Tng quan v virus cm A/H5N1: vn dch t hc, tin ha, hnh thnh genotype v tng ng khng nguyn-min dch-vaccine

Cm gia cm l bnh truyn nhim cp tnh ca gia cm, do nhm virus cm A, thuc h Orthomyxoviridae gy ra, c kh nng lan truyn t ng vt sang ngi. Nhm virus cm A c 16 phn type HA (H1 - H16) v 9 phn type NA (N1 - N9) c kh nng ti t hp to nn hng trm phn type khc nhau v c tnh v kh nng gy bnh. H5N1 th c lc cao (HPAI) vn ang l mi e da cho chn nui v sc khe cng ng. T cui nm 2003 n thng 6/2008, c 385 trng hp mc cm A/H5N1, trong , 243 trng hp t vong, chim 63,11%, trong Vit Nam v Indonesia l hai quc gia c s ngi nhim v t vong cao nht. Khng ging nh dch cm A/H5N1 giai on 1996 - 2002, c th khng ch bng cch tiu dit v loi tr loi gia cm trong vng dch ct t truyn ly, cm A/H5N1 th c lc cao giai on 2003 n nay, vi s xut hin nhiu genotype khc nhau, c bit l genotype Z, lan truyn t Nam Trung Quc n cc ni khc trn th gii. y vn l thch thc i vi cng ng v cn nghin cu ng dng cc phng php khc nhau khng ch hiu qu. Tnh gy bnh ca A/H5N1 th c lc cao khng ch gii hn chc nng im ct protease ca HA v hot tnh ca NA, m l hiu ng ca sn phm a gen v kh nng ti t hp to virus mi vi c tnh gy bnh v c lc khc nhau l vn cn tnh n. Hng ngn cng trnh nghin cu v cm A ni chung v cm A/H5N1 ni ring, c bit trong 5 nm gn y, trong c pht trin cng ngh v (cc loi) vaccine gy min dch cho gia cm v chun b cho i dch c th xy ra ngi. V phng din dch t, tin ha, to bin chng, bin i khng nguyn - min dch, tnh mn cm v khng vi dc liu, khc vi nhiu virus khc gia cm, virus cm A/H5N1 c xu hng t bin nhanh to nn nhiu bin chng phn chia thnh nhiu phn dng khc nhau v c tnh thch ng ph rng i vi loi mc. Vaccine c pht trin t nhiu chng virus cm A/H5N1 bng k thut hin i, trong c cng ngh di truyn ngc, c bit vaccine t chng NIBRG-14 ha hn tim nng phng chng. Trong bi tng quan ny, chng ti gii thiu nhng c im c bn ca vn dch t hc, virus hc, sinh hc phn t, qu trnh tin ha, s hnh thnh genotype v khi qut mi quan h khng nguyn - min dch - vaccine ca cm A/H5N1 Vit Nam v th gii. Tm lc nhng nghin cu ca cc nh khoa hc v t chc Vit Nam ti bi cnh Vit Nam v trn nn virus cm A/H5N1 ca Vit Nam, trong c ng gp ca Vin Cng ngh sinh hc v gen hc/protein hc v gen khng nguyn HA/NA v vaccine th h mi cng c gii thiu. T kha:Cm A/H5N1, dch t hc, genotype, Orthomyxoviridae, khng nguyn - min dch, phn type, th c lc cao (HPAI), tin ha, vaccine C IM DCH T HC BNH CM GIA CM Vi nt v dch t hc bnh cm gia cm A/H5N1 Khi qut bnh cm gia cm

Cm gia cm (Avian Influenza, AI) l bnh truyn nhim cp tnh ca gia cm, do nhm virus cm A, thuc h Orthomyxoviridae gy ra. y l nhm virus c bin ch rng, c phn chia thnh nhiu phn type khc nhau da trn khng nguyn HA v NA c trn b mt capsid ca ht virus (de Wit, Fouchier, 2008). Nhm virus cm A c 16 phn type HA (t H1 n H16) v 9 phn type NA (t N1 n N9), v s ti t hp (reassortment) gia cc phn type HA v NA, v mt l thuyt, s to ra nhiu phn type khc nhau v c tnh v kh nng gy bnh. Mt khc, virus cm A c c tnh quan trng l d dng t bin trong gen/h gen (c bit gen NA v HA), hoc trao i cc gen khng nguyn vi nhau, trong qu trnh xm nhim v tn ti ly truyn gia cc loi vt ch. H Orthomyxoviridae c pht hin bao gm 4 nhm virus, l: nhm virus cm A (Influenza A); nhm virus cm B (Influenza B); nhm virus cm C (Influenza C); v nhm Thogotovirus. Cc nhm virus khc nhau bi cc khng nguyn b mt capsid, virus cm A v B l Hemagglutinin (HA), virus cm C l Hemagglutinin Esterase Fusion (HEF), v Thogotovirus l Glycoprotein (GP) (Murphy, Webster, 1996; Ito et al., 1998). Tnh hnh dch bnh cm A/H5N1 trn th gii Cm A/H5N1 l mt virus c c lc cao, v gy bnh trn ngi trong cc v dch cm g nhng nm 1996 - 2008, c bit c lit l do virus cm A/H5N1 th c lc cao (HPAI, highly pathogenic avian influenza) gy ra k t nm 2003 cho n nay v pht sinh nhiu di dng (sublineage) v nhm/phn nhm (clade) c c lc rt cao (Horimoto, Kawaoka, 2001; Nicholson et al., 2003; Perkins, Swayne, 2003; Ligon, 2005) (Hnh 1). Chng virus cm A/H5N1 c pht hin ln u tin gy bnh dch trn g ti Scotland vo nm 1959. C th gi cm A/H5N1 phn lp nm 1959 ti Scotland l virus cm A/H5N1 c in (danh php: A-Ck-Scotland-(59)(H5N1) (s ng k: X07869). T cho n nay, H5 v N1 c thay i ln xt v cu trc thnh phn gen v khng nguyn min dch (Horimoto, Kawaoka, 2006). Sau gn 40 nm khng pht hin, cm A/H5N1 xut hin ti Qung ng (1996), v Hng Kng (1997) vi bin i su sc, khng nhng gy cht gia cm m cn thch ng v gy cht ngi bnh. C th coi dng virus cm A/H5N1 t 1996 n nay l cm A/H5N1 hin i mi xut hin (de Jong, Hien, 2006). c bit, t 2003 n nay, virus H5N1 gy ra dch cm trn gia cm ti Hng Kng, Trung Quc v ly lan sang hng chc quc gia trn th gii chu , chu u v chu Phi. Cm A/H5N1 giai on 2003 n nay, c bn v cu trc vn nh trc , nhng xt v c lc (tnh gy bnh), loi vt ch nhim bnh, tnh khng nguyn - min dch v mc truyn ly c nhiu nt c trng hn v khc vi nhiu bin chng H5N1 trc y (HulsePost et al., 2005; Subbarao, Luke, 2007; Zhao et al., 2008) (Hnh 1).

Hnh 1. Bn cc quc gia xy ra dch cm A/H5N1 (WHO, tnh n 15/09/2008). Phn bi m l vngdch cm xy ra trn gia cm; phn bi nht l vng dch cm ch xy ra trn chim hoang d.

Bng 1. Tng s trng hp nhim cm gia cm A/H5N1 ngi bo co cho WHO t thng 12/2003 n 19/6/2008.
Quc gia 2003 Mc Azerbaijan Bangladesh Cambodia Trung Quc Djibouti Ai Cp Indonesia Irak Lo Myanmar Nigeria Pakistan Thi Lan Th Nh K Vit Nam Tng 0 0 0 1 0 0 0 0 0 0 0 0 0 0 3 4 Cht 0 0 0 1 0 0 0 0 0 0 0 0 0 0 3 4 2004 Mc 0 0 0 0 0 0 0 0 0 0 0 0 17 0 29 46 Cht 0 0 0 0 0 0 0 0 0 0 0 0 12 0 20 32 2005 Mc 0 0 4 8 0 0 20 0 0 0 0 0 5 0 61 98 Cht 0 0 4 5 0 0 13 0 0 0 0 0 2 0 19 43 2006 Mc 8 0 2 13 1 18 55 3 0 0 0 0 3 12 0 115 Cht 5 0 2 8 0 10 45 2 0 0 0 0 3 4 0 79 2007 Mc 0 0 1 5 0 25 42 0 2 1 1 3 0 0 8 88 Cht 0 0 1 3 0 9 37 0 2 0 1 1 0 0 5 59 2008 Mc 0 1 0 3 0 7 18 0 0 0 0 0 0 0 5 34 Cht 0 0 0 3 0 3 15 0 0 0 0 0 0 0 5 26 Tng s Mc 8 1 7 30 1 50 135 3 2 1 1 3 25 12 106 385 Cht 5 0 7 20 0 22 110 2 2 0 1 1 17 4 52 243

Ghi ch: S ca mc bnh bao gm c cc ca t vong. WHO ch ghi nhn nhng ca c xc nhn bng xt nghim v c bo co chnh thc. Ngun: T chc Y t th gii - WHO (tnh n thng 6/2008).

T cui 2005, cm A/H5N1, ch yu l cc chng virus thuc phn dng Thanh Hi (ngun gc vng Bc Trung Quc) bt u lan sang mt s nc vng Trung , trong c Nga, ri trn ngp ng u v xm nhp vo cc nc vng Tiu , bao gm Th Nh K, cc nc Bc- Trung Phi, c bit Ai Cp v Nigeria l cc nc chu thit hi nhiu nht (Salzberg, 2007). Nhm ngn chn dch bnh ly lan, trong hn mi nm qua, trn th gii c hng trm triu gia cm b tiu hy, gy thit hi nng n cho ngnh chn nui v kinh t. c bit, s ngi nhim v t vong do virus cm A/H5N1, mi nm mt cao hn, theo thng k s ngi b nhim cm gia cm H5N1 bo co vi T chc Y t th gii (WHO), t nm 2003 n thng 6/2008, c ti 385 trng hp mc cm A/H5N1, trong , 243 trng hp t vong chim ti 63,11%. Vit Nam v Indonesia l cc 2 quc gia c s ngi nhim v t vong cao nht do virus cm A/H5N1 trn th gii (Bng 1). Trong s 16 nc c ngi cht do cm gia cm, Inonesia v Vit Nam c WHO xc nh l quc gia im nng c th cm A/H5N1 c c cc iu kin thun li tin ha thch nghi ly nhim v tr thnh virus ca ngi. Tnh hnh dch bnh cm A/H5N1 Vit Nam Dch cm gia cm A/H5N1 bng pht ti Vit Nam vo cui thng 12/2003 cc tnh pha Bc, sau nhanh chng lan ti hu ht cc tnh/thnh trong c nc ch trong mt thi gian ngn. y l ln u tin dch cm gia cm A/H5N1 xy ra ti Vit Nam, c ti hng chc triu gia cm b tiu hy, gy thit hi nng nn ti nn kinh t quc dn. Tnh n nay (thng 10/2008), dch cm gia cm lin tc ti bng pht hng nm ti nhiu a phng trong c nc, c th phn chia thnh cc t dch ln nh sau: - t dch th nht t thng 12/2003 v 30/03/2004, dch cm xy ra cc tnh H Ty, Long An v Tin Giang. Dch bnh ly lan rt nhanh, ch trong vng hai thng xut hin 57/64 thnh trong c nc. Tng s g v thy cm mc bnh, cht v thiu hy hn 43,9 triu con, chim 17% tng n gia cm. Trong , g chim 30,4 triu con, thu cm 13,5 triu con. Ngoi ra, c t nht 14,8 triu chim ct v cc loi khc b cht hoc thiu hu. c bit, c 3 ngi c xc nh nhim virus cm A/H5N1 v c 3 t vong trong t dch ny. (http://www.cucthuy.gov.vn). - t dch th 2 t thng 4 n thng 11/2004: dch bnh ti pht ti 17 tnh, thi gian cao im nht l trong thng 7, sau gim dn n thng 11/2004 ch cn mt im pht dch. Tng s gia cm tiu hy c thng k trong v dch ny l 84.078 con. Trong , c gn 56.000 g; 8.132 vt; v 19.950 con chim ct. V c ti 27 ngi mc bnh virus cm A/H5N1, trong c 9 ca t vong (http://www.cucthuy.gov.vn). - t 3 t thng 12/2004 cho n thng 15/12/2005: dch cm g xy ra trn 36 tnh thnh trong c nc. S gia cm b tiu hy c Cc Th y thng k l 1,846 triu con (gm 470.000 g, 825.000 thy cm v 551.000 chim ct). Vo nhng thng cui nm 2005, dch cm g xy ra trong thng 10/2005 lan nhanh trong gn 40 tnh thnh v gim dn trong thng 12/2005 (http://www.cucthuy.gov.vn). - Sau mt nm (2006), do p dng chng trnh tim chng rng ri cho cc n gia cm trong c nc, cng vi cc bin php phng chng dch quyt lit, dch cm A/H5N1 khng xy ra Vit Nam. Mc d vy, n 06/12/2006 dch cm gia cm A/H5N1 ti bng pht C Mau, sau lan sang cc tnh Bc Liu, Hu Giang, Vnh Long v Cn Th(http://www.cucthuy.gov.vn). - Trong nm 2007, dch bnh ti pht ti Hi Dng vo ngy 17/02/2007 v c khng ch sau 1 thng. Tuy nhin, n ngy 01/05/2007 dch bnh tip tc ti pht ti Ngh An, sau lan sang nhiu tnh, thnh ph trong c nc. Theo Bo co ca Cc Th y (B Nng nghip v Pht trin Nng thn) n ngy 10/06/2007 dch xy ra trn 16 tnh, thnh ph (Ngh An, Qung Ninh, Cn Th, Sn La, Nam nh, ng Thp, Hi Phng, Bc Giang, Ninh Bnh, Bc Ninh, Vnh Phc, H Nam, Qung Nam, Hng Yn, Thi Bnh v Ph Th), v ch c khng ch hon ton vo 8/2007. - Gn y, dch bnh li tip tc ti bng pht mt s tnh pha Bc vo thng 3/2008. Cho n thng 6/2008, dch cm gia cm A/H5N1 v c bn c khng ch trn ton

quc (http://www.cucthuy.gov.vn). Quan h ly nhim v biu hin bnh Loi mc Qun th chim hoang d (ch yu l vt tri) v gia cm (vt, g ty, g, ngan, ngng) l cc vt ch t nhin ca virus cm A. Do , virus cm A thng khng gy bnh hoc ch gy bnh nh cho chim v gia cm. Tuy nhin, mt s bin th cng c (H5, H7) xut hin gn y do ti t hp vi nhiu gen khc nhau, c th gy dch cm nguy him lm cht hng lot c chim v gia cm (Li et al., 2004; Hulse-Post et al., 2005; Alexander, 2007). Tuy nhin, virus cm A c bit l chng virus A/H5N1 hin ang lu hnh c th xm nhim gy bnh cc loi ng vt c v khc nh h, mo nga, ln v ngi (Murphy, Webster, 1996; Alexander, 2007). Bnh ly truyn nhanh chng t c th bnh sang c th khe mnh bng ng h hp thng qua cc ht aerosol c cha virus trong khng kh v/hoc qua ng tiu ha do tip xc trc tip vi cc b mt (khng kh, t, nc, phng tin vn chuyn, dng c chn nui b nhim (dch bi xut, xc vt bnh, cht thi) ca c th bnh (Silas et al., 2007; Alexander, 2007). Triu chng Thi gian bnh t vi gi n 21 ngy, c trng hp ko di n 28 ngy. Gia cm bnh st cao, chy nc mt, ng tm mt ch, lng x, ph u v mt, da tm ti, chn xut huyt, chy nc di m. Con vt khi st cao c biu hin khng bnh thng h thng tiu ha, h hp, sinh sn v thn kinh. Triu chng chung l gim hot ng, gim tiu th thc n, gy yu. Trng hp nng c biu hin ho, kh th, ri lon thn kinh, a chy, mt s con c biu hin co git hoc t th khng bnh thng. Nhng triu trng trn c th xy ra cng mt lc hoc ring r (T Long Thnh, 2004; L Vn Nm, 2005). i vi ngi, sau khi nhim, thi gian bnh t 1 n 5 ngy trung bnh l o khong 3 ngy. Lc u bnh nhn st cao 39 C v ko di t 1 n 3 ngy, bnh nhn cm thy kh chu, ton thn m, ho, s mi nhc u khng khip, c th tin ti kh th ri nght th, km theo cc ri lon v thnh gic v th gic. c bit, chng virus cm A/H5N1 gy t l t vong rt cao c gia cm v trn ngi, c ngi c th nhim c ng h hp trn v di (Claas et al., 1998; Hui, 2008). Trong trng hp khng xy ra nhng bin chng phc tp, s gy nhim t gii hn v bnh nhn t phc hi trong vng mt tun. Tuy nhin, nu trng hp din bin phc tp, bnh c th tr nn trm trng thm ch c th dn n t vong, l trng hp b bin chng i km vim phi do virus hoc do vi khun hoc c hai (Bauer et al., 2006; Gambotto et al., 2008). Bnh tch Bnh tch i th cc loi khc nhau nn bnh tch khc nhau. i vi gia cm, bnh tch thng gp gm mo, ym sng to, ph quanh m mt. Th nh bnh tch xoang c trng bi vim cata, lng ng fibrin, c m hoc hnh thnh casien. C th ph nim mc kh qun vi dch thm xut khc nhau t thanh dch n casein. Vim xoang bng cata hoc fibrin, c th vim dnh bung trng vi xoang bng. Xut huyt im trn b mt nim mc v tng mc ni tng. Xut huyt hu ht ton b ng tiu ha, c bit thy r manh trng, d dy tuyn. Ty thng sng to, c nhng vch vng hoc sm theo chiu dc. Ti Fabricius g xung huyt v xut huyt. Bnh tch ny rt ging vi bnh tch ca bnh Newcastle (L Vn Nm, 2004). ngi, virus xm nhp nim mc ph nang ph qun, nhn ln gy hy hoi biu m ti kh, gy xung - xut huyt v trn dch phi bnh nhn. Mt khc, virus c kh nng gy nhim kh qun, rut, no, v c th xuyn qua nhau thai xm nhp bo thai. Mt cn bng iu ha min dch do hin tng tng cytokine khng kim ch (cytokine storm) thng xy ra dn n c th b suy sp v t vong (Korteweg, Gu, 2008). Chn on, iu tr Chn on Chn on bnh cm gia cm v bnh cm ngi thng ch yu da vo cc du hiu lm sng, dch t v cc xt nghim pht hin s hin din ca virus cm trong c th bnh, l cc

phn ng ngn ngng kt hng cu (HA, heagglutination test), test chn on nhanh (Capillia Flu A/B test ca hng Nippon Becton Dickinson), hoc cc chn on xc nh da vo cc xt nghim huyt thanh min dch hc v phn lp virus (Nicholson et al., 2003; OIE, 2005). Gn y, cc phng php Real-Time PCR v reverse transcription-PCR (RT-PCR), k cPCR a gen (multiplex-PCR), trc tip s dng ngun gen ca virus cm A v cm A/H5N1 t mu bnh phm (tracheal or cloacal swab), cng c a vo ng dng, cho php chn on xc nh v phn bit s hin din ca cc chng virus cm A gy bnh, c chnh xc v tin cy cao ch vi mt lng nh mu bnh phm (OIE, 2005; Zou et al., 2007; Wu et al., 2008). Phng chng bnh bng s dng ha dc liu Mt s dng ha dc ang c dng trong cc thuc c ch virus khng c hiu tc dng ngn cn qu trnh gii phng virus ra khi t bo nhim (c ch NA), hoc c ch qu trnh thot v (ci o) v bao gi ca virus do c ch knh vn chuyn protein NP vo nhn t bo trong qu trnh nhn ln ca virus trong t bo nhim (Hayden et al., 1999; Basler, 2007). Cc ch phm ch yu l Rimantadine, Amantadine (A/H5N1 xut hin s khng thuc vi hai thuc ny (Arinaminpathy et al., 2008) v Oseltamivir (Tamiflu) hoc phc hn hp c hai loi (Ilyushina et al., 2007). Trong , Oseltamivir c ch NA ca mi type virus cm, c s dng ung d phng trong vng 48 h sau tip xc vi mm bnh, v c s dng l thuc d tr chin lc phng dch cm A/H5N1, tuy nhin thuc cng c mt vi tc dng ph trn ngi c ghi nhn nh gy ri lon tm thn v c th gy c sau iu tr liu cao (ACIP, 2000; Hayden et al., 1999; Aoki et al., 2007). C IM VIRUS HC CM A PHN TYPE H5N1 GY BNH GIA CM Cu to v danh php Virus cm A (cn gi l virus cm gia cm, virus cm g) c tn khoa hc l Avian Influenza (AI), thuc h Orthomyxoviridae trong h thng phn loi chung (Basic Taxomomy) (Murphy, Webster, 1996). Cc ht virus cm A (virion) c hnh cu hoc hnh khi a din, ng knh 80 -120 nm, i khi cng c dng hnh si, khi lng phn t khong 250 triu Da. Phn tch thnh phn ha hc mt virion c cha khong 0,8 - 1,1% RNA; 70 - 75% l protein; 20 - 24% lipid v 5 - 8% l carbonhydrate (Murphy, 1996). Ht virus c cu to n gin gm v (capsid), v bc ngoi (envelope) v li l RNA si n m - negative single strand (Murphy, Webster, 1996) (Hnh 2).

Hnh 2. nh chp knh hin vi in t (A), m hnh (B), v phc hp ribonucleoprotein RNP (C) ca virus cm A. Ghi ch: A: Cc dng hnh thi khc nhau ca virus cm A di knh hin vi in t; B: M hnh cu to ht virus cm A (Hemagglutinin: phn t khng nguyn HA, Neuraminidase: phn t khng nguyn NA; PB2, PB1, PA: ba di n v phc hp enzyme polymerase ca virus). C: Cu trc ca phc hp ribonucleoprotein RNP (Ngun: Paul Digard, Dept Pathology, University of Cambridge).

V virus c chc nng bao bc v bo v vt cht di truyn RNA ca virus, bn cht cu to l

mng lipid kp, c ngun gc t mng t bo nhim c c hiu ha gn cc protein mng ca virus. Trn b mt c khong 500 gai mu nh ra v phn b dy c, mi gai mu di khong 10 14 nm c ng knh 4 - 6 nm, l nhng khng nguyn b mt v virus, bn cht cu to l glycoprotein gm: HA, NA, MA (matrix) v cc du n khc ca virus (Bender et al., 1999; Zhou et al., 2007). C s phn b khng ng u gia cc phn t NA v HA (t l khong 1NA/4HA), y l hai loi protein khng nguyn c vai tr quan trng trong qu trnh xm nhim ca virus t bo cm nhim (Murphy, Webster, 1996; Wagner et al., 2002). Vt cht di truyn (cn gi l h gen) ca virus cm A l RNA si n m (vit tt l (-) ssRNA), gm 8 phn on ring bit (HA, NA, M, NS, NP, PA, PB1 v PB2) ni vi nhau thnh mt si duy nht bn trong v virus, m ha cho 11 protein tng ng ca virus, trong phn on M m ha cho 2 protein l M1 v M2; phn on NS m ha cho 2 protein l NS v NEP, phn on PB1 m ha cho 2 protein l PB1 v PB1-F2 (Ito et al., 1998; Conenello et al., 2007) (Hnh 2). V danh php, nhm virus cm A c phn chia thnh nhiu phn type (subtype), cc phn type ny c phn bit bi s khc nhau cc c tnh khng nguyn b mt (NA v HA), cho p ng min dch khc nhau gia cc chng virus c th b nhim (Murphy, Webster, 1996; Wagner et al., 2002). C 16 phn type HA v 9 phn type NA c pht hin, s t hp gia cc phn type ny, v l thuyt, c th to ra hn 254 bin chng khc nhau, tr chng ban u (WHO/OIE/FAO, 2008; Chen et al., 2008). Hin nay, d liu gen v h gen ca virus cm A c th c tm thy trong Ngn hng gen (GenBank: http://www.ncbi.nlm.nih.gov/ genomes/FLU/FLU.html), ti Mng li chuyn gia cm gia cm ca T chc Nng lng th gii (FAO) v T chc Th y th gii (OIE) (OIE-FAO network of expertise on avian influenza:http://www.offlu.net/); v ti trang web ca Trung tm d liu cc gen virus cm (Influenza Sequence Database, ISD: http://www.flu.lanl.gov/). T chc Y t th gii (WHO) quy nh thng nht danh php theo th t k hiu: Tn serotype - Loi ng vt b nhim Vng a l phn lp-S hiu ng k chng virus - Thi gian phn lp - Loi hnh phn type [HA(H) v NA(N)]; v d: A/Chicken/Vietnam/ HG4/2005(H5N1). i vi cc virus c phn lp trn ngi bnh, th khng cn ghi loi mc trong danh php, v d: A/Vietnam/1194/2004(H5N1) (WHO/OIE/FAO, 2008). c lc v tnh thch ng vt ch c lc gy bnh ca virus cm A Tnh gy bnh hay c lc ca virus cm A c chia lm hai loi: Loi c lc cao (HPAI Highly pathogenic avian influenza), v loi c lc thp (LPAI - Low pathogenic avian influenza), c hai loi u cng tn ti trong t nhin (Kelly et al., 2008). - HPAI: l loi virus cm A c kh nng gy tn thng nhiu c quan ni tng trong c th nhim, trn gia cm chng thng gy cht 100% s gia cm b nhim trong vng 48 h sau nhim. Loi ny rt nguy him gy lo ngi cho cng ng. Virus loi HPAI pht trin tt trn t bo phi g v t bo thn ch trong mi trng nui cy khng c trypsin (Webster, 1998; Alexander, 2007). - LPAI: l loi virus khi pht trin trong c th nhim, c th gy bnh cm nh khng c triu chng lm sng in hnh v khng lm cht vt ch. y l loi virus ly truyn rng ri v to nn cc bnh trong t nhin ca virus cm A, loi ny c th trao i gen vi cc chng virus c c lc cao ng nhim trn cng mt t bo, v tr thnh loi virus HPAI nguy him (Webster, 1998; Alexander, 2007). Tnh thch ng a vt ch ca virus cm A/H5N1 Vt ch t nhin ca tt c cc chng virus cm A/H5N1 l chim hoang d (ch yu l vt tri), y l nguyn nhn lan truyn virus trong t nhin rt kh kim sot. Virus cm A c kh nng gia tng bin vt ch ca chng trong qu trnh ly truyn t nhin (Kashet al., 2006; de Wit, Fouchier, 2008). Nh c tnh lun thay i khng nguyn trong t nhin, virus cm A c kh nng xm nhim nhiu loi vt ch trung gian khc nhau nh gia cm, mt s loi ng vt c v (hi cu, c voi, nga, ln) v c ngi, to nn tnh thch ng lan truyn ni loi nh g - g, hay ngoi loi nh g -

ln; g - ln - ngi (Hnh 3). Vt (vt tri) v mt s loi thu cm khc (ngng) lun lun l vt ch tng tr ngun virus gy nhim (Webster et al., 2002; Weiss, 2003; Chen et al., 2004; Hulse-Post et al., 2005). c im thch ng vt ch ny l iu kin thun li cho virus cm A trao i, ti t hp cc phn on gen, c bit l cc phn on gen khng nguyn (gen c HA v NA) gia cc chng, to ra mt chng virus cm mi c kh nng thch ng xm nhim loi vt ch mi ca chng c bit khi chng vt qua c ro cn loi d dng thch ng ly nhim gy bnh t gia cm sang ngi v gia ngi vi ngi (Horimoto, Kawaoka, 2001; Hilleman, 2002; Chen et al., 2004). Trong lch s cc i dch cm ngi, ln thng l vt ch trung gian chuyn tip gip cho virus cm A bin i d dng ly nhim sang ngi gy nn bnh dch (Ito et al., 1998; Zhu et al., 2008). V d: cm A/H3N2 l kt qu ti t hp t nhin ca virus cm A/H2N2 ca ngi v virus cha gen H3 trong t nhin thng qua ng nhim trn ln, gy nn i dch cm chu nm 1968 (Nicholson et al., 2003; Webster, 1998; Horimoto, Kawaoka, 2001). Sc khng Virus cm A tng i nhy cm vi cc tc nhn bt hot vt l hay ha hc. Cc ht virus tn ti thch hp trong khong pH t 6,5 n 7,9. pH qu acid hay qu kim, kh nng ly nhim ca virus b gim mnh (Fang et al., 2008). Lp v ngoi ca virus bn cht l lp lipid kp, c ngun gc t mng t bo nhim, d b ph hy bi cc dung mi ha tan lipid, cht ty ra v cc cht st trng: formaldehyde, phenol, -propiolacton, sodium hypochloride, acid long v hydroxylamine. Virus b bt hot di nh sng trc tip sau 40 gi, tn ti c 15 ngy nh sng thng, tia t ngoi bt hot c virus nhng khng ph hy c khng nguyn ca virus. Tuy nhin, virus cm A d dng b tiu o o dit hon ton 100 C v 60 C/30 pht, tn ti t nht 3 thng nhit thp (trong phn gia cm), o o v ti hng nm nhit bo qun (70 C). Trong ph tng gia cm (40 C), virus tn ti 25 - 30 o ngy, nhng ch tn ti 7 - 8 ngy nhit c th ngi (37 C); trong nc, virus c th sng ti 4 o ngy nhit 30 C (Murphy, Webster, 1996; Webster, 1998). Kh nng gy bnh ca virus cm A Virus cm A c tnh thch ng ly nhim cao vi biu m ng h hp, gy bnh ch yu ng h hp, v cng c th tc ng gy tn thng nhiu c quan khc trong c th ca cc ng vt cm nhim, do cn c gi l virus hng a ph tng (Webster, 1998; Nicholson et al., 2003; Phm Ngc nh et al., 2004). Kh nng gy bnh ca virus cm A ph thuc vo c lc v tnh thch nghi vt ch ca tng chng virus. Thng thng chng khng gy bnh hoc ch gy bnh nh gii hn ng h hp ca chim hoang d v gia cm nhim, nhng mt s chng cng c (H5, H7, v H1, H2, H3) c th gy bnh nng hu ht cc c quan trong c th, gy nn dch cm gia cm v ngi, c l do tnh thch ng th th sialic ca chng (Suzuki, 2005; Yamada et al., 2006). Hu ht cc chng virus cm A nhn ln rt tt trong phi g sau ln cy truyn th nht, tuy nhin cc chng cng c phn type H5, H7 gy cht phi g ngay sau vi gi, c khi hm lng virus rt thp cha c nhn ln nhiu, v c th gy bnh cm thc nghim trn chut lang, chut Hamster, chn t (Horimoto, Kawaoka, 1995; de Wit, Fouchier, 2008). Sau khi b nhim virus cm A, c th vt ch sinh ra p ng min dch chng li virus bo v c th, nhng p ng min dch ny c th khng c tc dng bo v hon ton cho nhng ln nhim sau, do virus cm A lun c s bin i khng nguyn ca n trong qu trnh lu hnh t nhin, v khng c p ng min dch cho gia cc chng virus cm A (Webster, 1998). Do , khi xut hin nhng bin chng virus cm A c c tnh khng nguyn khc vi cc chng virus trc , c th nhim s khng hoc t c p ng min dch bo h thch ng vi chng virus cm mi. y l nguyn nhn lm cho gia cm v con ngi thng b mc bnh cm nhiu ln trong nm, v cc t dch cm xy ra v sau thng nng n hn v c th gy nn i dch cm mi (Doherty et al., 2005). Kh nng gy bnh ca bin chng virus cm mi gim hoc bin mt, khi c th c c p ng min dch c hiu vi bin chng v chng tr nn thch nghi ly nhim loi vt ch mi (Weiss, 2003), v d: virus A/H1N1, A/H2N2, A/H3N2 l nguyn nhn ca cc i dch cm trn ngi trc y v thch nghi ly nhim ngi (Ito et al., 1998). Tuy nhin, cc chng ny vn thng gy ra

cc v dch cm tn pht hng nm ngi, do kh nng bin i khng nguyn ca chng (Hilleman, 2002). y cng chnh l ngun virus trao i gen vi cc chng virus cm ang lu hnh gia cm, thch ng ly nhim gy bnh cho nhiu loi khc ngay c trn ngi (Webster, 1998; Weiss, 2003; Weber, Stilianakis, 2007; Chen et al., 2008).

Hnh 3. Mi

quan h ly nhim v thch ng cc loi vt ch ca virus cm A.

C ch xm nhim gy bnh ca virus cm A trong t bo vt ch Virus cm A/H5N1 k sinh ni bo bt buc, qu trnh xm nhim v nhn ln ca virus xy ra ch yu cc t bo biu m ng h hp, ng tiu ha ca c th nhim (Murphy, Webster, 1996; Nicholson et al., 2003), c nhng nt c trng nh sau: - Qu trnh xm nhim ca virus cm A c m u bng s kt hp ca HA v th th thch ng ca n trn b mt cc t bo ny, v cui cng l gii phng h gen ca virus vo trong bo tng ca t bo nhim (Hnh 4). - Qu trnh nhn ln ca RNA virus cm A ch xy ra trong nhn ca t bo, y l c im khc bit so vi cc virus khc (qu trnh ny xy ra trong nguyn sinh cht), v cui cng l gii phng cc ht virus ra khi t bo nhim nh vai tr ca enzyme neuraminidase. Thi gian mt chu trnh xm nhim v gii phng cc ht virus mi ca virus cm ch khong vi gi (trung bnh 6 h). S to thnh cc ht virus mi khng ph tan t bo nhim, nhng cc t bo ny b ri lon h thng tng hp cc i phn t, v ri vo qu trnh cht theo chng trnh (apoptosis) lm tn thng m ca c th vt ch (Webster, 1998; Uiprasertkul et al., 2007). - Sau khi c gii phng vo trong bo tng t bo nhim, h gen ca virus s dng b my sinh hc ca t bo tng hp cc protein ca virus v cc RNA vn chuyn ph thuc RNA (RNAdependent RNA transcription). Phc hp protein RNA ca virus c vn chuyn vo trong nhn t bo (Basler, 2007). - Trong nhn t bo cc RNA h gen ca virus tng hp nn cc si dng t khun l si m ca h gen virus, t cc si dng ny chng tng hp nn RNA h gen ca virus mi nh RNApolymerase. Cc si ny khng c Adenine ha (gn thm cc Adenine - gn m) u 5- v 3-, chng kt hp vi nucleoprotein (NP) to thnh phc hp ribonucleoprotein (RNP) hon chnh v

c vn chuyn ra bo tng t bo. ng thi, cc RNA thng tin ca virus cng sao chp nh h thng enzyme tng phn on gen ca virus, v c enzyme PB2 gn thm 10 - 12 nucleotide Adenin u 5-, sau c vn chuyn ra bo tng v dch m ti li ni bo c ht tng hp nn cc protein ca virus (Hnh 4). - Cc phn t NA v HA ca virus sau khi tng hp c vn chuyn gn ln mt ngoi ca mng t bo nhim nh b my Golgi, gi l hin tng ny chi ca virus. NP sau khi tng hp c vn chuyn tr li nhn t bo kt hp vi RNA thnh RNP ca virus. Sau cng cc RNP ca virus c hp nht vi vng ny chi, to thnh cc chi virus gn cht vo mng t bo ch bi lin kt gia HA vi th th cha sialic acid. Cc NA phn ct cc lin kt ny v gii phng cc ht virus trng thnh tip tc xm nhim cc t bo khc (Murphy, Webster, 1996; Nayak et al., 2004).

Hnh 4.

M hnh c ch xm nhim v nhn ln ca virus cm A t bo ch. C IM SINH HC PHN T CA VIRUS CM A/H5N1 c im chung H gen virus cm A l RNA si n m (-)ssRNA, bao gm 8 phn on gen ring bit mang tn t 1-8 theo th t gim dn ca kch thc phn t (hay c gi theo tn protein m chng m ha tng hp), m ha cho 11 protein khc nhau ca virus gm: HA, NA, M (M1 v M2), NP, NS (NS1 v NS2), PA, PB1, PB1-F2 v PB2 (Murphy, Webster, 1996; Luong, Palese, 1992; Rabadan et al., 2006) (Hnh 2B). Mi phn on RNA ca virus cm A c cu trc xon bc 2 i xng di 50 - 100 nm, ng knh 9 - 10 nm, c bao bc bi nucleoprotein (NP) - bn cht l lipoprotein, to thnh cu trc ribonucleoprotein (RNP). Mi RNP kt hp vi 3 protein enzyme polymerase (PA, PB1 v PB2) chu trch nhim trong qu trnh phin m v sao chp RNA ca virus (Hnh 2C). Cc phn on ca h gen virus cm A ni vi nhau bng cc cu ni peptide to nn vm (loop) ti gii hn cui ca mi phn on, v to thnh mt si RNA duy nht c tng di t 10.000 - 15.000 bp (tu theo tng chng virus cm A), cha ng khong 13,5 kilobase thng tin di truyn v c cu trc xon (-helix) bn trong v virus. Hai u 5- v 3- si RNA h gen virus c gn thm chui nucleotide c tnh bo tn cao gia cc phn type l: 5-AGUAGAACAAGG v 3- UCG(U/C)UUUCGU CC (Luong, Palese, 1992; Murphy, Webster, 1996). Cc phn on gen ca virus cm A/H5N1 Cc phn on 1, 2 v 3 l nhng phn on m ha tng hp cc enzyme trong phc hp

polymerase (RNA transcriptase) ca virus, c di n nh v c tnh bo tn cao, bao gm: - Phn on 1 (gen PB2) c kch thc 2431 bp, m ha tng hp protein enzyme PB2, l tiu n v thnh phn trong phc hp enzyme polymerase ca virus, chu trch nhim khi u phin m 3 RNA virus. Protein PB2 c khi lng phn t theo tnh ton khong 84.10 Da (trn thc t l 3 87.10 Da) (Murphy, Webster, 1996). Tnh thch nghi nhit c th loi vt ch c cho l c lin quan n v tr amino acid 627 protein PB2 ( virus cm gia cm v tr ny l Glu - thch ng nhit o c th gia cm khong 40 C, cn virus thch nghi trn ngi l Lys - thch ng nhit c th ngi o khong 37 C) (Subbarao, 1993; Wasilenko et al., 2008). - Phn on 2 (gen PB1) cng c kch thc 2431 bp, m ha tng hp enzyme PB1 - tiu n v xc tc ca phc hp enzym polymerase trong qu trnh tng hp RNA virus, chu trch nhim gn m RNA (Murphy, Webster, 1996). Gn y, c pht hin thm mt protein (PB1-F2) c m ha bi mt khung c m khc ca PB1, c vai tr gy ra hin tng apoptosis (hin tng t bo cht theo chng trnh) (Uiprasertkul et al., 2007). - Phn on 3 (gen PA) c kch thc 2233 bp, l phn on gen bo tn cao, m ha tng hp 3 3 protein enzyme PA c khi lng phn t theo tnh ton khong 83.10 Da (trn thc t l 96.10 Da). PA l mt tiu n v ca polymerase chu trch nhim ko di s phin m RNA trong qu trnh tng hp RNA ca virus (Luong, Palese, 1992). Cc phn on 4 v 6 m ha cho cc protein (HA v NA) b mt capsid ca virus, c tnh khng nguyn c trng theo tng chng virus cm A, bao gm: - Phn on 4 (gen HA) c di thay i tu theo tng chng virus cm A ( A/H1N1 l 1778 bp, H9N1 l 1714 bp, H5N1 l khong 1704 - 1707 bp). y l gen chu trch nhim m ha tng hp protein HA - khng nguyn b mt virus cm, gm hai tiu phn l HA1 v HA2. Vng ni gia HA1 v HA2 gm mt s amino acid mang tnh kim c m ha bi mt chui oligonucleotide, l im ct ca enzym protease, v y l vng quyt nh c lc ca virus (Bosch et al., 1979; 3 Gambotto et al., 2008). Protein HA c khi lng phn t khong 63.10 Da (nu khng c glycosyl 3 3 3 ha) v 77.10 Da (nu c glycosyl ha, trong HA1 l 48.10 Da v HA2 l 29.10 Da) (Luong, Palese, 1992; Keawcharoen et al., 2005). - Phn on 6 (gen NA) l mt gen khng nguyn ca virus, c chiu di thay i theo tng chng virus cm A ( A/H6N2 l 1413 bp, A/H5N1 thay i khong t 1350 - 1410 bp). (Nguyn Th Bch Nga, 2006). y l gen m ha tng hp protein NA, khng nguyn b mt capsid ca virus, c 3 3 khi lng phn t theo tnh ton khong 50.10 Da (trn thc t l 50 - 60.10 Da). Cc nghin cu phn t gen NA ca virus cm cho thy phn u 5- ca gen ny (hay phn tn cng N ca polypeptide NA) c tnh bin i cao v phc tp gia cc chng virus cm A, s thay i ny lin quan n qu trnh thch ng v gy bnh ca virus cm trn nhiu i tng vt ch khc nhau (Castrucci, Kawaoka, 1993; Baigent, McCauley, 2001; Wagner et al., 2002). c trng bin i ca gen NA trong virus cm A l hin tng t bin trt-xa mt on gen l 57 nucleotide, ri sau l 60 nucleotide, lm cho di vn c trc y ca NA(N1) l 1410 bp cn 1350 bp (Castrucci, Kawaoka, 1993; L Thanh Ha, 2006; Keawcharoen et al., 2005). Cc phn on gen M, NP v NS m ha tng hp cc protein chc nng khc nhau ca virus, c di tng i n nh gia cc chng virus cm A, bao gm: - Phn on 5 (gen NP) kch thc khong 1556 bp, m ha tng hp nucleoprotein (NP) thnh phn ca phc h phin m, chu trch nhim vn chuyn RNA gia nhn v bo tng t bo ch. NP l mt protein c glycosyl ha, c c tnh khng nguyn biu hin theo nhm virus, tn ti trong cc ht virus dng kt hp vi mi phn on RNA, c khi lng phn t theo tnh ton 3 3 khong 56.10 Da (trn thc t l 50 - 60.10 Da) (Luong, Palese, 1992; Murphy, Webster, 1996;Salzberg, 2007). - Phn on 7 (gen M) c kch thc khong 1027 bp, m ha cho protein m (matrix protein M) ca virus (gm hai tiu phn l M1 v M2 c to ra bi nhng khung c m khc nhau ca

cng mt phn on RNA), cng vi HA v NA c khong 3000 phn t MP trn b mt capsid ca virus cm A, c mi quan h tng tc b mt vi hemagglutinin (Scholtissek et al., 2002). Protein M1 l mt protein nn, l thnh phn chnh ca virus c chc nng bao bc RNA to nn phc hp RNP v tham gia vo qu trnh ny chi ca virus (Luong, Palese, 1992; Murphy, Webster, 1996; Basler, 3 2007). Protein M2 l chui polypeptide b, c khi lng phn t theo tnh ton l 11.10 Da (trn thc 3 t l 15.10 Da), l protein chuyn mng - knh ion (ion channel) cn thit cho kh nng ly nhim ca virus, chu trch nhim ci o virus trnh din h gen bo tng t bo ch trong qu trnh xm nhim trn vt ch (Scholtissek et al., 2002). - Phn on 8 (gen NS), l gen m ha protein khng cu trc (non structural protein), c di n nh nht trong h gen ca virus cm A, kch thc khong 890 bp, m ha tng hp hai protein l NS1 v NS2 (cn gi l NEP, nuclear export protein), c vai tr bo v h gen ca virus nu thiu chng virus sinh ra s b thiu nng (Murphy, Webster, 1996; Sekellick et al., 2000). c tnh ca virus c s lin quan vi gen khng cu trc (non-structural gen) ny c tm thy bin chng A/H5N1/97 (Webster, 1998), trong t nhin, vic t bin xa i mt phn gen c lin quan n gim 3 c lc (Zhu et al., 2008). NS1 c khi lng phn t theo tnh ton l 27.10 Da (trn thc t l 3 25.10 Da), chu trch nhim vn chuyn RNA thng tin ca virus t nhn ra bo tng t bo nhim, v tc ng ln cc RNA vn chuyn cng nh cc qu trnh ct v dch m ca t bo ch. NEP hay NS2, l gen hnh thnh t hai on gen (30 bp v 336 bp) m ha loi protein c khi lng phn t 3 3 theo tnh ton khong 14.10 Da (trn thc t l 12x10 Da), ng vai tr vn chuyn cc RNP ca virus ra khi nhn t bo nhim lp rp vi capsid to nn ht virus mi (Sekellick et al., 2000; Zhu et al., 2008). Nh vy, virus cm A (c th: cm A/H5N1) c h gen c cu trc t 8 phn on ring bit v khng c gen m ha enzyme sa cha RNA, to iu kin thun li cho s xut hin cc t bin im trong cc phn on gen/h gen qua qu trnh sao chp nhn ln ca virus, hoc trao i cc phn on gen gia cc chng virus cm ng nhim trn cng mt t bo, rt c th dn n thay i c tnh khng nguyn to nn cc chng virus cm A mi (Suarez, Schultz-Cherry, 2000; Doherty et al., 2006). C IM TIN HA V HNH THNH GENOTYPE CA VIRUS CM A/H5N1 GIAI ON 1996 - 2008 V s tin ha cm A/H5N1 Nm 1996, H5N1 c phn lp t ngng ti mt dch Qung ng (Trung Quc) v chng ny c coi l chng nguyn thy to nn cc dng virus gy bnh cm gia cm trong 12 nm qua (Xu et al., 1999). Chng virus nguyn thy ny, lc cung cp ngun gen HA(H5) cho tin trnh ti t hp to nn cc bin chng gy dch bnh trn gia cm v ngi Hng Kng nm 1997, v ngun gen khung khc ca virus cm A/H5N1 Hng Kng c kin to t virus cm A c chim ct (Guan et al., 1999). Ring ngun gen NA(N1) cn cha bit c ly t u, nhng cu trc ca gen c hin tng xa i 57 nucleotide m ha cho 19 amino acid, ti vng u N ca protein neuraminidase, v t bin xa gen ny ca N1 c lin quan n tnh thch ng ca virus cm t thu cm ln gia cm trn cn v ngi (Matrosovich et al., 1999). i vi gen HA(H5) t bin gin n chui ni gia HA1 v HA2 m ha cho cc amino acid kim (Arginine v Lysine) c lin quan n tin trnh tng cng c lc, v cc chng thuc dng Qung ng (Guangdong-like sublineage), cc amino acid thng thng l -RRRKK(Claas et al., 1998; Matrosovich et al., 1999). Sau mt nm gy bnh ti Hng Kng, do ton b n gia cm b tiu dit, virus cm A/H5N1 nguyn thy gc Qung ng khng cn gia cm cn gy bnh, ngi ta tng chng bin mt, nhng thc t chng nguyn thy ny vn tip tc tn ti trong ngng vng Nam Trung Quc, tr thnh ngun gen ti t hp hnh thnh bin chng mi (Cauthen et al., 2000; Webster et al., 2002). Trong cc nm 1997 - 2002, nhiu bin chng virus cm A/H5N1 mang nhiu c tnh khng nguyn khc nhau ca phn type H5 c hnh thnh to nn nhm khng nguyn (clade) 1 c c lc cao vi g nhng thp i vi vt, ri b o thi trong nhng nm 2001 - 2002 (Guanet al., 2002; Li et

al., 2004). Tip tc, trong nm 2002 - 2003, gen HA(H5) c nhng t bin mi do hu qu ca hin tng lch khng nguyn (antigenic drift), ri to nn bin chng c tnh gy bnh cc k cao, c bit i vi vt, v ly sang ngi (Guan et al., 2004; Sturm-Ramirez et al., 2004). c tnh thch ng gy bnh trn ngi cng ngy cng cao dn, cng vi c lc tng cng i vi a vt ch bao gm vt, g, ngan, ngng, chim ct, chim hoang d v ngi, ri hnh thnh nhiu bin chng xm nhp cc nc pha Nam chu trong c Vit Nam, Thi Lan (Guan et al., 2004; Peiris et al., 2004) (Hnh 5).

Hnh 5. Phn nhm tin ha (clade) ca cm A/H5N1 bt ngun t chng nguyn thu A-Goose-Gd1-1996(H5N1) xut hin ti Qung ng (Trung Quc) nm 1996.

C th ni, sau giai on 1997 - 2003, virus cm A/H5N1 t n mc hon thin v c tnh gy bnh v thch ng a vt ch, tr nn mi nguy c gy bnh rt cao i vi gia cm v ngi trong cc nm 2004 - 2005 (Li et al., 2004; Smith et al., 2006). Tuy nhin, xt v di truyn hc v tnh khng nguyn, cc chng H5N1 giai on 1997 - 2002 vn mang tnh ng nht khng nguyn cng vi chng nguyn thu A/Gs/Gd/1/96 ca Qung ng (Chen et al., 2004), v bt u phn ha giai on dch cm c lit xy ra nm 2003 - 2005 (de Jong, Hien, 2006). S xut hin ca genotype Z vi

tnh gy bnh c lit trong nhng nm ny cc nc ng Nam l bng chng ca s t bin lch khng nguyn ca cm A/H5N1 (Li et al., 2004; Wu et al., 2008b). Cui nm 2005, c nhiu di dng ca virus cm A/H5N1 cng lc c hnh thnh, l s xut hin phn dng Thanh Hi (Qinghai and Qinghai-like sublineage) v phn dng Phc Kin (Fujian and Fujian-like sublineage), trn ngp chu bao gm Trung Quc, Hng Kng, Vit Nam, Indonesia, Thi Lan (Li et al., 2004; Sims et al., 2005; Smith et al., 2006), trn sang Trung , chu u v chu Phi c tnh gy bnh cao i vi ngi (Skeik, Jabr, 2008; Ducatez et al., 2007; Peiris et al., 2007; Zhao et al., 2008). Cc chng thuc di dng Phc Kin c cu trc gen NA(N1) khng thay i nhiu, nhng gen HA(H5) c motif amino acid vng ni ca im ct protease l -RRRK-, gim mt mt Lysine (K) so vi cc chng di dng Qung ng (Smith et al., 2006), v do vy k t 2006 n nay, nhiu chng/dng virus cm A/H5N1 cng tn ti gy bnh, trong c nc ta (Dung Nguyen et al., 2008). Trong cc nm 2006 2008, tuy bnh din dch cm gia cm khng c lit nh nhng nm 2003-2005, nhng do xut hin nhiu chng A/H5N1 c bin ng khng nguyn v c lc, vn dch t hc cm A/H5N1 c th tr nn phc tp hn (Zhao et al., 2008; Gambotto et al., 2008). V s hnh thnh genotype K t khi cm A/H5N1 hin i xut hin ti Qung ng (A/Gs/CN/Gd1/1996(H5N1) (Xu et al., + 1999), c tt c 12 genotype, bao gm GD, A, B, C, D, E, X(X0-X3), V, Y, W, Z(Z ) v G c pht hin, trong cc chng H5N1 ca Vit Nam l loi mi bin i, hu ht thuc genotype Z v G (Nguyn Tin Dng et al., 2004; Dung Nguyen et al., 2008). T nm 2002 tr li y, nhng genotype nguyn thy ca dng Qung ng khng cn tn ti ( l cc genotype A, C, D, E), nhng li tin ha xut hin 8 genotype ca H5N1 (V, W, X1, X2, X3, Y, Z v Z+) (Li et al., 2004; Macken et al., 2006). Cc chng H5N1 phn lp ti Vit Nam trong nhng nm 2004 - 2006, thuc dng Qung ng, tp trung ch yu l genotype Z, gn y xut hin thm genotype G, nhng c phn bit lm thnh 2 nhm: nhm N (North) ph bin Bc Vit Nam v nhm S (South) phn b min Nam (Smith et al., 2006; Dung Nguyen et al., 2008). Nm 2007, ngoi cc chng v genotype thuc phn dng Qung ng gy dch cm gia cm, cn c mt phn dng khc c phn lp v xc nh bng sinh hc phn t phn tch gen H5 v N1, l dng Phc Kin (Dung Nguyen et al., 2008 ; L Thanh Ha et al., 2008). Tin ha chng/type v dng ti t hp mi thng nh hnh v xut pht t cc tnh Nam Trung Quc (Chen et al., 2006; Smith 2006; Wang et al., 2008). Tr phn dng Thanh Hi (Qinghai-like sublineage) cha c pht hin ti Vit Nam, hai phn dng Qung ng (Guangdong-like sublineage) v Phc Kin (Fujian-like sublineage) v ang gy dch cm gia cm nc ta, lm cho tnh hnh dch t v phng chng c th tr nn phc tp trong nh hng i ph (Dung Nguyen et al., 2008). C IM KHNG NGUYN-MIN DCH Cu trc, chc nng ca HA v NA Hai protein HA v NA l cc protein chnh b mt capsid (major capsid protein) c trng cho bn cht ca tng chng virus cm A (Murphy, Webster, 1996; Keawcharoen et al., 2005). Protein HA (Hemagglutinin) Protein hemagglutinin l mt glycoprotein thuc protein mng type I (lectin), c kh nng gy ngng kt hng cu g trong ng nghim (in vitro), khng th c hiu vi HA c th phong ta s ngng kt , c gi l khng th ngn tr ngng kt hng cu (HI- Hemagglutinin Inhibitory antibody). C 16 phn type HA c pht hin (H1 - H16, phn type H16 mi c tm thy virus gy bnh cho Hi u u en - Thy in, 1999 L Thanh Ha, 2006)), ba phn type (H1, H2 v H3) thch ng ly nhim gy bnh ngi lin quan n cc i dch cm trong lch s (Murphy, Webster, 1996). C khong 400 phn t HA trn b mt capsid ca mt virus, c vai tr quan trng trong qu trnh nhn din virus v khi ng qu trnh xm nhim ca virus vo t bo ch (Bender et al., 2001; Wagner et al., 2002). Phn t HA c dng hnh tr, di khong 130 ngstron (), cu to gm 3 n phn (trimer), mi n phn (monomer) c to thnh t hai di n v HA1 (36 kDa) v HA2 (27 kDa), lin kt vi nhau bi cc cu ni disulfide (-S-S-). Cc n phn sau khi tng hp c

glycosyl ha (glycosylation) v gn vo mt ngoi capsid l di n v HA 2, phn u t do hnh chm cu c to bi di n v HA1 cha ng v tr gn vi th th thch hp ca HA trn b mt mng t bo ch (Bosch et al., 1979; 1981; Wagner et al., 2002). S kt hp ca HA vi th th c hiu (glycoprotein cha sialic acid) trn b mt mng t bo, khi u qu trnh xm nhim ca virus trn vt ch gip cho virus xm nhp, ha mng v gii phng RNA h gen thc hin qu trnh nhn ln trong t bo cm nhim. Qu trnh kt hp ph thuc vo s ph hp cu hnh khng gian ca th th cha acid sialic ca t bo ch vi v tr gn vi th th ny trn phn t HA ca virus cm, quyt nh s xm nhim d dng ca virus cc loi vt ch khc nhau (Webster, 1998; Wagner et al., 2002). V tr amino acid 226 (aa226) ca di n v HA1 c xc nh l v tr quyt nh ph hp gn HA vi th th c hiu ca n, hu ht cc chng virus cm A lu hnh trong t nhin v tr ny l Glycine, thch ng vi th th Gal -2,3 sialic acid (cha sialic acid lin kt vi nhm hydroxyl (4-OH) ca galactose gc quay -2,3) ca t bo biu m ng h hp ca chim v gia cm (vt ch t nhin ca virus cm A). Ngoi ra, mt s v tr amino acid khc: Glutamine 222, Glycine 224, hay cu trc SGVSS v NGQSGR cng c s lin quan cht ch n kh nng thch ng vi th th cha sialic acid b mt mng t bo ch (Keawcharoen et al., 2005). c bit, mt s chng virus cng c A/H5Nx; A/H7Nx lu hnh hin nay c th xm nhim trn ngi, khi chng c ti lng cao trong ng h hp (do tip xc trc tip vi cht thi hay gia cm nhim bnh) (Webster, 1998; Bauer et al., 2006; Hui et al., 2008). Trnh t m ha chui ni, v thnh phn chui ni trn protein HA cng nh cc v tr amino acid lin quan n kh nng gn vi th th thch ng, c coi l cc ch th phn t trong nghin cu phn tch gen khng nguyn HA (Keawcharoen et al., 2005). Protein HA cn l khng nguyn b mt quan trng ca virus cm A, kch thch c th sinh ra p ng min dch dch th c hiu vi tng type HA, v tham gia vo phn ng trung ha virus, c coi l protein va quyt nh tnh khng nguyn, va quyt nh c lc ca virus, l ch ca bo v min dch hc nhm ngn chn s xm nhim ca virus c th nhim, c s iu ch cc vaccine phng cm hin nay (Boschet al., 1982; Matrosovich et al., 1999; Keawcharoen et al., 2005 ; Horimoto, Kawaoka, 2006). Protein NA (neuraminidase) Protein neurominidase cn gi l sialidase (m s quc t l E.C 3.2.1.18), l mt protein enzyme c bn cht l glycoprotein c gn trn b mt capsid ca virus cm A, mang tnh khng nguyn c trng theo tng phn type NA (Baigent, McCauley, 2001; Wagner et al., 2002). C 9 phn type (t N1 n N9) c pht hin ch yu virus cm gia cm, hai phn type N1 v N2 c tm thy virus cm ngi lin quan n cc i dch cm trong lch s (Webster, 1998). C khong 100 phn t NA xen gia cc phn t HA trn b mt capsid ht virus. Phn t NA c dng nt li hnh nm, u t do (cha vng hot ng) gm 4 di n v ging nh hnh cu nm trn cng mt mt phng, v phn k nc gn vo v capsid (Castrucci, Kawaoka, 1993; Wagner et al., 2002). Protein NA c vai tr l mt enzyme ct t lin kt gia gc sialic acid ca mng t bo nhim vi phn t cacbonhydrate ca protein HA, gii phng ht virus ra khi mng t bo nhim, y nhanh s ly nhim ca virus trong c th vt ch, v ngn cn s tp hp ca cc ht virus mi trn mng t bo. Mt khc, NA tham gia vo phn ct lin kt ny trong giai on ha mng, y nhanh qu trnh ci o uncoating gii phng h gen ca virus vo trong bo tng t bo nhim, gip cho qu trnh nhn ln ca virus din ra nhanh hn (Wagner et al., 2002). Ngoi ra, NA cn phn ct cc lin kt glycoside, gii phng neuraminic acid lm tan long mng nhy b mt biu m ng h hp, to iu kin cho virus nhanh chng tip cn t bo biu m v thot khi cc cht c ch khng c hiu. Cng vi vai tr ca khng nguyn HA, c 3 khu tc ng trn ca NA u tham gia lm gia tng c lc gy bnh ca virus cm A c th vt ch. Do , NA l ch tc ng ca cc thuc, ha dc c ch virus khng c hiu hin nay, c bit l Oseltamivir (bit dc l Tamiflu) phong ta enzyme ny, ngn cn s gii phng ht virus mi khi cc t bo ch, bo v c th (Castrucci, Kawaoka, 1993; Aoki et al., 2007). Bn cnh , NA cn l mt khng nguyn b mt ca virus, tham gia kch thch h thng min

dch ca c th ch, sinh ra khng th c hiu vi khng nguyn NA ca cc chng virus ng nhim c tc dng phong ta protein NA (Doherty et al., 2006). Nh vy, khng nguyn NA cng vi khng nguyn HA ca virus l cc ch ch yu ca c ch bo h min dch ca c th vi virus cm A, v l c s iu ch cc vaccine phng cm hin nay cho ngi v gia cm, nhm ngn chn dch cm gia cm v hn ch ly truyn sang ngi (Suarez, Schultz-Cherry, 2000; Wu et al., 2008b). Cc phng thc bin i khng nguyn C ba phng thc ch yu lm bin i khng nguyn virus cm A (Wu et al., 2008b). Hin tng lch khng nguyn Lch khng nguyn (antigenic drift) thc cht l cc t bin im xy ra cc phn on gen/h gen ca virus. Do virus cm A k sinh ni bo bt buc, khng c c ch c v sa bn sao - proof reading trong qu trnh phin m v sao chp nhn t bo ch. S thiu ht enzyme sa cha RNA dn n cc enzyme sao chp ph thuc RNA s c th gi thm (t bin gin n), lm mt i hoc thay th (t bin trt-xa) (L Thanh Ha et al., 2006) mt hay nhiu nucleotide m khng c sa cha trong phn t RNA chui n mi ca virus (Murphy, Webster, 1996; Conenello et al., 2007). Tu thuc v tr xy ra cc t bin trong b ba m ha, m c th trc tip lm thay i cc amino acid trong trnh t ca protein c m ha biu hin, dn n thay i thuc tnh ca protein, hoc c tch ly trong phn on gen xy ra t bin (t bin im). Tn sut xy ra t bin im rt cao, c mi 10.000 nucleotide (tng ng vi di ca RNA h gen ca virus cm A) th c 1 nucleotide sai khc (Webster, 1998; Rabadan et al., 2006). Nh vy, gn nh mi ht virus mi c sinh ra u cha ng mt t bin im trong h gen ca n, v cc t bin ny c tch ly qua nhiu th h virus s lm xut hin mt phn type virus mi c nhng c tnh khng nguyn mi c th b sai lch (Hnh 6). Hin tng ny thng xy ra cc phn on gen khng nguyn NA v HA, to ra cc b m tng hp cc amino acid mi, hoc lm thay i cu trc dn n thay i c tnh ca protein , hoc c kh nng glycosyl ha rt cao trong cu trc chui polypeptide khng nguyn, to ra mt bin th virus mi thay i c lc gy bnh hay c tnh khng nguyn mi (Webster, 1998; Macken et al., 2006; Chen et al., 2008). Hin tng trn khng nguyn Hin tng trn khng nguyn (cn gi l trao i hay ti t hp) cc gen khng nguyn (antigenic shift) ch c virus cm, v rt t mt s virus RNA gy bnh gia cm khc, cho php virus c kh nng bin chng rt cao. H gen gm 8 phn on gen ring bit ca virus cm A c 2 chng virus cm A khc nhau khi ng nhim trong mt t bo trao i cho nhau, c th xy ra s ho trn (reassort) hoc trao i (swap) cc phn on gen ca hai chng virus trong qu trnh kt hp li RNA h gen, to ra cc trng thi khc nhau ca RNA h gen ca cc ht virus mi t hai RNA h gen ca nhng virus ban u. Kt qu l to ra th h virus mi c cc phn on gen kt hp, v i khi gip cho chng c kh nng ly nhim loi vt ch mi hoc gia tng c lc gy bnh (Hnh 7) (Hilleman, 2002; Mackenet al., 2006; Chen et al., 2008). Hin tng glycosyl ha Glycosyl ha (glycosylation) l s gn kt ca mt chui carbonhydrate (oligosaccharide) vo vi amino acid Asparagine (N) mt s v tr nht nh trong chui polypeptide HA hay NA, hay mt s polypeptide khc ca virus cm. Thng thng chui oligosaccharide c gn ti v tr N-X-S/T (N = Asparagine; X = amino acid bt k, tr Proline; S/T = Serine hoc Threonine) (Baigent, McCauley, 2001). y l nhng v tr c cho l gn kt vi cc khng th c c th sinh ra do kch thch ca khng nguyn, nhm bo v c th nhim. Hin tng lch khng nguyn sinh ra t bin im hnh thnh b m ca Asparagine, to tin cho hin tng glycosyl ha xy ra khi tng hp chui polypeptide HA hay NA, lm thay i biu hin c tnh khng nguyn ca HA v NA, gip cho virus thot khi tc ng min dch bo h ca c th ch v iu ho s nhn ln ca virus (Baigent, McCauley, 2001; Phan Vn Chi et al., 2008).

Hin tng lch khng nguyn v glycosyl ha xy ra lin tc theo thi gian, cn hin tng trn khng nguyn c th xy ra vi tt c cc chng ca virus cm A, khi ng nhim trong mt t bo tt c cc loi vt ch khc nhau. y cng chnh l vn ng lo ngi ca virus cm A/H5N1 hin nay, mc d virus ny cha c s thch nghi ly nhim d dng ngi, nhng n c kh nng gy bnh c cho ngi, v rt c th A/H5N1 ti t hp (vay mn) gen HA hay NA, hoc c hai gen ca cc chng virus cm A thch nghi ngi, to ra mt bin chng virus mi thch ng ly nhim d dng ngi, gy ra nguy c ca mt i dch cm mi v t ra mt nh hng mi trong phng chng (Hilleman, 2002; Guanet al., 2004; Li et al., 2004; Arinaminpathy et al., 2008; Korteweg, Gu, 2008)

Hnh 6. S minh ha t bin im ca hin tng khng nguyn (antigenic drift) (A) v t bin ti t hp ca hin tng trn khng nguyn (antigenic shift) virus cm A (B).

NGHIN CU PHT TRIN VACCINE CM A/H5N1 TRN TH GII V TI VIT NAM Cc chng virus cng c A/H5N1 sau 1996, qua thi gian tin ha, c xu hng bin i ni gen nhm tng tnh gy bnh, v thay i thnh phn ni gen khng nguyn lm mt tng quan min dch gia chng v cc chng vaccine c to ra. Do vy, vn ny phi c ht sc ch trong chin lc kin to vaccine, cng nh s dng vaccine c to ra (Horimoto, Kawaoka, 2006). Trong t nhin, lch khng nguyn v glycosyl ha l cc hin tng xy ra lin tc theo thi gian, cn trn khng nguyn ti t hp subtype Hemagglutinin (HA) v Neuraminidase (NA) thng nh k xy ra gia cc loi mc nhm to nn cc bin th virus cm mi m h min dch ca c th vt ch khng c kh nng nhn dng p ng v kp hnh thnh min dch. H5N1 s dng th th sialic xm nhp t bo (Yamada et al., 2006). Hemagglutinin cng vi protein enzyme neuraminidase, c xc nh l protein c vai tr khng nguyn v c lc, (Scholtissek et al., 2002; Wagner et al., 2002 ; Keawcharoen et al., 2005) v cc protein khng nguyn c ngun gen t cc chng H5N1 ca Vit Nam, cng c nghin cu sn xut bng k thut ti t hp v s dng vi cc mc ch khc nhau trong c th nghim lm khng nguyn kch thch min dch v chn on (Nguyn Tin Minh et al., 2004; Simmons et al., 2007). i vi bnh truyn nhim, vaccine c coi l bin php c tnh chin lc, nhm ngn chn ly lan, to bo h min dch (Subbarao, Luke, 2007). i vi dch cm A/H5N1 gia cm v d phng dch cm trn ngi, nghin cu pht trin vaccine khng nhng ngn nga lm gim c bnh gia cm, m cn khng ch ngun truyn ly ca loi virus nguy him ny sang ngi (Horimoto, Kawaoka, 2001; OIE, 2005; Subbarao, Luke, 2007). Khng th c hiu c th c c th sinh ra do kch thch ca khng nguyn trong vaccine, v l cc khng th khng HA, NA, MA v nhiu loi hnh khc ca virus ng nhim, gp phn v hiu ha virus cm ng i tng khi chng xm nhp vo. C nhiu loi khng th, nhng trc ht ch khng th khng HA(H5) c vai tr tin quyt quan trng trong qu trnh trung ha virus cho bo h min dch. Cc vaccine phng bnh hin nay da trn c s hai loi chnh: vaccine truyn thng v vaccine th h mi (Subarao, Luke, 2007; Capua, Alexander, 2008). Vaccine truyn thng: bao gm vaccine v hot ng chng v d chng. Vaccine v hot ng

chng (homologous vaccine), l cc loi vaccine c sn xut cha cng nhng chng virus cm g ging nh chng gy bnh trn thc a (Swayne, Suarez, 2000). Vaccine v hot d chng (heterologous vaccine) l vaccine s dng cc chng virus c khng nguyn HA ging chng virus trn thc a, nhng c khng nguyn NA d chng. Vaccine th h mi hay vaccine cng ngh gen: l loi vaccine c sn xut da trn s dng k thut gen loi b cc vng gen c ang c nghin cu v a vo s dng ph bin, bao gm: - Vaccine ti t hp c vector u gia cm dn truyn: s dng virus u gia cm lm vector ti t hp song gen H5 v N1 phng chng virus type H5N1 v H7N1 (Qiaoet al., 2006). V d, hng Merial ca Php sn xut vaccine TrovacAIV-H5 ly ngun gen H5 t chng A/Turkey/Ireland/83 (H5N2), s dng c cho gia cm lc 1 ngy tui. - Vaccine di nhm cha protein khng nguyn NA, HA ti t hp v tch chit lm vaccine (Peyre et al., 2008; Prel et al., 2008). - Vaccine ti t hp c vector dn truyn: s dng adenovirushoc Newcastle virus hoc virus u chim lm vector dn truyn, lp ghp gen khng nguyn H5 vo h gen ca adenovirus, to nn virus ti t hp lm vaccine phng chng virus cm A/H5N1 (Hoelscheret al., 2006; Gao et al., 2006; Rmer-Oberdrfer et al., 2008). - Vaccine DNA: sn phm DNA plasmid ti t hp cha gen HA, NA, NP, M2 n l hoc a gen (Kodihalli et al., 1999; Keawcharoen et al., 2005). - Vaccine nhc c virus cm nhn to: csn xut bng k thut di truyn ngc, l vic lp ghp virus cm nhn to cha y h gen, trong cc gen khng nguyn H5 c vng "c" c bin i bng k thut gen (Liu et al., 2003; Tian et al., 2005). C 3 loi vaccine c T chc Y t th gii (WHO) cng nhn v an ton v khuyn co a vo chng trnh sn xut vaccine trn th gii hin nay, l NIBRG-14 (NIBSC), VN/04xPR8-rg (SJCRH) v VNH5N1-PR8/CDC-rg (CDC). Hai chng cm A/H5N1 cung cp ngun gen H5 v N1 l A/Vietnam/1194/2004(H5N1) hoc A/Vietnam/1203/2004(H5N1) (Nicolson et al., 2005). Trung Quc cng l nc sn xut nhiu ging virus vaccine chng cm, v d, Vin Nghin cu Th y Harbin (Cp - Nh - Tn) thnh cng trong vic to ging vaccine v hot nh du n chng ly ngun gen H5 v N2 t chng A/Turkey/England/N-28/73(H5N2), loi subtype H5N2 c c lc yu; hay ging vaccine v hot nh du n chng ly ngun gen H5 v N1 t chng A/Goose/Guangdong/1996(H5N1), loi c c lc yu (Tian et al., 2005; Qiao et al., 2006). Cc loi vaccine ny c nhp v s dng ti Vit Nam t nm 2006 cho n nay. Vaccine th h mi chng NIBRG-14: Chng NIBRG-14 l ging virus vaccine nhc c (attenuated vaccine) th h mi, thuc loi hnh vaccine c xa gen bng cng ngh gen (genedeletion vaccines), c lp rp nhn to bng k thut di truyn ngc (reverse genetics based technology) v thch ng nhn ln khi nui cy trn phi g (Marsh, Tannock, 2005).Phng php di truyn ngc c s dng to ra chng virus nhn to nhc c lm vaccine, c th h gen ca chng nhn to NIBRG-14 ny c ti t hp gen trn c s s dng chng gc PR8/34 (A/Puerto Rico/8/34/Mount Sinai(H1N1)) cung cp 6 gen khung l PA, PB1, PB2, NP, MA, NS lm nn, cn cc gen khng nguyn HA(H5) v NA(N1) c ly t chng cm cng c gy bnh phn lp nm 2004 ti Vit Nam (A/Vietnam/1194/2004(H5N1)) (Tian et al., 2005). Bng thao tc k thut gen, gen H5 b t bin lm mt hn 4 amino acid RRRL, cng vi mt s t bin im cc b m hai u ca vng c, lm thay i 3 amino acid ti vng gy c. Nh vy v mt min dch hc, mc d virus c x l lm mt c tnh gy bnh nhng vn gi nguyn bn cht c tnh khng nguyn b mt ging ht nh virus cm A/H5N1 ly mu ban u, do vy, c kh nng to khng th khng li khng nguyn b mt loi virus H5N1 gy bnh trong t nhin (Doherty et al., 2006; Peyre et al., 2008). THAY LI KT LUN V NH HNG TNG LAI Nghin cu nh type, bin i di truyn v gen hc tin ha ca virus cm A/H5N1 c cc c

quan nghin cu ca Vit Nam tin hnh ngay t nhng thng u tin xy ra dch cm gia cm cui nm 2003. Nhng chui gen gip xc nh phn type H5, phn type N1 v cc gen cu trc c Vin Cng ngh Sinh hc, Vin Pasteur TP H Ch Minh, Vin V sinh dch t trung ng, Vin Th y gii m v cng b trn Ngn hng gen (Le et al., 2005; Le Thanh Hoa et al., 2006; Dung Nguyen et al., 2008). Trn c s phn tch trnh t gen khng nguyn H5 v N1, cc tc gi khng nh ngun gc ca virus cm A gy bnh trn gia cm v ngi ti Vit Nam cng nhm vi virus H5N1 phn lp ti Trung Quc (Nguyn Tin Dng et al., 2004; L Thanh Ha et al., 2006; Muramoto et al., 2006). Cc bin chng H5N1 ca Hng Kng, Trung Quc phn lp nhng nm 1997 - 2001 v Hn Quc, i Loan (phn lp nm 2003) u c ngun gc t chim ct v ngng (A/Goose/Guandong/1/96) vng Qung ng (Trung Quc), l cc bin chng thuc dng Qung ng (Nguyn Tin Dng et al., 2004; L Thanh Ha et al., 2006; L Trn Bnh et al., 2006). Nh vy, virus cm gia cm gy bnh gia cm v ngi ti Vit Nam l cm H5N1 type A thuc th h mi c bin i c bn v gen H5 v gen N1, nhng vn c cng ngun gc vi H5N1 t vng a l Nam Trung Quc v Hng Kng (Nguyn Tin Dng et al., 2004; 2008; Li et al., 2004; Smith et al., 2006). Cc chng phn lp nhng nm 2004-2006 c nghin cu kh chi tit v gc gen hc v quan h phn t vi cc chng trong vng v th gii, kt qu khng nh virus H5N1 vng Nam v ng Nam thuc nhm di truyn VTM (vit tt: Vietnam-Thailand-Malaysia), c nhng c tnh sinh hc nht nh khc vi cc nhm vng Trung Quc v Hng Kng (Li et al., 2004; Chen et al., 2006; 2008). Nm 2007, xut hin thm bin chng H5N1 di dng Phc Kin ti Vit Nam, v ang lm phc tp thm vn dch t hc v quan h khng nguyn v min dch, do t l tng ng khng nguyn HA(H5) v NA(N1) thp so vi cc chng phn dng Qung ng, tuy nhin vn cn c kh nng bo h min dch (Nguyn Mnh Kin et al., 2008; L Thanh Ha et al., 2008; Nguyn Tin Dng et al., 2008; Dung Nguyen et al., 2008 ; Le et al., 2008). Nghin cu vn gen hc khng nguyn lin quan n vaccine v min dch, c Vin Cng ngh sinh hc, Vin V sinh dch t Trung ng, Vin Pasteur TP H Ch Minh, Vin Th y trung ng tin hnh l vic thu thp gen khng nguyn H5 v N1 t cc chng phn lp trn g, vt, ngan ca Vit Nam cc nm 2004 - 2008, v so snh vi trnh t chui gen cm A/H5N1 ca cc chng cng c ng nhim v vaccine ca Vit Nam v th gii (L Thanh Ha et al., 2004; Nguyn Tin Dng et al., 2004; 2008; Nguyn Th Bch Nga, 2006; L Trn Bnh et al., 2006; Le Thanh Hoa et al., 2006; L Trn Bnh, 2007; Dung Nguyen et al., 2008 ; Le et al., 2008). Nm 2007, xut hin thm chng H5N1 di dng Phc Kin (clade 2.3.4) ti Vit Nam, chc chn lm phc tp thm vn quan h khng nguyn v min dch, do t l tng ng khng nguyn HA(H5) qua phn tch gen gia cc chng phn lp ti Ngh An (Vit Nam), A/Dk/Vietnam/NA114/2007(H5N1) v A/Dk/ Vietnam/NA72/2007(H5N1), thuc dng Phc Kin) vi cc chng H5N1 thuc phn dng Qung ng, bao gm mt s chng lm vaccine ang s dng, ch t 94% (L Thanh Ha et al., 2008 ; Nguyn Mnh Kin et al., 2008). Nhn nh hn hp virus gy bnh v phn ha khng nguyn ca virus cm A/H5N1 ti Vit Nam cng c xc nhn qua phn tch hng chc chng thu nhn t nhiu vng khc nhau trong c nc (Nguyn Tin Dng et al., 2008; Dung Nguyen et al., 2008; Le et al., 2008). iu ny nh hng n dch t, chn on, phng tr v quan h ly nhim trong t nhin (Smith et al., 2006b; Alexander, 2007), cng nh vai tr min dch ca cc chng c in ang lm vaccine ti Vit Nam v th gii (vaccine H5N1, chng gc: A-Gs-CN-Gd1(96)(H5N1); vaccine H5N2, chng gc: A-Turkey-ENG-N28(73)(H5N2); vaccine TrovacAIV-H5, chng gc: A-TkIRE-1378(83)(H5N8)); vaccine H5N2, chng gc: A-Ck-MEX-Hidalgo-232(94)(H5N2)) v vaccine H5N1 th h mi chng NIBRG-14, s dng chng gc: A/Vietnam/1194/2004(H5N1) hin nay ang c Vit Nam nghin cu sn xut (L Trn Bnh et al., 2006; L Trn Bnh, 2007; Nguyn Th Lan Phng, L Vn Hip, 2006). Vn chn on v xy dng phng php pht hin nhanh v phn bit cm A vi cc tc nhn gy triu chng h hp khc, cng nh phn bit cc phn type HA v NA c cc nh khoa hc Vit Nam quan tm, kt hp nghin cu vi cc t chc th gii. Pht hin nhanh H5N1 v cc phn type khc bao gm vic s dng khng nguyn hoc khng th, hoc sinh hc phn t c xy dng thnh phng php (Chan et al., 2007; Wu et al., 2008b). Nghin cu vaccine v min dch,

cc nh khoa hc Vit Nam cng c nhng ng gp nht nh v to ch phm khng nguyn, to vaccine di truyn ngc hoc vector ti t hp trn nn virus cm A/H5N1 ca Vit Nam (Gao et al., 2006; Bch Th Nh Qunh, 2006; Nguyn Th Bch Nga, 2006; Lu et al., 2006; Ge et al., 2007). Vnghin cu sn xut vaccine, Vin Cng ngh sinh hc c giao ti c lp cp Nh nc Nghin cu xy dng qui trnh sn xut vaccine cm A/H5N1 cho gia cm v nh gi cht lng vaccine phng bnh cm A/H5N1 cho gia cm ti Vit Nam bng chng NIBRG-14 trong giai on 2006 - 2008, kt hp vi Vin Th y, X nghip thuc Th y trung ng (VETVACO), Cng ty Thuc th y trung ng II (NAVETCO), Trung tm Kim nghim thuc th y trung ng, thc hin nghin cu c c vaccine sn xut t chng NIBRG-14 (L Trn Bnh, 2007). y l chng vaccine c to ra bng cng ngh di truyn ngc ti Vin Tiu chun v Kim nh sinh hc Quc gia (Vng quc Anh), thch ng trn phi g 10 ngy tui v vaccine to ra di dng v hot nh du. ti c thc hin trong 3 nm (2006 - 2008), kt qu l xy dng c cc quy trnh sn xut ging, sn xut vaccine, kim nghim v bo qun vaccine cm A/H5N1 v kim nghim min dch t cht lng bng phng php huyt thanh hc v th thch cng c (Nguyn Th Lan Phng, L Vn Hip, 2006; L Trn Bnh, 2007). Ngoi ra, mt s ti nh nc khc nghin cu dch t hc phn t, chn on, vector ti t hp dn truyn gen khng nguyn s dng adenovirus hoc LaSoTa virus, chuyn gen vo thc vt cng c tin hnh (trao i ring: Trng Nam Hi; Trng Vn Dung; L Trn Bnh; L Thanh Ha; Nguyn Tin Dng v nhiu ngi khc). Mt s nghin cu ng dng k thut/cng ngh cao (di truyn ngc, ti t hp vector) s dng ngun gen H5N1 ca Vit Nam, cng ngh ti t hp sn xut ch phm khng nguyn H5 trong thc vt v t bo, cng ang c tin hnh ti Vin Cng ngh sinh hc, Vin Th y, Vin Di truyn nng nghip v mt s c s nghin cu khc (L Trn Bnh, 2007; L Thanh Ha, 2006a; Nguyn Tin Dng et al., 2008). Song song vi nhng ni dung nghin cu v cm gia cm gia cm, cc c s y t gm bnh vin, vin nghin cu (Bnh vin Nhi trung ng, Vin V sinh dch t Trung ng, Vin Pasteur TP H Ch Minh, Vin Vaccine Nha Trang) u c nhng trin khai cc lnh vc nghin cu lin quan n cm A/H5N1 trn ngi (Le et al., 2005; 2008; Nguyn Th Kim Tin, 2005; Dinh et al., 2006; Chan et al., 2007; Hatta et al., 2007) Nhng kt qu nghin cu v cm A/H5N1 gia cm v ngi trn nn virus cm A/H5N1 ca Vit Nam v ang lm sng t thm v mi quan h tin trin bnh hc ly nhim, dch t hc phn t, pht trin tin ha v genotype v khng nguyn - min dch - vaccine ca cm gia cm ti Vit Nam. Li cm n:Chng ti cm n B Khoa hc v Cng ngh cp kinh ph cho ti c lp cp nh nc v nghin cu vaccine cm A/H5N1 do GS. TS. L Trn Bnh ch nhim v ti Ngh nh th Vit Nam - Thi Lan v sinh hc phn t cm A/H5N1 do PGS. TS. L Thanh Ha lm ch nhim. TI LIU THAM KHO 1THE A/H5N1 INFLUENZA VIRUS: INSIGHTS OF EPIDEMIOLOGY, EVOLUTION, GENOTYPE GENERATION AND COMPATIBILITY OF ANTIGENICITY - IMMUNOGENICITY - VACCINE Le Thanh Hoa*, Dinh Duy Khang, Phan Vn Chi, Nong Van Hai, Truong Nam Hai, Pham Viet Cuong, Nguyen Thi Bich Nga, Le Tran Binh Institute of Biotechnology SUMMARY Avian influenza is a highly pathogenic and contagious disease caused by influenza A viruses, of which the causative agent belongs to the family Orthomyxoviridae, which can be transmitted from the infected animals to humans. There are 16 subtypes HA (H1 - H16) and 9 subtype NA (N1 - N9) to be known to go through reassortment to generate hundreds of variants of different virulence and pathgenicity. H5N1 highly pathogenic avian influenza (HPAI) has continued to pose a significant threat to both animal and human health. From late 2003 to June 2008, accumulated number of infected human cases is 385 with 243 deaths, presenting 63.11% mortality rate, whereas Vietnamand Indonesia are the two countries of highest casualty. Unlike previous poultry outbreaks of highly pathogenic avian influenza due to H5 occurred from 1996 -2002 that they were

controlled by depopulation with or without vaccination, the presently circulating A/H5N1 viruses (from 2003 - present) with formation of different genotypes, particularly genotype Z, have since been spreading from Southern China to other parts of the world. This is still a great threat to public health and defferent measures are needed to be developed to control. The virulence determinants for highly pathogenic avian influenza viruses are not limited to the best characterized virulence factor, the hemagglutinin (HA) cleavage site and the enzymatic activity of neuraminidase (NA), but rather are considered to be multigenic and capable to reassort gene segments for new viruses to emerge with different virulence characteristics, evidently encountered. Tens of thousand papers presenting the research work related to avian influenza A, in general, and to A/H5N1 in particular, ie. in recent 5 years, have been generated of which many come from the development of new technologies and vaccine(s) for immunization of poultry and for preparedness of pandemic possibly to occur in humans. In the view of epidemiology, evolution, genotype formation, antigenic variation to alter immunogenicity, sensitivity and resistance to pharmaceuticals, unlike other avian viruses, A/H5N1 virus tends to mutate in a fast way to generate new variants of new sublineages and tends to search for wider host spectrum to infect. Vaccine(s) from different reference strains of H5N1 have been developed using high-technologies including reverse-genetics, particularly, those which have been based on NIBRG-14 promise a good potentiality in control in future. Here, we review the basic understandings of epidemiological aspects, virology, molecular biology, evolutionary ocurrence, genotype formation, and current strategies for the development of H5N1 influenza vaccines, and possibilities of antigenic-immunogenic-vaccine relationships in Vietnam and worldwide. Summarized research works of the Vietnamese scientists and institutions regarding to research on the H5N1 endemics in Vietnam and on the materials of Vietnam origin including the contribution of the Institute of biotechnology on genomics/proteomics of the gene(s) of HA/NA and new generation vaccines are also introduced. Keywords:A/H5N1, antigenicity-immunogenicity, epidemiology, evolution, genotype, highly pathogenic avian influenza (HPAI), Orthomyxoviridae, subtype, vaccine

*Author for correspondence: Tel/Fax: 84-4-37567297; E-mail: imibtvn@gmail.com

Ngy 07/10/2010 PGS.TS. L Thanh Ha Vin Cng ngh sinh hc, Vin Khoa hc v Cng ngh Vit Nam