OAc DI^M

. C 6 N G NGHE SINH HOC TOAN 0 U 6 C 2013

OAc DI^M GEN HEMAGGLUTININ {HA/H5) C G A HAI C H O N G VIRUS C O M A/H5N1 MOri P H A N LAP NAM 2013 TAI VIET NAM

Nguyin Thj Bich Nga, Lfi Thanh Hfla*

Vidn Cdng ngh$ sinh hgc, Vign Hdn Idm Khoa hgc vd Cdng nghS Vigt Nam T6M TAT

Hai chung virus cum A/H5N1 14 A/Chicken/Vietnam/KH23/2013(H5N1) vi A/Duck/VietnamflaJ24/2013(H5Nl) tfiu nh|n nam 2013 tai KMnh Hoa da dugc xac dinh thupc clade 2.3.2.1 va c6 s^ kbac biet vk m6i quan hf nguon g5c pha b? Su xuit hign clade 2.3.2.1 cia vims ciira A/H5NI d Viet Nam da duoc WHO/FAO/OIE khing dinh tii nam 2011. Tuy nhien trong qud trinh tiln h6a, gen HA/H5 da tgo thdnh cfic nhinh (clade) da dgng. Chiing A-Ck-VN-KH23-20i3(H5Nl) c6 m6i quan h§ gSn gui vi!ri cac chiing cua Vift Nam phan % nim 2012. bao g^m: A-Dk-VN-LBMl 39-2012; A-Dk-VN-LBM 132-2012; A-Dk-VN-LBM 136-2012; A-Md- VN-LBM258-20I2; A-Md-VN-LBM260-2012, A-Md-VN-LBM 133-2012. trong khi do, chung A-Dk-VN-KH24-2013 cung thu nhjn l?i ciing mpt dia phirong Iai tao thinh m$t nhanh khic hoan toin vi c6 m6i quan he g5n gai vdi cac chung cua Vi?t Nam khic, id la: A-Md-VN-LBM315-2012; A-Md-VN-LBM227-2012; A-Md-VN-LBM295-2012; A-Dk-VN-OIE-2236-2012; A-Dk-VN-OIE- 2202-2012; A-Dk-VN-OIE-2211-2012. Cung trong nhom nay con c6 svr xuit hi?n cua clade 2.3.2.1 i cac chung thu nh|n t?i Indonesia vao nam 2012, mi trade day quoc gia nay chi c6 sv luu hinh cua cac chiing thugc clade 2.1. Dilu niy lim cho vin dc dich te hpc dim A/H5N1 n p y cang dien bi6n phiic t?p Irong chin doin va phong ch^ng khong chl rieng 6 Vi^t Nam, mi con li vSn dk quan lam cua nhifiu quoc gia do da c6 sv da nhiSm nhik clade trong quan ttie gja cSm.

Tir khda: dim A/H5N1; hemagglutinin; clade; phi he; tiSn h6a

MdoAu

B$nh cijm gia cam do vims cilm A/H5N1 thugc hp Orthomyxovlridae gfiy ra (Guan ef al.. 2009). Bfinh khfing nhung chi xufit hifin b gia cam, thuy cam m^ c6n cfi the Ifiy nhiSm sang ngu-dn vtri t? lfi tt> vong cao. Tir cufii nSm 2003 d^n nay, dich cum oia cilm t h i dfic iyc cao (HPAI) da bOng phdt it nhieu nu^c trfin thi gi6i, trong do cfi \flfit Nam. Theo thfing bfio cCia T6 chtrc Nfing luwng the gidi (FAO) vd lb chCrc Y t4 the gifii (WHO), hien nay da phat hifin cfic nhfinh clade 1.1 vd 2.3.2.1 6- Dong A. Dfing Nam A vfi Vifit Nam (http://www.fao.orq/news/storv.en/item/87196/lcode/: WHO/OIE/FAO H5N1 Evolution Working Group. 2011). Trii qua tirng giai dogn ti4n hfia. cum A/H5N1 xuit hifin tgi Vifit Nam g6m nhieu genotype va clade khSc nhau, trong dfi cfi genotype G vfi Z duvc phan chia thfinh 9 genotype tir VN1-VN9: VN1 (2001), VN2-VN3 (2003). VN4-VN5 (2005) VN6-VN7-VN8-VN9 (2007) vfi cac clade chu yfiu lfi: clade 1. clade 2.3.2 vfi clade 2.3.4 (Nguyen et al., 2008; Wan et al.. 2008; Lfi QuSrnh Mai, 2011). Trong do, clade 1.1 (phfit sinh tir clade 1) tap tmng * phia Nam vfi vCing d6ng bing sfing Mfi Kfing; clade 2.3.2.1 phfin b6 tai m$t s6 tlnh phia Bkc, vimg duyfin hfii mlfin Tmng vd TSy Nguyfin; cfin clade 2.3.4 dfin nay khfing thiy xuat hifin. Tuy nhifin, d cfic dja phu'ang khfic nhau cfi cfic biln the CLia vims nay iiru hfinh vfi gfiy bfinh khfic nhau vfi cfi sg khfic bifit Ifi'n vk khang nguyen (Nguyln Ngpc Tiln sf al., 2011, Nguyen ef al., 2012; Van Dang Ky. 2012).

Theo cfing vfin s6 316/TY-DT ngfiy 28/02/2013 cOa Cyc Thu Y gCri Chi cue Thd y cfic tlnh, thfinh phfi trgc thupc Tmng uong v l tinh hinh lu-u hfinh vims cOm gia cam H5N1 thi ti> thang 10/2012 din thfing 2/2013, nhanh 2.3.2.1 da phan thfinh nhfinh virus 2.3.2.1, nh6m A (xuit hifin it cac tlnh Lgng Son. QuSng Nam, LSm Ding, Blnh Thufin, Tfiy Ninh.

thfinh ph6 Ho Chi Minh vfi Ca Mau), nhanh vims 2.3.2.1, nhfim B (Uing Son); nhanh vims 2.3.2.1, nhdm C (Difin Bifin.

Hfia Binh. So'n La, Hai Phfing, VTnh Phuc, Lgng Son, Thfii Nguyfin, Qufing Ninh, Ninh Blnh, Nghfi An. Thira Thifin-Hul.

Qufing Nam, Khfinh Hoa. Dfik LSk. Lfim Ding, Bfi Rja-Vung Tfiu. Sfic Trfing, Bgc Lifiu va Ca Mau).

Trong bfii bao nay chting tfli trinh bfiy kit qufi phfin tich gen HA/H5 cua hai chCing virus eiim A/H5N1 mfi'i phfin Ifip nam 2013 tgi Khanh Hfia vfi so sanh mfii quan hfi pha hfi vfi'i mot s6 chOng cua Vifit Nam vfi thk gidi.

NGUYEN LIEU VA PHlfONG P H A P Nguyfin lifiu

Miu bfinh phim thu nhfin thang 2/2013 tgi tlnh Khanh Hoa, du-crc cung d p is dang RNA ting s6 tach chilt, da kilm tra xfic djnh A/H5N1. du'p'c vilt tfit vfi ky hifiu theo danh phfip: A-Ck-VN-KH23-2013(H5N1):

,/VChickenAfletnam/KH23/2013(H5N1)vfiA-Dk-VN-KH24-2013(H5N1):AflDuckA/ietnam/KH24/2013(H5N1).

' Phuwng phfip

T/itre hi^n RT-PCR thu nh$n gen H5

Phfin ijng RT-PCR du'pc thyc hifin vfi'i cap moi H5F. 5'-TCTGTC/\AAATGGAG/W\ATAGTG-3' va HSR: 5'- r-TTAAATGCAAATTCTGCATTG-S' theo chu trinh nhifit: 1 chu ky d 42''C trong 60 phut, 1 chu ky d gS^C trong 15 phut, sau do 35 chu ky ir 94°C trong 1 phut, 55°C trong 1 phut; TZ^C trong 2 phut va cufii ciing 1 chu ky 6 72°C trong 10 phdt.

Tit ca cfic ttifinh phin (mli, khuon. thfinh phan phan dng) du-oc cho vao cung mpt luc va thyc hifin lifin tyc ca hai giai dogn; giai dogn chuyin dli (RT) chuyen RNA thfinh cDNA vfi giai doan nhfin gen (PCR). Difin di kilm tra tren thgch agarose 1%. sfin phim tofin bO gen H5 cfi dfi dfii khoang 1,7 kb, di^rc tinh sach bing bO sinh phim QIAquick PCR Punfication kit (QIAGEN), nli vfio vector pCR2.1 (Invitrogen), chuyen ngp vao t l bfio kha biln E coli. tach dong, chpn l9C DNA tfit to hpp inang gen H5 va gdi dpc trinh ty.

Xir ly chuoi gen vd phan tich so ligu

HOI NGHI KHOA HOC CONG N G H t SINH HOC TO/ "

Sau khi gifii hlnh ty gen H5, cfic chu& nucleotide thfi duvc xir ly bfing cac phin mem cua mfiy tinh Macintosh bao gom chiKmg trinh SeqEd1.03. so sfinh bing Asembl^lGNvl .9c vfi h$ chu-cng trinh MacVector 8.2 (Accelrys Inc). So sfinh dii chilu vdt cfic chuoi gen cfi trong Ngfin hfing gen (Genbank) bang chuang frinh GENED0C2i (http-7/w.w.w psc.edu/biomed/genedoc. Distributed by the author). Trlnh ty gen H5 cua cfic rfiung vims cum gia elm A/H5N1 trang nghiin a>u nfiy di«7C so s^nh vdi cac chiing thu thfip cfi frong Ngfin hfing gen ve thfinh phan nucleotide d l xem xfit mii quan hfi pha hfi nguln g l c bing chyong trinh MEGA5.0 phirong phfip "kit nil llln k l ' NJ (Neighbor- Joining) vin hfi s l kilm djnh tin hrong bootsfrap 1000 lln (Tamura ef al.. 2011).

KfiT QUA vA THAO LUAN

Thu th$p chuSi gen H5 sir dgng trong so sdnh phdn tich trinh tu- nucleotide

Thyc hifin phfin irng RT-PCR mflt birirc (one step RT-PCR cua hang Qiagen) tofin bO gen H5 cua vims cum A/H5N1 cda 2 Chung nghifin cdu duac thu nhfin. Gen H5 dirg'c nhfin len vdi cfip mli H5F - H5R. cho s^n phim RT-PCR cfi kich thudc khoang 1.7 kb dung nhu- dy kien. Cac sfin phim gen H5 dirge tinh sach vfi nil tn/c tilp vfio vector pCR2.1 cila bO kit tfich ddng fTA-cloning. hang Invitnsgen) d l liru gid gen vfi giai frinh ty. Sau khi cfi chuoi nucleotide ciJa gen H5, cfic chuli nfiy dugc sir dyng d i fruy cgp Ngfin hfing gen sd dyng chucng frinh blast nhfim thu thfip cfic chu5i da cfing b l de phfin tich so sfinh dfic dilm vfi mli quan hfi nguon glc phfi hfi dupc li$t kfi d Bfing 1.

o l nghiSn cdu v l mdc dfi biln doi thfinh phin nucleotide cOa gen H5, frinh ty 2 chting mdi phfin l§p nfim 2013 dypc so sfinh vdl mfit s l chung dgi difin cda Vifit Nam vfi m$t s6 quoc gia frong khu vyc da dang ky trong Ngfin hfing gen tgi (http://www.ncbi.nlm.nih.qov/) vfi Tmng tfim dd lifiu cfic chufii gen vims cijm (IDS = Influenza Sequence Database t^l website (httD://www.flu.lanl.QovA. D l cd the khfii qufit mOt phan dlin biln cOa vims cum gia d m tgi Vifit Nam, chiing tfli dfi thu thfip cfic chuoi gen H5 xuat hien Uidi gian gin dfiy nhlt va cfi lifin quan d i n 2 chiing nghifin cdu, bao gim 37 chiing (cfi 2 chiing frong nghifin cdu nay).

Bing 1. Oanh sfich cic chiing virus ciim A/H5N1 cOa Vl^t Nam vfi th4 gidl sir dyng Irong phdn tich so sfinh thinh phin gen HS v i mol quan hf nguin g&c pha h^

S6 T T 1 . 2 . 3 . 4 . 5 . fi 7 8 9.

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K y t i l ^ u d a n h p h a p

A / C h c k e n / V i e t n a n i f l < H 2 3 « l l 3 ( H 5 N 1 )•

AflDuck/Vlelnamfl<H24/2013(HSN1 )•

A / M u s c o v y • u c k A n e t n 3 m / L B M 2 2 7 / 2 0 l 2 ( H 5 N 1 } A / M u s c o w D t J C k / V i e t n a m / L B M 2 2 g r 2 a i 2 ( H 5 N 1 ) A / M u s c o v y D t i c k / V l e t n a m / t B M 2 9 5 / 2 0 1 2 ( H 5 N 1 ) A / M u s c o v y D u c k A / i e t n a m / L B M 3 1 5 / 2 0 1 2 ( H 5 N 1 ) A/hfeiscovy D u c k / V l e t n a m / L B M 1 4 / Z 0 1 2 ( H 5 N 1) A / M u s c o v y D o < * / V i e t n a m / L B M 5 7 / 2 0 1 2 { H 5 t ^ 1 ) A / M u s c o v y O u c k A / l e t n a m / L B M 1 1 3 / 2 0 1 2 ( H S N 1 ) A / M u s c o v y D u c k ; V i e t n a m / L B M 2 5 8 / 2 0 1 2 ( H S r i 1 ) A / D u c l O V i e t n a m / L B M 1 3 2 / 2 0 1 Z ( H 5 N 1) A m u c k f l r t e t n a m A - B I ^ I 3 6 / 2 0 1 2 ( H 5 N 1 ) A / D u c l ( / V i e t n a m / L B t u n 3 9 ^ 0 1 Z ( H 5 N 1 ) A / O i K k / \ r i e t n 3 [ n / L B M 2 6 a / 2 0 1 2 ( H 5 N 1 ) A / D i K : k / \ f i e [ n a m / O I E a j 0 2 / 2 0 1 2 ( H 5 N 1 ) A r O u c k A / l e l n B m / O I E / 2 2 1 1 / 2 0 1 2 ( H 5 N 1 ) A r o > i C k A f l e t n a m / O I E / 2 2 3 6 / 2 0 1 2 { H 5 N l ) A / D L c k / B a n t u l / B B V W / 1 4 4 3 - 9 / 2 0 1 2 { H 5 N 1 ) A/DiJcWSlemarVBBVW/1463/1(W2012(H5N1) A / O u ( * / B l r t a r / B B V W - I 7 3 1 - I 1 - 2 0 1 2 { H S N 1 ) A / M u s c o v y D o c k / T e g a y B B V W / 1 1 7 3 2 / 1 1 / 2 0 1 2 ( H 5 N 1 )

A - D u c l ( - S u k o t i a n a - B B V W - 1 4 2 8 - 9 - 2 0 1 2 ( H S N 1) A / D u c W V i e t n a m r a T 8 0 1 / 2 0 1 1 (HSN 1 )•

A / D u c k / V i 8 l n a m / Q T 8 0 2 / 2 0 1 1 (HSN 1 )*

A / D u c K A / i e t n a m / O I E / 2 5 3 3 / 2 0 1 1 (HSN 1) A / D u c l ( / V i e t n 3 m / Q u a n g n g a i / 1 0 3 7 / 2 0 1 1 ( H 5 N 1 ) A/snviron me n t/H u n a n / 3 / 2 0 1 1 ( H S N 1) A/Chickenf J r a n g s u / k 0 4 0 2 / 2 0 1 0 ( H S N 1)

A/Ouck/Laos/4e9/2010<H5N 1) A / e n v i t D n m e n t / C h a n 9 S h a / 2 5 / 2 0 0 9 ( H 5 N 1 } A / H u b e i / 1 / 2 0 1 0 ( H 5 N 1 f

A / B [ O w n - h e a d e < ] . g u l I / Q i n g t i ^ 1 / 2 0 0 9 ( H 5 N 1) A-g r e a l - c r a s l e d - g r e b e - Q i n g h a i - 1 - 2 0 0 9 ( H S N 1 )

K y t i i $ u c h ^ n g A - C k - V N - K H 2 3 - 2 0 1 3 ( H 5 N 1 ) A - D k - V N - K H 2 4 - 2 0 1 3 ( H 5 N 1 ) A - ^ M • V N - L B M 2 2 7 - 2 0 1 2 { H 5 N 1 ) A - t i M - V N - L B M 2 2 9 - 2 0 1 2 ( H 5 N 1 ) A M d . V N - L B M 2 9 5 - 2 0 1 2 ( H 5 N 1 ) A - t i M - V N - L B M 3 1 6 - 2 0 1 2 ( H 5 N 1 ) A - M d - V N - L B M 1 4 - 2 0 1 1 ( H 5 N 1 } A - M d - V N - L B I ^ 5 7 - 2 0 1 1 ( H 5 N 1 ) A - M d - V N - L B M I I 3 - 2 0 1 2 ( H 5 N 1 ) A - M d - V N - L B M Z 5 e - 2 0 1 2 ( H S N 1 ) A-DK.VN-LBIl/1132-2012(H5N1) A - D k - V N - L B M I 3 6 - 2 0 1 2 ( H 5 N 1 ) A - D k - V N - L B M I 3 9 - 2 0 1 Z ( H 5 N 1 ) A-lLld-VN-LBt\/l260-2012(H5N1) A - D k - V N - O I E - 2 2 0 2 - 2 0 1 2 ( H 5 N 1 ) A - D k - V N ' O I E - 2 2 1 1 - 2 0 1 2 ( H 5 N 1 ) A - D k - V N - O t E - 2 2 3 6 - 2 0 1 2 ( H S N l ) A - D k - B a n t u l - B B V W - 1 4 4 3 - 9 - 2 0 1 2 (HSN 1) A - D k - S l e m a n - B B V W - 1 4 6 3 - 1 0 - 2 0 1 2 ( H 5 N 1 ) A - D k - B I i l a r - B B V W - 1 7 3 1 - 1 1 - 2012(HSN1) A - M d . T e g a l - B B V W - 1 7 3 2 - 1 1 - 2 0 1 2 ( H 5 N n A - D k - S u k o h a r j o - B B V W - 1 4 2 8 - 9 - 2 0 1 2 ( H 5 N 1 J Afl)WVI4/QTB01/2011 (HSN 1) A / D k / V N / Q T a 0 2 / 2 0 1 1 (HSN 1) A - D k - V N - O I E - 2 5 3 3 - 2 0 1 1 ( H 5 N 1 ) A - D k - Q u a n g n g a i - 1 0 3 7 - 2 0 1 1 ( H S N I )

A - e n v i r o n m e n t - H u n a n 3 - 2 0 1 1 ( H 5 N 1 ) A-Clc-Jiangsu-k0402- 2 0 1 0 ( H 5 N 1 ) A - D k - L a o 8 - 4 6 3 - 2 0 1 0 ( H S N 1 ) A . D k - L a o s - 4 6 9 - 2 0 1 0 ( H 5 N 1 ] A - e n v l r o n m 8 n t - C h a n g S h a - 2 S - 2 0 0 9 ( H S N 1 ] A - H u t K i - 1 - 2 0 1 0 ( H S N 1 ) A - B r o w n - h e a d e d - g u t l - O m g h a i - l - 2009(HSN1) A - g r e a t - c r e s t e d - g r e b e - Q i n g h a i - 1-2009[H5N11

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»• ADk-VIMHG J236-20l2(H5N1)AB78D4t6

Hlnh 1; M6i quan he ngudn g6c pha h$ gida cfic chung cDm A/H5N1 xfic l^p d i ^ tr3n thfinh phan nucleotide gen H5. Sd dgng chu'ong trlnh MEGA5.0 (phucrng phfip k4t noi Hen ke NJ, d$ hn cay 1000 bootstrap) MQi t§n chl dan lfi cfic chOng do chung tfii phfin Ifip nam 2013 va phan tich trong nghifin cdu nfiy.

Gen H5cda2 chOng vims cum A/H5N1 nam 2013 cua Vifit Nam du'Q'c phfin tfch so sfinh vdi cfic chiing khfic da dfing ky trong Ngfin hfing gen dira fren frinh td chuoi nucleotide sd dgng chutrng frinh GeneDoc 2.7 vfi MEGA5.0. Trong tfing so 37 Chung so sanh, c6 21 chiing cua Vifit Nam (nfim 2011-2012); 7 chung cOa Tmng Qu6c (xu^t hifin cfic nam 2007, 2009, 2010, 2011); 5 chung cua Indonesia (nam 2012); 2 chung ciia M6ng Co (nam 2009) vfi 2 chung cua Lao (nam 2010), Cfic chiing du'Q'c phfin Ifip td nhilu nguon vat chd khfic nhau: gfi, v]t, ngan, ngSng, ngirdi, chim (iifit ke d Bfing 1).

Kit qufi xfic Ifip cfiy phfi hfi, du'Q'c trinh bfiy tgt HInli 1.

Kit qua d Hlnh 1 cho th^y: hai chCing mfi'i phfin Ifip nfim 2013 cua Vifit Nam cfi sy khfic bifit v l m6i tuwng quan quan hfi pha hfi. Chung A-Ck-VN-KH23-2013(H5N1) gan gui v6i cfic chung cQa Vifit Nam m6i phfit hifin nfim 2012, bao g6m: A- Dk-VN-LBM139-2012 (s6 dang ky trong Ngfin hfing gen: AB742287); A-Dk-VN-LBM132-2012 (AB742263); A-Dk-VN- LBM136-2012 (AB742279); A-Md-VN-LBM258-2012 (AB787201); A-Md-VN-LBU260-2012 (AB787209); A-Md-VN- LBMl 13-2012 (AB742255), trong khi chiing A-Dk-VN-KH24-2013 lai nfim i> mot nhanh khac hofin tofin vfi cfi m6i quan 111 gSn gui vtr\ cfic chung thu nhfin nfim 2012 ciia Vifit Nam khfic, do lfi: A-Md-VN-LBM315-2012 (AB807753); A-Md-VN- LBM227-2012 (AB786676); A-Md-VN-LBM295-2012 (AB807880); A-Dk-VN-OIE-2236-2012 (AB780486): A-Dk-VN-OIE- 2202-2012 (AB769252), A-Dk-VN-OIE-2211-2012 (AB780478). Bac bifit, chung A-Dk-VN-KH24-2013 tfip hgp tnang nhom cfic chijng ciJa Indonesia nfim 2012 thupc clade 2.3.2.1, mfi fru-o'c dfiy qu6c gia nay chi cd sij' t6n t^i vfi lu'u hanh cOa clade 2.1. dfiy 'a phfin nhfinh m6l nfiy sinh hay xfim nhfip lu-u hfinh trong cfic nircrc Dfing Nam A nam 2012-2013, cin dirge lam sfing tfi. Tuy nhien, dilu nfiy chu-ng tfi da co sg da nhiem cac clade tnang quan the virus cum gia d m H5N1 trong nhO'ng nfim gan day. Sg tiln hfia da dgng cua gen hemagglutinin (HA/H5) lfi tiln d l tao nen da d?ng cfic clade ir khu v i ^ Chfiu A vfi D6ng Nam A.

HOI NGHI KHOA HOC C O N G NGHE SINH HOC T O ^,

Nhl/ vfiy, h> khi phat hifin vims ciim gia d m H5N1 l l n d i u tifin thupc dade 0 (A-Gs-Gd-1996(H5N1) nfim 1996 cr Qufing

€)6ng (Tmng Quec) den nay. trong qufi frinh tien h6a cua gen hemagglutinin (HA/H5) da phat sinh them nhilu dade dirge coi la men xuit hifin trong khoang gan 10 nam (tir 2005) fror lai dfiy. Sg> xuit hifin ttifim dade mdi 2.3 2.1 p h i bid|

ir Dong Nam A va da dirge phat hifin d nhilu qulc gia tir nam 2009 dd id Bangladesh, Tmng Quic. Mong C6, Hfin Qulc. Nga, Hfin Quic, Nhfit BSn. An Do. Myamar, Lfio. Vifit Nam (Kang e(a/.. 2011; Reid ela/., 2010; Wan eta/., 2008) vfi nam 2012 ir lndor>esia (McGrane vfi Herianto, 2013). Phfin nhanh moi cr gia cam tnjng clade 2.3.2.1 hi^n nay ir Vifit Nam, gom dade 2.3.2.1a, b. c. lam cho chiFcng trinh tifim phfing vaodn si> dyng cac chiing thuOc dade 1 bj hgn c h l tgi nir6c ta vfi mfit s l quoc gia trong vimg.

Hai Chung vims aim A/H5N1 phfin lap nam 2013 tgi Khanh Hoa lfi A-Ck-VN-KH23-2013(H5N1) vfi A-Dk-VN-KH24- 2013(H5N1)dadugcxficd!nh nfim trong dade 2.3.21 thufic nhanh vims mcri xuat hifin nam 2012 tai Viet Nam. Sirkh^c bifit giOa 2 chiing nfiy cho thiy mirc dp t i l n hfia da dgng vfi bien d l i cua vims H5N1 d gia cam trong khi vacxin phong l>finh khong hifiu qua dang lam tfing nguy CCT vims biln chung va kha nang JSy nhiem trfin ngu-oi Vi vfiy. vific quan trpng c i n Ifim trong thcri gian toi lfi tang arimg cfing tfie giam sat virus cum d gia d m A/H5N1 de chil dOng fip dyng cfic bi§n phap phong ching didi thich tigp. ngfin chgn sg tfiy tmyfin djch ir gia d m va ngan chfin sg lay truyfin tir gia d m sang nguvri.

L v i c f i m o n

Cdm on Vi$n COng ngh$ sinh hgc ddh&trg hnh phi de tdi ca sd (2012-2013) vd Phdng Thi nghi$m trpng dlSm Cdng ngh$ gen v4 trang thdt W. d4 thi/c hidn nghidn cuu ndy. Cam on cdc cosdthuy da hgp lac cung cip mSu nghidn OJu.

l A l L i e u T H A M K H A O

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CHARACTERIZATION OF THE HEMAGGLUTININ (HA/H5) GENE OF TWO STRAINS OF A/H5N1 AVIAN INFLUENZA VIRUS ISOLATED IN 2013 IN VIETNAM

Nguyen Thi Bich Nga and Le Thanh Hoa*

InsUhJte of Bhtechnology, VAST

Two strains of A/H5N1 influenza vims, namely A/ChickenA/ietnam/KHZS/ZOia (H5NI) and A/Duck/Vtemam/KH24/2013 (HSNI) j collected in 2013 in Klianh Hoa was determined to clade 2.3.2.1 and to show difTerenccs in phyiogenetic relationship. The emergewe | ofdade2.3J.I in Vietnam has been reported by WHO / FAO / OIE since 2011. However, diiing evolution, the HA/H5 gene hu ^

C CONG NGHE SINH HOC TOAN QU(!)C 2013

generated a num))er of clades. The strain A-Ck-VN-KH23-2013 (H3N1) has a close relationship with die Vietnam strains emerged in 2012, including A-J)k-VN-LBM 139-2012; A-Dk-VN-LBJM132-20i2; A-Dk-VN-LBMI36-2012; A-Md-VN-LBM258-20I2; A-Md- VN-LBM260-2012; A-Md-VN-LBM 133-2012, whilist die Slram A-Dk-VN-KH24-2013 isolated in Ihe same geogr^hical locality (Khanh Hoa province) formed another group with those strains of Vicmam collected in 2012 such as A-Md-VN-LBM315-2012; A- Md-VN-LBM227-2012, A-Md-VN-LBM295-2012; A-Dk-VN-OIE-2236-2012; A-Dk-VN-OlE-2202-2012; A-Dk-VN-OIE-22U- 2012. Also in this group, there are a group of strains of clade 2.3.2.1 collected in 2012 in Indonesia, in which country there was only die circulation of clade 2.1 previously reported , This conq)licaies the epidemiological situation of A/H5N1 avian influenza in diagnosis and prevention not only in Viemam but also in many countries due to the multi-clade introduction in poultry populations.

Key words: A/H5N1 influenza, hemagglutinin; clade; phjdogcny; evolution.

* Authorfor corresspondence. Tel- +84-915079583/912336855; Email: tachnqal53(Sqmail.com: imibtvntawmail.com