2株鸭甲肝病毒的分离及其全基因组序列分析

Isolation and determination of complete genomic sequence of two duck hepatitis A virus

为深入研究鸭甲肝病毒(DHAV),本研究通过RT-PCR方法和血清中和法分离鉴定得到两株DHAV(命名为A株和H株)。采用RT-PCR方法分段克隆这两个分离株的全基因组序列。分析结果显示,A株的基因组全长为7 647 nt(不含polyA序列),5'和3'非编码区长度分别为583 nt和314 nt,单一ORF为6 759 nt,编码2 253个氨基酸;H株的基因组全长为7 648 nt(不含polyA序列),5'和3'非编码区长度分别为482 nt和314 nt,单一开ORF为6 852 nt,编码2 284个氨基酸。A和H分离株与DHAV-1参考株比较,其核苷酸同源性分别为93.4%～97%和92.7%～95.8%。遗传进化分析表明,DHAV-1型主要分为两个亚群,这两个病毒分离株分别属于不同的亚群。

Two duck hepatitis A virus were isolated and identified by RT-PCR and virus neutrlization test with antiserum, designated isolate A and H. The complete sequences of the isolates were amplified and analyzed. The results showed that the full genomic length of isolate A or H contained 7,647 nt or 7,648 nt except poly（A） sequence, and had 5＇-and 3＇-terminal non-coding regions of 583 nt and 314 nt or 482 nt and 314 nt respectively, including a single open reading frame （ORF） which encode a polypeptide of 2,253 or 2,284 amino acids. Comparative genomie sequence analysis with other DHAV-1 reference strains indicated that the nucleotide sequence homology was 93,4% to 97% or 92.7% to 95.8%, which were divided into 03D-like group （isolate A） and 5886-1ike group （isolate H）.