2株鸭甲肝病毒的分离及其全基因组序列分析

摘要

为深入研究鸭甲肝病(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 647nt(不合polyA序列),5＇和3＇非编码区长度分别为583nt和314nt,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 genomic 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-like group (isolate H).