鸭甲型肝炎病毒3型信阳株的分离鉴定及遗传演化分析

Amplification and phylogenetic analysis of duck hepatitis A virus type 3 Xinyang strain

为了解信阳地区鸭甲型肝炎病毒3型的遗传变异特性,对信阳市发生疑似肝炎的金定雏鸭进行9种常见病毒PCR/RT-PCR检测,同时采集病鸭肝脏组织无菌处理后,尿囊腔接种10日龄SPF鸭胚进行病毒分离,并对分离毒株进行全基因组和VP1基因序列分析。结果:成功分离到1株鸭甲型肝炎病毒3型(DHAV3),命名为HNXY23;接种病毒的鸭胚发育不良,死亡胚体全身呈出血水肿;序列比对和系统发育树表明,分离株HNXY23属于DHAV3 GⅠ基因型,与2022年分离株HB-1、HZ-1、HZ-2和HZ-3亲缘关系较近;利用DNAMAN软件比对31株DHAV3 VP1氨基酸位点变异情况,发现在184、187、195、206和209位这5个氨基酸位点有较为明显的变化差异。本研究结果丰富了信阳地区DHAV3的分子流行病学资料,为进一步研究DHAV3的致病机制奠定了基础。

In order to understand the genetic variation characteristics of duck hepatitis A virus type 3(DHAV3)in the Xinyang area,PCR/RT-PCR was used to detect 9 common viruses in Jinding ducklings with suspected hepatitis in Xinyang City.After aseptic treatment of the liver tissue samples of diseased ducks,10-day-old SPF duck embryos were inoculated in allantoic cavity for virus isolation,and whole genome and VP1 gene sequence analysis were carried out.The results showed that a DHAV3 strain was successfully isolated and named HNXY23.The duck embryos inoculated with the virus showed dysplasia,and the dead embryos showed hemorrhage and edema.Sequence alignment and phylogenetic tree showed that HNXY23 belonged to the DHAV3 GI genotype,and it had a close relationship with the isolates HB-1,HZ-1,HZ-2 and HZ-3 in 2022.Then,the DNAMAN software was used to compare the amino acid site variation of VP1 of 31 DHAV3 strains,and it was found that there were significant differences in five amino acid sites such as 184,187,195,206 and 209.The results of this study enriched the molecular epidemiological data of DHAV3 in the Xinyang area,and it laid a foundation for further research on the pathogenic mechanism of DHAV3.