Title 狂犬病抗体検査の性能向上に関する研究( 本文(Fulltext) ) Author(s) 白石, 力也 Report No.(Doctoral Degree) 博士(獣医学) 乙第145号 Issue Date 2016-09-26 Type 博士論文 Version ETD URL http://hdl.handle.net/20.500.12099/55522 ※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。
≬≟ᢠయ᳨ᰝࡢᛶ⬟ྥୖ㛵ࡍࡿ◊✲
㸰㸮㸯㸴ᖺ
ᒱ㜧ᏛᏛ㝔㐃ྜ⋇་Ꮫ◊✲⛉
ⓑ ▼ ຊ ஓ
┠
┠ḟ
⥴ゝ
1➨㸯❶ ≬≟ά࣡ࢡࢳࣥࡀ᥋✀ࡉࢀࡓࢾཬࡧࢿࢥ
࠾ࡅࡿ≬≟୰ᢠయ౯ࡢ᥎⛣
8 ᗎㄽ 9 ᮦᩱཬࡧ᪉ἲ 11 㸯㸬ᐇ㦂ື≀ 11 㸰㸬࣡ࢡࢳࣥ᥋✀ 11 㸱㸬᥇⾑ཬࡧ⾑Ύ᥇ྲྀ 12 㸲㸬≬≟ᢠయ᳨ᰝ 12 㸳㸬࣮ࣝࢳ᳨ࣥయ 14 㸴㸬⤫ィゎᯒ 14 ⤖ᯝ 15 㸯㸬ࢾ࠾ࡅࡿ୰ᢠయ౯ࡢ᥎⛣ 15 㸰㸬ࢿࢥ࠾ࡅࡿ୰ᢠయ౯ࡢ᥎⛣ 16 㸱㸬࣮ࣝࢳࣥࢾ᳨య࠾ࡅࡿ୰ᢠయ౯ཬࡧᢠయ᳨ᰝྜ᱁≧ἣ 17 㸲㸬࣮ࣝࢳࣥࢿࢥ᳨య࠾ࡅࡿ୰ᢠయ౯ཬࡧᢠయ᳨ᰝྜ᱁≧ἣ 18 ⪃ᐹ 19 ᅗ⾲ 24
➨
➨㸰❶ ≬≟ᢠయ᳨ᰝἲ㸦FAVN ἲ㸧࠾ࡅࡿ⣽⬊ẘᛶ
Ⓨ⏕せᅉཬࡧࡑࡢ㝖ཤ᪉ἲ
40 ᗎㄽ 41 ᮦᩱཬࡧ᪉ἲ 43 㸯㸬≬≟࢘ࣝࢫᑐࡍࡿ୰ᢠయࡢ᳨ᰝ㸦FAVN㸧 43 㸰㸬⣽⬊⥅௦ࡼࡿ⣽⬊ࡢᙳ㡪 43 㸱㸬⣽⬊ẘᛶࡢศ㢮ཬࡧホ౯᪉ἲ 44 㸲㸬ᐇ㦂ື≀ 44 㸲㸬㸯㸬ᐇ㦂⏝ࢾ 44 㸲㸬㸰㸬ᐇ㦂⏝ࣔࣝࣔࢵࢺ 45 㸳㸬ᐇ㦂1㸸⾑⢾ࠊ⾑Ύ⬡㉁ཬࡧ pH ࡢ ᐃ 45 㸳㸬㸯㸬࣮ࣝࢳ᳨ࣥయA 45 㸳㸬㸰㸬⾑⢾ཬࡧ⾑Ύ⬡㉁ࡢ ᐃ 45 㸳㸬㸱㸬୰ヨ㦂࠾ࡅࡿᇵ㣴ୖΎࡢpH ᐃ 46 㸴㸬ᐇ㦂2㸸⣽⬊ẘᛶ㝖ཤ᪉ἲࡢ᳨ウ 46 㸴㸬㸯㸬࣮ࣝࢳ᳨ࣥయB 46 㸴㸬㸰㸬⾑Ύࡢ⣽⬊྾ฎ⌮ 46 㸴㸬㸱㸬⾑Ύࡢࢺࣜࣉࢩࣥ-EDTA ฎ⌮ 47 㸴㸬㸲㸬⾑Ύࡢ࢝࢜ࣜࣥฎ⌮ 47 㸴㸬㸳㸬⣽⬊ᇵ㣴⏝ฎ⌮ࣉ࣮ࣞࢺ 47 㸵㸬ᐇ㦂 3㸸FAVN ᢠయ౯ࡢᙳ㡪 47 㸵㸬㸯㸬࣮ࣝࢳ᳨ࣥయC 47 㸵㸬㸰㸬⣽⬊ẘᛶ㝖ཤἲࡼࡿ≬≟୰ᢠయ౯ࡢᙳ㡪 48㸶㸬⾑ᾮจᅛ㛵㐃≀㉁ࡢῧຍཬࡧ⾑Ύศ㞳 48 㸷㸬⁐⾑⾑Ύࡢㄪ〇 49 㸯㸮㸬౮㞧≀ྵ᭷⾑Ύࡢㄪᩚ 49 㸯㸯㸬⿕㦂᳨యࡢฎ⨨ཬࡧಖ⟶ 50 㸯㸰㸬ᢠBHK-21 ⣽⬊ IgG ࢆ⏝࠸ࡓ FAVN ἲ 50 㸯㸱㸬⤫ィゎᯒ 51 ⤖ᯝ 51 㸯㸬⣽⬊ࡢ⥅௦ࡀ⣽⬊࠼ࡿᙳ㡪 52 㸰㸬⾑ᾮจᅛ㛵㐃≀㉁⣽⬊ẘᛶࡢ㛵ಀ 52 㸱㸬⾑Ύࡢ⁐⾑ཬࡧ⾑Ύዃ㞧≀⣽⬊ẘᛶࡢ㛵ಀ 53 㸲㸬⿕㦂᳨యࡢฎ⨨ཬࡧಖ⟶᪉ἲ⣽⬊ẘᛶࡢ㛵ಀ 53 㸳㸬ᢠ BHK-21 ⣽⬊ IgG ⣽⬊ẘᛶࡢ㛵ಀ 53 㸴㸬⾑⢾ཬࡧ⾑Ύ⬡㉁⣽⬊ẘᛶࡢ㛵ಀ 54 㸵㸬୰ヨ㦂࠾ࡅࡿᇵ㣴ୖΎࡢ pH ⣽⬊ẘᛶࡢ㛵ಀ 54 㸶㸬ቑṪ㜼ᐖᆺ⣽⬊ẘᛶᑐࡍࡿ⣽⬊ẘᛶ㝖ཤἲࡢຠᯝ 54 㸷㸬᥋╔㜼ᐖᆺ⣽⬊ẘᛶᑐࡍࡿ⣽⬊ẘᛶ㝖ཤἲࡢຠᯝ 55 㸯㸮㸬FAVN ୰ᢠయ౯ᑐࡍࡿ⣽⬊ẘᛶ㝖ཤἲࡢᙳ㡪 56 ⪃ᐹ 57 ᅗ⾲ 63
➨
➨㸱❶ ᐊ ಖ⟶ࡋࡓ⿕㦂⾑ΎⓎ⏕ࡍࡿ⣽⬊ẘᛶࡢண㜵
ཬࡧ㝖ཤ᪉ἲ୪ࡧࡑࡢⓎ⏕ᶵᗎ
77 ᗎㄽ 78 ᮦᩱཬࡧ᪉ἲ 80 㸯㸬⣽⬊ẘᛶヨ㦂 80 㸰㸬⣽⬊ẘᛶࡢホ౯ᇶ‽ 80 㸱㸬⾑Ύࡢ᥇ྲྀㄪᩚ 81 㸲㸬ྛ✀⾑Ύฎ⌮᮲௳ཬࡧಖ⟶ᮇ㛫⣽⬊ẘᛶࡢ㛵㐃ᛶࡢゎᯒ 81 㸲㸬㸯㸬ᐇ㦂 1㸸⾑Ύࡢຍ⇕ཬࡧ PI ฎ⌮ಖ⟶ᮇ㛫ࡢ㛵ಀ 82 㸲㸬㸰㸬ᐇ㦂2㸸↓ฎ⌮ㄪᩚ⾑Ύ࠾ࡅࡿಖ⟶ᮇ㛫ຍ⇕ฎ⌮ࡢ㛵ಀ 82 㸲㸬㸱㸬ᐇ㦂3㸸ㄪᩚ⾑Ύࡢྛ✀ࣉࣟࢸ࣮ࢮฎ⌮⣽⬊ẘᛶࡢ㛵ಀ 83 㸲㸬㸲㸬ᐇ㦂4㸸⾑Ύࡢಖ⟶᮲௳㸦 ᗘ࣭ᮇ㛫㸧⣽⬊ẘᛶࡢ㛵ಀ 83 㸳㸬ຍ⇕ฎ⌮ཬࡧ PI ฎ⌮ࡼࡿ≬≟୰ᢠయ౯ࡢᙳ㡪 84 ⤖ᯝ 85 㸯㸬⾑Ύࡢຍ⇕ཬࡧ PI ฎ⌮ಖ⟶ᮇ㛫ࡢ㛵ಀ㸦ᐇ㦂 1㸧 85 㸰㸬↓ฎ⌮ㄪᩚ⾑Ύ࠾ࡅࡿಖ⟶ᮇ㛫ຍ⇕ฎ⌮ࡢ㛵ಀ㸦ᐇ㦂 2㸧 86 㸱㸬ㄪᩚ⾑Ύࡢྛ✀ࣉࣟࢸ࣮ࢮฎ⌮⣽⬊ẘᛶࡢ㛵ಀ㸦ᐇ㦂 3㸧 87 㸲㸬⾑Ύࡢಖ⟶᮲௳㸦 ᗘ࣭ᮇ㛫㸧⣽⬊ẘᛶࡢ㛵ಀ㸦ᐇ㦂 4㸧 87 㸳㸬ຍ⇕ฎ⌮ཬࡧ PI ฎ⌮ࡼࡿ≬≟୰ᢠయ౯ࡢᙳ㡪 87 ⪃ᐹ 89 ᅗ⾲ 93
⤖ㄽ
118ㅰ㎡
120ᘬ⏝ᩥ⊩
1211 ⥴ ⥴ゝ ≬≟ࡣࠊ㔜⠜࡞⚄⤒≧ࢆᚩࡍࡿ࢘ࣝࢫᛶࡢே⋇ඹ㏻ឤᰁ࡛࠶ࡿࠋ ≬≟ࡢⓎ⏕ࡣࠊୡ⏺ⓗ☜ㄆࡉࢀ࡚࠾ࡾࠊࢪࠊࣇࣜ࢝࡞ࡢⓎᒎ㏵ୖ ᅜࢆ୰ᚰࠊẖᖺ55,000 ே௨ୖࡀ≛≅࡞ࡗ࡚࠸ࡿ㸦11, 20, 70, 74㸧ࠋ2016 ᖺ 4 ᭶⌧ᅾࠊ≬≟ࡢΎίᆅᇦࡣࠊ᪥ᮏࠊ࣮ࣚࣟࢵࣃࠊ࢜ࢭࢽཬࡧࣁ࣡࡞ 㝈ᒁࡉࢀࡿ㸦ཌ⏕ປാ┬㸸 http://www.mhlw.go.jp/bunya/kenkou/kekkaku-kansenshou10/㸧ࠋ≬≟ࢆᘬࡁ㉳ࡇࡍཎయࡣࠊࣛࣈࢻ࢘ࣝࢫ⛉ࣜࢵࢧ࢘ ࣝࢫᒓࡢ㑇ఏᏊᆺ 1 ᒓࡍࡿ≬≟࢘ࣝࢫ࡛࠶ࡾࠊ⾑Ύᆺࡀ༢୍࡛࠶ࡿࡇ ࡀ▱ࡽࢀ࡚࠸ࡿࠋ≬≟࢘ࣝࢫࡢᐟᇦࡣᗈࡃࠊࣄࢺࢆྵࡴ့ங㢮ឤᰁࡍ ࡿ⪃࠼ࡽࢀ࡚࠸ࡿ㸦WHO㸸http://www.who.int/mediacentre/factsheets/ fs099/en/㸧ࠋ㏻ᖖࡣࠊညയࢆࡍࡇࡼࡾࠊឤᰁື≀ࡢၚᾮ୰Ꮡᅾࡍࡿ≬ ≟࢘ࣝࢫࡀࡢಶయఏࡍࡿࠋᐟឤᰁࡋࡓ≬≟࢘ࣝࢫࡣࠊᮎᲈ⚄ ⤒ධᚋࠊ୰ᯡ⚄⤒⮳ࡾࠊ⚄⤒≧ࢆ≉ᚩࡍࡿ㔜⠜࡞⬻⬨㧊⅖ࢆᘬࡁ㉳ࡇ ࡍࡀࠊࡑࡢ₯అᮇ㛫ࡣ10 ᪥⛬ᗘࡽ 1 ᖺ௨ୖࡲ࡛ከᒱࢃࡓࡾࠊᐃ࡛࠶ࡿࡇ ࡀࡁ࡞≉ᚩ࡛࠶ࡿ㸦WHO㸸http://www.who.int/mediacentre/factsheets/ fs099/en/㸧ࠋ≬≟ࢆⓎࡋࡓࣄࢺཬࡧື≀ࡢ⮴Ṛ⋡ࡣࠊࡰ 100㸣࡛࠶ࡿࠋ ࣄࢺ࠾࠸࡚ࡣࠊ≛≅⪅ࡢ99㸣ࡀ≬≟ࢆⓎࡋࡓࢾࢆឤᰁ※ࡋ࡚࠸ࡿࠋ ᚑࡗ࡚ࠊᮏὶ⾜ᅜ࠾ࡅࡿࣄࢺࡢ≬≟ᑐ⟇ࡋ࡚ࡣࠊࢾࢆᑐ㇟ࡋࡓ࣡ ࢡࢳࣥ᥋✀ࡼࡿண㜵ࠊࡍ࡞ࢃࡕឤᰁ※ᑐ⟇ࡀ᭱Ⰻࡢᡭẁ࡛࠶ࡿࠋࣄࢺᑐࡋ࡚ ࡣࠊ≬≟࢘ࣝࢫᭀ㟢௨๓࣡ࢡࢳࣥ᥋✀ࢆᐇࡍࡿࡇࡀ᭷ຠ࡛࠶ࡿࠋࡲࡓࠊ ≬≟≉ᚩⓗ࡞ண㜵᪉ἲࡋ࡚ࠊᭀ㟢ᚋචࡀᣲࡆࡽࢀࡿࠋ≬≟࢘ࣝࢫ ᭀ㟢ࡉࢀࡓࣄࢺᑐࡋࠊ㏿ࡸ࣡ࢡࢳࣥࢆ᥋✀ࡋࠊ㛗࠸₯అᮇ㛫ࡢ㛫චࢆ ࡍࡿࡇࡼࡾࠊⓎࢆᢚ࠼ࡿࡇࡀྍ⬟࡛࠶ࡿࠋ୍᪉ࠊ᪥ᮏࡢࡼ࠺࡞≬≟
2 Ύίᅜ࡛ࡣື≀᳨ࢆᚭᗏࡋࠊᮏឤᰁື≀ࡢධࢆ㜵ࡄࡇࡀ᭱ࡶ㔜せ࡛ ࠶ࡿࠋࡋࡋ࡞ࡀࡽࠊ≬≟ࡢ₯అᮇࡀᐃ࡛࠶ࡿࡇࡽࠊ࡞᳨ࡣᅔ㞴 ࡛࠶ࡿࠋࡇࡢࡓࡵࠊᡃࡀᅜ࡛ࡣࠊ≬≟ࡢධഛ࠼ࡿࡓࡵࠊ㍺ධࡉࢀࡿࢾࠊ ࢿࢥ➼ᑐࡍࡿ๓ࡢ࣡ࢡࢳࣥ᥋✀ࡀ⩏ົࡅࡽࢀ࡚࠸ࡿ㸦69㸧ࠋ ⌧௦♫࠾࠸࡚ࡣࠊࢢ࣮ࣟࣂࣝࡀᛴ㏿㐍ࢇ࡛࠾ࡾࠊ᪥ᮏࡶࡇࡢ₻ὶࡢ୰ࠊ ᾏእࡢ㛫࡛ࣄࢺཬࡧ≀㈨ࡢ ᮶ࡀάⓎࡋ࡚࠸ࡿࠋࢢ࣮ࣟࣂࣝࣅࢪࢿࢫక ࠺㛗ᮇᾏእฟᙇཬࡧᾏእ᪑⾜࡞࡛ࡣࠊ࣌ࢵࢺື≀ࡢྠకࡢቑຍࡸ࣌ࢵࢺ ࣅࢪࢿࢫࡢࢢ࣮ࣟࣂࣝࡼࡾࠊື≀ࡢ㍺ฟධࡀάⓎࡋ࡚࠸ࡿ㸦㎰ᯘỈ⏘┬㸸 http://www.maff.go.jp/aqs/tokei/toukeinen.html㸧ࠋࡇࡢࡼ࠺࡞≧ἣୗ࡛ࡣࠊᾏእ ⏤᮶ࡢᵝࠎ࡞ཎయࡀ᪥ᮏධࡍࡿࣜࢫࢡࡀୖ᪼ࡋ㸦18, 29, 46, 62㸧ࠊ↓ㄽࠊ ≬≟ࡶእ࡛ࡣ࡞࠸ࠋཌ⏕ປാ┬ࡼࢀࡤࠊᡃࡀᅜࡢࢾ࠾ࡅࡿ≬≟ࡢண 㜵᥋✀⋡ࡣࠊᖺ70㸣௨ୖ࡛᥎⛣ࡋ࡚࠸ࡿ㸦ཌ⏕ປാ┬㸸http://www.mhlw.go.jp/ bunya/kenkou/kekkaku-kansenshou10/01.html㸧ࠋࡋࡋ࡞ࡀࡽࠊᮍⓏ㘓ಶయ ࢆྵࡵࡿࠊ50㸣௨ୗ␃ࡲࡿࡢ᥎ィࡶ࠶ࡿ㸦9, 56㸧ࠋ᪥ᮏ࠾࠸࡚ࡣࠊ1957 ᖺࢆ᭱ᚋᮏࡢⓎ⏕ࡀ࡞ࡃࠊ௨᮶ࠊ≬≟Ύίᅜࡢ୍ࡘࡋ࡚⥔ᣢࡉࢀ࡚ࡁ ࡓࠋࡋࡋ࡞ࡀࡽࠊୖグࡢㅖ≧ἣࢆ⪃៖ࡍࢀࡤࠊ᪥ᮏࡢ≬≟ࡢධࡘ࠸࡚ ྰᐃࡍࡿࡇࡣ࡛ࡁ࡞࠸ࠋ ≬ ≟ ࡢ Ⓨ ⏕ ࡣ ୡ ⏺ ྛ ᅜ ཬ ࡪ ࡇ ࡽ ࠊ ᅜ 㝿 ⋇ ົ ᒁ 㸦World
Organisation for Animal Health㸸OIE㸧ࡣࠊࡢື≀ྠᵝࠊᮏ㛵ࡍ ࡿሗ㞟ཬࡧศᯒࠊྛᅜᑐࡍࡿሗᥦ౪ࠊᢏ⾡ⓗᨭཬࡧຓゝ୪ࡧ⾨⏕ᇶ
‽ࡢ⟇ᐃ࡞ࢆᐇࡋ࡚࠸ࡿࠋOIE ࡣࠊ≬≟ࡀே⋇ඹ㏻ឤᰁ࡛࠶ࡿほⅬ
ࡽࠊୡ⏺ಖᶵ㛵㸦World Health Organization㸸WHO㸧࡞ࡶሗࢆඹ᭷
ࡋࠊ㐃ᖏࡋ࡚άືࢆ⾜࡞ࡗ࡚࠸ࡿࠋࣇࣛࣥࢫᅜ㣗ရ⎔ቃປാ⾨⏕Ᏻᗇࢼࣥࢩ࣮
3
Occupational Health & Safety㸸ANSES㸧ࡣࠊOIE ࡼࡾᣦᐃࡉࢀࡓ≬≟ 㛵ࡍࡿࣜࣇࣞࣥࢫࣛ࣎ࣛࢺ࣮࡛ࣜ࠶ࡿࠋᮏ◊✲ᡤࡣࠊ≬≟㛵ࡍࡿㄪᰝࠊ◊ ✲ཬࡧ᳨ᰝ᪉ἲࡢᵓ⠏࡞ࢆᢸᙜࡋ࡚࠸ࡿࠋࡑࡢᴗົෆᐜࡋ࡚ࠊ≬≟ᢠయ᳨ ᰝ㛵ࡍࡿྛ✀つ᱁್ࡢ⟇ᐃࠊࣜࣇࣞࣥࢫ⾑Ύࡢ౪⤥ཬࡧᅜ㝿ᢏ⬟ヨ㦂㸦እ㒊 ⢭ᗘ⟶⌮㸧ࡢദ࡞ࡀᣲࡆࡽࢀࡿࠋᅜ㝿ᢏ⬟ヨ㦂ࡣࠊྛᅜ࠾ࡅࡿ≬≟ᢠయ ᳨ᰝタࢆᑐ㇟ࡋ࡚࠾ࡾࠊᮏ᳨ᰝ⢭ᗘࡢ⥔ᣢࠊྥୖࢆ┠ⓗࡋ࡚࠸ࡿࠋ≬≟ ࠾ࡅࡿື≀᳨࡛ࡣࠊ᪥ᮏࢆྵࡵࠊ≬≟ᢠయ᳨ᰝࡢᡂ⦼ࢆᚲ㡲ࡍࡿᅜࡀከ ࠸ࠋࡑࡢ㝿ࡣࠊᅜ㝿ᢏ⬟ヨ㦂ྜ᱁ࡋࡓ᳨ᰝタࡼࡿ᳨ᰝᡂ⦼ࡢࡳࡀ᭷ຠ࡛ ࠶ࡿࠋ ᪥ᮏ࠾࠸࡚≬≟ࡣࠊឤᰁࡢண㜵ཬࡧឤᰁࡢᝈ⪅ᑐࡍࡿ་⒪ 㛵ࡍࡿἲᚊ㸦௨ୗࠊឤᰁἲ␎ࡍ㸧ࠊᐙ␆ఏᰁண㜵ἲཬࡧ≬≟ண㜵ἲࡢつ ไࢆཷࡅࡿࠋឤᰁἲࡣࠊࣄࢺ࠾ࡅࡿ≬≟ᑐ⟇ཬࡧ≬≟࢘ࣝࢫࡢ⟶⌮᪉ ἲࡘ࠸࡚ࠊᐙ␆ఏᰁண㜵ἲࡣࠊᐙ␆㸦࢘ࢩࠊ࣐࢘ࠊࣄࢶࢪࠊࣖࢠཬࡧࣈࢱ㸧 ࠾ࡅࡿ≬≟ᑐ⟇ࡘ࠸࡚ࠊ≬≟ண㜵ἲ࡛ࡣࠊឤᰁ⎔ࢆ⥔ᣢࡋࠊࣄࢺࡢឤ ᰁࣜࢫࢡࡀ㧗࠸ື≀✀ࡋ࡚ࠊࢾࠊࢿࢥࠊ࢟ࢶࢿࠊࣛࢢ࣐ཬࡧࢫ࢝ࣥࢡࢆ ᳨ᑐ㇟ࡋ࡚࠸ࡿࠋ≬≟㛵ࡍࡿື≀᳨ࡣࠊ≬≟ண㜵ἲᇶ࡙ࡃ≟➼ࡢ ㍺ฟධ᳨つ๎㸦௨ୗࠊ᳨つ๎␎ࡍ㸧ࡀ㐺⏝ࡉࢀࠊ㎰ᯘỈ⏘┬ື≀᳨ᡤ࡛ ᐇࡉࢀ࡚࠸ࡿࠋ2004 ᖺ௨๓ࡢᪧつ๎࠾࠸࡚ࡣࠊ≬≟Ⓨ⏕ᅜࡽࢾཬࡧ ࢿࢥࢆ᪥ᮏ㍺ධࡍࡿሙྜࠊ≬≟࣡ࢡࢳࣥࡀ᥋✀῭ࡳ࡛࠶ࡗ࡚ࡶ 14 ᪥௨ୖ 180 ᪥௨ୗࡢಀ␃ࡀ⩏ົࡅࡽࢀ࡚࠸ࡓࠋࡋࡋࠊ᪥ᮏ࠾࠸࡚≬≟ࡢⓎ⏕ࡀ 㛗ᮇ㛫ㄆࡵࡽࢀ࡞࠸ࡇࠊࢾཬࡧࢿࢥࡢឡ⋵ື≀ཬࡧື≀⚟♴࡞ࡢほⅬ ࡽࠊಀ␃ᮇ㛫▷⦰㛵ࡍࡿ♫ⓗせㄳࡀᙉࡲࡗࡓࠋ୍᪉ࠊ㜵ᥐ⨨ࡢほⅬࡽ ࡣࠊୗグ㡯ࡀᠱ࡞ࡗࡓࠋࡍ࡞ࢃࡕࠊ㸦ϸ㸧ಀ␃ᮇ㛫ࡀ14 ᪥㛫ࡢሙྜࠊ≬≟ Ύίᅜࡢ୰࡛ࡣ᭱▷࡞ࡾࠊᅜ㝿Ỉ‽ࡢ㛫㞳ࡀ࠶ࡿࡇࠊ㸦Ϲ㸧
4 WHO ཬࡧ OIE ࡼࡾࠊಀ␃▷⦰ࡢಖドࡋ࡚≬≟࣡ࢡࢳࣥ᥋✀ᚋ୰ᢠయ ౯ࢆ ᐃࡋࠊࡑࡢຠᯝࢆ☜ㄆࡍࡁࡢ່࿌ࡀ࡞ࡉࢀࡓࡇࠊ㸦Ϻ㸧ᑠᆺ≟ࣈ࣮ ࣒ࡼࡾࠊ2002㹼2003 ᖺࡅ࡚ண㜵᥋✀ࡢຠᯝࡀᮇᚅ࡛ࡁ࡞࠸Ꮚ≟ࡀከᩘ㍺ ධࡉࢀࠊಀ␃ᮇ㛫▷⦰ࡢุ᩿ཬࡧಶయ㆑ูࡀᅔ㞴࡞ࡗࡓࡇ࡞ࡀᣲࡆࡽࢀ ࡓࠋࡇࡢࡓࡵࠊ᳨つ๎ࡢᅾࡾ᪉ࡘ࠸࡚ࠊ᭷㆑⪅ࡼࡿ᳨ウࡀ㔜ࡡࡽࢀࡓ㸦4, 5㸧ࠋ2004 ᖺࡣࠊ᳨つ๎ࡢ୍㒊ࡀᨵṇࡉࢀࡿ⮳ࡗࡓࠋࡇࡢᨵṇࡼࡾࠊ ࢾཬࡧࢿࢥࢆᾏእࡢ≬≟Ⓨ⏕ᆅᇦࡽ᪥ᮏ㍺ධࡍࡿ㝿ࡣࠊ᪂ࡓࠊ㸦ϸ㸧 ≬≟࣡ࢡࢳࣥࡢ᥋✀๓ࠊయෆ࣐ࢡࣟࢳࢵࣉࢆᇙࡵ㎸ࡳࠊಶయ㆑ูࢆྍ⬟ ࡍࡿࡇࠊ㸦Ϲ㸧࣐ࢡࣟࢳࢵࣉ╔ᚋࠊ≬≟ά࣡ࢡࢳࣥཪࡣ≬≟ ⤌࠼࣡ࢡࢳࣥࢆ30 ᪥௨ୖࡢ㛫㝸࡛ 2 ᅇ௨ୖ᥋✀ࡍࡿࡇࠊ㸦Ϻ㸧ᣦᐃࡉࢀࡓ ᳨ᰝタ࠾࠸࡚≬≟࢘ࣝࢫᑐࡍࡿᢠయ౯ࢆ ᐃࡋࠊࡑࡢ್ࡀ 0.5 International Unit㸦IU㸧/mL ௨ୖ࡛࠶ࡿࡇࡢ 3 㡯┠ࡀㄢࡏࡽࢀࡿࡇ࡞ ࡗࡓࠋࡇࡢ᪂ࡋ࠸᳨つ๎ࡼࡾࠊಀ␃ᮇ㛫ࡀ12 㛫௨ෆ▷⦰ࡉࢀࡿࡇ ࡞ࡾࠊ᳨యไࡢṇࡀᅗࡽࢀࡓࠋ 2004 ᖺࡢ᳨つ๎ᨵṇక࠸ࠊ㎰ᯘỈ⏘┬ࡣࠊ≬≟᳨ࢆ┠ⓗࡋ࡚᪥ᮏ ᅜෆࡢタࡀᮏᢠయ᳨ᰝࢆᐇࡍࡿሙྜࡢᐇつ᱁ࢆྲྀࡾỴࡵࡓࠋࡑࡢ⤖ᯝࠊ
⺯ගᢠయ࢘ࣝࢫ୰ヨ㦂ἲ㸦Fluorescent Antibody Virus Neutralization㸸
FAVN ἲ㸧ཪࡣ㎿㏿⺯ග↔Ⅼᢚไヨ㦂ἲ㸦Rapid Fluorescent Focus Inhibition Test㸸RIFFT ἲ㸧ࢆ᳨ᰝ᪉ἲࡍࡿࡇࡀỴᐃࡉࢀࡓࠋFAVN ἲཬࡧ RIFFT ἲ
ࡣࠊOIE ࡼࡾつᐃࡉࢀࡓ᳨ᰝ᪉ἲ࡛࠶ࡿ㸦26, 57㸧ࠋ୧ἲࡣࠊ≬≟࢘ࣝࢫ
ᑐ ࡍ ࡿ ୰ ᢠ య ࢆ ᐃ ࡍ ࡿ ࡇ ࡽ ࠊ ୰ ᢠ య ௨ እ ࡢ ᢠ య ࡶ ᐃ ࡍ ࡿ Enzyme-linked immuno-sorbent assay㸦ELISA㸧ἲẚࠊ≬≟࢘ࣝࢫ
ᑐࡍࡿ㜵ᚚ⬟ࡢ┦㛵ࡀ㧗࠸ࡉࢀࡿ㸦50㸧ࠋ
5
◊✲ᡤ㸧㸦௨ୗࠊᏳ◊␎ࡍ㸧ࡀࠊ᪥ᮏ࠾ࡅࡿ≬≟ᢠయ᳨ᰝタࡋ࡚
㎰ᯘỈ⏘┬ࡼࡾᣦᐃࡉࢀࠊ2016 ᖺ 1 ᭶⌧ᅾࠊࢪᆅᇦ࠾ࡅࡿ၏୍ࡢ≬≟ ᢠయᣦᐃ᳨ᰝタ࡛࠶ࡿࠋᮏ᳨ᰝࡣࠊ㎰ᯘỈ⏘┬ື≀᳨ᡤࡢ⟶⌮┘╩ࡢࡶࠊ Good laboratory practice㸦GLP㸧‽ࡌࡓ㐠Ⴀయไࡢࡶ࡛ᐇࡉࢀࠊ᳨ᰝ᪉
ἲࡋ࡚ FAVN ἲࡀ᥇⏝ࡉࢀ࡚࠸ࡿࠋᮏ᳨ᰝࡣࠊᏳᛶࡢほⅬࡽࠊᮏ᳨ᰝࡣ Biosafety level㸦BSL㸧3 ᐇ㦂ᐊ࡛ᐇࡉࢀࡿࠋࡲࡓࠊ᳨ᰝ⢭ᗘࡢ⥔ᣢࢆ┠ⓗ ࡋ࡚ࠊ๓㏙ࡢᅜ㝿ᢏ⬟ヨ㦂ࢆᖺ1 ᅇཷ㦂ࡋ࡚࠾ࡾࠊẖᅇྜ᱁ࡋ࡚࠸ࡿࠋ㏆ᖺ࠾ ࡅࡿᖺ㛫᳨ᰝ௳ᩘࡣࠊࢾࡀ⣙3,000 ௳ࠊࢿࢥࡀ⣙ 1,000 ௳࡛࠶ࡾࠊᢠయ᳨ᰝ⏦ ㄳ⪅ࡢᒃఫᆅࡣࠊ᪥ᮏࠊ୰ᅜࠊྎ‴ཬࡧ㡑ᅜࡀ㒊ศࢆ༨ࡵࡿࠋ 2005 ᖺᮏ᳨ᰝࡀ㛤ጞࡉࢀ࡚௨᮶ࠊᏳ◊ࡣࠊᢠయ᳨ᰝ⏦ㄳ⪅ࡽࡢᵝࠎ ࡞ၥ࠸ྜࢃࡏࡀᐤࡏࡽࢀࡿࠋ≉ࠊᢠయ౯ࡀྜ᱁್ᮍ‶࡞ࡗࡓ⌮⏤㛵ࡍࡿෆ ᐜࡀከ࠸ࠋࡇࡢࡼ࠺࡞ಶయࡣࠊᗄᗘࡶ᥇⾑ࢆ⧞ࡾ㏉ࡋࠊᢠయ᳨ᰝࢆ」ᩘᅇཷࡅࡿ ᳨ᰝࡢࡀᑡ࡞ࡃ࡞࠸ࠋࡲࡓࠊᢠయ᳨ᰝ⏦ㄳ⪅ࡽ㏦ࡉࢀࡿࢾཬࡧࢿࢥ ⾑Ύࡢ≧ែࡣከᵝ࡛࠶ࡿࠋ⾑Ύࡢ≧ែࡼࡗ࡚ࡣヨ㦂ࢆవ࡞ࡃࡉࢀࠊሙྜ ࡼࡗ࡚ࡣ᳨ᰝྍ⬟࡞ࡿࡶ࠶ࡿࠋᢠయ᳨ᰝ⏦ㄳ⪅ࡽࡢၥ࠸ྜࢃࡏཬࡧ ᳨ᰝᐇົ㛵ࡍࡿሗࡀ✚ࡉࢀࡿࡘࢀࠊᮏ᳨ᰝࢆᐇࡍࡿୖ࡛ᨭ㞀࡞ࡿ ㇟ࡀ᫂ࡽ࡞ࡗࡓࠋࡑࡢ⤖ᯝࠊゎỴࡉࢀࡿࡁୗグࡢⅬࡀㄆ㆑ࡉࢀࡿ⮳ ࡗࡓࠋ ➨୍Ⅼࡋ࡚ࠊ๓㏙ࡢ᳨つ๎ᚑࡗ࡚≬≟ά࣡ࢡࢳࣥࢆ᥋✀ࡉࢀࡓ ࢾཬࡧࢿࢥࡘ࠸࡚ࠊ᭶༢࡛ᢠయ౯ ᐃࡋࡓሗ࿌ࡣ࠶ࡿࡶࡢࡢ㸦2, 4, 5㸧ࠊ ࡑࡢ᥎⛣ࢆヲ⣽ㄪᰝࡋࡓሗ࿌ࡀ࡞࠸ࡇࡀᣲࡆࡽࢀࡿࠋ≉ࠊࢿࢥࡘ࠸࡚ ࡣࠊỤࡽࡢሗ࿌㸦4㸧ࡀ၏୍࡛࠶ࡿࠋࡑࡢࡓࡵࠊ᳨つ๎ᚑࡗ࡚≬≟ά ࣡ࢡࢳࣥࢆ 2 ᅇ᥋✀ᚋࠊࡢ⛬ᗘࡢᮇ㛫ࢆ✵ࡅ࡚᥇⾑ࡋࠊᢠయ᳨ᰝࢆཷࡅࡿ ࡁ࡛᫂࠶ࡿࠋᢠయ౯ࡢ᥎⛣ࡀ᫂ࡽ࡞ࢀࡤࠊ᭱㐺࡞᥇⾑ᮇࢆᥦࡍࡿ
6 ࡇࡀ࡛ࡁࠊ↓⏝࡞᳨ᰝࢆᅇ㑊ࡍࡿࡇࡀྍ⬟࡞ࡿࠋࡲࡓࠊᡃࡀᅜࡢ᳨つ ๎࡛ࡣࠊ≬≟ᢠయ᳨ᰝྜ᱁ᚋࡢ᭷ຠᮇ㛫ࢆ 2 ᖺつᐃࡋ࡚࠸ࡿࡀࠊࡑࡢ᰿ ᣐࢆ♧ࡋࡓࢹ࣮ࢱࡣබ⾲ࡉࢀ࡚࠸࡞࠸ࠋ ➨Ⅼࡋ࡚ࠊᮏ᳨ᰝࡢ࢘ࣝࢫ୰ヨ㦂㐣⛬࡛Ⓨ⏕ࡍࡿࠊ⿕㦂⾑Ύ⏤᮶ࡍ ࡿ⣽⬊ẘᛶࡀᣲࡆࡽࢀࡿࠋ⣽⬊ẘᛶࡢ㝖ཤἲࡘ࠸࡚ࡣࠊ✀ࠎࡢ᪉ἲࡀᏑᅾࡍࡿ ࡀ㸦19, 30㸧ࠊࡇࢀࡽࡢ᪉ἲࢆయ⣔ⓗẚ㍑ࠊ᳨ドࡋࡓሗ࿌ࡣ࡞࠸ࠋࡲࡓࠊ୍⯡ ⓗࠊ⿕㦂⾑Ύ⏤᮶ࡍࡿ⣽⬊ẘᛶࡘ࠸࡚ࡣࠊࡑࡢ㝖ཤ᪉ἲ║ࡀ⨨ࢀࠊ Ⓨ⏕ண㜵ࡢほⅬࡽ᳨ウࡉࢀࡓሗ࿌ࡣᏑᅾࡋ࡞࠸ࠋ⣽⬊ẘᛶࢆᣢࡘ⾑Ύ࡛ࡣࠊ ࢘ࣝࢫ୰ヨ㦂⣽⬊ࡢจ㞟ཪࡣ㞳ࢆᘬࡁ㉳ࡇࡍࡇࡽࠊ୰ヨ㦂 ࡑࡢࡶࡢࡀᡂ❧ࡏࡎࠊุᐃ⬟࡞ࡿࠋ௨ୖࡢ≧ἣࡽࠊ⣽⬊ẘᛶ㛵ࡍࡿⓎ⏕ せᅉࡢ✲᫂ཬࡧ㝖ཤ᪉ἲࡢ☜❧ࡼࡾࠊ࠸࡞ࡿ≧ែࡢ⿕㦂⾑Ύࡶᑐฎྍ⬟ ࡞デ᩿యไࢆᵓ⠏ࡍࡿࡇࡀᮃࡲࢀࡿࠋ ୖグࡢㅖၥ㢟ࡣࠊᮏ᳨ᰝἲࢆᏳ◊ᑟධࡍࡿẁ㝵࡛ࡣணࡉࢀ࡚࠾ࡽࡎࠊ㐠 ⏝㛤ጞᚋ᳨ᰝᐇົ࡛ࣞ࣋ࣝㄆ㆑ࡉࢀࡓࡶࡢ࡛࠶ࡿࠋᚑࡗ࡚ࠊࡇࢀࡽࡢㅖၥ㢟ࡢ ゎỴࡣࠊ᳨ᰝࡢᐇ⢭㏻ࡋࡓ᳨ᰝᐇົ⪅ࡼࡗ࡚⾜ࢃࢀࡿࡇࡀጇᙜ⪃࠼ ࡽࢀࡿࠋᐇ㝿ࠊࡑࡢᑐฎἲཬࡧ㐠⏝᪉ἲ㛵ࡍࡿᶆ‽᧯సᡭ㡰᭩㸦Standard Operating Procedure㸸SOP㸧ࡣࠊཎ๎ࡋ᳨࡚ᰝᐇົ⪅ࡼࡗ࡚సᡂࡉࢀ࡚࠸ ࡿࠋ ๓㏙ࡢ࠾ࡾࠊࢢ࣮ࣟࣂࣝ࡞ࡼࡗ࡚ࠊ᪥ᮏ࠾ࡅࡿ≬≟ධࣜࢫࢡࢆ ྰᐃࡍࡿࡇࡣ࡛ࡁ࡞࠸ࠋࡇࡢࡓࡵࠊᮏ᳨ᰝᛶ⬟ࡢྥୖࡣᚲ㡲ㄢ㢟࡛࠶ࡿࠋࡑࡇ ࡛ᮏ◊✲࡛ࡣࠊୖグၥ㢟ⅬࡢゎỴࢆ┠ⓗࡋ࡚ࠊ≬≟࣡ࢡࢳࣥ᥋✀ᚋࡢࢾཬ ࡧࢿࢥ࠾ࡅࡿᢠయ౯᥎⛣ࢆᢕᥱࡍࡿࡶࠊ⿕㦂⾑Ύ⏤᮶ࡍࡿ⣽⬊ẘᛶ ࡢⓎ⏕せᅉཬࡧࡑࡢ㝖ཤἲࡢ☜❧ࢆヨࡳࡓࠋ ➨ 1 ❶࡛ࡣࠊᮏά࣡ࢡࢳࣥࢆ᥋✀ࡋࡓࢾཬࡧࢿࢥࡢᢠయ౯᥎⛣ࡘ
7 ࠸࡚ࠊヲ⣽ㄪᰝࡋࠊᢠయ᳨ᰝ࠾࠸࡚᭱㐺࡞᥇⾑ᮇࢆ᳨ウࡋࡓࠋࡲࡓࠊᮏ࣡ ࢡࢳࣥࡢᣢ⥆ᮇ㛫ࢆㄪᰝࡋࠊ᳨つ๎࡛つᐃࡉࢀࡓ᭷ຠᮇ㛫ࡢጇᙜᛶࡘ࠸࡚ ᳨ドࡋࡓࠋ➨2 ❶࡛ࡣࠊ⿕㦂⾑Ύ⏤᮶ࡍࡿ⣽⬊ẘᛶࡢⓎ⏕せᅉࡘ࠸࡚ࠊ᳨ᰝ ⾜⛬ࡈヲ⣽᳨ドࡋࡓࠋࡲࡓࠊຠᯝⓗ࡞⣽⬊ẘᛶ㝖ཤἲࡘ࠸᳨࡚ウࡋࡓࠋ ➨ 3 ❶࡛ࡣࠊ⿕㦂⾑Ύࢆᖖ ಖ⟶ࡋࡓ㝿࠾ࡅࡿ⣽⬊ẘᛶࡢⓎ⏕᮲௳ࢆࡑࡢⓎ ⏕ᶵᗎࡶ᳨ウࡋࠊࡇࡢ⤖ᯝࢆࡶຠᯝⓗ࡞⾑ΎಖᏑ᪉ἲཬࡧ⣽⬊ẘᛶ Ⓨ⏕㜵Ṇἲࡢ☜❧ࢆヨࡳࡓࠋࡉࡽࠊ⿕㦂⾑Ύࡢᖖ ಖ⟶ࢆཎᅉࡍࡿ⣽⬊ẘᛶ ࡢⓎ⏕ࡑࡢண㜵࣭㝖ཤ᪉ἲࡀࠊ✀ࠎࡢື≀⾑Ύཬࡧ᳨ฟ⣽⬊⣔࠾࠸࡚ඹ㏻ࡍ ࡿ⌧㇟ཬࡧᑐᛂἲ࡛࠶ࡿࢆ᳨ドࡋࠊᅇᚓࡽࢀࡓ▱ぢࡀࡢ࢘ࣝࢫ୰ヨ 㦂ࡶᛂ⏝ྍ⬟ྰࡘ࠸࡚ࡶ᳨ウࡋࡓࠋ
8
➨
➨ ❶
≬≟ά࣡ࢡࢳࣥࡀ᥋✀ࡉࢀࡓࢾཬࡧࢿࢥ
࠾ࡅࡿ≬≟୰ᢠయ౯ࡢ᥎⛣
9 ᗎㄽ ᪥ᮏ࠾࠸࡚ࠊࢾཬࡧࢿࢥࢆ㍺ධࡍࡿ㝿ࡣࠊ≬≟ண㜵ἲᇶ࡙ࡃ᳨つ ๎ࡼࡿつไࢆཷࡅࡿࠋ᳨つ๎ࡼࢀࡤࠊ⏕ᚋ91 ᪥㱋௨ୖࡢಶయᑐࡋࠊ≬ ≟ά࣡ࢡࢳࣥཪࡣ≬≟⤌࠼࣡ࢡࢳࣥࢆ30 ᪥௨ୖࡢ㛫㝸࡛ 2 ᅇ௨ୖ᥋ ✀ࡍࡿࡇࡀつᐃࡉࢀ࡚࠸ࡿࠋࡑࡢୖ࡛ࠊ⾑Ύ୰ࡢ≬≟୰ᢠయ౯ࡀ 0.5 International Unit㸦IU㸧/mL ௨ୖ࡛࠶ࡿࡇࡀせồࡉࢀ࡚࠸ࡿࠋࡇࡢ್ࡣࠊ≬ ≟࢘ࣝࢫࡢᭀ㟢ᑐࡋࠊ༑ศ࡞ឤᰁ㜵ᚚࡀᡂ❧ࡍࡿࡇࢆ᰿ᣐࡋ࡚タᐃ ࡉࢀࡓ㸦24㸧ࠋ Ᏻ◊ࡣࠊ㎰ᯘỈ⏘┬ࡼࡾᣦᐃࡉࢀࡓ≬≟ᢠయ᳨ᰝタ࡛࠶ࡿࠋᮏタ ࠾ࡅࡿ㏆ᖺࡢᖺ㛫᳨ᰝ௳ᩘࡣࠊࢾࡀ⣙3,000 ௳ࠊࢿࢥࡀ⣙ 1,000 ௳㐩ࡍࡿࠋ ࡑࡢ୰࡛ࠊᢠయ᳨ᰝ⏦ㄳ⪅ࡽࠊᮏ᳨ᰝ㛵ࡍࡿၥ࠸ྜࢃࡏࡀከᩘᐤࡏࡽࢀࡿࠋ ၥ࠸ྜࢃࡏࡢ୰࡛᭱ࡶከ࠸ෆᐜࡣࠊ᳨ᰝ࡛ྜ᱁࡞ࡗࡓ⌮⏤㛵ࡍࡿࡶࡢ࡛ ࠶ࡿࠋᐇ㝿ᮏタ࡛ࡣࠊࢾࠊࢿࢥࢆၥࢃࡎࠊ⣙5㸣ࡢ᳨య⾑Ύࡀᢠయ౯༑ ศࡢࡓࡵྜ᱁࡞ࡗ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊࡑࡢཎᅉࡘ࠸࡚ࡣ࡛᫂࠶ࡾࠊ ⤖ᯝࡋ࡚ᢠయ᳨ᰝࢆ⧞ࡾ㏉ࡍࡀከ࠸ࠋࡲࡓࠊᢠయ᳨ᰝ⏦ㄳ⪅ࡢ୰ࡣࠊࣅ ࢪࢿࢫࡸ᪑⾜࡞ࡢ᪥⛬ࡽࠊ᳨ᰝࡀྍ⬟࡞ࡿሙྜࡀᑡ࡞ࡃ࡞࠸ࠋᚑࡗ࡚ࠊ ≬≟ᢠయ᳨ᰝ࠾ࡅࡿࡾࢆᅇ㑊ࡍࡿࡓࡵࡣࠊᢠయ᳨ᰝ⏦ㄳ⪅ᑐࡋ࡚ࠊ㐺 ษ࡞࣡ࢡࢳࣥ᥋✀᪉ἲཬࡧ᥇⾑ᮇࢆᥦ♧ࡍࡿࡇࡀồࡵࡽࢀ࡚࠸ࡿࠋ ୍᪉ࠊ࢘ࣝࢫᏛࡸ࣡ࢡࢳࣥ㛤Ⓨࡢศ㔝࠾࠸࡚ࡶࠊ≬≟ά࣡ࢡࢳࣥࢆ ࢾཬࡧࢿࢥ᥋✀ࡋࡓ㝿ࡢᢠయ᥎⛣㛵ࡍࡿሗ࿌ࡣᑡ࡞࠸㸦2, 4, 5㸧ࠋ≉ࠊ ࢿࢥ㛵ࡍࡿሗࡣỤࡽ㸦4㸧ࡢሗ࿌௨እබ⾲ࡀ࡞࠸ࠋࡲࡓࠊᡃࡀᅜࡢ᳨ つ๎࠾ࡅࡿᢠయ౯ࡢ᭷ຠᮇ㛫ࡣ 2 ᖺ㛫࡛࠶ࡿࡀࠊࡑࡢタᐃ᰿ᣐ㛵ࡍࡿࢹ ࣮ࢱࡣබ⾲ࡉࢀ࡚࠸࡞࠸ࠋ
10
ࡑࡇ࡛ᮏ❶࡛ࡣࠊ᪥ᮏࡢ᳨つ๎࡛つᐃࡉࢀࡓ᪉ἲࡼࡾࠊࢾཬࡧࢿࢥ≬ ≟ά࣡ࢡࢳࣥࢆ᥋✀ࡋࠊࡑࡢᚋࡢᢠయ᥎⛣ࢆ㐌༢ཪࡣ᭶༢࡛ㄪᰝࡋ ࡓࠋࡲࡓࠊᮏタ㏦ࡉࢀࡓ⿕㦂⾑Ύ㸦࣮ࣝࢳ᳨ࣥయ㸧ࡘ࠸࡚ࠊ࣡ࢡࢳࣥ᥋
11 ᮦᩱཬࡧ᪉ἲ 1. ᐇ㦂ື≀ ࢾࡣࠊ13㹼24 ࣨ᭶㱋ࡢ࢜ࢫཬࡧ࣓ࢫࡢᐇ㦂⏝ࣅ࣮ࢢࣝ✀㸦ᰴᘧ♫ࢼࣝࢡࠊ ᪥ᮏ㸧ࢆ⏝ࡋࡓࠋࢿࢥࡣࠊ6㹼12 ࣨ᭶㱋ࡢ࢜ࢫཬࡧ࣓ࢫࡢᐇ㦂⏝㞧✀㸦ࣜࣂࢸ ࣮࣭ࣜࢧ࣮ࢳ♫ࠊ࣓ࣜ࢝㸧ࢆ⏝ࡋࡓࠋࡇࢀࡽࡢື≀ࡣࠊ≬≟࣡ࢡࢳࣥᮍ ᥋✀࡛ࠊࢾཬࡧࢿࢥࡶ1 㢌ࡎࡘࢣ࣮ࢪ࡛㣫⫱ࡉࢀࠊヨ㦂ᮇ㛫୰ࡣࠊᕷ㈍ࡢ 㣫ᩱ㸦ࢾ㸸ࣛ࣎D ࢫࢺࢵࢡ㸹᪥ᮏ㎰⏘ᕤᴗᰴᘧ♫㸹᪥ᮏࠊࢿࢥ㸸CF6㸹࢜ࣜ ࢚ࣥࢱࣝ㓝ẕᕤᴗᰴᘧ♫㸹᪥ᮏ㸧ཬࡧỈ㐨Ỉࢆ࠼ࡽࢀࡓࠋᮏヨ㦂ࡣࠊᏳ◊ ࠾ࡅࡿື≀ᐇ㦂ጤဨࡢᢎㄆࢆᚓ࡚ᐇࡉࢀࡓ㸦ᢎㄆ␒ྕ㸸071009-2㸧ࠋ 2. ࣡ࢡࢳࣥ᥋✀ 1 ⩌࠶ࡓࡾ 5 㢌࡛ᵓᡂࡋࡓࢾཪࡣࢿࢥᑐࡋ࡚ࠊ᪥ᮏ࡛ᕷ㈍ࡉࢀ࡚࠸ࡿ≬ ≟⤌⧊ᇵ㣴ά࣡ࢡࢳࣥ㸦A ♫〇ཪࡣ B ♫〇㸧ࢆࠊࢾ࡛ࡣ 33 ᪥㛫㝸ࠊࢿ ࢥ࡛ࡣ35 ᪥㛫㝸࡛ࡑࢀࡒࢀ 2 ᅇࠊᣦ♧᭩ᚑࡗ࡚ 1 ࢻ࣮ࢬ㸦1mL/㢌㸧ࡎࡘ㢁 㒊⓶ୗ᥋✀ࡋࡓࠋ࡞࠾ࠊ㛗ᮇⓗ࡞ᢠయ᥎⛣ࢆㄪࡿ┠ⓗ࡛ࠊ᪥ᮏ࡛ࡢ≬≟᳨ ไᗘᚑ࠸ࠊࡇࢀࡽࡢ࣡ࢡࢳࣥࡢ᭷ຠᮇ㝈࠶ࡓࡿ 2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 1 ᖺᚋ㸦365 ᪥ᚋ㸧ࠊྠᵝࡢ᪉ἲࡼࡾ 3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࢆᐇࡋࡓࠋ࡞ ࠾ࠊ᪥ᮏ〇࣡ࢡࢳࣥ⏝ࡉࢀࡿ࢘ࣝࢫࡣRC࣭HL ᰴ࡞ࡗ࡚࠾ࡾ㸦10㸧ࠊA ♫ B ♫ࡢ㛫〇㐀࢘ࣝࢫࡢ㐪࠸ࡣ࡞࠸㸦1, 10㸧ࠋࡲࡓࠊ᪥ᮏ〇࣡ࢡࢳࣥ ࡣࠊࢪࣗࣂࣥࢺࡀῧຍࡉࢀ࡚࠸࡞࠸ࠋ
12 3. ᥇⾑ཬࡧ⾑Ύ᥇ྲྀ ࢾཬࡧࢿࢥࡢಶయࡘ࠸࡚ࠊ࣡ࢡࢳࣥ1 ᅇ┠᥋✀┤๓ࠊ1 ᅇ┠᥋✀ 7 ᪥ᚋ 㸦ࢿࢥࡢࡳ㸧ࠊྠ14 ᪥ᚋࠊྠ 21 ᪥ᚋࠊྠ 28 ᪥ᚋࠊ2 ᅇ┠᥋✀ 7 ᪥ᚋࠊ14 ᪥ᚋࠊ 21 ᪥ᚋࠊ28 ᪥ᚋ᥇⾑ࢆ⾜࠸ࠊ⾑Ύࢆ᥇ྲྀࡋࡓࠋࡉࡽࠊ࣡ࢡࢳࣥ 1 ᅇ┠᥋✀ ࡢ25 ࣨ᭶ᚋࡲ࡛ 1 ࣨ᭶㛫㝸࡛⾑Ύࢆ᥇ྲྀࡋࡓࠋಶయู๓⭎㒊㟼⬦㸦ࢧࣇ࢙ࢼ㸧 ࡽὀᑕჾ࡛᥇⾑ࡋࠊ┿✵᥇⾑⟶㸦ࣉ࣮ࣞࣥࢱࣉࠊࢸࣝࣔᰴᘧ♫ࠊ᪥ᮏ㸧 ⛣ࡋࡓࠋࡇࡢ┿✵᥇⾑⟶ࢆ25Υ࡛㸯㛫㟼⨨ࡋࡓᚋࠊ500g ࡛㐲ᚰศ㞳ࡋࠊ⾑ Ύࢆ᥇ྲྀࡋࡓࠋ᥇ྲྀᚋࡢ⾑Ύࡣࠊヨ㦂ࡲ࡛ࡢ㛫ࠊѸ80Υ࡛ಖ⟶ࡉࢀࡓࠋ 4. ≬≟ᢠయ᳨ᰝ ≬≟ᢠయ౯ࡢ ᐃࡣࠊOIE ࡢ࣐ࢽࣗࣝ㸦57㸧つᐃࡉࢀ࡚࠸ࡿ FAVN ἲ
ࡼࡾ⾜࡞ࡗࡓࠋ∵⫾ඣ⾑Ύ㸦Fetal calf serum㸸FCSࠊࢧ࣮ࣔࣇࢵࢩ࣮ࣕࢧ
࢚ࣥࢸࣇࢵࢡ♫ࠊ⡿ᅜ㸧ࢆ10 vol 㸣ῧຍࡋࡓ Dulbecco’s modified Eagle
medium㸦DMEMࠊࢧ࣮ࣔࣇࢵࢩ࣮ࣕࢧ࢚ࣥࢸࣇࢵࢡ♫ࠊ⡿ᅜ㸧ࡼࡾࠊ ⿕㦂⾑Ύࠊཧ↷㝧ᛶ⾑Ύ㸦0.5IU/mL㸧ཬࡧཧ↷㝜ᛶ⾑Ύࢆ 3 ಸᕼ㔘ࡋࡓࠋ࡞࠾ࠊ ཧ↷㝧ᛶ⾑Ύཬࡧཧ↷㝜ᛶ⾑Ύࡣࠊࣇࣛࣥࢫᅜ㣗ရ⎔ቃປാ⾨⏕Ᏻᗇࢼࣥࢩ
࣮◊✲ᡤ㸦Nancy Laboratory, French Agency for Food, Environmental and
Occupational Health & Safety㸸ANSESࠊࣇࣛࣥࢫ㸧ࡽ㉎ධࡋࡓࠋࡇࢀࡽࡢ
ᕼ㔘⾑Ύࡘ࠸࡚ࠊࢾࡣ56Υࠊࢿࢥࡣ 65Υ࡛ࠊࡑࢀࡒࢀ 30 ศ㛫㠀ാࡋࡓࠋ ࢿࢥ⾑Ύࡘ࠸࡚ࡣࠊᡃࠎࡢ⤒㦂ୖࠊ56Υࡼࡿ㠀ാ࡛ࡣ㠀≉␗ⓗ࡞㝧ᛶ ᛂࡀⓎ⏕ࡍࡿྍ⬟ᛶࡀ࠶ࡿࡇࡽࠊࡑࡢゎỴ⟇ࡋ࡚65Υࡼࡿ㠀ാࢆᐇ ࡋࡓࠋ96 ࢙࣐࢘ࣝࢡࣟࣉ࣮ࣞࢺࡢ 2㹼8 ิ┠ࡢ࢙࢘ࣝࠊ10 vol 㸣FCS ῧຍDMEM ࢆ 100μL ࡎࡘศὀࡋࡓࠋ⾑Ύ 1 ᳨యࡘࡁࠊ1 ᕼ㔘࠶ࡓࡾ 4 ࢙࢘ ࣝࢆ⏝ࡋࠊ1 ิ┠ࡢ 4 ࢙࢘ࣝิࡣࠊ㠀ാ῭ࡳࡢ⿕㦂⾑Ύࢆ 150μL ࡎࡘศ
13
ὀࡋࡓࠋḟ࠸࡛ࠊ1 ิ┠ࡽ 8 ิ┠ࡲ࡛ 3 ಸ㝵ẁᕼ㔘ิ㸦3㹼6,561 ಸ㸧ࢆసᡂ ࡋࡓࠋཧ↷㝧ᛶ⾑Ύཬࡧཧ↷㝜ᛶ⾑Ύࡘ࠸࡚ࡶࠊ⿕㦂⾑Ύྠᵝᕼ㔘ࡋࡓࠋ 10 vol 㸣FCS ῧຍ DMEM ࡼࡾ 100 Median tissue culture infectious dose
㸦TCID50㸧/mL ㄪ〇ࡋࡓ≬≟࢘ࣝࢫ㸦CVS-11, ATCC number: VR 959㸧
ᾮࢆ࡚ࡢ࢙࢘ࣝ50μL ࡎࡘศὀࡋࠊ37Υࠊ5㸣CO㸰ࠊ95㸣✵Ẽࡢ᮲௳ୗ࡛ 1
㛫࣮ࣥ࢟ࣗ࣋ࢺࡍࡿࡇࡼࡾ࢘ࣝࢫ୰ᛂࢆ⾜࡞ࡗࡓࠋḟ࠸࡛ࠊ10
vol 㸣FCS ῧຍ DMEM ࡛ㄪ〇ࡋࡓ Baby Hamster Kidney 㸦BHK㸧-21 ⣽⬊
ᾋ㐟ᾮ㸦4×105ಶ/mL㸧ࢆ࡚ࡢ࢙࢘ࣝ 50μL ࡎࡘศὀࡋࠊ37Υࠊ5㸣CO2ࠊ
95㸣✵Ẽࡢ᮲௳ୗ࡛ 48 㛫ᇵ㣴ࡋࡓࠋᇵ㣴⤊ᚋࠊ࡚ࡢ࢙࢘ࣝ 80㸣ࢭ
ࢺࣥ㸦ᅛᐃᾮ㸧ࢆ150ȣL ຍ࠼ࠊᐊ ୗ㸦25Υ㸧࡛ 30 ศ㛫㟼⨨ࡋࡓࠋᅛᐃᾮࢆ
ᗫᲠᚋࠊᐊ ࡛1 㛫⇱ࡉࡏࠊ⣽⬊ࢆᅛᐃࡋࡓࠋ࡚ࡢ࢙࢘ࣝᢠ≬≟࢘
ࣝࢫfluorescein isothiocyanate (FITC) ࣔࣀࢡ࣮ࣟࢼࣝᶆ㆑ᢠయᾮ㸦ᐩኈࣞࣅ
࢜ࠊ᪥ᮏ㸧ࢆ 70ȣL ຍ࠼ࠊ⺯ගᰁⰍࡋࡓࠋ⺯ග㢧ᚤ㙾㸦BX60Mࠊ࢜ࣜࣥࣃࢫࠊ
᪥ᮏ㸧ࡼࡾ⺯ගࢆㄆࡵࡓ࢙࢘ࣝᩘࡼࡾྛ⿕㦂⾑Ύཬࡧཧ↷㝧ᛶ⾑Ύࡢ50㸣
᭷ຠ㔞㸦50㸣Effective Dose㸸ED50㸧್ࢆồࡵࠊ୧⪅ࡢẚ㍑ࡽIU/mL ࢆồࡵ
ࡓࠋIU ࡢ⟬ฟᘧࡣୗグࡢ࠾ࡾ࡛࠶ࡿࠋ
[10 (serum log D50 value)) × theoretical titer of OIE serum 0.5IU/mL]
ᢠయ౯㸦IU/mL㸧㸻
10 (log D50 of OIE serum 0.5 IU/mL)ͤ
14 5. ࣮ࣝࢳ᳨ࣥయ 2005 ᖺࡽ 2011 ᖺࡅ࡚ࠊᏳ◊࠾࠸࡚≬≟ᢠయ᳨ᰝࢆᐇࡋࡓ ࢾ⾑Ύ㸦12 ࣨ᭶㱋௨ୗ㸧ཬࡧࢿࢥ⾑Ύ㸦ᖺ㱋ၥ㸧ࢆᑐ㇟ࡋࡓࠋ௨ୗࠊ࣮͆ࣝ ࢳࣥࢾ᳨య͇ཬࡧ࣮͆ࣝࢳࣥࢿࢥ᳨య͇⾲グࡍࡿࠋࡇࢀࡽࡢ࠺ࡕࠊ᪥ᮏ〇ཬ ࡧእᅜ〇ࡢ≬≟ά࣡ࢡࢳࣥࢆ30 ᪥௨ୖࡢ㛫㝸࡛ 2 ᅇ᥋✀ࡉࢀࡓ 5,079 ᳨ య㸦ࢾ㸸1,795 ᳨యࠊࢿࢥ㸸3,284 ᳨య㸧ࢆヨ㦂⏝࠸ࡓࠋᮏ࣡ࢡࢳࣥྵࡲ ࢀࡿࢪࣗࣂࣥࢺࡢᙳ㡪ࢆ⪃៖ࡋࠊࢪࣗࣂࣥࢺ↓ῧຍࡢ᪥ᮏ〇࣡ࢡࢳࣥࢆ᥋ ✀ࡋࡓಶయཬࡧࢪࣗࣂࣥࢺῧຍ῭ࡢእᅜ〇࣡ࢡࢳࣥࢆ᥋✀ࡋࡓಶయ༊ศࡋ ࡚ゎᯒࡋࡓࠋ௨ୗࠊࡑࢀࡒࢀࡢ༊ศࢆ͆᪥ᮏ〇࣡ࢡࢳࣥ⩌͇ཬࡧ͆እᅜ〇࣡ࢡࢳ ࣥ⩌͇⾲グࡍࡿࠋᑐ㇟ಶయ࠾ࡅࡿ࣡ࢡࢳࣥ᥋✀ᅇᩘࡀ 2 ᅇࡢࡳ࡛࠶ࡿࡇ ࢆ☜ᐇࡍࡿࡓࡵࠊ࣮ࣝࢳࣥࢾ᳨యࡘ࠸࡚ࡣࠊ⏕ᚋ12 ࣨ᭶㱋௨ୗࡢಶయࢆ ᑐ㇟ࡋࡓࠋ୍᪉ࠊࢿࢥ࡛ࡣࠊ≬≟᳨௨እࡢ┠ⓗ࡛࣡ࢡࢳࣥࡀ᥋✀ࡉࢀࡿ ࡣ⛥⪃࠼ࡽࢀࡿࡓࡵࠊ࣮ࣝࢳࣥࢿࢥ᳨యࡘ࠸࡚ࡣࠊᖺ㱋ࡢಶయࢆᑐ㇟ ࡋࡓࠋ࣡ࢡࢳࣥࡢ᥋✀㛫㝸ཬࡧ࣡ࢡࢳࣥࡢ 2 ᅇ┠᥋✀ࡽ᥇⾑ࡲ࡛ࡢ㛫㝸ᇶ ࡙ࡁࠊྛ࣮ࣝࢳ᳨ࣥయࡢ୰ᢠయ౯ཬࡧᢠయ᳨ᰝྜ᱁⋡ࡘ࠸࡚ゎᯒࡋࡓࠋ 6. ⤫ィゎᯒ ࣮ࣝࢳ᳨ࣥయ㸦ࢾཬࡧࢿࢥ㸧ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚ࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࠊ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡽ᥇⾑ࡲ࡛ࡢ㛫㝸ࠊ᳨ᰝྜ᱁⋡ࡢ㛵ಀࢆㄪࡿࡓࡵࠊᚲせ ᛂࡌ࡚࢝2 ᳨ᐃࢆᐇࡋࡓࠋp㸺0.05 ࡢࡁ⤫ィᏛⓗ᭷ពᕪࡀ࠶ࡿࡶࡢᐃ ⩏ࡋࡓࠋ
15 ⤖ᯝ 1. ࢾ࠾ࡅࡿ୰ᢠయ౯ࡢ᥎⛣ ࢾ5 㢌 A ♫〇ࡢ࣡ࢡࢳࣥࢆ᥋✀ࡋࡓ⩌㸦ࢾ࣡ࢡࢳࣥ A ⩌㸧࠾ࡅࡿ୰ ᢠయ౯ࡢ᥎⛣ࡘ࠸࡚ࠊヨ㦂㛤ጞ㸦1 ᅇ┠࣡ࢡࢳࣥ᥋✀㸧ࡽ 61 ᪥┠㸦2 ᅇ ┠࣡ࢡࢳࣥ᥋✀ᚋ 28 ᪥┠㸧ࡲ࡛ࡢ⤖ᯝ㸦㐌༢㸧ࢆᅗ 1㸫1A ࠊヨ㦂ᮇ㛫 㸦ヨ㦂㛤ጞᚋ25 ࣨ᭶┠ࡲ࡛㸧ࡢ⤖ᯝ㸦᭶༢㸧ࢆᅗ 1㸫2A ࠊ3 ᅇ┠࣡ࢡࢳࣥ ᥋✀ࡽ28 ᪥┠ࡲ࡛ࡢ⤖ᯝ㸦㐌༢㸧ࢆᅗ 1㸫3A ࡑࢀࡒࢀ♧ࡋࡓࠋྠᵝ B ♫〇ࡢ࣡ࢡࢳࣥࢆ᥋✀ࡋࡓ⩌㸦ࢾ࣡ࢡࢳࣥ B ⩌㸧ࡢ⤖ᯝࡘ࠸࡚ࡶࠊᅗ 1㸫 1Bࠊᅗ 1㸫2B ཬࡧᅗ 1㸫3B ࡑࢀࡒࢀ♧ࡋࡓࠋࢾ࣡ࢡࢳࣥ A ⩌ཬࡧ B ⩌ ࡶࠊ1 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋ 14 ᪥┠ࡲ࡛ࠊ࡚ࡢಶయࡀ 0.5IU/mL ௨ୖࡢ ᢠయ౯ࢆ♧ࡋࡓ㸦1.5ࠥ17.7IU/mL㸧㸦ᅗ 1㸫1A ཬࡧ 1㸫1B㸧ࠋࡑࡢᚋࠊ28 ᪥┠ ࡅ࡚ྠ⛬ᗘ࡛᥎⛣ࡍࡿࠊⱝࡋࡃࡣࡸࡸపୗࡋࡓࠋ2 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ ࡼࡾࠊࢾ࣡ࢡࢳࣥA ⩌ཬࡧ B ⩌ࡶಶయ࡛ࡁࡃᢠయ౯ࡀୖ᪼ࡋࠊ ࢇࡢಶయ࠾࠸࡚2 ᅇ┠᥋✀ᚋ 7 ᪥┠ཪࡣ 14 ᪥┠ࡀࣆ࣮ࢡ࡞ࡗࡓ㸦53.3ࠥ 1094.0IU/mL㸧ࠋࡑࡢᚋ 28 ᪥┠ࡲ࡛ࠊከࡃࡢಶయ࠾࠸࡚⦆ࡸ࡞ୗ㝆ࢆ♧ࡋ ࡓࠋ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 2 ࣨ᭶┠ࡽ 4 ࣨ᭶┠ࡅ࡚ࡣࠊࢾ࣡ࢡࢳࣥ A ⩌ཬࡧB ⩌ࡶከࡃࡢಶయ࠾࠸࡚ୗ㝆ࡢ⛬ᗘࡀࡁࡃ࡞ࡗࡓ㸦ᅗ 1㸫2A ཬ ࡧ1㸫2B㸧ࠋࡋࡋ࡞ࡀࡽࠊ3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࡲ࡛ࡢ㛫ࡣࠊከࡃࡢಶయ࡛ྠ ⛬ᗘࡢᢠయ౯㸦0.5㹼40.5IU/mL㸧ࢆ⥔ᣢࡋࠊ0.5IU/mL ࢆୗᅇࡿಶయࡣ࡞ࡗ ࡓࠋ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 1 ᖺᚋ 3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࢆᐇࡋࡓࡇࢁࠊ ࢾ࣡ࢡࢳࣥA ⩌ཬࡧ B ⩌ࡶࠊ14 ᪥┠ࡲ࡛ࡢ㛫ከࡃࡢಶయ࡛ᢠయ౯ࡀୖ᪼ ࡋࡓ㸦17.8㹼631.0IU/mL㸧㸦ᅗ1㸫3A ཬࡧ 1㸫3B㸧ࠋ3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋࡣࠊ ࢾ࣡ࢡࢳࣥA ⩌ཬࡧ B ⩌ࡶࠊ2 ᅇ┠᥋✀ᚋẚ࡚ᢠయ౯ࡢୗ㝆ࡀᑡ࡞
16 ࠸ಶయࡀከࡃࠊࡑࡢᚋ1 ᖺ㛫ࠊࡰྠ⛬ᗘࡢᢠయ౯㸦2.0㹼831.0IU/mL㸧ࢆ⥔ᣢ ࡍࡿಶయࡀከࡗࡓ㸦ᅗ1㸫2A ཬࡧ 1㸫2B㸧ࠋࢾ࣡ࢡࢳࣥ A ⩌ཬࡧ B ⩌ࡶ ࠊ1 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋࡽヨ㦂⤊㸦1 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 25 ࣨ᭶┠㸧 ࡲ࡛ࡢⅬࡘ࠸࡚ࠊ0.5IU/mL ࢆୗᅇࡗࡓಶయࡣ࡞ࡗࡓࠋ 2. ࢿࢥ࠾ࡅࡿ୰ᢠయ౯ࡢ᥎⛣ ࢿࢥ5 㢌ᑐࡋ࡚ A ♫〇ࡢ࣡ࢡࢳࣥࢆ᥋✀ࡋࡓ⩌㸦ࢿࢥ࣡ࢡࢳࣥ A ⩌㸧࠾ ࡅࡿ୰ᢠయ౯ࡢ᥎⛣ࡘ࠸࡚ࠊヨ㦂㛤ጞ㸦1 ᅇ┠࣡ࢡࢳࣥ᥋✀㸧ࡽ 63 ᪥┠ 㸦2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 28 ᪥┠㸧ࡲ࡛ࡢ⤖ᯝ㸦㐌༢㸧ࢆᅗ 1㸫4A ࠊヨ㦂 ᮇ㛫㸦ヨ㦂㛤ጞᚋ25 ࣨ᭶┠ࡲ࡛㸧ࡢ⤖ᯝ㸦᭶༢㸧ࢆᅗ 1㸫5A ࠊ3 ᅇ┠࣡ࢡ ࢳࣥ᥋✀ࡽ28 ᪥┠ࡲ࡛ࡢ⤖ᯝ㸦㐌༢㸧ࢆᅗ 1㸫6A ࡑࢀࡒࢀ♧ࡋࡓࠋྠᵝ ࢿࢥ5 㢌ᑐࡋ࡚ B ♫〇ࡢ࣡ࢡࢳࣥࢆ᥋✀ࡋࡓ⩌㸦ࢿࢥ࣡ࢡࢳࣥ B ⩌㸧ࡢ⤖ ᯝࡘ࠸࡚ࡶࠊᅗ1㸫4Bࠊᅗ 1㸫5B ཬࡧᅗ 1㸫6B ࡑࢀࡒࢀ♧ࡋࡓࠋࢿࢥ࣡ࢡ ࢳࣥA ⩌ཬࡧ B ⩌ࡶࠊ1 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋ 14 ᪥┠ࡲ࡛ࠊ࡚ࡢಶయ
ࡀ 0.5IU/mL ௨ୖࡢᢠయ౯ࢆ♧ࡋࡓ㸦0.66㹼23.4IU/mL㸧㸦ᅗ 1㸫4A ཬࡧ 1㸫
4B㸧ࠋࡑࡢᚋࠊ21 ᪥┠ࡲ࡛ୖ᪼ࡋࠊ28 ᪥┠ࡅ࡚ྠ⛬ᗘ࡛᥎⛣ࡍࡿࠊࡸࡸ పୗࡋࡓࠋ2 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࡼࡾࠊࢾ࣡ࢡࢳࣥ A ⩌ཬࡧ B ⩌ࡶ ಶయ࡛ࡁࡃᢠయ౯ࡀୖ᪼ࡋࠊࢇࡢಶయ࠾࠸࡚ 7 ᪥┠ࡀࣆ࣮ࢡ࡞ࡗ ࡓ㸦10.3㹼480.0IU/mL㸧ࠋࡑࡢᚋ 28 ᪥┠ࡲ࡛ࠊከࡃࡢಶయࡀྠ⛬ᗘࡢᢠయ౯࡛ ᥎⛣ࡋࡓ㸦2.6㹼277.0IU/mL㸧ࠋ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 2㹼3 ࣨ᭶┠ࡅ࡚ࡣࠊ ࢿࢥ࣡ࢡࢳࣥA ⩌ཬࡧ B ⩌ࡶከࡃࡢಶయ࠾࠸࡚ᢠయ౯ࡀపୗࡋࡓ㸦0.5㹼 276.0IU/mL㸧㸦ᅗ1㸫5A ཬࡧ 1㸫5B㸧ࠋࡋࡋ࡞ࡀࡽࠊ3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࡲ ࡛ࡢ㛫ࡣࠊከࡃࡢಶయ࡛ྠ⛬ᗘࡢᢠయ౯ࢆ⥔ᣢࡋࡓ㸦0.29㹼160.0IU/mL㸧ࠋࡓࡔ ࡋࠊࢿࢥ࣡ࢡࢳࣥB ⩌ࡢ 1 ಶయ㸦No.2㸧ࡘ࠸࡚ࡣẚ㍑ⓗప್࡛᥎⛣ࡋࠊ0.5IU/mL
17 ࢆୗᅇࡿⅬ㸦2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 4 ࣨ᭶┠㸸0.38IU/mLࠊྠ 5 ࣨ᭶┠㸸 0.29IU/mL㸧ࡀ࠶ࡗࡓࠋ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 1 ᖺᚋ 3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࢆ ᐇࡋࡓࡇࢁࠊࢿࢥ࣡ࢡࢳࣥA ⩌ཬࡧ B ⩌ࡶࠊ14 ᪥┠ࡲ࡛ࡢ㛫ከࡃࡢ ಶయ࡛ᢠయ౯ࡀୖ᪼ࡋࡓ㸦13.5㹼277.0IU/mL㸧㸦ᅗ1㸫6A ཬࡧ 1㸫6B㸧ࠋẚ㍑ⓗ ప್࡛᥎⛣ࡋ࡚࠸ࡓࢿࢥ࣡ࢡࢳࣥB ⩌ࡢ 1 ಶయ㸦No.2㸧ࡘ࠸࡚ࡶࠊୖ᪼ࡀࡳ ࡽࢀࡓ㸦30.8IU/mL㸧ࠋࢿࢥ࣡ࢡࢳࣥ A ⩌ཬࡧ B ⩌ࡶ 3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋ ✀ᚋࡣࠊ2 ᅇ┠᥋✀ᚋྠᵝࠊࡑࡢᚋ 1 ᖺ㛫ࡰྠ⛬ᗘࡢᢠయ౯ࢆ⥔ᣢࡍࡿಶ యࡀከࡗࡓ㸦2.0㹼480.0IU/mL㸧㸦ᅗ 1㸫5Aࠊ1㸫5B㸧ࠋ1 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ ᚋࡽヨ㦂⤊㸦1 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋ 25 ࣨ᭶┠㸧ࡲ࡛ࡢⅬࡘ࠸࡚ࠊࢿ ࢥ࣡ࢡࢳࣥB ⩌ࡢಶయ No.2 ࢆ㝖࠸࡚ࡣࠊ0.5IU/mL ࢆୗᅇࡗࡓಶయࡣ࡞ࡗࡓࠋ 3. ࣮ࣝࢳࣥࢾ᳨య࠾ࡅࡿ୰ᢠయ౯ཬࡧᢠయ᳨ᰝྜ᱁≧ἣ ࣡ࢡࢳࣥ᥋✀㛫㝸୰ᢠయ౯㸦ᗄఱᖹᆒ್㸧ࡢศᕸཬࡧᢠయ᳨ᰝྜ᱁⋡ ࠾ࡅࡿゎᯒ⤖ᯝࢆ⾲1㸫1A㸦᪥ᮏ〇࣡ࢡࢳࣥ㸧ཬࡧ⾲ 1㸫1B㸦እᅜ〇࣡ࢡࢳࣥ㸧 ♧ࡋࡓࠋ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ30㹼90 ᪥࡛ࡣࠊ᪥ᮏ〇ཬࡧእᅜ〇࣡ࢡࢳࣥ⩌ ࡶࠊ࣡ࢡࢳࣥ2 ᅇ┠᥋✀ᚋ 8㹼14 ᪥㏆ᢠయ౯ࡢࣆ࣮ࢡࡀࡳࡽࢀࡓࠋ୍᪉ࠊ ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ91 ᪥௨ୖ࡛ࡣࠊ᪥ᮏ〇࣡ࢡࢳࣥ⩌࠾࠸࡚ࠊᢠయ౯ࡢࣆ࣮ ࢡࡣ15㹼21 ᪥㏆ࡳࡽࢀࠊእᅜ〇࣡ࢡࢳࣥ⩌࡛ࡣࠊᢠయ౯ࣆ࣮ࢡࡣ 8㹼21 ᪥ ࡳࡽࢀࡓࠋ ᢠయ᳨ᰝࡢྜ᱁⋡ࡘ࠸࡚ࡣࠊ᪥ᮏ〇࣡ࢡࢳࣥ⩌࠾࠸࡚ࠊ࣡ࢡࢳࣥ 2 ᅇ ┠᥋✀┤ᚋࡀࡑࢀ௨㝆ࡢ᥇⾑᪥ẚ࡚ 5.4㹼29.0%᭷ព㧗ࡃ㸦p㸺0.01㸧ࠊ 1㹼7 ᪥ᚋ࠾࠸࡚ࡶ 5㸣๓ᚋ࡛࠶ࡗࡓࠋ≉ࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ 41 ᪥௨ୖ࡛ ࡣࠊྜ᱁⋡ࡀୖ᪼ࡍࡿഴྥࡀࡳࡽࢀࡓࠋ୍᪉ࠊ࣡ࢡࢳࣥ2 ᅇ┠᥋✀ᚋ 15㹼28 ᪥࡛ࡣࠊྜ᱁࡞ࡿࡣ࡞ࡗࡓࠋ࡞࠾ࠊእᅜ〇࣡ࢡࢳࣥ⩌ࡘ࠸࡚ࡣࠊ᥇⾑
18 ᮇྜ᱁⋡ࡢ㛫┦㛵ᛶࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ 4. ࣮ࣝࢳࣥࢿࢥ᳨య࠾ࡅࡿ୰ᢠయ౯ཬࡧᢠయ᳨ᰝྜ≧ἣ ࣡ࢡࢳࣥ᥋✀㛫㝸୰ᢠయ౯㸦ᗄఱᖹᆒ್㸧ࡢศᕸཬࡧᢠయ᳨ᰝྜ᱁⋡ ࠾ࡅࡿゎᯒ⤖ᯝࢆ⾲1㸫2A㸦᪥ᮏ〇࣡ࢡࢳࣥ㸧ཬࡧ⾲ 1㸫2B㸦እᅜ〇࣡ࢡࢳࣥ㸧 ♧ࡋࡓࠋ᪥ᮏ〇࣡ࢡࢳࣥ⩌ࡘ࠸࡚ࡣࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ 30㹼90 ᪥࡛ࡣࠊ ࣡ࢡࢳࣥ2 ᅇ┠᥋✀ᚋ 8㹼14 ᪥㏆ᢠయ౯ࡢࣆ࣮ࢡࡀㄆࡵࡽࢀࡓࠋ࣡ࢡࢳࣥ ᥋✀㛫㝸ࡀ91 ᪥௨ୖ࡞ࡿࠊࣆ࣮ࢡࡣ 22㹼28 ᪥㏆㐜ࡃ࡞ࡿഴྥࡀࡳࡽ ࢀࡓࠋ እᅜ〇࣡ࢡࢳࣥ⩌ࡘ࠸࡚ࡣࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ30㹼90 ᪥࡛ࡣࠊ࣡ࢡࢳࣥ 2 ᅇ┠᥋✀ᚋ 8㹼28 ᪥㏆ᢠయ౯ࡢࣆ࣮ࢡࡀㄆࡵࡽࢀࡓࠋ࣡ࢡࢳࣥ᥋✀㛫㝸 ࡀ91 ᪥௨ୖ࡛ࡣࠊࣆ࣮ࢡࡣ 29㹼49 ᪥㏆㐜ࡃ࡞ࡿഴྥࡀࡳࡽࢀࡓࠋ ᪥ᮏ〇࣡ࢡࢳࣥ⩌࠾ࡅࡿᢠయ᳨ᰝྜ᱁⋡ࡣࠊ࠸ࡎࢀࡢ࣡ࢡࢳࣥ᥋✀㛫㝸 ࡶ࣡ࢡࢳࣥ2 ᅇ┠᥋✀┤ᚋࡀ 7.9㹼30.6㸣㧗ࡃ㸦᥋✀㛫㝸 30㹼40 ᪥ཬࡧ ྠ91 ᪥௨ୖ㸸p㸺0.01㸧ࠊ1㹼7 ᪥ᚋ࠾࠸࡚ࡶ 4.0㹼8.6㸣࡛࠶ࡗࡓࠋ࡞࠾ࠊእᅜ 〇࣡ࢡࢳࣥ⩌ࡘ࠸࡚ࡣࠊ᥇⾑ᮇྜ᱁⋡ࡢ㛫┦㛵ᛶࡣㄆࡵࡽࢀ࡞ࡗ ࡓࠋ
19 ⪃ᐹ ᪥ᮏࡢ≬≟᳨ไᗘᚑ࠸ࠊࢾཬࡧࢿࢥࡢ୰ᢠయ᥎⛣ࢆㄪᰝࡋࡓᩥ⊩ ࡣࠊỤࡽ㸦4, 5㸧ࡢሗ࿌ࡢࡳ࡛࠶ࡿࠋᮏヨ㦂ࡽᚓࡽࢀࡓᐇ㦂⏝ࢾཬࡧᐇ㦂 ⏝ࢿࢥ࠾ࡅࡿᢠయ᥎⛣ࡣࠊࡇࢀࡽࡢሗ࿌ᴫࡡ୍⮴ࡍࡿࡇࡀ☜ㄆࡉࢀࡓ㸦ᅗ 1㸫2Aࠊ1㸫2Bࠊ1㸫5A ཬࡧ 1㸫5B㸧ࠋࡓࡔࡋࠊỤࡽ㸦4, 5㸧ࡢሗ࿌࡛ࡣ 3 ᅇ┠ ࡢ࣡ࢡࢳࣥ᥋✀ᚋ 1 ࣧ᭶┠ࡢ⤖ᯝࢆࡶࡗ࡚⤊ࡋ࡚࠸ࡿࡓࡵࠊࡑࢀ௨㝆ࡢᢠయ ᥎⛣ࡘ࠸࡚ࡣࠊᮏヨ㦂࡛ᚓࡽࢀࡓ⤖ᯝࡀ၏୍࡛࠶ࡿࠋᐇ㦂ື≀ࢆ⏝࠸ࡓᮏヨ㦂 ⤖ᯝࡽࠊࢾཬࡧࢿࢥࡶ2 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋࠊᴫࡡ 7㹼14 ᪥㏆ ᢠయ౯ࡢࣆ࣮ࢡࡀ࠶ࡿࡇࡀุ᫂ࡋࡓ㸦ᅗ1㸫1Aࠊ1㸫1Bࠊ1㸫4A ཬࡧ 1㸫4B㸧ࠋ ࡞࠾ࠊB ♫〇ࡢ࣡ࢡࢳࣥࡀ᥋✀ࡉࢀࡓࢿࢥ㸦ಶయ No.2㸧࠾࠸࡚ࠊ2 ᅇ┠ࡢ࣡ ࢡࢳࣥ᥋✀ᚋ4 ཬࡧ 5 ࣨ᭶┠ࡢⅬ࡛ 0.5IU/mL ࢆୗᅇࡗࡓ㸦ᅗ 1㸫4B㸧ࠋࡇࡢ ⌮⏤ࡣ᫂ࡽ࡛ࡁ࡞ࡗࡓࡀࠊᙜヱಶయࡣࠊࡢࢿࢥẚࠊ1 ᅇ┠ཬࡧ 2 ᅇ ┠࣡ࢡࢳࣥ᥋✀ᚋࡢᢠయୖ᪼ࡀపࡗࡓࡇࡽࠊච⬟ࡢప࠸ಶయ࡛࠶ࡗࡓ ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽࠊឤᰁ㜵ᚚ᭱ప㝈ᚲせ࡞ᢠయ౯ࡀ0.1IU/mL ௨ୖ ࡛࠶ࡿࡇ㸦24㸧ཬࡧ 3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࡼࡾ༑ศ࡞ᢠయୖ᪼ࡀㄆࡵࡽࢀ ࡓࡇࡽࠊಶయNo.2 ࡢࢿࢥࡘ࠸࡚ࡶ㜵ୖࡢၥ㢟ࡣ࡞࠸ࡶࡢ⪃࠼ࡽࢀࡓࠋ Kennedy ࡽ㸦38㸧ࡢሗ࿌ࡼࢀࡤࠊࢾ࠾࠸࡚ࠊ≬≟ᢠయ᳨ᰝࡢྜ᱁⋡ ࡀ᭱ࡶ㧗ࡃ࡞ࡿ᥇⾑ᮇࡣࠊ≬≟࣡ࢡࢳࣥ᥋✀ᚋ 30 ᪥┠㏆ࡉࢀ࡚࠸ࡿࠋ ࡲࡓࠊMansfield ࡽ㸦45㸧ࡢሗ࿌࡛ࡣࠊྠᵝࡢ᥇⾑ᮇࡘ࠸࡚ࠊࢾ࡛ࡣ≬≟ ࣡ࢡࢳࣥ᥋✀ᚋ1㹼4 㐌┠ࠊࢿࢥ࡛ࡣ 4㹼5 㐌┠ࢆ᥎ዡࡋ࡚࠸ࡿࠋᅇࡢヨ㦂 ࡛ࡣࠊࢾཬࡧࢿࢥࡶ1 ᅇ┠᥋✀ᚋ 2㹼3 㐌ࠊ2 ᅇ┠᥋✀ᚋ 1㹼2 㐌࡛᭱ࡶ ᢠయ౯ࡀ㧗ࡃ࡞ࡿഴྥࡀࡳࡽࢀ㸦ᅗ1㸫1Aࠊ1㸫1Bࠊ1㸫4A ཬࡧ 1㸫4B㸧ࠊ᪤ሗ ࡢ◊✲⤖ᯝࡢ㛫࡛㞳ࡀ࠶ࡗࡓࠋࡇࡢཎᅉࡣࠊࡇࢀࡽࡢሗ࿌࠾࠸࡚ࠊ౪ヨ
20 ື≀ࡢᵓᡂᖺ㱋ཬࡧ≬≟࣡ࢡࢳࣥࡢ᥋✀ᅇᩘࡀ⤫୍ࡉࢀ࡚࠸࡞࠸ࡇ㉳ᅉ ࡍࡿ⪃࠼ࡽࢀࡿࠋᅇࠊᐇ㦂⏝ࢾཬࡧࢿࢥࡶ≬≟࣡ࢡࢳࣥࡢ᥋✀㛫㝸 ཬࡧ᥋✀ᅇᩘࢆ㝈ᐃࡋࡓࡇ࡛ࠊࡼࡾಙ㢗ᗘࡢ㧗࠸ࢹ࣮ࢱࢆᥦ౪࡛ࡁࡓ⪃࠼ ࡿࠋ ᐇ㦂⏝ࢾཬࡧᐇ㦂⏝ࢿࢥ࠾ࡅࡿᢠయ᥎⛣ࡢ⤖ᯝࡽࠊ᳨つ๎ᚑࡗ࡚ 30 ᪥௨ୖࡢ㛫㝸࡛≬≟ά࣡ࢡࢳࣥࢆ 2 ᅇ᥋✀ࡋࠊࡑࡢᚋ 1 ᖺ௨ෆ 3 ᅇ ┠ࡢ࣡ࢡࢳࣥ᥋✀ࢆࡍࡿࡇࡼࡾࠊ᥇⾑᪥ࡽ 2 ᖺ௨ୖࠊឤᰁ㜵ᚚᚲせ࡞ ᢠయ౯ࡀ⥔ᣢࡉࢀࡿࡇࡀ♧ࡉࢀࡓ㸦ᅗ 1㸫2Aࠊ1㸫2Bࠊ1㸫5A ཬࡧ 1㸫5B㸧ࠋ ࡇࡢࡇࡣࠊ᳨つ๎࠾࠸࡚ᚲせ࡞ࡿ≬≟ᢠయ౯ࢆᜏᖖⓗ☜ಖࡍࡿୖ ࡛ࠊᖺ 1 ᅇࡢ࣡ࢡࢳࣥ᥋✀ࡀ㐺ษ࡛࠶ࡿࡇࡢጇᙜᛶࢆࡅࡿࡶࡢ࡛࠶ࡿࠋ ࡲࡓࠊỤࡽ㸦4, 5㸧ࡢሗ࿌ྠᵝࠊ3 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ࡼࡾࠊᢠయ౯ࡀ ẚ㍑ⓗప್࡛᥎⛣ࡋ࡚࠸ࡓಶయࡘ࠸࡚ࡶ༑ศ࡞ࣈ࣮ࢫࢱ࣮ຠᯝࡀᚓࡽࢀࡓ 㸦ᅗ1㸫3Aࠊ1㸫3Bࠊ1㸫6A ཬࡧ 1㸫6B㸧ࠋࡇࡢ⌧㇟ࡣࠊ≬≟࣡ࢡࢳࣥ࠾ࡅ ࡿ㏣ຍ᥋✀ࡢ㔜せᛶࢆㄆ㆑ࡉࡏࡿࡶࡢ࡛࠶ࡿࠋ ࣮ࣝࢳࣥࢾཬࡧ࣮ࣝࢳࣥࢿࢥ᳨యࢆ⏝࠸ࡓゎᯒ⤖ᯝࡽࠊ≬≟ά࣡ ࢡࢳࣥࡢ᥋✀㛫㝸୰ᢠయ౯ࡢࣆ࣮ࢡᮇࡣḟࡢ㛵ಀࡀ࠶ࡿࡶࡢ⪃࠼ࡽ ࢀࡓࠋࡍ࡞ࢃࡕࠊࢾཬࡧࢿࢥࡶ᪥ᮏ〇ࠊእᅜ〇࣡ࢡࢳࣥ⩌ࢆၥࢃࡎࠊ࣡ࢡ ࢳࣥ᥋✀㛫㝸ࡀ30㹼90 ᪥࠾ࡅࡿᢠయ౯ࣆ࣮ࢡࡣࠊ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 8 ᪥ ┠௨㝆Ꮡᅾࡋࡓ㸦⾲1㸫1Aࠊ1㸫1Bࠊ1㸫2A ཬࡧ 1㸫2B㸧ࠋࡲࡓࠊࢾ㸫እᅜ 〇࣡ࢡࢳࣥ⩌࡛ࡣࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ91 ᪥௨ୖ࠾࠸࡚ࡶࠊྠᵝࡢഴྥࡀࡳ ࡽࢀࡓࠋࡋࡋ࡞ࡀࡽࠊࢾ㸫᪥ᮏ〇࣡ࢡࢳࣥ⩌࠾࠸࡚ࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ 91 ᪥௨ୖࡢሙྜ࡛ࡣࠊ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 15 ᪥௨㝆ᢠయ౯ࣆ࣮ࢡࡀᏑᅾࡋ ࡓࠋ୍᪉ࠊࢿࢥ࠾࠸࡚࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ91 ᪥௨ୖࡢሙྜ࡛ࡣࠊ᪥ᮏ〇ࠊእ ᅜ〇࣡ࢡࢳࣥ⩌ࢆၥࢃࡎࠊ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ 22 ᪥┠௨㝆ᢠయ౯ࡢࣆ࣮ࢡ
21 ࡀᏑᅾࡋࡓࠋࢾཬࡧࢿࢥࡶࠊᢠయ౯ࡢࣆ࣮ࢡࡽ1 㐌ᚋࡅ࡚ࡣࠊᢠయ ᳨ᰝࡢྜ᱁⋡ࡀᴫࡡప࠸ഴྥࡀ♧ࡉࢀࡓࠋࡋࡋ࡞ࡀࡽࠊKennedy ࡽ㸦38㸧 ࡢሗ࿌ྠᵝࠊ2 ᅇ┠࣡ࢡࢳࣥ᥋✀ࡢ┤ᚋ᥇⾑ࢆᐇࡋࡓሙྜࡣࠊྜ᱁⋡ ࡀ㧗࠸ࡇࡀ♧ࡉࢀࡓࠋ࣮ࣝࢳࣥࢾཬࡧ࣮ࣝࢳࣥࢿࢥ᳨యࡶࠊ࣡ࢡࢳࣥ2 ᅇ┠᥋✀┤ᚋ࠾ࡅࡿᢠయ౯ࡣࠊ᪥ᮏ〇ࡼࡾࡶእᅜ〇࣡ࢡࢳࣥ᥋✀⩌࠾࠸࡚ 㧗࠸್ࡀ♧ࡉࢀࡓࠋࡇࡢせᅉࡋ࡚ࡣࠊ࣡ࢡࢳࣥ〇㐀⏝࢘ࣝࢫᰴࡢ㐪࠸እᅜ 〇࣡ࢡࢳࣥྵࡲࢀࡿࢪࣗࣂࣥࢺࡢస⏝ࡢ㸰ࡘࡀ⪃࠼ࡽࢀࡿࠋ≬≟࢘ࣝ ࢫࡣ⾑Ύᆺࡀ༢୍࡛࠶ࡿࡇࡽࠊ୧࣡ࢡࢳࣥ᥋✀⩌㛫࡛ࡳࡽࢀࡓᢠయ౯ࡢᕪ ࡀ࣡ࢡࢳࣥ〇㐀ᰴࡢᢠཎᛶࡢ㐪࠸ࡼࡿࡣ⪃࠼ࡃ࠸ࠋᚑࡗ࡚ࠊእᅜ〇࣡ࢡࢳ ࣥࡢ࠺ࡀ᪥ᮏ〇࣡ࢡࢳࣥ᥋✀⩌ࡼࡾ㧗࠸ᢠయ౯ࢆ♧ࡋࡓഴྥࡢ⌮⏤ࡋ࡚ࠊ ࣓࣮࣮࢝㛫࠾ࡅࡿ࣡ࢡࢳࣥ〇㐀ᕤ⛬ࡢᕪ␗࡞ࡶྰᐃ࡛ࡁ࡞࠸ࡶࡢࡢࠊእᅜ 〇࣡ࢡࢳࣥྵࡲࢀࡿࢪࣗࣂࣥࢺࡢస⏝ຠᯝࡼࡿࡶࡢࡀࡁ࠸᥎ ࡉࢀ ࡓࠋ ࣮ࣝࢳࣥࢾཬࡧ࣮ࣝࢳࣥࢿࢥ᳨య࠾࠸࡚ࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ30㹼40 ᪥ ࡢሙྜࡳࡽࢀࡿᢠయ౯ࡢࣆ࣮ࢡ㸦2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋࠊᴫࡡ 8㹼14 ᪥㏆㸧 ࡣࠊᐇ㦂⏝ࢾཬࡧᐇ㦂⏝ࢿࢥ࠾ࡅࡿᢠయ౯ࡢࣆ࣮ࢡ㸦2 ᅇ┠࣡ࢡࢳࣥ᥋✀ᚋࠊ ᴫࡡ 7㹼14 ᪥㏆㸧ࡰ୍⮴ࡋ࡚࠾ࡾࠊᐇ㦂ᐊෆ࡛ࡢヨ㦂⤖ᯝࡀ㔝እ࡛ࡢᢠ య᳨ᰝᡂ⦼ࡽࡶ☜ㄆࡉࢀࡓࠋ୍᪉ࠊ࣮ࣝࢳࣥࢾཬࡧ࣮ࣝࢳࣥࢿࢥ᳨యࡢ⤖ᯝ ࡽࠊ࣡ࢡࢳࣥ᥋✀㛫㝸ࡀ91 ᪥௨ୖ࡛ࡣᢠయ౯ࡢࣆ࣮ࢡࡀᑡ࡞ࡃࡶ 1 㐌⛬ᗘ 㐜ࡃ࡞ࡿഴྥࡀㄆࡵࡽࢀࡓࡀࠊ㐌༢࡛ᢠయ౯ࢆ㏣㊧ࡋࡓྠᵝࡢሗ࿌ࡀ࡞࠸ ࡓࡵࠊࡑࡢ⌮⏤ࡘ࠸࡚ࡣ࡛᫂࠶ࡿࠋ ௨ୖࠊ≬≟ᑐࡍࡿᢠయ᳨ᰝ࠾࠸࡚㐺ṇ࡞᥇⾑ᮇࢆ᳨ウࡋࡓ⤖ᯝࠊ㸦ϸ㸧 ࢾཬࡧࢿࢥࡶࠊ≬≟࣡ࢡࢳࣥࡢ᥋✀㛫㝸ࡀ 30㹼90 ᪥⛬ᗘࡢሙྜࡣࠊ2 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋࠊ7㹼21 ᪥ࢆ┠Ᏻ᥇⾑ࡍࡿࡇ㸦Ϲ㸧ࢾཬࡧࢿࢥࡶ
22 ࠊ≬≟࣡ࢡࢳࣥࡢ᥋✀㛫㝸ࡀ91 ᪥௨ୖࡢሙྜࡣࠊ2 ᅇ┠ࡢ࣡ࢡࢳࣥ᥋✀ᚋࠊ 14㹼21 ᪥ࢆ┠Ᏻ᥇⾑ࡍࡿࡇࡀ᥎ዡࡉࢀࡿࠋᮏヨ㦂ࡢ࠺ࡕࠊᢠయ᥎⛣ヨ㦂࡛ ࡣࠊỤࡽ㸦4, 5㸧ࡢヨ㦂ྠᵝࠊࢾࡢᐇ㦂ࣔࢹࣝࡋ࡚ࣅ࣮ࢢࣝ✀ࠊࢿࢥࡢ ᐇ㦂ࣔࢹࣝࡋ࡚㞧✀ࢆ⏝࠸ࡓࠋࡇࢀࡽࡢᐇ㦂⏝ື≀ࡣࠊᗈࡃࢾཬࡧࢿࢥヨ㦂 ࠾ࡅࡿᶆ‽ࣔࢹࣝࡋ࡚⏝ࡉࢀ࡚࠸ࡿࡇࡽࠊᮏヨ㦂࡛ᚓࡽࢀࡓ⤖ᯝࡣࠊ ᵝࠎ࡞≟✀ཬࡧ⊧✀ᑐࡋ࡚㐺⏝ྍ⬟࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋ ୍⯡ࠊࢿࢥࡣࢾẚ࡚ᨷᧁᛶࡀప࠸ࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋᐇ㝿ࠊ㝔ࡢ ညയᨾࡼࡾ㝔᮶㝔ࡍࡿᝈ⪅ࡢෆヂࡣࠊࢾࡼࡿࡶࡢࡀ 8 ࠊḟ࠸࡛ ࢿࢥࡀ 1 ࡞ࡗ࡚࠸ࡿ㸦ᅜ❧㝔ᶵᵓ⇃ᮏ་⒪ࢭࣥࢱ࣮㸸http://www.nho-kumamoto.jp/kusu-press/kusu-169-01.html㸧ࠋ୍᪉࡛ࠊࢿࢥࡢ∳ࡣ⣽ࡃ㗦࠸ࡓ ࡵࠊឤᰁࢆᘬࡁ㉳ࡇࡍ☜⋡ࡣࢿࢥࡢ࠺ࡀࢾࡼࡾ10 ಸ⛬ᗘ㧗࠸⪃࠼ࡽࢀ ࡚࠸ࡿࠋࡲࡓࠊእᅜ࠾࠸࡚ࢥ࢘ࣔࣜࢆࡋࠊࢿࢥ≬≟࢘ࣝࢫࡀఏࡋࡓ ࡀᏑᅾࡍࡿ㸦14, 32, 51㸧ࠋᚑࡗ࡚ࠊὶ⾜ࡢほⅬࡽࠊࢿࢥࡣࢾࡼࡾࡶࣜࢫ ࢡࡀప࠸ࡶࡢࡢࠊࣄࢺࡢ㌟㏆Ꮡᅾࡍࡿࡇࡽࠊࢿࢥࡀ≬≟ࢆࣄࢺఏࡍ ࡿࣜࢫࢡࡣྰᐃ࡛ࡁ࡞࠸ࠋࡇࡢࡓࡵ࣓ࣜ࢝࡞࡛ࡣࠊࢾࡔࡅ࡛࡞ࡃࢿࢥࡶ࣡ ࢡࢳࣥ᥋✀ࡢᑐ㇟ື≀࡞ࡗ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊࢿࢥᑐࡍࡿ≬≟࣡ࢡࢳ ࣥࡢ᭷ຠᛶࢆホ౯ࡋࡓሗ࿌ࡣࢇ࡞࠸ࠋࡇࡢࡓࡵࠊᮏヨ㦂࠾࠸࡚ࠊ≬≟ ά࣡ࢡࢳࣥᑐࡍࡿࢿࢥࡢᢠయᛂ⟅ࢆヲ⣽ㄪᰝࡋࠊࢾࡰྠࡌ࡛࠶ ࡿࡇࢆ♧ࡋࡓࡇࡣព⩏῝࠸⪃࠼ࡽࢀࡿࠋ ᮏ❶࡛ࡣࠊࢾࢿࢥ࠾࠸࡚ࠊࡇࢀࡲ࡛᫂࡞Ⅼࡀከࡗࡓ≬≟ά࣡ ࢡࢳࣥᑐࡍࡿᢠయᛂ⟅ࡘ࠸࡚ࠊᐇ㦂ື≀ཬࡧᐇ㝿ࡢ᳨యࢆ⏝࠸࡚ヲ⣽᳨ ウࡋࡓࠋ≉ࢿࢥࡘ࠸࡚ࡣࠊ≬≟࣡ࢡࢳࣥᑐࡍࡿᢠయᛂ⟅ࢆㄪᰝࡋࡓ㈗㔜 ࡞ሗ࿌࡛࠶ࡿࠋᮏ❶࡛ᚓࡽࢀࡓ▱ぢࡣࠊ≬≟࣡ࢡࢳࣥࡢ◊✲࣭〇㐀㛵ಀ⪅ࡔ ࡅ࡛࡞ࡃࢾࡸࢿࢥࢆక࠺ᵝࠎ࡞ே㛫άື࠾࠸࡚ࠊᴟࡵ࡚᭷⏝࡞ሗ࡞ࡿ
23
24 ㄪᰝ㡯┠ཬ䜃 ᳨యᩘ 0 1 䡚 78 䡚 14 15 䡚 21 22 䡚 28 29 䡚 49 50 䡚 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 7. 0 ± 6. 9 23. 9 ± 6. 0 41. 3 ± 3. 9 27. 5 ± 4. 8 26. 5 ± 3. 9 21. 2 ± 4. 3 19. 8 ± 5. 4 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 5 .4 a 3. 6 1. 3 0. 0 0. 0 1. 3 3. 3 ᳨యᩘ 93 140 224 95 53 75 61 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 1. 9 ± 6. 5 12. 2 ± 7. 1 39. 0 ± 4. 8 31. 3 ± 4. 4 20. 9 ± 6. 2 23. 2 ± 5. 1 14. 3 ± 5. 4 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 28. 9 a, b 5. 9 3.4 0. 0 0.0 2. 6 0.0 ᳨యᩘ 45 34 59 36 15 39 23 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 1. 5 ± 6. 3 19. 6 ± 7. 2 24. 1 ± 6. 1 52. 3 ± 2. 8 33. 4 ± 4. 4 34. 1 ± 3. 8 7. 4 ± 8. 9 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 29. 0 a, b 4. 2 2. 7 0. 0 0. 0 0. 0 11. 1 ᳨యᩘ 31 24 37 28 7 16 9 a 䠖 䝽 䜽 䝏 䞁 㻞 ᅇ ┠᥋✀᪥䇲 ᥇ ⾑᪥㛫㝸䛜㻝 ᪥௨ୖ䛾᳨య䛻 䛚 䛡 䜛 ྜ᱁⋡䛸 䛾㛫䛻 ᭷ ពᕪ䛒 䜚 䠄 㼜 䠘 㻜 㻚㻜㻝䠅 b 䠖 䝽 䜽 䝏 䞁 ᥋✀㛫㝸䛜㻟 㻜 䡚 㻠 㻜 ᪥䛾᳨య䛻 䛚 䛡 䜛 ྜ᱁⋡䛸 䛾㛫䛻 ᭷ ពᕪ䛒 䜚 㻔 㼜 䠘 㻜 㻚㻜㻝䠅 91 ᪥௨ୖ ⾲ 1-1A 䚷᪥ᮏ〇≬≟䝽 䜽䝏 䞁䜢 ᥋✀䛥 䜜 䛯 䝹 䞊 䝏 䞁䜲 䝚 ᳨ య䛻 䛚 䛡 䜛 ≬≟䜴 䜲 䝹䝇 ୰ᢠయ౯ཬ䜃 ≬ ≟ᢠయ᳨ᰝྜ᱁⋡ ≬≟䝽 䜽 䝏 䞁 2 ᅇ┠᥋✀᪥䠉᥇⾑᪥㛫㝸䠄 ᪥䠅 䝽䜽䝏 䞁 ᥋✀㛫㝸 䠄㻝ᅇ ┠ 㻙㻞ᅇ ┠ 䠅 30 䡚 40 ᪥ 41 䡚 90 ᪥
25 ㄪᰝ㡯┠ཬ䜃 ᳨యᩘ 0 1 䡚 78 䡚 14 15 䡚 21 22 䡚 28 29 䡚 49 50 䡚 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 18. 1 ± 2. 7 18. 7 ± 7. 1 49. 8 ± 3. 0 36. 7 ± 3. 9 17. 8 ± 4. 7 26. 7 ± 3. 5 9. 0 ± 4. 6 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 0. 05 .0 0. 01 .1 4. 01 .5 7. 1 ᳨యᩘ 14 140 121 93 25 69 42 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 15. 9 ± 9. 1 4. 8 ± 7. 4 70. 2 ± 2. 1 27. 9 ± 5. 7 10. 6 ± 14. 8 19. 0 ± 3. 5 17. 4 ± 4. 3 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 0. 0 10. 5 0. 0 5. 6 20. 0 0. 0 0. 0 ᳨యᩘ 5 19 8 18 15 12 16 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 9. 4 ± 7. 3 30. 8 ± 4. 2 44. 1 ± 4. 1 43. 9 ± 5. 4 24. 5 ± 4. 7 11. 0 ± 4. 6 11. 5 ± 3. 1 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 0. 0 0.0 0. 0 0.0 0. 0 0.0 0. 0 ᳨యᩘ 74 16 7677 30 䡚 40 ᪥ 91 ᪥௨ୖ 41 䡚 90 ᪥ ⾲ 1-1B 䚷እᅜ〇≬≟䝽 䜽䝏 䞁䜢 ᥋✀䛥 䜜 䛯 䝹 䞊 䝏 䞁䜲 䝚 ᳨ య䛻 䛚 䛡 䜛 ≬≟䜴 䜲 䝹䝇 ୰ᢠయ౯ཬ䜃 ≬ ≟ᢠయ᳨ᰝྜ᱁⋡ 䝽䜽䝏 䞁 ᥋✀㛫㝸 䠄㻝ᅇ ┠ 㻙㻞ᅇ ┠ 䠅 ≬≟䝽 䜽 䝏 䞁 2 ᅇ┠᥋✀᪥䠉᥇⾑᪥㛫㝸䠄 ᪥䠅
26 ㄪᰝ㡯┠ཬ䜃 ᳨యᩘ 0 1 䡚 78 䡚 14 15 䡚 21 22 䡚 28 29 䡚 49 50 䡚 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 4. 1 ± 6. 8 13. 4 ± 5. 2 25. 7 ± 4. 8 20. 2 ± 5. 7 13. 1 ± 6. 0 17. 1 ± 5. 1 7. 1 ± 8. 2 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 16. 6 a 8. 6 2. 7 5. 6 4. 6 3. 4 14. 5 ᳨యᩘ 145 256 475 214 87 176 62 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 8. 4 ± 6. 6 17. 2 ± 6. 7 34. 7 ± 4. 2 26. 2 ± 5. 8 29. 6 ± 4. 2 21. 9 ± 6. 1 13. 5 ± 5. 2 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 7. 9 7. 0 1. 2 3. 7 3. 3 3. 6 2. 7 ᳨యᩘ 38 43 81 54 30 55 37 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 3. 3 ± 8. 9 20. 8 ± 6. 9 24. 9 ± 6. 5 27. 2 ± 5. 5 32. 5 ± 6. 9 26. 6 ± 4. 7 13. 2 ± 7. 3 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 3 0. 6 a 4. 0 6.1 4. 6 5.9 0. 0 8.7 ᳨యᩘ 36 25 66 22 17 17 46 a 䠖 䝽 䜽 䝏 䞁 㻞 ᅇ ┠᥋✀᪥䇲 ᥇ ⾑᪥㛫㝸䛜㻝 ᪥௨ୖ䛾᳨య䛻 䛚 䛡 䜛 ྜ᱁⋡䛸 䛾㛫䛻 ᭷ ពᕪ䛒 䜚 䠄 㼜 䠘 㻜 㻚㻜㻝䠅 ⾲ 1-2A 䚷᪥ᮏ〇≬≟䝽 䜽䝏 䞁䜢 ᥋✀䛥 䜜 䛯 䝹 䞊 䝏 䞁䝛 䝁᳨య䛻 䛚 䛡 䜛 ≬≟䜴 䜲 䝹䝇 ୰ᢠయ౯ཬ䜃 ≬ ≟ᢠయ᳨ᰝྜ᱁⋡ 41 䡚 90 ᪥ 91 ᪥௨ୖ 30 䡚 40 ᪥ ≬≟䝽 䜽 䝏 䞁 2 ᅇ┠᥋✀᪥䠉᥇⾑᪥㛫㝸䠄 ᪥䠅 䝽䜽䝏 䞁 ᥋✀㛫㝸 䠄㻝ᅇ ┠ 㻙㻞ᅇ ┠ 䠅
27 ㄪᰝ㡯┠ཬ䜃 ᳨యᩘ 0 1 䡚 78 䡚 14 15 䡚 21 22 䡚 28 29 䡚 49 50 䡚 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 20. 7 ± 4. 9 22. 3 ± 5. 1 34. 9 ± 3. 9 38. 7 ± 3. 5 33. 7 ± 3. 7 30. 2 ± 3. 6 18. 7 ± 4. 2 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 4. 71 .1 1. 10 .8 1. 20 .9 2. 8 ᳨యᩘ 64 88 179 127 84 116 144 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 26. 4 ± 4. 9 23. 4 ± 3. 6 53. 0 ± 3. 1 35. 7 ± 3. 0 48. 1 ± 2. 8 31. 0 ± 3. 4 19. 9 ± 5. 4 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 0. 0 0. 0 0. 0 0. 0 0. 0 0. 0 4. 7 ᳨యᩘ 21 20 58 35 16 39 42 ୰ᢠయ౯䠄 IU /m L 䠅 䠄ᗄ ఱ ᖹ ᆒ ± ᗄఱᶆ‽೫ᕪ䠅 7. 6 ± 5. 6 20. 4 ± 4. 0 27. 2 ± 4. 1 23. 4 ± 3. 9 34. 8 ± 2. 7 39. 0 ± 3. 2 21. 5 ± 3. 8 ᢠయ᳨ᰝྜ᱁⋡䠄 䠂䠅 5. 0 0. 0 0. 0 0. 0 0. 0 0. 0 0. 0 ᳨యᩘ 20 20 33 35 27 41 93 91 ᪥௨ୖ 41 䡚 90 ᪥ ⾲ 1-2B 䚷እᅜ〇≬≟䝽 䜽䝏 䞁䜢 ᥋✀䛥 䜜 䛯 䝹 䞊 䝏 䞁䝛 䝁᳨య䛻 䛚 䛡 䜛 ≬≟䜴 䜲 䝹䝇 ୰ᢠయ౯ཬ䜃 ≬ ≟ᢠయ᳨ᰝྜ᱁⋡ 䝽䜽䝏 䞁 ᥋✀㛫㝸 䠄㻝ᅇ ┠ 㻙㻞ᅇ ┠ 䠅 ≬≟䝽 䜽 䝏 䞁 2 ᅇ┠᥋✀᪥䠉᥇⾑᪥㛫㝸䠄 ᪥䠅 30 䡚 40 ᪥
28 ᅗ $ $ ♫ 〇 ࡢ ≬ ≟ ά ࣡ࢡ ࢳࣥ ࢆ᥋ ✀ࡋ ࡓ ࢾ 㢌 㸦 ࢾ ࣡ ࢡ ࢳ ࣥ $ ⩌ 㸧 ࠾ࡅ ࡿ୰ ᢠ య౯ࡢ ᥎⛣ 㸦 ࣡ ࢡࢳࣥ ᅇ ┠ ᥋ ✀ ┤ ๓ ࣡ࢡ ࢳࣥ ᅇ ┠᥋ ✀ ᪥ ᚋ㸧
29 ᅗ % % ♫ 〇 ࡢ ≬ ≟ ά ࣡ࢡ ࢳࣥ ࢆ᥋ ✀ࡋ ࡓ ࢾ 㢌 㸦 ࢾ ࣡ ࢡ ࢳ ࣥ % ⩌ 㸧 ࠾ࡅ ࡿ୰ ᢠ య౯ࡢ ᥎⛣ 㸦 ࣡ ࢡࢳࣥ ᅇ ┠ ᥋ ✀ ┤ ๓ ࣡ࢡ ࢳࣥ ᅇ ┠᥋ ✀ ᪥ ᚋ㸧
30 ᅗ $ ࢾ ࣡ࢡ ࢳࣥ $ ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ヨ 㦂ᮇ 㛫 ࣨ ᭶ 㛫 㸧
31 ᅗ % ࢾ ࣡ࢡ ࢳࣥ % ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ヨ 㦂ᮇ 㛫 ࣨ ᭶ 㛫 㸧
32 ᅗ $ ࢾ ࣡ࢡ ࢳࣥ $ ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ࣡ࢡ ࢳ ࣥ ᅇ ┠᥋ ✀┤๓ ᥋ ✀ ᪥ ᚋ 㸧
33 ᅗ % ࢾ ࣡ ࢡ ࢳ ࣥ % ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ࣡ࢡ ࢳ ࣥ ᅇ ┠᥋ ✀┤๓ ᥋ ✀ ᪥ ᚋ 㸧
34 ᅗ $ $ ♫ 〇 ࡢ ≬ ≟ ά ࣡ࢡ ࢳࣥ ࢆ᥋ ✀ࡋ ࡓࢿ ࢥ 㢌 㸦 ࢿ ࢥ ࣡ ࢡ ࢳ ࣥ $ ⩌ 㸧 ࠾ࡅ ࡿ୰ ᢠ య౯ࡢ ᥎⛣ 㸦 ࣡ ࢡࢳࣥ ᅇ ┠ ᥋ ✀ ┤ ๓ ࣡ࢡ ࢳࣥ ᅇ ┠᥋ ✀ ᪥ ᚋ㸧
35 ᅗ % % ♫ 〇 ࡢ ≬ ≟ ά ࣡ࢡ ࢳࣥ ࢆ᥋ ✀ࡋ ࡓࢿ ࢥ 㢌 㸦 ࢿ ࢥ ࣡ ࢡ ࢳ ࣥ % ⩌ 㸧 ࠾ࡅ ࡿ୰ ᢠ య౯ࡢ ᥎⛣ 㸦࣡ࢡ ࢳ ࣥ ᅇ ┠᥋ ✀┤ ๓࣡ ࢡ ࢳ ࣥ ᅇ ┠ ᥋ ✀ ᪥ ᚋ 㸧
36 ᅗ $ ࢿࢥ ࣡ࢡ ࢳࣥ $ ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ヨ 㦂ᮇ 㛫 ࣨ ᭶ 㛫 㸧
37 ᅗ % ࢿࢥ ࣡ࢡ ࢳࣥ % ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ヨ 㦂ᮇ 㛫 ࣨ ᭶ 㛫 㸧
38 ᅗ $ ࢿࢥ ࣡ࢡ ࢳࣥ $ ⩌ ࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ࣡ ࢡ ࢳ ࣥ ᅇ ┠ ᥋ ✀ ┤๓ ᥋ ✀ ᪥ ᚋ 㸧
39 ᅗ % ࢿࢥ ࣡ࢡ ࢳࣥ % ⩌࠾ ࡅࡿ ୰ ᢠయ ౯ࡢ ᥎⛣㸦 ࣡ࢡ ࢳ ࣥ ᅇ ┠᥋ ✀┤๓ ᥋ ✀ ᪥ ᚋ 㸧
40
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≬≟ᢠయ᳨ᰝἲ㸦)$91 ἲ㸧࠾ࡅࡿ⣽⬊ẘᛶ
Ⓨ⏕せᅉཬࡧࡑࡢ㝖ཤ᪉ἲ
41 ᗎㄽ ᪥ᮏ࠾࠸࡚ࠊࢾཬࡧࢿࢥࢆ㍺ධࡍࡿ㝿ࡣࠊ≬≟ண㜵ἲᇶ࡙ࡃ᳨ つ๎ࡼࡿつไࢆཷࡅࡿࠋᮏつ๎ࡼࢀࡤࠊ≬≟࣡ࢡࢳࣥࢆ᥋✀ᚋࡢಶయ ࡽ⾑Ύࢆ᥇ྲྀࡋࠊ≬≟୰ᢠయ౯ࢆ ᐃࡍࡿࡇ࡞ࡗ࡚࠸ࡿࠋᏳ◊ࡣࠊ ㎰ᯘỈ⏘┬ࡼࡾᣦᐃࡉࢀࡓ≬≟ᢠయ᳨ᰝタ࡛࠶ࡾࠊᖺ㛫⣙4,000 ௳ࡢ⿕ 㦂⾑Ύࡘ࠸࡚ᢠయ౯ࢆ ᐃࡋ࡚࠸ࡿࠋᏳ◊࡛ࡣࠊOIE ࡢつᐃ㸦57㸧ᇶ࡙ ࡁࠊFAVN ἲࡼࡾᢠయ᳨ᰝࢆᐇࡋ࡚࠸ࡿࠋ Ᏻ◊ࡣࠊࢪࢆ୰ᚰࡋࡓᵝࠎ࡞ᆅᇦࡽ⿕㦂⾑Ύࡀ㏦ࡉࢀࡿࠋࡑ ࢀࡽࡢ⾑Ύࡢ୰ࡣࠊ⣽⳦ቑṪࡸ⁐⾑ࡀㄆࡵࡽࢀࡿࡶࡢࠊ⾑Ύศ㞳ࡀ༑ศ࡞ ࡶࡢ࡞ࠊ≧ែࡢᝏ࠸ࡶࡢࡀᩓぢࡉࢀࡿࠋࡇࡢࡓࡵࠊᏳ◊฿╔ࡋࡓ⿕㦂⾑ Ύࡘ࠸࡚ࡣࠊ᳨ᰝ๓㐲ᚰศ㞳ࢆᐇࡋࠊΰධ⣽⳦ࡸዃ㞧≀ࢆ㝖ཤࡋ࡚࠸ ࡿࠋࡋࡋ࡞ࡀࡽࠊ⣙30㸣ࡢ⿕㦂⾑Ύࡀࠊᢠయ᳨ᰝ㐣⛬ࡢ࢘ࣝࢫ୰ヨ㦂 ࠾࠸࡚ࠊ⣽⬊ẘᛶࢆᘬࡁ㉳ࡇࡍࠋලయⓗࡣࠊ⿕㦂⾑Ύ⃰ᗘࡀ㧗࠸࢙࢘ࣝ࠾ ࠸࡚ࠊ⣽⬊ࡢ༢ᒙᙧᡂࡀ㜼ᐖࡉࢀࡿࡇࡀཎᅉ࡛࠶ࡿࠋᮏ᳨ᰝࡢྜྰุᐃࡣࠊ ⾑Ύᕼ㔘ಸ⋡ࡀẚ㍑ⓗప࠸ࠊ9 ಸ㏆࠾࠸࡚ᐇࡉࢀࡿࠋࡇࡢࡓࡵࠊ⾑Ύ୰ ࡢᢠయ౯ࡀప࠸⿕㦂⾑Ύࠊᚲ↛ⓗ㧗࠸⾑Ύ⃰ᗘ࡛ྜྰุᐃࢆ⾜࠺ࡇ ࡞ࡿࠋ⣽⬊ẘᛶࡀᙉ࠸ሙྜࡣࠊ⣽⬊ࡢቑṪ㜼ᐖࠊจ㞟ཪࡣ㞳ࡀ㉳ࡇࡾࠊ୰ ヨ㦂ࡑࡢࡶࡢࡀᡂ❧ࡋ࡞࠸ࠋࡑࡢ⤖ᯝࠊࢾཬࡧࢿࢥࡶࠊ⣙2㸣ࡢ⿕㦂⾑ Ύ࠾࠸ุ࡚ᐃ⬟࡞ࡗ࡚࠸ࡿࠋࡲࡓࠊFAVN ἲࡢ࢘ࣝࢫ୰ヨ㦂㐣⛬ ࠾࠸࡚ࠊᇵ㣴ࣉ࣮ࣞࢺࡽ⣽⬊ࡀ㞳ࡋࡓሙྜࡣࠊ≬≟࢘ࣝࢫࡢቑṪࡀ ☜ㄆ࡛ࡁ࡞࠸ࠋࡇࡢࡓࡵࠊ࢘ࣝࢫ㝜ᛶุᐃࡉࢀࠊࡑࡢ⤖ᯝࠊᢠయ౯ࡀ㐣 ⟬ฟࡉࢀࡿྍ⬟ᛶࡀ࠶ࡿࠋࡇࢀ㛵㐃ࡋ࡚ࠊ࢘ࣝࢫ୰ヨ㦂᪉ἲࡢᨵⰋ ࡼࡿ⣽⬊ẘᛶ㛵ࡍࡿ᳨ウࡘ࠸࡚ࡣࠊBedeković ࡽ㸦12㸧ࡢሗ࿌ࡀ࠶ࡿࡢࡳ
42 ࡛࠶ࡿࠋᮏሗ࿌࡛ࡣࠊ⣽⬊⿕㦂⾑Ύࢆྠᇵ㣴ࡍࡿ᪉ἲࢆᨵࡵࠊணࡵ⣽⬊ࡢ ࡳࢆࣉ࣮ࣞࢺ࡛ᇵ㣴ࡋ࡚ࣉ࣮ࣞࢺ᥋╔ᛶࢆ༑ศ☜ಖࡋࡓࡢࡕࠊ⿕㦂⾑Ύࢆຍ ࠼ࡿ᪉ἲࢆᥦࡋ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊ⿕㦂⾑Ύ⏤᮶ࡍࡿ⣽⬊ẘᛶࡘ࠸ ࡚ࠊⓎ⏕ண㜵ࡢほⅬࡽ᳨ウࡋࡓሗ࿌ࡣ࡞࠸ࠋ ࡑࡇ࡛ᮏ❶࡛ࡣࠊ≬≟ᢠయ᳨ᰝࡢᐇ⾜⛬ࡈ⣽⬊ẘᛶࡢⓎ⏕せᅉࢆ᳨ ドࡋࠊ⣽⬊ẘᛶࢆᮍ↛㜵Ṇࡍࡿᑐ⟇ࡘ࠸᳨࡚ウࡋࡓࠋࡲࡓࠊᮏ᳨ᰝࡢᐇ ༶ࡋࡓࠊຠᯝⓗ࡞⣽⬊ẘᛶ㝖ཤἲࡘ࠸᳨࡚ウࡋࡓࠋ
43
ᮦᩱཬࡧ᪉ἲ
1. ≬≟࢘ࣝࢫᑐࡍࡿ୰ᢠయࡢ᳨ᰝ㸦FAVN㸧
➨ 1 ❶ྠᵝࡢ᪉ἲࡼࡾࠊFAVN ἲࢆᐇࡋࡓࠋᮏヨ㦂࡛⏝࠸ࡓ BHK-21
⣽⬊ࡣࠊAmerian Type Culture Collection㸦ATCCࠊ⡿ᅜ㸧ࡽ㉎ධࡋࡓࠋ㉎ධ
ᚋࡢ⣽⬊ࡣࠊFCS ࢆ 10 vol 㸣ῧຍࡋࡓ DMEM ࢆ⏝࠸࡚ 37Υࠊ5㸣CO2ࠊ95㸣
✵Ẽࡢ᮲௳ୗ࡛ 5 ᅇ⥅௦ࡋࠊቑṪࡉࡏࡓࠋቑṪࡉࡏࡓ⣽⬊ࡣࠊ⣽⬊⤖ಖᏑᾮ 㸦ࢭࣝࣂ࣮ࣥ࢝㸸ࢱ࢝ࣛࣂ࢜ᰴᘧ♫ࠊ᪥ᮏ㸧ࢆ⏝࠸࡚ࠊᾮయ❅⣲୰࡛ಖ⟶ࡉ ࢀࡓࠋ 2. ⣽⬊⥅௦ࡼࡿ⣽⬊ࡢᙳ㡪 ㏻ᖖࠊ◊✲⏝࠸ࡽࢀࡿᰴ⣽⬊ࡣࠊ୍ᐃࡢ⥅௦㝈ᗘᩘࢆつᐃࡋࠊ⟶⌮ࡉࢀࡿ ࡇࡀồࡵࡽࢀࡿࠋ୍᪉ࠊᮏヨ㦂࠾࠸࡚ࡣࠊBHK-21 ⣽⬊࠾ࡅࡿ㐺ṇ࡞⥅௦ ᩘࡀ᫂☜࡞ࡗ࡚࠸࡞࠸ࠋࡑࡇ࡛ࠊBHK-21 ⣽⬊࠾ࡅࡿ⥅௦ᩘࡑࡢ㝿ࡢ≧ែ ࡢ㛵ಀࡘ࠸᳨࡚ウࢆ⾜࡞ࡗࡓࠋ๓グ1 ࡛⤖ಖᏑࡋࡓ⣽⬊ࢆ⼥ゎᚋࠊ10 vol 㸣 FCS ῧຍ DMEM ࢆ⏝࠸ࠊ37Υࠊ5㸣CO2ࠊ95㸣✵Ẽࡢ᮲௳ୗ࡛ 4 ௦⥅௦ࡋࡓ Ⅼࢆ⥅௦ᩘ1 ᐃ⩏ࡋࡓࠋ75cm2ࡢ⣽⬊ᇵ㣴ࣇࣛࢫࢥ㸦ࣇࣝࢥࣥࣇࣛࢫࢥࠊ࣋ ࢡࢺࣥࢹࢵ࢟ࣥࢯࣥ♫ࠊ⡿ᅜ㸧ࠊ4105ಶ/mL ㄪᩚࡋࡓ BHK-21 ⣽⬊ᾋ㐟 ᾮࢆ22mL ศὀࡋࠊ3 ཪࡣ 4 ᪥ࡈ⥅௦ࡋࡓࠋࡇࡢ⥅௦ࢆ 100 ௦ࡲ࡛⾜࡞࠸ࠊ 5 ௦ࡈᚋ㏙ࡍࡿ⣽⬊ẘᛶࢫࢥ⏝࠸࡚⣽⬊ࡢ≧ែ㸦⣽⬊ྠኈࡢจ㞟ࠊࣉ࣮ࣞ ࢺࡽࡢ⣽⬊ࡢ㞳㸧ࢆุᐃࡋࡓࠋヨ㦂ࢆ3 ᅇ⧞ࡾ㏉ࡋࡓࠋ
44 3. ⣽⬊ẘᛶࡢศ㢮ཬࡧホ౯᪉ἲ FAVN ἲ࡛⿕㦂⾑Ύࢆ ᐃࡋࡓ㝿Ⓨ⏕ࡍࡿ⣽⬊ẘᛶࢆࠊࡑࡢᙧែⓗ≉ᚩ ࡽ2 ࡘࡢᆺศ㢮ࡋࡓ㸦ᅗ 2㸫1㸧ࠋ⣽⬊ࡀࢇቑṪࡋ࡞࠸ᆺࢆቑṪ㜼ᐖᆺẘ ᛶࡋࠊ⣽⬊ࡣቑṪࡍࡿࡶࡢࡢࣉ࣮ࣞࢺ᥋╔⬟ࢆḞࡁࠊ⣽⬊ྠኈࡀจ㞟ሢࢆᙧᡂ ཪࡣ㞳ࡍࡿᆺࢆ᥋╔㜼ᐖᆺẘᛶᐃ⩏ࡋࡓࠋ⣽⬊ẘᛶࡢ⛬ᗘࡣࠊᇵ㣴ࣉ࣮ࣞࢺ ࠾࠸࡚⣽⬊ẘᛶࡀ⏕ࡌࡓ㠃✚ᛂࡌࠊୗグࡢ࠾ࡾつᐃࡋࡓ㸦ᅗ2㸫2㸧ࠋ⣽⬊ ẘᛶࢫࢥ[㸫]㸸⣽⬊ẘᛶ㠃✚ 0㸣ࠊྠࢫࢥ[+]㸸ྠ㠃✚ 0㹼25㸣ࠊྠࢫࢥ [++]㸸ྠ㠃✚ 25㹼50㸣ࠊྠࢫࢥ[+++]㸸ྠ㠃✚ 50㹼75㸣ࠊྠࢫࢥ[++++]㸸 ྠ㠃✚ 75㹼100㸣ࠋᮏヨ㦂࡛ࡣࠊ⿕㦂⾑Ύ㸦⾑₢㸧ࢆ 9 ಸᕼ㔘ࡋࡓ㝿ࡢ⣽⬊ẘ ᛶࢫࢥࡀ[+]௨ୖ࡛࠶ࡿࡁࠊ⣽⬊ẘᛶࡢⓎ⏕ᐃ⩏ࡋࡓࠋࡲࡓࠊ⣽⬊ẘᛶࡢ 㝖ཤᥐ⨨ࡼࡾẘᛶࡀ㍍ῶࡉࢀࠊ୰ヨ㦂ࡋุ࡚ᐃྍ⬟࡞[+]ࢆྵࡴྠࢫࢥ ࡀ[+]ཪࡣ[㸫]࡞ࡗࡓሙྜࢆࠊ⣽⬊ẘᛶࡢ㝖ཤຠᯝᐃ⩏ࡋࡓࠋ 4. ᐇ㦂ື≀ 4.1. ᐇ㦂⏝ࢾ 18㹼24 ࣨ᭶㱋ࡢ࢜ࢫཬࡧ࣓ࢫ㸦ྛ 15 㢌㸧ࡢᐇ㦂⏝ࣅ࣮ࢢࣝ✀㸦ᰴᘧ♫ࢼ ࣝࢡࠊ᪥ᮏ㸧㸦௨ୗࠊ౪ヨࢾ⾲グࡍࡿ㸧ࢆ⏝ࡋࡓࠋࡇࢀࡽಶయࡣࠊ≬≟ ࣡ࢡࢳࣥࡢ᥋✀Ṕࢆ᭷ࡋ࡚࠸࡞ࡗࡓࠋᛶẚཬࡧᖺ㱋ࡘ࠸࡚ࡣࠊヨ㦂⩌㛫ࡢ ᕪࡀᴟຊᑠࡉࡃ࡞ࡿࡼ࠺ㄪᩚࡋࡓࠋ᳨つ๎ᚑ࠸ࠊヨ㦂㛤ጞ๓ಶయࢆᑐ ㇟ࡋ࡚ࠊ≬≟ά࣡ࢡࢳࣥ㸦A ♫〇㸧ࢆ 30 ᪥㛫㝸࡛ 2 ᅇ᥋✀ࡋࡓࠋᮏヨ 㦂ࡣࠊᏳ◊࠾ࡅࡿື≀ᐇ㦂ጤဨࡢᢎㄆࢆᚓ࡚ᐇࡉࢀࡓ㸦ᢎㄆ␒ྕ㸸 071009-2㸧ࠋ
45 4.2. ᐇ㦂⏝ࣔࣝࣔࢵࢺ 10 㐌㱋ࡢ࣓ࢫ㸦3 ༉㸧ࡢᐇ㦂⏝ࣔࣝࣔࢵࢺ㸦Hartley ⣔ࠊ㸧㸦᪥ᮏ࢚ࢫ࢚ࣝࢩ࣮ ᰴᘧ♫ࠊ᪥ᮏ㸧ࢆ⏝ࡋࡓࠋᮏヨ㦂ࡣࠊᏳ◊࠾ࡅࡿື≀ᐇ㦂ጤဨࡢᢎㄆ ࢆᚓ࡚ᐇࡉࢀࡓ㸦ᢎㄆ␒ྕ㸸071009-2㸧ࠋ 5. ᐇ㦂 1㸸⾑⢾ࠊ⾑Ύ⬡㉁ཬࡧ pH ࡢ ᐃ 5.1. ࣮ࣝࢳ᳨ࣥయ A ≬≟ᢠయ᳨ᰝࡢ౫㢗ࢆ┠ⓗࡋ࡚Ᏻ◊㏦ࡉࢀࡓࢾ⾑Ύࡢ୰ࡽࠊ ୰ヨ㦂࠾࠸࡚ 9 ಸᕼ㔘ࡋࡓ㝿ࡢ⣽⬊ẘᛶࢫࢥࡀ[+++]௨ୖࢆ♧ࡋࡓ⾑Ύ 230 ࢆᢳฟࡋࡓ㸦⾲ 2㸫1㸧ࠋෆヂࡣࠊቑṪ㜼ᐖᆺࡀ 104 ᳨యࠊ᥋╔㜼ᐖᆺࡀ 126 ᳨య࡛࠶ࡗࡓࠋࡲࡓࠊ⣽⬊ẘᛶࢫࢥࡀ[㸫]࡛࠶ࡗࡓ 133 ᳨యࢆᢳฟࡋࠊ⣽ ⬊ẘᛶ㝜ᛶ᳨యࡋࡓࠋࡇࢀࡽࡢ⾑Ύࡘ࠸࡚ࡣࠊ⾑⢾ཬࡧ⾑Ύ⬡㉁ࡢ ᐃ㸦ୗ グ5.2㸧୪ࡧ pH ࡢ ᐃ㸦ୗグ 5.3㸧౪ࡋࠊ࣮ࣝࢳ᳨ࣥయ A ࡋࡓࠋヨ㦂 ⏝ࡍࡿࡲ࡛ࡢ㛫ࠊ㸫30Υ࡛ಖ⟶ࡋࡓࠋ 5.2. ⾑⢾ཬࡧ⾑Ύ⬡㉁ࡢ ᐃ ࣮ࣝࢳ᳨ࣥయ A ࢆ⏝࠸࡚ࠊ⾑⢾ࠊ୰ᛶ⬡⫫ཬࡧ㐟㞳⬡⫫㓟ࡢྵ᭷㔞ࢆࡑࢀࡒ ࢀ ᐃࡋࡓࠋ⾑⢾ࡘ࠸࡚ࡣࠊ⾑⢾ ᐃჾ㸦࣓ࢹࢭ࣮ࣇ࣑ࢽ㸸ࢸࣝࣔᰴᘧ♫ࠊ ᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ୰ᛶ⬡⫫ࡘ࠸࡚ࡣࠊ୰ᛶ⬡⫫ ᐃ࢟ࢵࢺ㸦ࢺࣜࢢࣜ ࢭࣛࢻ E-ࢸࢫࢺ࣡ࢥ࣮㸸ග⣧⸆ᕤᴗᰴᘧ♫ࠊ᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ 㐟㞳⬡⫫㓟ࡘ࠸࡚ࡣࠊ㐟㞳⬡⫫㓟 ᐃ࢟ࢵࢺ㸦NEFA-HA ࢸࢫࢺ࣡ࢥ࣮㸸ග ⣧⸆ᕤᴗᰴᘧ♫ࠊ᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ
46 5.3. ୰ヨ㦂࠾ࡅࡿᇵ㣴ୖΎࡢ pH ᐃ ࣮ࣝࢳ᳨ࣥయ A ࢆ⏝࠸࡚ FAVN ἲࢆᐇࡋࠊ୰ヨ㦂⤊Ⅼࡢᇵ㣴ୖΎ 1mL ࢆ᳨యࡈᅇࡋࡓࠋྛᇵ㣴ୖΎࡢ pH ࡘ࠸࡚ࠊpH ィ ჾ㸦F-52㸸ᰴ ᘧ♫ᇼሙ〇సᡤࠊ᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ 6. ᐇ㦂 2㸸⣽⬊ẘᛶ㝖ཤ᪉ἲࡢ᳨ウ 6.1. ࣮ࣝࢳ᳨ࣥయ B ๓㡯ྠᵝᏳ◊㏦ࡉࢀࡓࢾ⾑Ύࡢ୰ࡽࠊ୰ヨ㦂࠾࠸࡚ 9 ಸ ᕼ㔘ࡋࡓ㝿ࡢ⣽⬊ẘᛶࢫࢥࡀ[++]㹼[++++]ࢆ♧ࡋࠊቑṪ㜼ᐖᆺẘᛶཪࡣ᥋╔ 㜼ᐖᆺẘᛶࢆ♧ࡋࡓ⾑Ύࢆࡑࢀࡒࢀ50 ᳨యᢳฟࡋࡓ㸦⾲ 2㸫1㸧ࠋ⣽⬊ẘᛶࢫࢥ ᳨యᩘࡢෆヂࡣࠊ⣽⬊ẘᛶࢫࢥ[++]㸸10 ᳨యࠊྠࢫࢥ[+++]㸸15 ᳨యࠊ ྠࢫࢥ[++++]㸸25 ᳨య࡛࠶ࡗࡓࠋࡇࢀࡽࡢ᳨యࡘ࠸࡚ࡣࠊྛ✀⣽⬊ẘᛶ㝖 ཤἲࡢ᳨ウ㸦ୗグ6.2㹼6.5㸧౪ࡋࠊ࣮ࣝࢳ᳨ࣥయ B ࡋࡓࠋヨ㦂⏝ࡍࡿ ࡲ࡛ࡢ㛫ࠊ㸫30Υ࡛ಖ⟶ࡋࡓࠋ 6.2. ⾑Ύࡢ⣽⬊྾ฎ⌮ 75 ੌࡢ⣽⬊ᇵ㣴ࣇࣛࢫࢥ㸦ࣇࣝࢥࣥࣇࣛࢫࢥ㸸࣋ࢡࢺࣥࢹࢵ࢟ࣥࢯࣥ♫ࠊ
⡿ᅜ㸧࠾࠸࡚ࠊ10 vol 㸣FCS ῧຍ DMEM ࢆ⏝࠸ࠊ37Υࠊ5㸣CO2ࡢ᮲௳ୗ
࡛BHK-21 ⣽⬊ࢆ༢ᒙᙧᡂࡉࡏࡓࠋྛࣇࣛࢫࢥࡽᇵᆅࢆ㝖ཤᚋࠊ࣮ࣝࢳ᳨ࣥ
యB ࢆࡑࢀࡒࢀ 1mL ῧຍࡋࠊ37Υࠊ5㸣CO2ࠊ95㸣✵Ẽ࡛ 24 㛫ឤసࡋࡓࠋ
ࡲࡓࠊୖグ10 ࡛ㄪᩚࡋࡓᢠ BHK-21 ⣽⬊ IgG ᑐࡋ࡚ࡶྠᵝᐇࡋࡓࠋᇵ
㣴⤊ᚋࠊᇵ㣴ୖΎࢆ10,000×g ࡛ 10 ศ㛫㐲ᚰศ㞳ࡋࠊࡑࡢୖΎࢆᅇࡋ᳨࡚
47 6.3. ⾑Ύࡢࢺࣜࣉࢩࣥ-EDTA ฎ⌮ ࣮ࣝࢳ᳨ࣥయ B ࠊ᭱⤊⃰ᗘࡀ 0.02 vol 㸣࡞ࡿࡼ࠺ࢺࣜࣉࢩࣥ-EDTA ⁐ ᾮ㸦ࣛࣇࢸࢡࣀࣟࢪ࣮♫ࠊ⡿ᅜ㸧ࢆῧຍࡋࡓࠋࡇࢀࡽࡢ⾑Ύࢆ37Υࠊ95㸣✵ Ẽ࡛ 4 㛫ࠊෆᐜࢆᨩᢾࡋ࡞ࡀࡽឤసࡋࠊ᳨యࡋࡓࠋFAVN ἲ࡛ ᐃࡍࡿࡲ ࡛ࡢ㛫ࠊ4Υ࡛ಖ⟶ࡋࡓࠋ 6.4. ⾑Ύࡢ࢝࢜ࣜࣥฎ⌮ ࣮ࣝࢳ᳨ࣥయB ࠊ࢝࢜ࣜࣥ㸦ࢧ࣮ࣔࣇࢵࢩ࣮ࣕࢧ࢚ࣥࢸࣇࢵࢡ♫ࠊ ⡿ᅜ㸧ࢆ10 vol 㸣࡞ࡿࡼ࠺ຍ࠼ࡓࠋࡇࢀࡽࡢ⾑Ύࢆ 25Υ࡛ 4 㛫ࠊෆᐜࢆᨩ ᢾࡋ࡞ࡀࡽឤసࡋࡓࠋឤస⤊ᚋࠊ10,000×g ࡛ 10 ศ㛫㐲ᚰศ㞳ࡋࠊࡑࡢୖΎ ࢆ᳨యࡋࡓࠋFAVN ἲ࡛ ᐃࡍࡿࡲ࡛ࡢ㛫ࠊ4Υ࡛ಖ⟶ࡋࡓࠋ 6.5. ⣽⬊ᇵ㣴⏝ฎ⌮ࣉ࣮ࣞࢺ FAVN ἲࡢ⣽⬊ᇵ㣴⏝࠸ࡿ 96 ࢙࢘ࣝࣉ࣮ࣞࢺࡘ࠸࡚ࠊ㏻ᖖࡢ㠀ฎ⌮ࣉ࣮ࣞ ࢺ㸦࣋ࢡࢺࣥࢹࢵ࢟ࣥࢯࣥ♫ࠊ⡿ᅜ㸧ࡢࠊࢥ࣮ࣛࢤࣥฎ⌮ࣉ࣮ࣞࢺ㸦࣋ࢡࢺ ࣥࢹࢵ࢟ࣥࢯࣥ♫ࠊ⡿ᅜ㸧ཬࡧࢮࣛࢳࣥฎ⌮ࣉ࣮ࣞࢺ㸦࣋ࢡࢺࣥࢹࢵ࢟ࣥࢯࣥ♫ࠊ ⡿ᅜ㸧ࢆ⏝ࡋࠊ⣽⬊ẘᛶࡢⓎ⏕≧ἣࢆẚ㍑ࡋࡓࠋ⿕㦂⾑Ύࡋ࡚ࠊ࣮ࣝࢳ᳨ࣥ యB ࢆ⏝࠸ࡓࠋFAVN ἲ࡛ ᐃࡍࡿࡲ࡛ࡢ㛫ࠊ4Υ࡛ಖ⟶ࡋࡓࠋ 7. ᐇ㦂 3㸸FAVN ᢠయ౯ࡢᙳ㡪 7.1. ࣮ࣝࢳ᳨ࣥయ C ๓㡯ྠᵝᏳ◊㏦ࡉࢀࡓࢾ⾑Ύࡢ࠺ࡕࠊ⣽⬊ẘᛶࢫࢥࡀ[㸫]࡛࠶ ࡿ᳨యࢆᢳฟࡋࡓ㸦⾲2㸫1㸧ࠋࡇࢀࡽࡢ⾑Ύࡣࠊྛ✀⣽⬊ẘᛶ㝖ཤἲࡼࡿ≬≟ ୰ᢠయ౯ࡢᙳ㡪ホ౯㸦ୗグ7.2㸧౪ࡍࡿࡇࡽࠊ≬≟ᢠయ᳨ᰝࡢྜ
48 ᱁್㏆㸦≬≟୰ᢠయ౯ࡀ0.5IU/mL㸧ࡢ⾑Ύࢆ୰ᚰ㑅ࢇࡔࠋ0.29 ௨ୗࠊ 0.29㹼7.9ࠊ7.9 ௨ୖヱᙜࡍࡿࡶࡢࢆྛࠎ30 ᳨యࡎࡘࠊྜィ 90 ᳨యࢆᢳฟࡋ ࡓࠋࡇࢀࡽࡢ᳨యࡘ࠸࡚ࡣࠊ࣮ࣝࢳ᳨ࣥయ C ࡋࡓࠋヨ㦂⏝ࡍࡿࡲ࡛ࡢ 㛫ࠊ㸫30Υ࡛ಖ⟶ࡋࡓࠋ 7.2. ⣽⬊ẘᛶ㝖ཤἲࡼࡿ≬≟୰ᢠయ౯ࡢᙳ㡪 ࣮ࣝࢳ᳨ࣥయC ᑐࡋࠊࡑࢀࡒࢀ 5 ✀㢮ࡢ⣽⬊ẘᛶ㝖ཤἲ㸦ୖグ 6.2㹼6.5㸧 ࢆᐇࡋࡓࠋྛ⾑Ύࡘ࠸࡚ࠊ⣽⬊ẘᛶ㝖ཤἲᐇᚋࡢ≬≟୰ᢠయ౯ 㸦IU/mL㸧ࢆ ᐃࡋࠊᐇ๓ẚ㍑ࡋࡓࠋ 8. ⾑ᾮจᅛ㛵㐃≀㉁ࡢῧຍཬࡧ⾑Ύศ㞳 ௨ୗࡢ11 ✀㢮ࡢ⾑ᾮจᅛ㛵㐃≀㉁ࢆ⿕㦂≀㉁ࡋࡓࠋ⾑ᾮจᅛಁ㐍ࡋ࡚ࠊ ࢘ࢩࢺࣟࣥࣅࣥ㸦ࢩࢢ࣐ࣝࢻࣜࢵࢳ♫ࠊ⡿ᅜ㸧ࠊࣄࢺࢺࣟࣥࣅࣥ㸦ࢩࢢ࣐ࣝ ࢻࣜࢵࢳ♫ࠊ⡿ᅜ㸧ࠊ࢘ࢩࣉࣟࢳࢽࣥ㸦ࢩࢢ࣐ࣝࢻࣜࢵࢳ♫ࠊ⡿ᅜ㸧ཬࡧ◲ 㓟ࣉࣟࢱ࣑ࣥ㸦࣓ࣝࢡ♫ࠊࢻࢶ㸧ࢆ⏝ࡋࡓࠋࡲࡓࠊ⾑ᾮจᅛ㜼Ṇࡋ࡚ࠊ ࣊ࣃࣜࣥࢼࢺ࣒ࣜ࢘㸦ᣢ⏣〇⸆ᰴᘧ♫ࠊ᪥ᮏ㸧ࠊࢡ࢚ࣥ㓟୕ࢼࢺ࣒ࣜ࢘㸦ග ⣧⸆ᕤᴗᰴᘧ♫ࠊ᪥ᮏ㸧ࠊࣇࢵࢼࢺ࣒ࣜ࢘㸦ග⣧⸆ᕤᴗᰴᘧ♫ࠊ᪥ᮏ㸧ࠊ EDTA-2 ࢼࢺ࣒ࣜ࢘㸦ᰴᘧ♫ྠோᏛ◊✲ᡤࠊ᪥ᮏ㸧ࠊEDTA-4 ࢼࢺ࣒ࣜ࢘㸦ᰴ ᘧ♫ྠோᏛ◊✲ᡤࠊ᪥ᮏ㸧ࠊEDTA-2 ࣒࢝ࣜ࢘㸦ᰴᘧ♫ྠோᏛ◊✲ᡤࠊ ᪥ᮏ㸧ཬࡧ EDTA-3 ࣒࢝ࣜ࢘㸦ᰴᘧ♫ྠோᏛ◊✲ᡤࠊ᪥ᮏ㸧ࢆ⏝ࡋࡓࠋ ࡇࢀࡽࡢ⿕㦂≀㉁ࡣࠊ࠸ࡎࢀࡶ᥇⾑୍⯡ⓗ⏝ࡉࢀࡿࡶࡢ࡛࠶ࡿࠋ⿕㦂≀ ㉁ࡈࠊୗグࡢࡼ࠺ࣄࢺ࠾ࡅࡿ⾑ᾮ⏕Ꮫ᳨ᰝ࡛ࡢ᥎ዡ⃰ᗘ㏆࡞ࡿ ࡼ࠺⛗㔞ࡋࠊ15mL ࢳ࣮ࣗࣈධࢀࡓࠋ⾑ 1mL ᑐࡋ࡚ྛ⿕㦂≀㉁⃰ᗘ[࢘ࢩ ࢺࣟࣥࣅࣥ㸦10U㸧ࠊࣄࢺࢺࣟࣥࣅࣥ㸦10U㸧ࠊ࢘ࢩࣉࣟࢳࢽࣥ㸦500KIU㸧ࠊ◲
49 㓟ࣉࣟࢱ࣑ࣥ㸦0.2mg㸧ࠊ࣊ࣃࣜࣥࢼࢺ࣒ࣜ࢘㸦20U㸧ࠊࢡ࢚ࣥ㓟୕ࢼࢺ࣒ࣜ࢘(4mg)ࠊ ࣇࢵࢼࢺ࣒ࣜ࢘㸦5mg㸧ࠊEDTA-2 ࢼࢺ࣒ࣜ࢘㸦1mg㸧ࠊEDTA-4 ࢼࢺ࣒ࣜ࢘ 㸦1mg㸧ࠊEDTA-2 ࣒࢝ࣜ࢘㸦1mg㸧ࠊEDTA-3 ࣒࢝ࣜ࢘㸦1mg㸧]ࢆῧຍࡋࡓࢳ ࣮ࣗࣈࡑࡢ10 ಸ㔞ࢆῧຍࡋࡓࢳ࣮ࣗࣈࢆ‽ഛࡋࡓࠋࡇࢀࡽࡢࢳ࣮ࣗࣈࠊ౪ ヨࢾ㸦30 㢌㸧ࡽ᥇ྲྀࡋࡓ⾑ᾮ 1mL ࢆಶయูຍ࠼ࠊࡼࡃΰࡋࡓࠋ25Υ࡛ 1 㛫㟼⨨ᚋࠊ500×g ࡛ 10 ศ㛫㐲ᚰศ㞳ࡋࠊ⾑Ύ㸦⾑₢㸧ࢆ᥇ྲྀࡋࡓࠋࡇࢀࡽ ࡢ⿕㦂⾑Ύࡘ࠸࡚ࠊFAVN ἲࢆᐇࡋࡓࠋ⿕㦂⾑Ύࡣࠊヨ㦂⏝ࡍࡿࡲ࡛ࡢ 㛫ࠊ4Υ࡛ಖ⟶ࡋࡓࠋ 9. ⁐⾑⾑Ύࡢㄪᩚ ౪ヨࢾ㸦࢜ࢫ10 㢌ࠊ࣓ࢫ 10 㢌㸧ࡘ࠸࡚ࠊ23 ࢤ࣮ࢪὀᑕ㔪ࢆ╔ࡋࡓὀ ᑕჾ㸦ࢸࣝࣔᰴᘧ♫ࠊ᪥ᮏ㸧ࢆ⏝࠸ࠊಶయู 10mL ᥇⾑ࡋࡓࠋࡇࢀࡽࡢὀ ᑕჾ㧗࠸ᅽຊࢆࡅ࡚⾑ᾮࢆฟ࣭྾ᘬࡉࡏࡿ᧯సࢆ 5 ᅇ⛬ᗘ⧞ࡾ㏉ࡋࠊ㉥ ⾑⌫ࢆ◚ࡉࡏࡓࠋ25Υ࡛ 1 㛫㟼⨨ᚋࠊ500×g ࡛ 10 ศ㛫㐲ᚰศ㞳ࡋࠊ⾑Ύࢆ ᥇ྲྀࡋࡓࠋࡇࢀࡽࡢ⿕㦂⾑Ύࡘ࠸࡚ࠊFAVN ἲࢆᐇࡋࡓࠋ⿕㦂⾑Ύࡣࠊヨ㦂 ⏝ࡍࡿࡲ࡛ࡢ㛫ࠊ4Υ࡛ಖ⟶ࡋࡓࠋ 10. ౮㞧≀ྵ᭷⾑Ύࡢㄪᩚ ౪ヨࢾ㸦30 㢌㸧ࡘ࠸࡚ࠊ┿✵᥇⾑⟶㸦ࣉ࣮ࣞࣥࢱࣉࠊࢸࣝࣔᰴᘧ♫ࠊ ᪥ᮏ㸧࡛ಶయู10 mL ᥇⾑ᚋࠊ25Υ࡛ 1 㛫㟼⨨ࡋࡓࠋ500×g ࡛ 10 ศ㛫㐲 ᚰศ㞳ࡋࠊ⾑Ύ⾑⌫ࡢቃ⏺㏆Ꮡᅾࡍࡿ౮㞧≀㸦ⓑ⾑⌫࡞㸧ࢆྵࡴࡼ࠺⾑ Ύࢆ᥇ྲྀࡋࡓࠋࡇࢀࡽࡢ⿕㦂⾑Ύࡘ࠸࡚ࠊFAVN ἲࢆᐇࡋࡓࠋ⿕㦂⾑Ύࡣࠊ ヨ㦂⏝ࡍࡿࡲ࡛ࡢ㛫ࠊࢆ4Υ࡛ಖ⟶ࡋࡓࠋ
