我国鸡群禽戊型肝炎病毒RT-PCR检测及部分ORF2基因序列分析

Detection of avian hepatitis E virus RNA by RT-PCR

为从核酸水平证实我国鸡群中禽戊型肝炎病毒(HEV)的存在,本实验从山东省某鸡场患有肝脾肿大综合征的病鸡中采集10份粪便和8份胆汁样品,利用RT-PCR方法检测其中禽HEV ORF2基因片段,并将阳性PCR产物克隆测序。结果显示:18份粪便和胆汁样品中,13份为禽HEV RNA阳性;其序列间的同源性为97%～99%,与GenBank中登录的参考序列同源性为76.6%～98.1%;进化树显示与欧洲地区的禽HEV在同一分支,属于禽HEV基因3型。禽HEV ORF2基因的检出为进一步了解禽HEV对我国家禽养殖业的危害以及在我国的流行情况奠定了基础。

In China,the avian hepatitis E virus （HEV） infection in chicken flocks has been identified by serological studies,however,the detection of avian HEV RNA has not been reported. In this study,a total of 94 serum samples,10 fecal and 8 bile samples were collected from boilers of breeding stock suffered from the big liver and spleen disease in Shandong area. The fecal and bile samples detected by RT-PCR for the avian HEV ORF2 gene. The positive PCR products were cloned and sequenced. Thirteen of 18 fecal and bile samples were positive for avian HEV ORF2 RNA. Sequences analyses revealed that all 13 ORF2 RNA positive samples shared 97% to 99% nucleotide sequence identities to each other,and 76.6% to 98.1% to avian HEV isolates reported in NCBI. Phylogenetic analysis showed that these avian HEVs are closely related to the Europe isolates and belonged to avian HEV genotype 3. The results confirmed the existence of avian HEV infection in China.