Ứng dụng kỹ thuật NAT (nucleic acid) phát hiện sớm sự có mặt của vi rút HIV, HBV, HCV ở người cho máu

I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - Trn Võn Chi NG DNG K THUT NAT (NUCLEIC ACID) PHT HIN SM S Cể MT CA VI RT HIV, HBV, HCV NGI CHO MU LUN VN THC S KHOA HC H Ni - 2015 Trần Vân Chi Luận văn Thạc sĩ Khoa học I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - Trn Võn Chi NG DNG K THUT NAT (NUCLEIC ACID) PHT HIN SM S Cể MT CA VI RT HIV, HBV, HCV NGI CHO MU Chuyờn ngnh: Mó s: Vi sinh vt hc 60420107 LUN VN THC S KHOA HC Ngi hng dn khoa hc: PGS.TS Bch Khỏnh Hũa TS Trn Vn Tun XC NHN HC VIấN CHNH SA THEO GểP í CA HI NG Giỏo viờn hng dn Ch tch hi ng chm lun thc s khoa hc TS Trn Vn Tun PGS.TS Bựi Th Vit H H Ni - 2015 Trần Vân Chi Luận văn Thạc sĩ Khoa học LI CM N Li u tiờn, cho phộp tụi c by t lũng bit n sõu sc nht ti PGS.TS Bch Khỏnh Hũa; ngi thy ó tn tỡnh hng dn, giỳp tụi lm lun Ngi thy a tụi n vi Huyt hc Truyn mỏu, ngi thy ó truyn cho tụi nim am mờ vi Huyt hc Truyn mỏu Ngi thy ó bi dng cho tụi nhng c tớnh trung thc, cn mn ca mt ngi lm cụng tỏc nghiờn cu t nhng ngy u tiờn bc vo ngh Tụi xin by t lũng cm n sõu sc n BSCKII Phm Tun Dng ngi ó ng viờn cho tụi i hc cao hc v sut quỏ trỡnh hc Ngi ó tn tỡnh giỳp , dỡu dt tụi cụng tỏc Tụi by t lũng cm n sõu sc n TS Trn Vn Tun ngi ó hng dn ch dn, giỳp tụi lm lun Tụi cng xin chõn thnh cm n s giỳp on kt ca anh ch em ng nghip Khoa Xột nghim sng lc mỏu ó giỳp tụi hon thin s liu, mi iu kin thun li nht cho tụi sut quỏ trỡnh hc v lm lun Tụi xin gi li cm n ti: Ban lónh o Vin HHTMTW ó cp kinh phớ cho tụi i hc, cỏc khoa phũng hnh chớnh ó giỳp tụi hon thnh th tc hc tp, Khoa Di truyn v sinh hc phõn t, Khoa Thalassemia, Khoa Hemophilia, khoa HIV vin v sinh dch t ó cho phộp v giỳp tụi xột nghim nh lng virus, ton th Khi truyn mỏu ó to iu kin thun li cho tụi thc hin ti Tụi xin gi li cm n ti Phũng sau i hc, Khoa Sinh hc, B mụn Vi sinh hc ca trng i hc Khoa hc T nhiờn ó giỳp tụi hon thnh cỏc th tc lun Tụi xin by t lũng bit n ti cỏc bc b m , b m chng, chng ó giỳp ng viờn, dnh tỡnh cm v iu kin thun li cho tụi hon thnh lun Tụi xin by t lũng cm n n ngi hin mỏu ó cho tụi nhng s liu quý bỏu Hc viờn Trn Võn Chi Trần Vân Chi Luận văn Thạc sĩ Khoa học M U Vi rỳt l mt nhng nguyờn nhõn gõy mt s bnh nghiờm trng ngi núi chung Riờng vi rỳt viờm gan B gi tt l HBV (Hepatitis B virus ), vi rỳt viờm gan C gi tt l HCV (Hepatitis C virus), vi rỳt gõy suy gim dch ngi HIV (Human immunodeficiency virus) l vi rỳt gõy bnh thụng qua ng mỏu Cỏc vi rỳt ny bin i liờn tc v cu trỳc v h gen trn thoỏt cỏc loi sinh phm xột nghim hu qu l tng t l lõy nhim cng ng Phng phỏp xột nghim dựng cho sng lc HIV, HBV v HCV lõy qua ng truyn mỏu l xỏc nh tỏc nhõn gõy bnh giỏn tip da trờn kt qu ca phn ng dch khỏng nguyờn-khỏng th u im ca xột nghim l n gin, tn ớt thi gian, giỏ thnh hp lý, song phng phỏp ny cú nhc im l giai on ca s ú khỏng th hoc khỏng nguyờn cha t ngng phỏt hin, vy xột nghim s cho kt qu õm tớnh Bờn cnh ú, tng tỏc khụng c hiu ca khỏng th hoc khỏng nguyờn vi nhng bt thng protein khỏc mỏu cú cu trỳc gn ging vi khỏng nguyờn hoc khỏng th cú th dn n kt qu dng tớnh gi Cựng vi s phỏt trin ca k thut sinh hc phõn t, k thut NAT ó c a vo s dng, k thut ny cú nhy cao cho phộp phỏt hin v nhõn bn c hiu theo hm m cỏc trỡnh t ớch ca tỏc nhõn gõy bnh t mt lng nh vi rỳt, ú, cho phộp phỏt hin sm v chớnh xỏc cỏc tỏc nhõn gõy bnh Hn th na, NAT cú th c s dng phỏt hin ng thi HIV, HBV v HCV thụng qua mt xột nghim thi gian l 4-5 gi, m bo an ton cho n v mỏu truyn Trờn Th gii, cỏc nc phỏt trin, m bo sng lc mỏu Anh, Phỏp, M, c thc hin xột nghim song song gia phng phỏp giỏn tip khỏng nguyờn khỏng th v xột nghim NAT cho n v mỏu t cui nhng nm 1990 u nhng nm 2000 Nm 1997, Hi ch thp c ó s dng PCR cho mc ớch sng lc HCV RNA cho ngi hin mỏu, tip theo l Nht Bn v Scotland Sau ú vo thỏng 3/1998 ngi ta ó gii thiu NAT cho xột nghim HCV RNA[50] Trần Vân Chi Luận văn Thạc sĩ Khoa học Vit Nam, hu ht cỏc phũng xột nghim sng lc HIV, HBV v HCV u s dng phng phỏp xột nghim dch khỏng nguyờn khỏng th hay cũn gi l xột nghim huyt hc, xột nghim NAT cha c s dng Nm 2008 ti cp B v ng dng NAT cho xột nghim sng lc mỏu t kt qu tt [3] n nm 2015, Vin HHTMTW thc hin theo Thụng t 26 TT-BYT ngy 16/9/2014 Hng dn hot ng truyn mỏu, trin khai xột nghim NAT m bo an ton truyn mỏu Xut phỏt t thc t trờn, chỳng tụi tin hnh nghiờn cu ng dng k thut NAT (nucleic acid) phỏt hin sm s cú mt vi rỳt HIV, HBV, HCV ngi cho mỏu nhm hng ti cụng tỏc xột nghim sng lc sm cỏc tỏc nhõn lõy qua ng truyn mỏu ny mt cỏch chớnh xỏc, m bo an ton, cht lng cho mỏu v ch phm s dng iu tr Mc tiờu nghiờn cu ca ti - Kho sỏt t l HBsAg, anti HCV, HIVAgAb phn ng trờn n v mỏu hin c xột nghim dch bng k thut CMIA, ECLIA T l nhim HBV, HCV, HIV bnh nhõn truyn mỏu nhiu ln - T l HBV, HCV, HIV phn ng phỏt hin bng k thut NAT (nucleic acid) trờn n v mỏu hin m xột nghim dch khụng phỏt hin c, theo dừi ngi hin mỏu cú phn ng k thut NAT, phỏt hin sm h gen vi rỳt nhm b tr cho xột nghim dch khỏng nguyờn- khỏng th Trần Vân Chi Luận văn Thạc sĩ Khoa học KT LUN Xột nghim sng lc khỏng nguyờn khỏng th bng k thut CMIA, ECLIA vi t l HBsAg l 0,75%; 0,79%; anti HCV l 0,43%, 0,24%, HIV AgAb l 0,1%, 0,23% Bnh nhõn nhn mỏu nhiu ln cú t l nhim HBV l 5,9% nhim HCV l 23,89% Xột nghim NAT (nucleic acid) cú t l HBV, HCV, HIV tng ng l 1:1086 ; 1:192221; 2:192221 (HBV 0,92, HCV 0,0052, HIV 0,01) Nghiờn cu theo dừi c ngi hin mỏu mi nhim HIV giai on ca s huyt thanh, ngi mi nhim HCV giai on ca s, ngi nhim HBV giai on ca s Trần Vân Chi Luận văn Thạc sĩ Khoa học KIN NGH Thc hin xột nghim sng lc mỏu bng k thut NAT b tr cho k thut dch khỏng nguyờn- khỏng th Nghiờn cu mt s trng hp nhim HBV th n OBI Trần Vân Chi Luận văn Thạc sĩ Khoa học TI LIU THAM KHO TI LIU TING VIT V Thựy An v cng s (2012), Tỡnh hỡnh sng lc cỏc bnh nhim trựng qua ng mỏu ngi hin mỏu tỡnh nguyn khu vc ụng-Nam b ti Trung tõm truyn mỏu Ch Ry t 2009-2011, Tp Y hc Vit Nam, 396, 272-280 Phm Tun Dng v cng s (2014), Kt qu xột nghim sng lc HBV, HCV, HIV, giang mai ngi hin mỏu ti Vin Huyt hc Truyn mỏu TW nm 20122013, Tp Y hc Vit Nam, thỏng 10 s c bit, 45-50 Bch Khỏnh Hũa v cng s (2007), Xột nghim 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