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^4 ^ r -ì 'n ( _ V i :-_ i i\u ié^* ,.1fV '-v ^ •V'S V •••''V.'- ,• *f i; ^ %• J^~ - Wv?c.V{^ '- -t^^>• * ^ ' l ' ^ - ^ ^ - j : : ' :>C C S V^^' >^:% f V - •.'^^*- - M^'r^ , '• , -*»JK t*^- ( >.j v\l^ V -A > BO CIAO DUC VA DAO TAO TOUONG DAI HOC TONO HOF HÀ NÓI *=!:* Lai Minh Hién ^? mm V^ l'i/Q^ NGHIÉN CLfU LfNG DUNG CÀC PHUONG PHÀP HUYÉT THANH HOC DÙNG TRONG CHAN DỒN BÉNH SĨT Q DO Coxiella burnetii CHUYEN NGÀMH : V! 5INH WOC MA s6 :1.05.12 LUAN AN PHO TIEN SI KHOA HOC SINII HOC c o VAN KìlOAFipC: POS ITS PIIAM VÀN1\ PPS VVQUAWGIJIA' \'^nM\ iik;^*L a: HANOI " 1995 MUC LUC Trang - Dat vàn de - Tóng quan tài Uéu — 2.1 - Lugc sur nghièn culi sò't Q 2.2 - Dac diém sinh hoc cùa màm bènh 2.2.1 - Dac diém hình thài va tinh chà't ni cà^ 2.2.1.1 - Dac diém hình thài ' 10 2.2.1,2-Tinh bàtmàu 12 2.2.1.3 - Tinh chà't nuòi cày Coxiella 13 2.2.2 -Dac diém mièn dich hoc 2.2.2.1 - Càu trùc khàng nguyèn cùa C.bumetu 21 2.2.2.2 -Dac diém mièn dich hoc 27 2.2.3 - Tinh de khàng 29 2.3 - Chin doàn phòng nghiém 32 2.3.1 - Phàn làp màm bénh 32 2.3.2 - Phucfng phàp huyè't hoc 33 - Vàt héu va phuang phàp 3.1 - Vàtliéu 44 3.2-Phucrng phàp 45' 3.2.1 - Diéu che'khàng nguyèn 45 3.2.2 - Diéu che"khàng thè' dac hiéu 46 3.2.3 - Diéu chè'hóng càu gàn khàng nguyèn C.bumetii dùng ngimg kè't hóng càu thu dòng 47 3.2.4 - Diéu che"càc còng hop 51 3.2.4.1 - Diéu che" còng hop khàng thè' gàn FITC 51 3.2.4.2 - Diéu che"còng hgrp khàng thè' gàn POD 53 3.2.5 - Phucmg phàp khuéch tàn trèn thach 55 3.2.6 - Phuomg phàp mièn dich dién di doi lini 56 3.2.7 - Càc phucmg phàp khàc 57 - Kè"t qua va bàn luàn • 59 4.1 - Cày trun C.bumetii long dò phói gà 59 4.2 - Diéu che'va kiè'm tra boat tinh KN 63 4.3 - Diéu che" huyè"t mièn dich 68 4.3.1 - Kè"t qua diéu che'dich bang mièn dich 68 ^— A32 - Diéu chè'huyét mièn dich thò 68 4.4 - Diéu che" còng hop khàng thè' gàn FITC va POD 71 4.4.1 - Diéu che" còng hgp khàng thè' gàn FITC va POD 72 4.4.2 - Dành già chà"t lirong còng hop FITC va POD 4.4.3 - Nghièn cuti su phàt trién cùa C.bumetii tè'bào dich bang Eurhch bang KTHQ va KT enzim 75 4.5 - Kè"t qua diéu che'hóng càu gàn KN C.bumetii phàn ùng NKHCro 80 4.5.1 - Ành hucfng cùa càc chà"t xù ly hóng càu dè"n qua trình gàn 80 4.5.2 - Ành huomg cùa nóng dò KN C.bumetii dè'n qua trình _gàn 81 4.5.3 - Xàc dinh dò nhày va tinh dac hièu cùa chè'ph£m ch^n dồn 4.5.4 - Tnòi han bào quàn cùa chè'phàm chàn doàn 82 83 4.5.5 - Kiém tra mùc dò gàn khàng nguyèn C.bumetu lèn bé mat hóng càu bang KTHQ va KT enzim 84 4.6 - Xàc dinh KT dac hièu vód C.bumetii bang càc phucfng phàp huyét hoc 4.6.1 - Xàc dinh KT bang NKHCTO 88 4.6.2 - Xàc dinh KT bang MDDDDL 89 4.6.3 - Xàc dinh KT bang KTTT 89 4.7 - Diéu tra khàng thè Imi hành khàng C.bumetii huyét nguòi 92 - Kè"t luàn 100 - Tài Uéu tham khào ^Q2 NHCNG CH0 VIET TAT KHBT : Kèt hop bò thè Khàng thè huynh quang KTHQ Kl'iT Khuèch tàn trèn thach : Khàng nguyèn KN KT : Khàng thè' EUSA : Hà"p thu mièn dich co gàn enzim RIA : Mièn dich phóng xa FITC : Fluorescein isothiocyanat POD : Peroxidaza MDDDDL : Mièn dich dién di ddi luu NKHCTD : Ngung kèt hóng càu thu dòng irCNKHCTD : Ùc che ngung kèt hóng càu thu dòng LPS : Lipopolisacarit UCNKHC : Uc che" ngung két hóng càu I - DÀT VAN DE Sot Q Coxiella burnetii gay khong phài bénh nhiém trùng bien co ty le mac cao, song mot bénh truyen nhièm càn dilòc quan tàm dùng mùc nhtìng dac diém sau: Là mot bénh lày nhiém phàn bò' trén tồn the' giòi, ké cà mot so' niióc can vói Viet Nam nhif Trung Quò'c, N h a t Bàn, Malaixia, An Do (32, 87, 112, 120, 152, 157) Bénh lày truyén qua nhiéu duòng khàc n h a u nhil diiòng tiéu hoà, màu, trùng trung gian va dac biet qua dng ho hap dilói dang dung co khà nàng gay bénh cho nhiéu ngilòi mot tliòi gian ngan va lan truyén di r a t xa (42, 112, 152) Màm bénh sot Q, khàc v6i nhiéu loai Rickettsia khàc, co sue de khàng cao va co doc \\Lc m a n h (109, 112, 152) Bénh cành làm sàng cùa nhiém C.bumetii r a t da dang, khòng the hồc rat khó chuan dồn xàc dinh bénh bang * càc bieu hién làm sàng, hon ntìa co nhitng the bénh dién bién nang dàn den tu vong (32, 112, 160, 157, 163, 174) •2 Mam bénh sot Q co the dUdc su dung làm vù sinh vàt chién tranh sinh hoc (112, 152) Càc tài liéu nghièn cùu ve bénh sot Q rat it òi ò nxiàc ta Tuy nhién, càc so' liéu cong bò' thày bénh co lifu h n h ò ngxlòi va dòng vàt nuòi tai Viet Nam (5, 112, 119, 120) Do vay tièp tue nghièn cùu diéu tra bénh càn thiét De giài quyé't vàn de này, chùng ta càn co diltìc càc phiidng tien, bao gom sinh pham chan doàn, svi hoàn thien càc phiidng phàp de thUc hién càc xét nghiém phòng thi nghiém Y thùc diiòc nhtìng vàn de:, trén de tài nghièn cùu cùa chùn^ tòi n h m càc muc tiéu sau: Diéu che' so' che' pham sinh hoc dùng chan dồn sò't Q Dành già chat IvLóng càc sinh pham che" tao va luta choh phiidng phàp thìch hòp de xàc dinh m m bénh, xàc dinh KN, KT Ung dung càc che phàm va phiidng phàp dà nghièn cùu dtfdc de bifòc dàu diéu tra so' nguòi co k h n g C.bumetii the .3 •a .Ci • •CP H: "5 ri •I-4 i' HO P ' o ":'"H :/ì P*: : :Sr- vivC::::; :;:^:;:;:;pf:;: Ti cac ph • -• t - M H Đ 7^ ^ 5r fi m •0 • • * 2: 05 o o>, o X a- X gi pr^ o o rs g rtằo ô: >;- C/5;; o r o> P^ S SS xa •p 'K ^ liii 25 n * o a> H a T e* «• 3;; - TĨNG QUAN TÀI LIÈU J - LUDC SlTNGHIÈN c ù u SOT Q Bénh sóì Q Rickettsia, dàu tién durgfc Derrick xàc dinh ve mat làm sàng ò Uc (mien nam Queensland) nàm 1937, nhimg tnxòc dò, nàm 1935 tàc già dà phàl hién ihà'y càc trirìnig hop bénh tircfng tu ò Brisben (trong so càc còng nhàn xi nghiép bào quàn thit), vi vày tàc già goi bénh sot "Q" ("Q" tijr tiéhg Anh "Querf nghTa chua xàc dinh) (29) CQng nàm 1937 Derrick dà phàn làp trén chuol lang duttc so' màu màu bénh nhàn bi bénh vimt ma Bumcl va Frecman xé'p vào Rickettsia dang qua loc Nàm 1939 Deirick goi mam bénh tìm dirofc "Rickettsia humetif (30) Cuoi nàm 1942 càc nhà nghièn curu Uc dà làp lai thi nghiém gay nhiém bé va v'é màt huyét hoc dà xàc dinh d\xac su' co mat cùa bénh nhiém trùng dà mó tà truac Trén ca so càc nghièn ciai cùa càc tàc già Ùc, nguòfi ta dà di dèh già thiét bò màc bénh sò't Q qua ve bj nhièm Nguòi bi nhiém bénh qua ve va dac bièt hit phài bui co chira phàn ve hồc tié'p xùc vai bò bi bénh qua vé't xc Nói chung bénh thucmg gap a nòng dàn va còng nhàn lo mó De)c làp vai càc nhà nghièn cùu nguòi Uc, a My vào nàm 1938 Davis va Cox dà phàn làp duac lù ve mot loai virut qua loc, sau dò Cox xàc dinh Rickettsiavh dat tén "Rickettsia diapotica" (27) Nàm 1938 tai Hoc vién y khoa Washington (My), Dyer xàc dinh rang chuòt lang dà truyen bénh Dira trén làm sàng va dich té hoc, Dyer dà xàc dinh su giòng giùa nói Uc R.bumetii cùo Deirick nàm 1939 va ActaVu"oL,797S, 3, 309-315 50-Kazar J X ' a l y s ^ m l ' ' ' T^^T^ ' " ' i-munoglobulins in sera of rabbits and guinea pigs immunized with Coxiella bumeth Acta Virol , 7977, 2, 246-255 51 - 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180 3MJIHHA, M.A B03M07KHOCTM MCnOJIbSOBaHVlH iJ^aSBot^ M3MeHMMB0CTM pMKKeTCMW B e p n e T a B cepojiorwHecKot^ ;i.MarHocTMKe jiMxopaj^KM Ky Kyp MwKpMOJi onw^eMMoji M MMMynMOJi., 9 , , - 3MJIMHA, M.A PeaKUMH yrHCTeHMn cBH3tiBaHMH KoMnjieMenra MSTO^ jiaóopaTopHoti ;^Ma^HOOTMKM Jiwxopa^^Ka Ky % p MuKpMOJi, Qnw^eMMoji M MMMyHMoji., , , - T GMCBEPr, B M B03MAH, ^ JlaòopaTopHaH ^MarHocTMKa BPipycHUx M pMKKeTCMosHUx sojiesaHMtì M T974 - 7 T8? ICTPEBOB, B.K OnHT npMMeHem^H peaKUMw nenpHMoR reMMarrjiioTMHauMM JIJIH M3yHeHVi5 KOHxaKTOB e Bosóy^MTejieM KjiemeBoro pMKKeTCMosa cKBepnoti A3MM Kyp MMKpoóMoji QnM^eMMoji M MMMyHoóMoji., , T I , I - T ... HOC TONO HOF HÀ NÓI *=!:* Lai Minh Hién ^? mm V^ l'i /Q^ NGHIÉN CLfU LfNG DUNG CÀC PHUONG PHÀP HUYÉT THANH HOC DÙNG TRONG CHAN DỒN BÉNH SĨT Q DO Coxiella burnetii CHUYEN NGÀMH : V! 5INH WOC MA s6... hop bénh tircfng tu ò Brisben (trong so càc còng nhàn xi nghiép bào quàn thit), vi vày tàc già goi bénh sot "Q" ( "Q" tijr tiéhg Anh "Querf nghTa chua xàc dinh) (29) CQng nàm 1937 Derrick dà phàn... giàu khàng nguyén thucmg co 500 D.AM Vi c làm giàu Coxiella h rat quan beri lé mite de) giàu khàng nguyèn sé quyé't dinh vi c pha lồng nòng de) Coxiella dùng de gay nhiém cho phòi gà Co thè dành