences, Tokyo, Japan. ⾨⏕ື≀, 65, 4, 2014, 183-188. ࠕࡘࡘࡀࠖ ࡘࡘࡀࣜࢣࢵࢳ Orientia tsutsugamushi ࡢ Shimokoshi ᆺࡼࡿឤᰁࡣ⛥ࡉࢀ㸪ࡑࡢ బ⸨ᐶᏊ ⮫ᗋീࡸᏛⓗ࡞≉ᚩࡘ࠸࡚ࡣ᫂࡞Ⅼࡀከ ࠸ ࠋ ࢃ ࢀ ࢃ ࢀ ࡣ 㸪 ⛅ ⏣ ┴ ࠾ ࠸ ࡚ Shimokoshi ྾, 33, 12, 2014, 1249-1255. ᆺࢆᢠయ᳨ᰝᑐ㇟ྲྀࡾධࢀࡓ 2009 ᖺ௨㝆ࡢ 4 tsutsugamushi ᖺ㛫 5 ࡢ Shimokoshi ᆺࡘࡘࡀᝈ⪅ࡀ 㸦O.tsutsugamushi㸧ࢆಖ᭷ࡍࡿࢶࢶ࣒࢞ࢩࡢᗂ ☜ㄆࡉࢀࡓࠋࡇࢀࢆዎᶵ㸪㐣ཤࡢ࠸ࢆྵ ྾╔ࡉࢀࡿࡇࡼࡾ㸪ឤᰁ࣭Ⓨࡍࡿ⇕ ࡵࡓࢆ᳨ウࡋࡓࡇࢁ㸪⛅⏣┴࠾࠸࡚ ᛶⓎ⑈࡛㸪ឤᰁἲᇶ࡙ࡃឤᰁⓎ⏕ືྥ 1992 ᖺ㹼2012 ᖺ 15 ࡢ Shimokoshi ᆺࡘࡘ ㄪᰝ࡛ࡣ㸪4㢮ឤᰁศ㢮ࡉࢀࡿᩘᒆฟᝈ ࡀࡀⓎ⏕ࡋ࡚࠸ࡓࡇࡀ☜ㄆࡉࢀࡓࠋࡇࢀ ࡛࠶ࡿࠋᝈ⪅Ⓨ⏕ࡣすࢪ㹼ᴟᮾࣟࢩ㹼ኴ ࡽࡣ㸪㍍ࡽ㔜ࢆྵࡴከᙬ࡞⮫ᗋീࢆ ᖹὒᆅᇦࡅࡓᗈᇦΏࡿࠋ᪥ᮏᅜෆ࠾࠸ ♧ࡋ࡚࠸ࡓࠋⓎ⏕ᮇࡣ㸦4 ᭶ࡽ 6 ᭶㸧 ࡚ࡣ㸪ᖺ㛫400๓ᚋࡢᝈ⪅Ⓨ⏕ሗ࿌ࡀ࠶ࡿࡀ㸪 ⛅㸦10 ᭶ 11 ᭶㸧࡛㸪ᆅᇦࡣ┴ࡢ㒊ࡽ༡ Ⓨ⏕ᮇࡣ࣋ࢡࢱ࣮ࡢ✀㢮ࡸࡑࡢᗂάືᮇ 㒊ࡲ࡛࠾ࡼࢇ࡛࠸ࡓࠋࡲࡓ㸪Shimokoshi ᆺ᳨ ࡘ ࡘ ࡀ ࡣ Orientia ࡼࡾᆅᇦ≉ᛶࡀ࠶ࡿࠋࡘࡘࡀࡣ㸪ᛴ㏿㔜 ฟࢆ┠ⓗࡋࡓ PCR primer ࢆタィࡋ㸪3 ᳨య ࡋ⬻⅖㸪DICేⓎࡼࡾṚ⮳ࡿሙྜࡀ࠶ ࡽࡇࡢᆺࡢ DNA ࢆ᳨ฟࡋࡓࠋࡉࡽ㸪1 ࡢᝈ ࡿࡓࡵ㸪᪩ᮇデ᩿㐺ṇ࡞⒪ࡀᚲ㡲࡛࠶ࡿࠋ ⪅ࡽᅜෆ࡛ࡣ 2 ᰴ┠࡞ࡿ Shimokoshi ᆺࣜࢣ - 81 - 秋田県健康環境センター年報 第10号 2014 ࢵࢳࢆศ㞳ࡋࡓࠋࡲࡓ㸪15 ࡢ࠺ࡕ㸪2 ࡢDNAࡸRNAࡀྵࡲࢀࡿࡀ㸪ࡑࢀࡽࡀ㏫㌿ ࡣᙜᢠయ㝜ᛶุᐃࡉࢀ㸪ࡘࡘࡀࢆྰᐃ ᛂ࠾ࡼࡧPCRᛂ➼ᙳ㡪ࡋ㸪ࡑࡢ⤖ᯝࡋ࡚ ࡉࢀ࡚࠸ࡓࠋࡇࢀࡣᙜᢠయ᳨ᰝ⏝ࡋ࡚࠸ ᳨ฟ⋡ࡸ㑇ఏᏊᐃ㔞್ᙳ㡪ࢆཬࡰࡍࡢ࡛㸪᰾ ࡓᶆ‽ 3 ᢠཎ㸦Gilliam㸪Karp㸪Kato㸧 Shimokoshi 㓟ᢳฟ௨㝆ࡢ᧯సࡘ࠸࡚ᮏἲ࡛ᚓࡓヨᩱ࡛ࡢ ᢠཎࡣᕪᛂᛶࡀ㠀ᖖప࠸ࡇࡀせᅉ⪃ ᭷⏝ᛶࢆ᳨ドࡋ࡚࠾ࡃᚲせࡀ࠶ࡿࠋ *1 ࠼ࡽࢀࡿࠋ⌧ᅾࡢᡤ㸪ከࡃࡢ᳨ᰝᶵ㛵࠾࠸࡚ 㸸ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Shimokoshi ᆺࡣ᪥ᖖ᳨ᰝྲྀࡾධࢀࡽࢀ࡚࠸࡞ ࠸ࠋᚋࡣ Shimokoshi ᆺឤᰁ࠾࠸࡚ࡶ㸪✚ ୍ᮏ㙐㧗ḟᵓ㐀ከᙧ㸦66&3㸧ゎᯒἲ ᴟⓗ࡞ᐇ㦂ᐊデ᩿యไࡢᩚഛࡀᮃࡲࢀ㸪ࡲࡓࣄ ࢺᑐࡍࡿཎᛶࡢホ౯࣋ࢡࢱ࣮ゎ᫂ྥ ࡅࡓㄪᰝࡀㄢ㢟࡞ࡿࠋ *1 ᩪ⸨༤அ 㸸⛅⏣Ꮫ་Ꮫ㒊ྡᩍᤵ 㣗ရ⾨⏕᳨ᰝᣦ㔪 2015㸦ᚤ⏕≀⦅㸧, 2015, 648-656㸬 㣗ရ࣭⮫ᗋᮦᩱ࣭ࡩࡁྲྀࡾࡢ๓ฎ⌮ἲ 㣗 ရ ᳨ య ࡸ ⮫ ᗋ ᳨ య ࡽ RT-PCR ࡼ ࡗ ࡚ ᳨ ฟࡉࢀࡓ࢘ࣝࢫ㑇ఏᏊࢆゎᯒࡋ㸪ཎᅉ✲᫂ ᩪ⸨༤அ㸪㔝⏣ ⾨ *1 ᚲせ࡞⛉Ꮫⓗሗࢆᚓࡿࡓࡵ㸪PCRቑᖜ⏘≀ ࡢሷᇶ㓄ิࢆỴᐃࡍࡿ᪉ἲࡀ⏝࠸ࡽࢀࡿࡇࡀ 㣗ရ⾨⏕᳨ᰝᣦ㔪 2015㸦ᚤ⏕≀⦅㸧, 2015, ከ࠸ࠋ୍᪉࡛㸪ゎᯒࡍࡁPCR⏘≀ࡢᩘࡀከ࠸ 607-617㸬 ሙྜࡣ㸪࡚ࡢ⿕᳨యࡢሷᇶ㓄ิࢆỴᐃࡍࡿ ࡢ㛫ࢆせࡍࡿࡓࡵ㸪⾜ᨻୖࡢᑐᛂࡢ ࣃࣥࢯࣝࣅ࣭ࣥࢺࣛࢵࣉἲࡣ୍⯡㣗ရࡽࡢ 㐃ᦠࡀ㞴ࡋ࠸ࡇࡀᐃࡉࢀࡿࠋࡇࡇ࡛ゎㄝࡍ ࢘ࣝࢫ᳨ฟἲ࡛࠶ࡾ㸪↝ࡁࡑࡤ➼ࡢἜᡂศࡀ ࡿ୍ᮏ㙐㧗ḟᵓ㐀ከᆺ㸦Single Strand Conformation ከ࠸㣗ရ㸪࣏ࢸࢺࢧࣛࢲ➼ࡢⅣỈ≀ࡀከ࠸㣗 Polymorphism : SSCP㸧ゎᯒἲࡣ㸪ሷᇶ㓄ิࡑࡢ ရ࡞ከ✀㸪ከᵝ࡞㣗ရ㐺ᛂࡍࡿࡇࡀ࡛ࡁ ࡶࡢࢆ▱ࡿࡇࡣ࡛ࡁ࡞࠸ࡀ㸪୍ሷᇶࡢ㐪࠸࡛ ࡿࠋᮏἲࡣ࢘ࣝࢫࢆᤕᤊࡍࡿࡓࡵᢠయࢆ 㟁ẼὋືࣂࣥࢻࡢ⨨ࡀ㗦ᩄኚືࡍࡿ࠸࠺ ⏝ࡍࡿࡀ㸪ከࡃࡢ᳨ᰝᐊ࡛ࡣ࢘ࣝࢫ≉␗ⓗᢠ ≉㛗ࡀ࠶ࡿࠋࡲࡓ㸪」ᩘ✀㢮ࡢሷᇶ㓄ิࡀΰᅾ యࡣධᡭ࡛ࡁ࡞࠸ࡇࡽ㸪࢘ࣝࢫ≉␗ⓗᢠ ࡋ࡚࠸࡚ࡶࡑࡢࡲࡲゎᯒ࡛ࡁࡿ࠸࠺Ⅼࡶ࠶ యࡢ᭰ࡾ㸪࣐࢞ࣥࢢࣟࣈࣜࣥ〇ࢆ⏝ࡍࡿ ࡿࠋࡇࢀࡽࡢᛶ㉁ࢆࡶከᩘࡢPCR⏘≀ࡢሷ ࡇ࡞ࡿࠋࡑࡢሙྜ㸪ᝈ⪅౽➼⏤᮶ࡢ࢘ࣝ ᇶ㓄ิࡢ␗ྠࢆྠ↷ྜࡍࡿࡇࡀ࡛ࡁࡿࡢ ࢫࢆ⏝࠸࡚㸪ᮏἲ࡛࢘ࣝࢫࡀᅇ࡛ࡁࡿࡇ ࡛㸪㞟ᅋឤᰁࡢయീࢆ᪩ᮇᢕᥱࡍࡿࡢ 㸦ᢠయ㝧ᛶ࡛࠶ࡿ㸧ࢆ☜ㄆࡋ࡚࠾ࡃᚲせࡀ࠶ࡿࠋ 㐺ࡋ࡚࠸ࡿࠋ ᮏἲ࡛ᚓࡽࢀࡓヨᩱࡣ㔞ࡢࣈࢻ࢘⌫⳦⏤᮶ - 82 - ⛅⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ᖺሗ ➨ 10 ྕ 2014 Ⓨ⾜᪥ ᖹ ᡂ 27 ᖺ 12 ᭶ Ⓨ⾜ᡤ ⛅⏣┴ᗣ⎔ቃࢭࣥࢱ࣮ ࠛ 010-0874 ⛅ ⏣ ᕷ ༓ ⛅ ஂ ಖ ⏣ ⏫ 6 ␒ 6 ྕ TEL: 018-832-5005 FAX: 018-832-5938