CAJ | 학술논문

目的分析2003-2008年云南边境禽流感H5N1亚型病毒血凝素(HA)基因分子结构特征.方法对2003-2008年在云南边境采集境外禽类样品420份,做H5/N1亚型特异性RT-PCR及多重RT-PCR检测,对阳性样品中H5N1病毒HA 基因进行RT-PCR扩增,克隆至pMD18-T载体测序,并与国内外已知参考毒株序列进行比对及系统发育分析.结果 21份代表性病毒样品HA裂解位点序列存在4种不同排列方式,均具有高致病性禽流感病毒分子结构特征;HA受体结合位点、糖基化位点及表位关键性氨基酸位点存在变异;系统发育分析表明可划分为5个不同进化分枝(1、2.4、2.3.2、2.3.4、7).结论 2003-2008年云南边境外H5N1病毒间具有遗传差异,进化分枝2.3.4毒株为当地流行的优势毒株,不同进化分枝或同一进化分枝不同毒株HA基因存在变异.
목적 분석2003-2008년운남변경금류감H5N1아형병독혈응소(HA)기인분자결구특정.방법 대2003-2008년재운남변경채집경외금류양품420빈,주H5/N1아형특이성RT-PCR급다중RT-PCR검측,대양성양품중H5N1병독HA 기인진행RT-PCR확증,극륭지pMD18-T재체측서,병여국내외이지삼고독주서렬진행비대급계통발육분석.결과 21빈대표성병독양품HA렬해위점서렬존재4충불동배렬방식,균구유고치병성금류감병독분자결구특정;HA수체결합위점、당기화위점급표위관건성안기산위점존재변이;계통발육분석표명가화분위5개불동진화분지(1、2.4、2.3.2、2.3.4、7).결론 2003-2008년운남변경외H5N1병독간구유유전차이,진화분지2.3.4독주위당지류행적우세독주,불동진화분지혹동일진화분지불동독주HA기인존재변이.
To elucidate the molecular characteristics of the hemagglutinin (HA) genes of H5N1 subtype of avian influenza viruses in the boundary region of Yunnan province. Of 420 samples were collected from the foreign poultry in boundary region of Yunnan province during 2003 to 2008 and these samples were subjected to screening by H5/N1 subtype-specific and multiplex RT-PCR. testing. The HA genes of H5N1 viruses from positive samples were amplified by RT-PCR and cloned into vector pMD18 T for subsequent sequencing. The alignment with sequences of the known reference strains and phylogenetic analysis were then performed. The genes from 21 representative positive samples with 4 different sequences at the cleavage site were obtained and all of them possessed the molecular characteristic of highly pathogenic avian influenza virus. The mutation of key amino acids had been found among receptor-binding sites, potential glycosylation sites and neutralizing epitopes.-Phylogenetic analysis showed those positive samples could be divided into 5 distinct clades, including clade 1, 2.4. 2.3.2, 2.3.4 and 7. It is evident that H5N1 viruses from the foreign boundary region of Yunnan province in 2003 to 2008 show genetic divergence and clade 2,3,4 is the dominant clade in this region.