TY LE NHliM HDVVA P H A N B 6 KIEU GEN HDV dBENH NHAN Dl/ONGTINH HBsAg DIEU TRj TAI MOT SO BENH VIEN PHIA BAC
Bui Tiin Sy', Nguyin LTnh ToanS Nguyen Minh Hung^, Nguyen Hdng ThangS Phan Quoc Hoan", Le Hfru Song'.
Nhilm vi nit viem gan D (HDV) dupe xem la nguyen nhan dan den cac bien chung nghiem trpng tren benh nhan nhilm vi nit viem gan B (HBV) so vdi chi don nhilm HBV. Vdi sd lupng khoang 9,5 trieu ngudi bi nhilm o Viet Nam thi HBV da, dang va se trd thanh ganh nang rat 1cm ve van de sue khde cho toan xa hdi. Chiing tdi tien hanh nghien cuu vdi muc tieu danh gia ty le nhilm HDV tren benh nhan nhidm HBV d miln Bac Viet Nam va xac dinh su phan bd kieu gen HDV. 266 mau huyet thanh cua cac benh nhan duong tinh HBsAg o mien Bac Viet Nam da dupe phan tich ve sy cd mat HDV su dyng ki thuat RT - PCRs dac hieu cho HDV va xac dinh kieu gen bang phuong phap giai trinh ty va cay phan tich loai.
Ket qua cho thay ty le nhiem HDV d nhimg benh nhan duong tinh HBsAg tai Viet Nam la 15,4%, trong dd cao nhat d nhdm viem gan cap voi ty le 43,3%. Kieu gen HDV - 1 chiem uu the, tuy nhien kieu gen HDV - 2 cung dupe phat hien trong nhdm benh itiian nghien cuu. Ndng dp huyet thanh cua aminotransferase (AST, ALT) ciing nhu la birilibin toan phan va birilibm tryc tiep tang dang ke d nhdm benh nhan duong tinh vdi HDV (p < 0,05).
TCr khoa; Viem gan vi nit D, xac dinh kieu gen.
DAT VAN DE
Nhiem vi nit viem gan D (delta vims, HDV) diTdc xem la mot trong cac yeu to dan tdi cac bien chiing nghiem trpng cua viem gan vi nit do thuc day nhanh qua trinh ti&n tdi xd gan, tang nguy cd xd gan mat bu va tir vong so vdi chi ddn nhiem HBV. HDV sir dung khang nguyen vd cua VI nit viem gan B (HBsAg) de xam nhap va thoat khdi te bao gan, do vay viem gan do HDV chi xay ra tren benh nhan dUdng tinh vdi HBsAgi'i. Viem gan do HDV vi vay co 2 the hoac la dong nhiem hoac la bdi nhiem vdi viem gan do HBV
HDV virion la phan tii' hinh cau khoang 36mm dddng
"'Bfnh vifn Trung uung Quan dqi 108. BQ QUOC phong: '"Hpc Kpn Qudn Y. BQ Quae phong. "'Dai Hpc Duy Tdn Dd Ndng.
NgAy nh$n b^l: 17/6/20/5 N g i y phan bi^n xong: 27/6/2015.
N g i y duy^t ding: 31/7/2015.
Chju Irilch ohi^m ngi dung khoa hgc: Biii Tiin Sy. B4nh vi$n Thing mrng Qudn dgt 108. BQ Qudc phong.
Difn ihoQi: 097455294. E-mail Iiensy20l5@yahoa com
kinh, dypc tao thanh bdi HBsAg va thanh phan lipids ciia te bao chii. Phi'a trong la nudeocapsid bao gom 2 dong phan cua khang nguyen vi rut viem gan D (HDAg nho - sHDAg va Idn - IHDAg) va chuoi RNA vdng mach ddn 1^ bp gen cua vi rut viem gan D (HDV-RNA) vdi chieu dai khoang l,7kbf*]. Trong khi sHDAg dypc yeu cau cho qua trinh tong hdp HDV-RNA thi IHDAg LTC che qua frinh tong hdp HDV - RNA va thuc day hinh thanh HDV virioni'i.
Cac nghien cijy da trung tam lien tiep gan day cho thay ty le nhiem HDV da giam xudng ro ret d cac nUdc trydc day cd ty le cao nhd ty le nhiem HDV giam tir 23%
nam 1983 xudng 8,7% nam 1997, tai Italia, Dai Loan (23,7% xudng 4,2%), Tay Ban Nha (15,1% xuong cdn 7,1%) va Tho NhT Ky (29% xuong cdn 21,1%).
Chydng trinh tiem chiing md rpng vac xin HBV b i t dau thyc hien tai Viet Nam nam 2003 da cd tac dung giiJ ty le nhiem HBV khdng tang len trong nhifng nam gan day. Tuy nhien vdi sd Iddng dan sd cao, ty le nhiem HBV khoang 10,7% dan sd, ddc ti'nh cd khoang 9,5 trieu
2 - T R U Y ^ N H l i M V f $ T N A M * s 6 3 ( 1 1 ) - 2 0 1 5
ngddi nhiem HBV d Viet Namt^i va trd thanh ganh nang cho xa hdi trong nhiing thap nien tdi day do cac bien chiing hj" nhiem HBV Mac dii HDV lam hen trien nhanh cac bien chiing do nhiem HBV va lam nang them tinh trang benh nhuhg nghien ciili ve HDV van cdn rat it duUc tien hanh d Viet Nam. Mdt nghien ciiy tai tinh Thai Binh nam 2007 cdng bo ty le dddng tinh rat thap vdi HDAg, khoang l,3%i^l Mdt nghien ciili khac tren cac benh nhan nhiem HBV d mien Nam Viet Nam da khdng phat hien mpt trydng hdp nao nhiem HDV trong 73 tru'dng hdp nhiem HBV bang ky thuat RT - PCR'^"]. Tuy nhien mac dii chddng trinh tiem chiing md rpng vac xin HBV dypc tien hanh sdm hdn d Thai Lan va Malaysia nhy ty le nhiem HDV dddc cdng bo cao hdn tai Malaysia vdi ty le 4,9%, ty le nay cao hdn nhieu tren cac doi typng nghien hiit tiem chich ty le nay la 20 - 34% va tai Thai Lan cd the len den 21,8%.
8 kieu gen khac nhau ciia HDV da duUc bao cao trong cac nghien ciTu trddc day vdi ty le khac biet nucleotide giiJa cac kieu gen la 20 - 36%, trong khi dd trong cung mpt kieu gen ty le nay cd the len den 16%[*i. Kieu gen HDV - 1 phan bo toan cau va la kieu gen chiem uli the d chau Au, HDV - 2 chu yeu phat hien d chau A, HDV - 3 hau nhd chi dypc xac djnh d phia Nam cua khu vyc Nam My, HDV - 4 dddc bao cao luU hanh d Dai Loan, Nhat Ban va kieu gen HDV - 5 den HDV - 8 chi phat hien thay d chau Phi. s y lien quan giu^ kieu gen HDV va mirc do ciia benh viem gan dypc bao cao trong mpt so nghien ciiU, HDV - 1 dddc cho la cd the lien quan den ca miic dp nhe va nghiem trpng cua benh, trong khi do HDV - 2 chii yeu phat hien d benh nhan mirc dp nhe.
Trong nghien ciTu nay chiing tdi tien hanh nghien CLIIJ tren mgt quan the benh nhan nhiem HBV cd day du cac thdng tin ve lam sang va can lam sang de khao sat ty le nhiem HDV va djch te hoc phan tu" ciia HDV tren benh nhan Viet Nam.
D O I T U O N G V A PHl/ONG PHAP
Ddi tUdng: 266 benh nhan nhiem HBV dddc dieu tri tai Benh vien Bach Mai, Benh vien Quan Y 103 va Benh vien Trung udng Quan dpi 108 giai doan 2003 - 2009 dong y tham gia nghien CUIJ. Tat ca cac benh nhan deu am tinh vdi HCV, HIV va khong cd tien sir nghien rypu va tiem chich. Trong sd dd 30 benh nhan viem gan B cap tinh (AHB), 62 viem gan B man tinh (CHB), 84 benh nhan xd
gan do HBV (LC) va 90 benh nhan ung thd gan (HGC).
Phu'dng phap: Nghien CUIJ md ta cat ngang.
Phat hien HDV - RNA trong huyet thanh Axit nucleic tdng sd d\Sdc tach tir huyet tiianh benh nhan nhiem HBV bang bp kit High Pure Viral Nucleic Acid Kit (Roche, Grenzach-Wyhlen, Germany), chia nhd thanh cac tube lOpI luti giLJ" d -80°C cho den khi sir dung. Hai phan iihg nested/semi - nested PCR dddc phat trien de phat hien, khang dinh HDV - RNA trong huyet tiianh. Cac san pham PCR dddng ti'nh du*dc diing lam trinh ti/ nguon cho ky thuat giai trinh tu" de phan tich kieu gen ciia HDV.
Moi mpt mau dddc tien hanh lap lai it nhat trong 1 lan chay. Nhihig mau nghi ngd ddpc chay lai PCR bang mdt ky thuat vien khac.
Chu trinh nhiet ciia phan iing nested/semi - nested PCR nhy sau: Tong hdp cDNA axit nucleic tong so difdc thdc hien tai 50°C/30 phiit, sau dd 95°C/15 phut. Chu trinh khuech dai PCR gom 35 chu ky: 94<'C/30s, 56°C/30s va 72°C/45s, bddc keo dai cuoi ciing trong 10 phut tai 72°C. Vdng nested/semi - nested cd chu trinh nhiet 95°C/2 min, 30 chu ky vdi 9 4 ^ /30s, 58''C/30s, 72°C/45s va bydc keo dai cuoi ciing trong 5 phiit tai 72°C, thap nhat d 72''C. Trinh t y va vj tri moi dUdc bleu hien d Bang 1 va Hinh 1.
Bang 1 : Trinh tU cac primer
Primer
HDV-04 HDV-05 HDV-05 HDV-07 HDV-19 HDV-20 HDV-22
Sequences
GGATGC0CAGGTCG6ACCG AAGAAGAGRAGCCGGCCCGY ATGCCATGCCGACCCGAAGA GGGGAGCGCCCGGDGGCGG GGACCCGTCAGOJAACAG GGCCATCAGGTAAGAAAGGA CAaCGGATGGCTAAGGGAG
Position
856 - 874 1159-1179
888 - 907 1104 - IIZZ
316 - 334 672 - 691 819 - 838
Nested/semi - nested RT - PCR
I'PCR l^PCR 2'«'PCR Z'-PCR I'&a™" PCR
1»PCR Z'-'PCR W tri gin moi di/a vao trinti tif tham chieu cua dimg HDV NC10D1653 (geneBank)
T R U Y ^ N H 6 M V 1 | T N A M * S 6 3(11) - 2015 - 3
Hinh 1 : Chien thuat thiet ke primer va cac phan uhg RT - PCR
Hinh 1: (A)(^c primer tJuVc thiet ke rao viing bao tan cua bd gen HDV dwa vao trinh tU tham chieu cua 38 bo gen HDV duVc cong bo tren geneBank. RI, R2 = Vung Ribozyme, C = vung keo dai amino adde dau C (C - terminal amino aade extension), HDAg = khang nguyen HDV. (B)Vi tri gSn primer cua cac phan ijtig nested/semi - nested RT - PCR tuifng ung tren bo gen HDV, vi tn dude danh theo Chung HDV NC1001653. (C) Kit qua dien di san pham nested RT- KR phat hien HDV-RfM. M=marker, K=d^lihg duUng, NC=ch({ng im, cac so tit 1' 41 la vi tri dien di aia san pham PCR tit 21 mau binh phim (moi miu difdc chay lip lai 2 tan)
Giai trinh ti/ va xac djnh kieu gen
San pham PCR dypc tinh sach Exo SAP - IT kit (USB Corporation, Ohio, USA). Giai ti-inh tu" bang he thong ABI3100. Trinh t d dddc phan tich va xay ddng cay phat sinh loai bang phan mem BioEdit 9,7 va MEGA 5.
Phan tich va xuT ly so' lieu
Phan tfch thdng ke dddc thyc bang phan mem SPSS phien ban 20 (IBM Corporation, Armonk, NY, USA) va phan mem PrismS (phien ban 5.01, GraphPad Software, San Diego California USA, www.graphpad.com). Cac diJ lieu phan loai dddc phan tich bang thuat toan Fisher's exact test. Cac dU lieu phi tham sd dddc so sanh bang thuat toan Mann - Whitney U test, vdi gia tri p < 0,05 dddc xem la cd y nghTa ve mat thdng ke.
KET QUA
Dac diem lam sang, can lam sang cua benh nhan nhiem HBV
266 benh nhan dddng ti'nh HBsAg d Viet Nam dddc duS vao nghien CLUJ nay, ti'ong dd cd 48/266 (18%) la NU va 218/266 (82%) la Nam. Khdng ai trong sd cac benh nhan dddc dieu tri thuoe khang vi rut. Op tuoi tmng binh ciia benh nhan la 48 tuoi (15 - 79 tudi). Tai Iddng HBV trung binh ciia cac benh nhan la 3,92 loglO HBV ban sao/ml (2,46 - 6,37 loglO HBV ban sao/ml). Dac diem cd ban ciia cac benh nhan trong cac nhdm dUdc tdm tat d Bang 2. Benh nhan khong dong nhiem HCV va, hoac HIV.
Benh nhan trong nhdm AHB cd dd tuoi tre hdn (tmng binh 35 tudi) so vdi nhung ngddi trong LC (trung binh 51 tuoi, p < 0,001), HCC (tnjng binh 53 tuoi, p < 0,001) va CHB (tmng binh 40 tudi, p < 0,05) (Bang 2). Nong dp alanine aminotransferase huyet thanh (ALT), aspartate amino- transferase (AST), cQng nhu" bilirubin toan phan, va biliai- bin trdc tiep trong nhdm AHB tang cao hdn han so vdi cac nhdm khac (p < 0,001) (Bang 2).
Dang chu y, khdng cd sy" khac biet Idn trong HBV loads giti^ cac nhdm benh nhan dieu tra (AHB, CHB, LC, va HCC) (ANOVA - test, p = 0,70). Trong 30 benh nhan dddng ti'nh HDV xac djnh dddc kieu gen HBV 3, kieu gen chiem ud the la HBV - A (6/30; 20%), C (5/30; 16,7%), va D (8/30; 26,6%). Cac kieu gen HBV khac nhd HBV - B, E, G chi chiem ty le nhd (1/30; 3,3%) va khong phat hien dddc HBV-H. Dang chii y, cd 26,6% (8/30) benh nhan dddng tinh HDV nhiem nhieu kieu HBV khac nhau (Du" lieu khdng trinh bay).
Tyle nhiem HDV d benh nhan dttdng tinh HBsAg
HDV - RNA dddc phat hien trong 41/266 benh nhan dddng tinh HBsAg vdi la 15,4% (CI95[11,1 -19,8], hinh IC va 2). Trong sd 41 benh nhan dddng tinh HDV cd 10/41 la NiJ (24,4%), 31/41 la Nam (75,6%). Ty le nhiem HDV d nhdm benh nhan AHB la 43,3% (13/30) chiem ty le cao cd y nghTa so vdi 11,3% (7/62) nhdm CHB, 11,9%
(10/84) nhdm LC va 12,2% (11/90) nhdm HCC vdi gia trj p < 0,01 (Bang 2).
4 -TRliY^NHliMVl|TNAM*sd3(11)-2015
Bang 2. Dac diem can lam s^ng benh nhan nhiem HBV
Tuoi (nam) Gidi b'nh (Nam/NuO ALT (lU/L) AST (lU/L) Bilirubin toan phan (mg/dl) Bilirubin tru'c tiep (mg/dl) AI=P (mg/dl) Tai IWdng HBV (ban sap/ml)
AHB n = 30
35*
24/6 969,0***
1128,5***
123,9***
9 2 , 7 * "
ND 3,77
CHB n = 62
40 52/10 147,5 156,5 31,5 17,0 ND 4,05
LC n = 84 5 1 * * * 66/18 63,0 67,5 29,4 16,0 ND 3,98
HCC n = 90 53**«
76/14 60,5 59,5 17,5 8,2 87,1 3,86 Gia tri dddc bSng gia tn twng binh; lU: Ddn vi quoc te; ND: Khong thirtr hien
* SI/ khac biet tuoi benh nhin nhom AHB vdi nhom CHB c6 y nghia thong ke vdip < 0,05
* * * SI/khac biet cac thong so can lam sang cua cac nhom khac vdi nhom AHB co y nghia thong kevdip< 0,001
' Gia tii trung binh duVc tinh bang loglO ban sao/ml, p = 0,70 (Anova test)
AHB: Acute HepatiSs B; CHB: Oironic Hepabtis B; LC: Uver Cirrtio- sis; HCC: Hepatocellular carcinoma; ALT: Alanine aminotransferase;
ALT: Aspartate aminotransferase; AFP: Alpha - fetoprotein.
Hinh 2: Ty 1^ nhiem HDV 6 benh nhan dydng tinh HBsAg
Dac diem can lam sang cua binh nhan nhiem HDV Cac dac diem can lam sang ciia benh nhan HDV da dddc md ta trong Bang 3. Ket qua cho thay cac benh nhan dddng tinh HDV tre hdn so vdi nhdm am tinh vdi HDV (tiling binh 41 so vdi 49 tudi, p < 0,05). ALT va AST d nhdm HDV dddng tinh tuUng u^g la 213,0 va 223,0 lU/l cao hdn so vdi nhdm benh nhan am tinh HDV vcfi ket qua tddng irtig 82,0 va 88,0 lU/l (p < 0,001). Tddng t d nong
dp bilirubin toan phan (59,0 so vdi 23,9 mg/dl) cung nhd bilirubin tn/c tiep (34,3 so vdi 13,1 mg/dl) cao hdn dang ke d benh nhan dudng tinh HDV so vdi benh nhan am tinh HDV (p < 0,001) (Bang 3). Vdi nhung benh nhan am ti'nh vdi HDV, tai Iddng HBV d mu'c vula phai, khdng cao hdn nhieu so vdi benh nhan ddng nhiem HBV/HDV (3,98 so vcfi 3,76 loglO ban sao/ml, p = 0,60) (Bang 3).
Bang 3: Dac diem can lam sang nhom benh nhan HBsAg dUdng tinh nhiem va khong nhiem HDV
HDV- negatrve {n = 225)
HDV- posibve (n = 41)
Age (years)
49
41- ALT ( l U / L )
82,0
213,0**' AST ( W / t )
»
223,0*"
Total Billfubin (mg/dl)
23,9
59,0***
Direct Bi lira bin (mg/dl)
13,1
34,3***
HBV load' ban sao/ml
3,98
3,75
Gia tri difdc bang gia trj trung binh; lU: Ddn vi quoc te; ND: Khong thWc hien
*su'khac biet coy nghia thong ke vdi p< 0,05
*'* sW khac biet nghia thong kevdip < 0,001 ' Gia tri trung binh tft/tfc b'nh bang loglO ban sao/ml, p = 0,6
Phan bo cac kieu gen HDV d benh nhan Viet Nam nhiem HBV
De xac i^nh kieu gen HDV tren benh nhan nhiem HBV d khu vdc mien Bac Viet Nam, chung tdi phan ti'ch 21 mau HDV - RNA tach dddc tii' benh nhan nhiem dong tiidi HBV va HDV. San pham RT - PCR khuech dai mdt phan bp gen HDV nt888 -1122 (vung ma hda HDAg) dddc giai trinh tij"
tn/c tiep bang phddng phap giai trinh t d Sanger. De Wiang djnh cac chung HDV rieng biet dac hieu cho tiJTig benh nhan, mot viing khac ciia bd gen HDV dddc khuech dai va giai trinh td. Ket qua d Hinh 3A khang djnh cac chiing HDV dac hieu cho tiitig benh nhan va khac biet vdi cac chiing d cac benh nhan khac. Ket qua nay cho khang dinh khdng cd qua trinh nhiem cheo vat chat di truyen trong qua trinh xu"
ly benh pham va khuech dai vat chat di truyen. Cac trinh t d thu dddc sau khi giai trinh t d mdt phan viing HD/\g dddc xii" ly va phan tich phat sinh chiing loai ciing vdi cac trinh t d HDV tham chieu lay tir NCBI - GenBank (Hinh 38). Ket qua phan tich cho thay hai kieu gen HDV dddc tim thay trong nhdm benh nhan ngddi Viet Nam, trong do kieu gen HDV - 1 la chiem ud die (90,5%, 19/21), kieu gen HDV - 2 da dddc phat hien d hai benh nhan (9,5%, 2/21).
TRUYtNNHiwIVI&TNAM* s o 3(11) - 2015 - 5
BAN LUAN
Nhiem vi rut viem gan B la mdt trong nhDtig van de ve sifc khde nghiem ypng tai Viet Nam vdi ty le 10 - 20%
ngddi dan. Mac dii chddng tiinh tiem chiing md rdng vac- cine HBV da dddc pho cap toan qudc nam 2003 tai Viet Nam, nhutig sd Iddng ngdcfl bl nhilm HBV van tang do dan sd ngay cang tSng. Ty le nhiem HBV cd the se giam trong tddng lai gan, hjy nhien ty le HBV lien quan LC, HCC cd the tang tiong hai thap ky tdi do sd keo dai ciia benh viem gan B man ti'nh .
HDV chi CO tiie lay nhiem va lan duyen khi cd mat ciia HBsAg. Cac bao cao gan day cho tiiay nhiem HDV da va dang quay trd lai d chau Au vdi ty le 8,1%, 1 1 % , 8,5%, 20,4% tddng iiYig d cac nddc Y, Ddc, Anh, Romani. Ty le nhiem HDV cao cung dddc bao cao d mdt so nddc chau A, nhd Pakistan (35,2%), Mdng Co (13,6%) va d mpt so tinh Trung Qudc. Ngddc lai, ty le nhiem HDV thap cung dddc ghi nhan d cac nddc chau A nhd Han Qudc (0,32%), Malaysia (4,9%) va Indonesia (< 0,5%).
Nghien ciid ve HDV chda dddc thdc hien nhieu d Viet Nam. Hai nghien ciid trddc day cho thay ty le nhiem HDV rat thap tren benh nhan dddng tinh HBsAg d Viet Nam chi tif 0% den 1,3% '^•"". Ngddc lai mdt bao cao gan day md ta t / le HDV la 10,7% dan so Viet Nam, tixing khi dd ty le nay d ngddi tiem chich ma tiiy la 25,6% va 17,8% trong nhom quan nhan^^i.
De xac dinh ty le HDV d Viet Nam, chung tdi xay duhg phddng phap RT-PCR dac hieu cho HDV cd do nhay cao (Hinh 1). Tren 266 benh nhan nhiem HBV vdi day du cac du' lieu can lam sang, t / le nhiem HDV la 15,4 % ( 0 9 5 : 11:1 - 19,8) (Hinh 2). Ket qua nay cung phu hdp vdi mdt nghien ciili tiudc day ciia Dunford cho thay 10,7% dan sd Viet Nam nhiem HDV (095: 7,3 - 14,1). Nhiem HDV cao nhat d nhdm benh nhan AHB vdi 43,3%, sau dd la HCC 12,2%, LC 11,9% va CHB 11,3%. Mac du phddng phap va cac ddi tddng nghien cutJ dddc diing de xac dinh ty le HDV trong vi§m gan B cap tinh la khac nhau, nhdng ty le HDV cao d benh nhan AHB cung phu hdp vdi cac nghien cud trddc dd d Nga va cac Idu vdc sdng Amazon d Peru vdi ty le dddng tinh HDV tddng iitig la 39% va M%^^'^\
Cac chi tieu can lam sang ciia cac benh nhan Viet Nam dddng tinh vdi HBsAg nhd ALT va AST, bilirubin toan phan va bilirubin trdc tiep tang gap 2 lan tix)ng benh nhan nhiem HDV so vdi benh nhan khdng nhiem HDV (p <
0,001). Ket qua nay cung da dddc Wiang dinh d nhijtig nghien ciiU trddc day^^'. Dac biet frai ngddc vdi nhuhg mo ta trddc, benh nhan viem gan B man tinh d Italia thi tai Iddng virus HBV d nhiJng benh nhan ddng nhilm HBV/HDV thap hdn so vdi benh nhan ddn nhiem HBV, vdi ndng do tddng dng d hai nhdm nay la 641 [70 - 9,4 x lo' ban sao/ml] so vdi 1,6 x 10' ban sao/ml [3,1 x lo^ - 6,5 X 10« ban sao/ml], chiing tdi quan sat tiiay khdng cd sd khac biet nhieu ve tai Iddng HBV giife benh nhan dudng tinh va am tinh HDV Ket qua nay cd the phu hdp vdi mot nghien cud cho thay d benh nhan nhiem 2 loai vi nit nay 23% benh nhan ca hai loai vi rut ciing nhan len, 70% la vi riit HBV bi canh tranh va so it 4 % HDV bi ii'c che.
Hinh 3: Phan bo kieu gen HDV d nhdm benh nhan dUdng tinh HBsAg
(A) Trinh ti/HDV ciia benh nhan HBsAg dudng tinh va trinh ti/^am chieu aJa Mng NC001653 tuang uhg vdi vdng HDV- RNA tifnt335
• 635. (B) Phan ti'ch cay phat chiing loai vdi mo hinh Kimura 2 pa- rameters model va mo hinh ndghbor - pining vdi trinh ti/DNVcua benh nhan va cac trinh ti/chung HDV tham chieu tUdng utig tif nt888 -1122. Cac trinh ti/HDVttiam djiai vdi8 kieu gen khacnhau thu thap tit GeneBank tfiAfc danh so tit 1 den 8 kem theo so tniy dp cua GeneBank, vidu 2 - AF104624_Taiwa la chiing HDVAF104624 kieu gen HDV - 2 phan lap tai Dai Loan.
6 -TRUYENNHliMVirrNAM*s63(11)'2015
Rieng chiing HDV B171 co lien quan mat tliiet v f l cliung HDV - 1 Trung Phi (A]000558) vcli ilo tin cay la 9 1 % . Ket qua ciia chung toi mgt lan n i h cho thay kieu HDV - 1 phan bo rong Idiap tren the gidi nhu* cac nghien CCHJ tru'6c day da bao cao. Kieu gen HDV - 2 cung du'dc phat hien trong nhom benh nhan nghien cu\j ciia chung toi. HDV - 2 du*cfc cong bo iu\j hanh pho bien Dai Loan, Nhat Ban, Nga.
KET LUAN
Ket qua cua nghien cutJ cho thay:
Ty le HDV cao (15,4%) va pho bien nhat trong nhom viem gan giai doan cap tinh (43,3%), trong do kieu gen HDV - 1 chiem uli the cl benh nhan du'dng b'nh HBsAg.
Nhiem dong thSi HBV/HDV co the lien quan vdi sy tang cao cac chi so can lam sang lien quan den chii'c nang gan tren cac nhom benh nhan viem gan man b'nh (CHB, LC, HCC).
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TRUYiwrjHltM V I E T N A M * SO 3(11) - 2015 - 7
PREVALENCE AND GENOTYPE DITRIBUTION OF HEPATIHS D VIRUS AMONG HBsAg - POSITIVE PATIENTS I N NORTHERN VIETNAM
Summary
Hepatitis D virus (HDV) infection is considered to cause more severe hepatitis than hepatitis B virus (HBV) monoinfection. With more than 9.5 million HBV - infected people, Vietnam will face an enormous health burden.
Therefore, we assessed the prevalence of HDV in Viet- namese patients, determined the HDV - genotype distn- bution and compared the findings with the clinical outcome. 266 sera of well - characterized HBsAg-positive patients in Northern Vietnam were analysed for the pres- ence of HDV using newly developed HDV - specific RT -
Key words: Hepatitis D virus, determination of genotype.
PCRs. Sequencing and phylogenetic analysis were per- formed for HDV - genotyping. The HDV - genome preva- lence observed in the Vietnamese HBsAg - positive patients was high with 15.4% while patients witii acute hepatitis showed 43.3%. Phylogenetic analysis demon- strated a predominance of HDV-genotype 1 clustering in an Asian clade while HDV - genotype 2 could be also de- tected. The serum aminotransferase levels (AST, ALT) as well as total and direct bilirubin were significantly elevated in HDV - positive individuals (p < 0.05).
- TRUYfeu NHilUI VI|T NAM * s o 3(11)
