VIRUS HANTA TREN DONG VAT GAM NHAM O TAY NGUYEN VA MIEN NAM VIET NAM, 2006 - 2009
Vu Dinh Luan, Kumiko Yoshimatsu**, Le Nhi, Midori Taruishi**, Vo Thi Huong*, Rika Endo**, Cao Minh Thing, Hoang Kim Loan, Jiro Arikawa** , Vu Thj Que Huong
Vien Pasteur thanh phd Ho Chi Minh
* Vien Ve sinh Dich te Tiiy Nguyen
** Vien Thirc nghiem Dpng vat, Trir&ng 1 - Dai hoc Hokkaido, Nhat bdn
TOM TAT
Giam sat tinh hinh nhiim virus hanta d khu vuc Tay Nguyen va mien Nam Viet Nam dupe thuc hien dua tren ket qua xet nghiem 1.066 mlu huyet thanh thu thap tu 6 loai dpng vat gam nham va 245 mlu huyet
thanh tir Suncus murinus. Ket qua xet nghiem cac mau huyet thanh thu thap tir dpng vat gam nham bang ky thuat ELISA cho thay ty le phat hien IgG khang virus hanta la 16,76 % a Rattus norvegicus va 13,1 % d S.murinus. Cac mau huyet thanh xac dinh duong tinh sang lpc bang ELISA, khang dinh lai bang IFA va WB, dupe luu giu de djnh type huyet thanh. Ket qua FRNT (Focus Reduction Neutralization Test) xac djnh dupe type huyet thanh ciia virus hanta luu hanh d mien Nam Viet Nam la type huyet thanh Seoul (SEOV) va Thottopalayam (TPMV) Bang ky thuat RT-PCR da phat hien duoc vat lieu di truyen ciia virus hanta tir 3 mau md phoi co ky hieu CSG5. SCG11 va 24D12 - day la nhirng mau dupe thu thap d thanh pho Hd Chi Minh (Cang Sai Gdn va Quan 12). Phan tich trinh tu va xay dung cay phat sinh loai tir nhung doan gen dupe phat hien, xac djnh genptype ciia virus hanta luu hanh tren R norvegicus ct thanh phd Ho Chi Minh (Cang Sai gdn va quan 12) cd mdi lien quan chat che vdi cac virus Seoul dai dien d Nhat, Indonesia, Singapore va mien Bac Viet Nam Ket qua huyet thanh hpc che th ay cd su luu hanh ciia virus hanta bao gom ca chung SEOV tir Rattus.sp, va chung TPMV tir S.murinus d mien Nam va Tay Nguyen, Viet Nam Ngoai ra, vpi su phat hien IgM dac hieu va khang thi trung hoa khang SEOV a benh nhan ket qua nghien ciru da cung cap bang chirng dau tien ve sir luu hanh va su lay truyen SEOV tir dpng vat gam nham sang ngudi.
Tir khoa: virus hanta, ELISA, RT-PCR, Sequencing I. DAT VAN DE
Virus hanta thudc ho Bunyaviridae la virus hinh cau, cd vd, bo gen cua virus gdm ba phan doan spi don RNA am dupe gpi ten theo kich thudc: doan S (Small) la doan nhd nhat dai 1.6 Kb ma hda cho nucleocapsid pro- tein, doan M (Medium) la doan trung binh dai 3.6 Kb ma hda cho cac protein vd glycopro-
tein va doan L (Large) dai 6,3 Kb ma hda cho cac polymerase [1]. Virus hanta gay ra hai hpi chirng benh ly chinh 0 ngudi: hpi chimg sot xuat huyet kem vdi trieu chiing than (HFRS
Hemorrhagic Fever with Renal Syndrome) va hpi chiing viem phoi (HPS - Hantavirus Pulmonary Syndrome). Vat chii truyen benh la mpt sd loai dpng vat gam nham, virus lay nhiem thdng qua dudng ho hap khi tiep xuc Tac gia : Vu Dinh Luan
Dia chi co quan: Vien Pasteur thanh phd Ho Chi Minh. 167 Pasteur Quan 3, TP HCM Dien thoai: 08 38 29 63 5 1. 0903 678 809
Email: [email protected]
Tap chi Y hoc du phdng. Tap XX, so 6(1 14) 171
voi chat thai hay nude tieu co mang virus tir dong vat gam nham. Virus hanta co nhieu type huyet thanh khac nhau. moi nhom co mot titc chii gam nham rieng biet.
Theo thong ke ciia Td chirc Y te The gioi (WHO), hang nam co khoang 500.000 ca benh duoc xac djnh do nhiem virus hanta. Cac ca benh HFRS chu yeu gap a khu vuc Chau A va cac nude Balkan voi ty le tu vong khoang 4 - 15 %, con cac ca benh HPS thuong gap 6' Chau My voi ty le tii' vong khoang 40 %.
Hien co 5 loai chung virus hanta luu hanh ciia Chau A do la: virus Seoul (SEOV), virus Thai Land (THAIV). virus Puumala (PUUV). vi- rus Hantaan (HTNV) v a v irus Thottopalayam (TPMV) [1]. Gan day. co thong bao ve su luu hanh ciia SEOV va TPMV o mien Bac Viet Nam, cung nhu theo bao cao ciia WHO ty le luu hanh huyet thanh tren nguoi d nuoc ta la 5.4 %, chu veu 0 mien Bac Viet Nam [2], [3].
Cho den nay. chua co thong tin ve tinh hinh nhiem v irus hanta 6' mien Nam va Tay Nguyen cua Viet Nam. Tren thuc te, viec lay nhiem v irus len cong dimg co lien he mat thiet voi su luu hanh cua virus tren quan the gam nham. Do vay. giam sat tinh hinh nhiem virus hanta tren quan the gam nham la dieu cap thiet de tao tien de phiit trien nhung nghien cuu tren cong dong.
II. PHUONG PHAP NGHIEN CUU 2.1. Vat lieu:
Te bao va virus: Cac chung SR-11 (SEOV). HTNV 76-118. THAIV va TPMV duoc nuoi cay tren dong te bao Vero E6 (ma so CI008. do Trung tam djnh danh te bao Hoa Kv cung cap).
Mau benh phiim vii thdi diem thu thap:
Co 1.066 huyet thanh cua Rutins.sp va 245 huyet thanh cua Suncus murinus thu thap 0 Dong Nai, Cang Sai Gdn. VTnh Long va Tra Vinh vao cac thai diem khac nhau, 2007 - 2008;
0 Tav Nguven (Dak Lak. Gia Lai. Kon Turn
va Pleiku) hang thang tir 3 2007 den 12/2009.
Mau tim va mo phoi ciia dong vat gam nham duoc thu thap tai thuc dia va bao quan o -70"C tai phong thi nghiem Vien Pasteur thanh pho Ho Chi Minh cho den khi xet nghiem.
Sinh pham va hda chat: Khang nguv en tai to hop HS103 vii TPMV [4]. Cac sinh pham hoa chat can thiet cho ky thuat ELISA.
IFA, WB. RT-PCR v a giai trinh gen. Cap moi dac hieu viing gen S cho san pham 1.000 bp co ky hieu MurS 11 OF (TGG TCC ACT TOT ATR CCC AT) va MurSl I60R (CAG AAG GTI A1G GAT GCAGA); trinh tir cap moi dac hieu viing gen M co ky hieu VNM41F (AGG ATC ATG TGG AG T TTG CTA) va VNSEOM-2060R (GGC AAA GAA AAG TCT A AT TCG AGG TCT GTA TGC AT) khuech dai viing gen khoang 2 Kbp.
Trang thiet hi vd vat lieu: Cac trang thiet bj va vat lieu can thiet de thuc hien k"\
thuat ELISA, ky thuat huynh quang. WB.
FRNT, PCR va giai trinh tu gen.
2.2. Phuong phap:
Ky thuat HI.ISA (Enzyme Linked Im- munosorbent Assay): Su' dung khang nguyen HS103 va TPMV cho ky thuat ELISA.
Ky thuat huynh quang gian tiep (IFA - Indirect Fluorescent Antibody Test): []]. IgG khang virus hanta trong mau duoc phat hien bang cong hop gan I-TTC (Fluorescein isothiocyanate), doc ket qua bang kinh hien vi huynh quang.
Ky thuat Western Blot (WB): Sir dung khang nguyen HTNV va TPMV tuong ling cho huyet thanh Rutins.sp va S.murinus.
Khang nguyen tai to hop gan tren mang nitro- cellulose se duac tiep xuc vai huyet thanh thii nghiem va cong hap, tuong tu nhu phan ling ELISA [5],
Ky thuat trung hda giam do tap trung (FRNT - Focus Reduction Neutralization Test): Phat hien khang the trung hoa khang virus: SEOV va THAIV (kiem tra vdi huyet thanh tir Rattus.sp) va TPMV (kiem tra voi huyet thanh tir S. murinus) |6|.
172 Tap chi Y hoc du phong, Tap XX. so 6 (1141
Ky thuat RT-PCR va phan tich di iruycn phan tu: Sir dung mot so doan moi chuyen biet khuech dai viing gen phan doan S va M cho ky thuat RT-PCR de phat hien ARN cua virus hanta tir mot so mau mo phoi tir dong vat da co ket qua xac dinh huyet thanh duong tinh vor El.ISA. IFA va WB; San pham khuech dai duoc giai trinh tu bang he thong ABI PRISM.
III. KET QUA
Mot so phan mem ho trtr: Phan tich dich te hoc phan tir duoc tien hanh dua tren ket qua giai trinh tir cua san pham khuech dai.
Trinh tu gen duoc phan tich bang phan mem Genetyx. CLC Workbench va Blast (NCBI) [6]: Server ESyPrcd3D 1.0. NCBI Protein Structure database duoc sir dung de mo hinh hoa va phan tich can true khong gian ciia cac protein duoc ma hoa.
l ong cong co 1.3 11 man huyet thanh thu duoc tir dong vat gam nham o I 1 dia diem khac nhau. Thanh phan IgG khang virus hanta trong cac man huyet thanh sau do dupe sang loc bang ky thuat liLISA voi khang nguyen I FI N V tai to hop; Cac ket qua tir ky thuat ELI-
SA deu duoc killing d|nh lai bang IFA va WB.
Ket qua cho thay ty le huyet thanh duong tinh vai IgG khang virus hanta tren tong so dong vat gam nham thu thap lit 4,7% (62/1311) va ty le huyet thanh duong tinh thay doi tiiy theo tung loai dong vat. Co 16.76% huyet Tap chi Y hpc du phong. Tap XX, so 6 (114) 173
thanh duong tinh vai SEOV tren quan the R.
norvegicus (chupt cong) chii yeu o Tp.HCM (22% = 28/127) va Ddng Nai (4,3% = 2/46).
Ket qua phat hien IgG khang virus hanta tir loai S1. murinus trong so 245 mau huyet thanh duac thu thap tir 8 dja diem ciia cac tinh Dong
Nai. Tra Vinh, Tay Nguyen va cang Sai Gon tu nam 2006 - 2009 la 13.10% (32/245) trong do 0 Dong Nai phat hien co 2 mlu duong tinh (6,45%) trong tong s6 3 1 mau, 0 Tay Nguy en phat hien co 30 mlu duong tinh (14,5%) trong tong so 207 mau (hinh 1).
Bang 1. Kit qua FRNT xac djnh type huyet thanh cua virus hanta Imj hanh cr dpng vat gam nham
HGTH HGTH Ma s6 Loai Ma s6
SR-11 THAIV TPMV CSG5 R. norvegicus 80 <20 312 160
CSG11 R. norvegicus 80 20 324 80 CSG41 R. norvegicus 320 20 46TV 20 CSG45 R. norvegicus 320 20 141 80 CSG47 R. norvegicus <20 20 276 <20 CSG58 R. norvegicus 640 40 300 80 116 R. argentiventer <20 <20 306 160 131 R.exulans <20 <20 310 160 24D12 R. norvegicus 80 Khong lam Anti-TPMV 80 2008.5877 Benh nhan 80 Khong lam
Anti-SR11 80 <20 Anti-THAIV <20 80
HGTH: hieu gia trung hoa. (&) Huyet thanh chirng duong cua chiing virus & huyet thanh benh nhan. xac dinh nhiem virus hanta 0 quan 12 TP.HCM vao thang 10/2008; Cac mlu duoc xac djnh hieu gia trung hoa vai TPMV deu la cac mau S. murinus.
Su dung ky thuat FRNT vai cac chiing SEOV (SR-tl), THAIV va TPMV (VRC- . 6641 2). tren cac mau huyet thanh duong tinh,
xac djnh duac ty pe huy et thanh ciia virus han- ta luu hanh 0 mien Nam Viet Nam thuoc type huyet thanh SEOV va TPMV.
Chon cac mau mo phoi ciia dpng vat gam nham co ket qua huy et thanh hpc duong
tinh dupe khang djnh bang ky thuat FRNT, chiet xuat RNA virus, khuech dai bang cap moi dac hieu cho viing gen S va M, xac djnh co ba mau: CSG5, CSG11 va 24D12 duong tinh. Cac mau nay sau do duac giai trinh tu va phan tich. Toan bp thong tin di truyen tren doan gen S va M cua hai chiing CSG da dupe xac djnh. Rieng mau 24D12, chi xac djnh duoc mpt viing gen khoang 2,2 Kbp tren doan M.
De xac djnh genptype virus hanta luu hanh 0 Viet Nam. trinh tu nucleotide ciia cac chung CSG5. CSG11 va 24D12 dupe phan tich. sp sanh moi lien quan di truyen voi cac chiing SEOV tren the gioi.
174 Tap chi Y hpc du phong-. Tap XX, so 6(1 141
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Hinh 2. Phan tich so sanh trinh ty nucleotide doan S (1.7Kb).
Chung virus Sin Nombre thuoc phu ho Sigmpdpntinae ciia hantavirus dupe diing nhu nhom rieng biet. So ghi o nirt cuoi moi nhanh bieu hien phan tram ket qua lap lai voi do tin cay Bootstrap 1000. chi bieu hien boot- strap '-70%. Cac chung su dung de so sanh deu dupe tham chieu tren GenBank: RN41 (G0274944), RN46 (GQ274945). HuBJ7 (G()27938l). HuB.122 (GQ279379). HuBJ16 (GC>279380). HuBJ 19 (GU279389). Rn- SI1Y17 (GQ279388). HuBJIS (GQ279390).
SD201 (GQ279385). Rn-Dl 127 (GQ279393) Rn-Mll (GQ279383), Rn-DC8 (GQ279386) HuBJ3 (GQ279391). 7.1711 (AY750171) ZT10 (AY706368), 93HBX12 (HP 192308) Pf26(,YY()06465). SEOV 80-39 (NC 005236).
1R461 (AF329388). K24-v2 (AP288655) K24-c7 (AF288653). R22 (AF288295) Hb8610 (AF288643). Gou3 (AF184988) Thailand 741 (ABI 86420). Hantaan TJJ6 (AY839871), Sin Nombre (NC 005216).
Tap chi Y hoc dir phong. Tap XX. so 6 (114) 175
Tuong tir nhu ket qua phan tich so sanh tren trinh tir nucleotide ciia doan S ciia hai chiing CSG5 \a CSG11 voi 32 chiing virus hanta tham chilu, ket qua phan tich so sanh tren doan M ciia cac chiing CSG va 24D12
voi trinh tu gen M cua 34 chung virus hanta da dang ky trong ngan hang gen cung cho thay virus hanta luu hanh o V iet Nam thuoc geno- type SEOV.
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Hinh 3. Phan tich so sanh trinh tir nucleotide mot phan doan M (vj tri 810 - 2355) 176 Tap chi Y hoc du phong, Tap XX. so 6(114)
Chung virus Sin Nombre thuoc ho phu Sigmodontinae ciia hantavirus duoc dung nhu nhom rieng biet: So ghi 6 nut cudi moi nhanh bieu hien phan tram ket qua lap lai voi do tin cay Bootstrap 1000, chieu dai moi nhanh bieu hien 0 dang thap phan chi bieu hien bootstrap >70%. KVPN: khu vuc phia Nam. Cac chiing sir dung de so sanh deu dupe tham chieu tren GenBank: SEOV KI-88-15 (D17594). KI-85-1 (D17593). K.I- 83-262 (D17592), SR11 (M34882). 80- 39 (S47716). Jakartal37 (AJ620583): Cang Haiphong7 (AB355728), 20 (AB355730).
16 (AB355729). 28 (AB355731). Ha Noi 25 (AB355733). 9 (AB355732): B-l (X53861),
Bj H DO 1 (DQ133505), ZT71 (EE117248), ZT10(DO 159911). K24-e7(AF288652). K24-
\2 (AF288654). HB55 (AF035832). IR461 (AE458104). Gou3-e5 (AE288650). va ZJ5 (EJ811839); THAIV 749 (E08756); HTNV 76 -11 8 (Ml4627).Hantaan (NC005219). LEE (D00377) va Hojo (D00376); Dobrava virus (DOBV) chung Dobrava (L33685); PUUV chung Sotkamo (X6I034); Tula virus (TUV) chung Tula/Moravia/5302v/95 (Z69993).
Ket qua trong nghien cuu nay, da xac dinh virus hanta luu hanh 6' mien Nam Viet Nam co do tuong dong cao vdi cac cluing virus Seoul phan lap d Nhat Ban. Indonesia.
Singapore va mien Bac Viet Nam.
B
eo 100
CSG 11 AASIQSKIDE LKRQLAORIA AGKNIGQDRD PTGVEPGDHL KERSALSYGN 100
CSG 5 100 1AA47826-SEOV SR-11 LQ Q . RT S 100
AAP3349B-SEOV 80-39 100 Consensus AASIQSKIDE LKRQLADRIA AGKNIGQDRD PTGVEPGDHL KERSALSYGN
D CSG 5 IWVSLLLLAAL AGQGFALKNV FDMRIOCPHS VNFGETSVSG YTELPPLSLQ 50
CSG 11 50 GNVUSR-SEOV SR-11 V L F 50
AAB19434 - SEOV 80-39 V K 50 Consensus MWSLLLLAAL XGOGFALKNV FDMRIOCPHS VNFGETSVSG YTELPPLSLQ
Hinh 4. So sanh trinh ty acid amin cac doan gen (S, M) va md hinh hoa cau triic protein ciia hai miu phat hien duoc ARN tir virus hanta liru hanh o Viet Nam
(A. B) Cau triic khdng gian nucleo- acid amin hai protein (nucleocapside protein capsid protein ma hda bdi doan gen S vii gly- va glycoprotein) cua hai cluing SliOV (CSG5 coprotein G l ma hda bdi doan gen M. va 11) vdi cac cluing dai dien. Cac dlu "." bieu
(C, D) Mot phan ket qua so sanh trinh tir thi acid amin tuong ddng.
Tap chi V hoc dp phdng. Tap XX. so'6 (114) 177
Trinh tu nucleotide cua hai viing gen S va M ciia hai chiing SKOV Viet Nam (CSG 5.
CSG 11) duoc phien ma thanh trinh tu acid amin. de so sanh va mo hinh hoa can true khong gian ciia protein. Ket qua cho thay virus hanta luu hanh d mien Nam Viet Nam co do tuong dong cao ve trinh tu acid amin va cau triic khong gian so uYi cac cluing virus Seoul khac trong khu vuc.
IV. BAN U AN
Ket qua phan tich huyet thanh hpc tren 1.066 Rallus.sp va 245 S.murinus tai cac diem thu bat cho thay ti le nhiem chung cho toan bp khu vuc mien Nam va Tiiy Nguyen la 4.7%.
Trong do. cac mau huyet thanh duong tinh vdi SEOV deu duoc phat hien tren quan the R.norvegicus va chu yeu tap trung tai khu vuc do thi. cu the la khu vuc quan 12 vii khu vuc cang Sai Gdn. thanh pho Ho Chi Minh (TP.
1 ICM) vdi ti le nhiem la 22%. Dieu niiy ciing phii hop vdi thuc te lii R.norvegicus dupe xac dinh la quan the uu the p cac khu dd thi cd mat dp dan cu cao. Trong khi do, TPMV dupe xac dinh luu hanh vdi ty le nhiem loan bp tai khu v uc Tay Nguy en lit 14.5 % vii tai Ddng Nai la 6.45 % tren S.murinus - mpt loai ddng vai gam nham an cdn trung tap trung nhieu d nhirng khu vuc ban rung ram, da khiing djnh sir phan bd dac thii vc mat dja ly vii vat chu trung gian truyen benh cua SEOV vii TPMV.
Ii le luu hanh khang the khang vi- rus hanta tren quan the dpng vat gam nham d mien Bac Viet Nam la 5.4 % |2J, trong nghien ciru niiy. xiic djnh o mien Nam va Tay Nguyen la 4.7 %. Dac thu cua v irus hanta cd lien quan mat thiet vdi vat chu trung gian truyen benh, nhu chimg SEOV luu hanh tren R.norvegicus.
cdn TPMV tren S.murinus. Do dd. ty le huyet thanh duong tinh vdi v irus hanta thay ddi khac
nhau tuy tirng loai vat chu truyen benh. dac biet ddi vdi loai vat chii tirong ling. Vi du: cd 16.76 % huydt thanh duong tinh vdi SEOV tren quan the R.norvegicus (chupt cdng) va
13.1 % huyet thanh duong tinh vdi TPMV tren quan the S.murinus. Dieu niiy cho thay cd su luu hanh thudng xuy en ciia virus hanta trong quan the ddng v at gam nham o nude ta.
Day la ngudn lily nhiem cho ngudi trong cdng ddng rat dang lo ngai.
Nghien ciru. phan tich di truyen tren doan gen S cho thay ca vat lieu di truy en ciia virus hanta phiit hien to mini md phoi cd ky hieu CSG5 va CSG 11 deu thudc genotype SEOV. khdng cd su khac biet diing ke so vdi cac chung SEOV' khac trong khu vuc. Dac biet. khoang each giira trinh tu nucleotide viing gen S cua cac cluing CSG so sanh vdi chiing SR-I I (la cluing SEOV phan lap d Sap- poro. Nhat Ban. dai dien dac trung cho SEOV o khu vuc Chau A) rat gan nhau vdi dp tin cay Bpptstrap 1 000 (hinh 2) va dp tuong ddng nucleotide cao 97.1%. Tuy nhien, cac cluing CSG5 va CSG11 lai tach han thanh mpt nhdm rieng so vdi cac chung dai dien khac. cho thay virus hanta luu hanh d mien Nam Viet nam cd the la mpt nhdm phu ciia SEOV.
Phan tich so sanh trinh tir nucleotide tren doan M ciia 34 chiing virus hanta (hinh 3). cho thay trinh tu nucleotide ciia cac mau CSG 5, CSG 11 va 24D12 tach ra thanh hai nhdm phu rieng biet (bootstrap 1000). Khoang each khac biet vc nucelotide giua man CSG va 24D12 vdi cac chung dai dien tang dan theo tlui tu sau: Seoul 80 - 39 (4 %). SR-1 1 (4 %). 13-1 (5
%)... .Ddng thdi. ket qua phan tich cung cho thay virus hanta luu hanh d mien Nam Viet Nam nam trong mpt cum (cluster) tach biet SP vdi cac chiing dai dien d khu vuc Trung Qudc va Nhat Ban. Mat khac khi md rdng so sanh 17S Tap chi Y hpc du phdng. Tap XX. sd 6(1 14)
vdi cac chung SF.OV gan day cua khu vuc Ddng Nam A nhu Indonesia (chiing Jarkata 137) va Singapore (chiing RN4I va RN46).
ket qua cho thay chiing nam ciing nhdm, mac dii chua hoan toan tach biet han nhung da hinh thanh mot nhdm rieng dac trung cho khu vuc Ddng Nam A. Han nua, chung Indonesia da duoc khang djnh la lien quan chat che vdi virus hanta luu hanh d mien Biic Viet Nam khi phan tich so sanh 283 nucleotide d nam tai viing gen tuong ddi bao ton ciia cac chung SEOV (hinh 3).
Phan tich trinh tu acid amin cua hai protein Nucleocapsid (do gen S ma hda) va glycoprotein (do gen M ma hda) tir vat lieu di truyen ciia hai man CSG5 va CSG 11 cho thay cd sir luong ddng rat Idn ve cau triic so vdi cac chung SHOV khac trong khu vuc: Cd sir tirong ddng 100 % ve trinh tu acid amin vii cau triic khdng gian. Ddi vdi chung SR- 11 (la cluing SHOV sir dung trong ky thuat trung hda), nucleocapsid protein cua cac mau CSG5 va CSG I I tuy cd sir khac biet d vj tri 69 - 71. 72 - 74 va 251 d dau NHf nhung van bao ton can triic khdng gian. khdng anh hudng den tinh khang nguy en va chiic nang ddng gdi RNA virus (hinh 4). Ket qua tuong tir ciing duoc ghi nhan tren trinh tir amino acid ciia gly coprotein.
Dua tren dac diem dja ly cac man duong tinh dupe xac djnh o cang Sai Gdn va quan 12. TP.HCM. ciing vdi ket qua phan tich, so sanh trinh tir doan gen cho thay virus hanta luu hanh d mien Nam vii mien Bac Viet Nam tao thanh mpt nhdm phu ciia SHOV.
Day la nghien cuu phat hien dau tien ve sir hien dien ciia virus hanta thudc nhdm phu cua SHOV tai mien Nam Viet Nam. Hon the nua. ca nhiem v irus hanta dien hinh tren ngudi dupe xac djnh dau tien tai quan 12, TP.HCM
nam 2008 (bang 1) va tiep thep la mpt sd trudng hpp nhiem virus hanta dupe chan doan xac djnh trong khu vuc npi thanh ciia TP.HCM da chp thay su lay nhiem virus hanta tren cdng dong [7], [8].
Dieu nay dupe ciing cd them vdi viec virus hanta phiit hien tir ddng vat truyen benh thu thap tai khu vuc sinh song ciia benh nhan nhiem hanta, cung dupe xac djnh la ciing sero- type - SHOV (bang 2) bang ca PRNT va giai trinh tir (mau cd ky hieu 24D12). Til dd. chp thay nguy co1 lay truyen virus tir vat chii sang cdng ddng can dupe quan tam nhieu hon trong cdng tac phdng benh. Do vay, can duy tri hoat ddng giam sat chu dpng ve mat djch te hpc, virus hpc va djch te hpc phan tu virus hanta luu hanh d nude ta.
V. KET LUAN
Xac djnh ty le ddng vat gam nham nhiem viurs hanta la 16,76 % a R.norvegicus va 13,1
% a S.murinus bang cac ky thuat ELISA, IFA, WB trong tdng sd 1.3 11 mini huyet thanh ddng vat thu thap 0 Tay Nguyen va mien Nam Viet Nam trong cac nam 2006 - 2009.
Type huyet thanh cua virus hanta luu hanh d mien Nam Viet Nam thupc type huyet thanh Seoul va TPMV dupe xac djnh bang ky thuat FRNT. Phat hien trudng hop sot xuat huyet the than tren ngudi dau tien d mien Nam ciing do virus Seoul gay ra, la bang chirng ve sir lan truyen ciia virus Seoul tir dpng vat sang ngudi.
Phat hien dupe vat lieu di truyen cua vi- rus hanta luu hanh d mien Nam Viet Nam bang ky thuat RT-PCR tu 3 mau md phoi cd ky hieu CSG5. CSG 11 va 24D12. Vice giai trinh tu gen.
xac djnh va phan tich genotype cho thiiy mdi lien quan chat che cua virus hanta luu hanh tren R.nor\'egicus d'TP.HCM (Cang Sai gdn va quan Tap chi Y hpc du phdng. Tap XX. so 6(114) 179
12) voi cac virus Seoul dai dien 6 Nhat, Indone- sia, Singapore va mien Bac Viet Nam.
TAI LIFT THAM KHAO
1. Lee H. W., Calisher C Schmaljohn C.
(1998). Manual of Hemorrhagic Fever with Renal Syndrome and Hantanvirus Pulmonary Syndrome. WHO Collaborating Center for Virus Reference and Research (Hantaviruses) Asan Institute for Life Sci- ences, Seoul: 1 8 - 33.
2. Truong liven Ninh (2004). Mot vdi nhan xet ve sty cd mat cua virus Hantaan luu hanh tren chudi vd ngudi tai mdi sd dia phuong mien Bdc Viet Nam. Tap Chi Nghien ciru y hpc (ISSN 0868-202X), Phu truong so 5: 22 - 27.
3. Bi Z., Formenty P. B. H„ Roth C. K. (2008).
Hantavirus Infection: a review and global update. W IIO. J. Infect Developing Coun- tries, 2 (1): 3 - 23.
4. Elgh F.. I.undk\ist A.. Alexeycv O. A..
Stcnlund PL. Hjelle B., l ee H. L., Smith K. J., Vainionpaa R.. Wadell G. R. and Juto P. (1997). Serological Diagnosis of Han- tavirus Infections hy an Enzyme-Linked Immunosorbent Assay Based on Detection ol Immunoglobulin G and M Responses
to Recombinant Nucleocapsid Proteins of Five Viral Serotypes. Journal of Clinical Microbiology. 35: I 122 1130.
5. Morii M.. Yoshimatsu K.. Arikawa J..
Zhou G., Kariwa IE, Takashima I. (1998).
Antigen Characterization of Hantaan and Seoul Virus Nucleocapsid Protein Express by Recombinant Baculovirus: Application of a Truncated Protein. Lacking an Anti- genic Region Common to the Two I 'iruses.
as a Serolyping Antigen. Journal of Clini- cal Microbiology. 36: 25 14 2512.
6. Wang IE, et al (2000). Genetic Dtvesiiv of Hantaviruses Isolation in China and Char- acterization of Novel Haulaviruse Isola- tion from Niviyenrer confucianus and Rat- tus rattus. Virology. 278: 332 - 345.
7. Huong V. T. Q. Yoshimatsu K... Luan V. D., Tuan L. V. Nhi I.., Arikawa L. and Nguyen T. M. N. (2010). Hemorrhagic Fever with Renal Syndrome. Vietnam. Emerging Infec- tious Diseases, www.cdc.gov/eid, 16(2):
363 - 365.
8. Luan V. D., Yoshimatsu K, Endo R., Thang C. M., Arikawa J., Huong V. T. Q (2010).
Hantavirus Hemorrhagic /-'ever with Re- nal Syndrome in Ho Chi Minh City. Y I lpc Tp.Ho Chi Minh* Vol. 14 Supplement of No 2: 267 - 273.
180 Tap chi Y hpc du phong. Tap XX. so' 6 (1 14)
HANTAVIRUS INFECTION AMONG RODENTS IN HIGHLANDS AND SOUTHERN VIETNAM, 2006 - 2009
Vu Dinh Luan, Kumiko Yoshimatsu**, Midori Taruishi**, Le Nhi, Endo Rika**, Vo Thi Huong*,Cao Minh Thang, Hoang Kim Loan, Jiro Arikawa *, Vu Thi Que Huong
Pasteur Institute of Ho Clu Minh City, Vietnam j
* Tay Nguyen Institute of Hygiene and Epidemiology, Vietnam;
** Institute for Animal Experimentation, Graduate School of Medicine, Hokkaido University, Japan.
From 2006 to 2009, a surveillance of the situation of hantavirus infection in Southern part and Highlands of Vietnam was carried out based on the tested results of serum samples from 1.066 rodents representing 6 species and 245 Suneits murinus.
The results of tested rodent sera sam- ples by ELISA. IFA and confirmed by West- ern Blot showed that the prevalence antibody to hantavirus was 16.76% from R.norvegicus, and 13.1% from S.murinus. The serotyping result by FRNT revealed that hantaviruses which circulate in Southern Vietnam belong to Seoul and TPMV seroty pe. RNA of hanta- v irus was detected from 3 lung tissues of Rat- tus norvegicus samples which are coded as CSG5. CSG11 and 24D12, all collected in Ho Chi Minh City (Sai Gon Habor and District
12). The sequencing and phylogenetic analy- sis on detected genes (partial small and medi- um segments) demonstrated the close genetic relationship with SEOV representatives found in Japan. Indonesia Singapore and Northern Vietnam.
The serological analysis revealed the circulation of hantavirus including Seoul vi- rus (SEOV) from Rattus.sp. and Thottopa- layam virus from 51. muriiuts in Southern and Highlands of Vietnam. Besides, the detection of specific IgM and the neutralizing antibody against SEOV in patient indicated the first evi- dence of the circulation and transmission of SEOV from rodent to human.
Key word: Hantavirus. ELISA, RT- PCR, Sequencing
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