通用引物鉴定野鸟源禽流感病毒HA亚型

准确、方便快捷的禽流感病毒亚型鉴定方法对于野鸟流感病毒感染与流行状况的调查监测是十分必要的。本研究参照Phipps L P等人建立的通用引物RT-PCR HA亚型鉴定方法,分别对已知为H1、H2、H3、H4、H6和H11亚型的22株野鸟流感病毒毒株以及6株未知亚型的野鸟流感病毒毒株进行了HA亚型分型的验证性试验。即利用1对通用引物对所有病毒样品进行RT-PCR扩增,将各PCR产物进行核苷酸序列测定并与GenBank上参考毒株序列进行同源性比较,以确定其HA亚型。BLAST分析显示,22株已知亚型毒株的PCR扩增产物的序列测定结果与GenBank公布的相应亚型毒株序列比较其核苷酸序列同源性均在99%～87%之间,而6株未知亚型的野鸟流感病毒毒株测得的核苷酸序列经BLAST比对分别确立为1株H4、1株H5、2株H7、2株H11亚型。利用该通用引物HA亚型鉴定方法对于已知亚型所获得的鉴定结果均与实验室原亚型鉴定结果一致,对于未知亚型能准确的确定其HA的亚型。因此相较于其他HA亚型鉴定方法,该方法简便易行,更适合于野鸟流感病毒的亚型鉴定。

Identification of HA Subtype Influenza Virus Strains Isolated from Wild Birds Through Universal Primers

Accurate and convenient methods are needed to identify influenza virus subtypes and quantify their prevalence to monitor influenza viruses among wild birds.Following the method to identify HA subtype using the universal primer RT - PCR, we verified the subtype classification of six influenza viruses previously identified as H1,H2,H3,H4,H6,and H11,and identified six additional but previously unknown HA subtypes.We used a pair of universal primers and the RT - PCR procedure to amplify the HA gene of all virus...