摘要
为了解广东省某两个养殖场发生的疑似鸭病毒性肝炎病例感染的病原体,试验对两个养殖场送检的2份病料采用RT-PCR、病毒分离和序列分析等方法对病毒进行鉴定;病料处理后接种10日龄SPF鸭胚、9日龄SPF鸡胚尿囊腔进行病毒分离,并扩增病毒全基因组及VP1基因,利用DNAStar 7.0和MEGA Ⅹ对分离株与GenBank中公布的相关病毒株序列进行同源性比对、氨基酸序列比对并构建系统进化树。结果表明:从病料中成功分离并鉴定2株3型鸭甲肝病毒(DHAV-3),分别命名为GD18株和GD21株;2份病料攻毒均未引起鸡胚的死亡与明显病变,但引起了鸭胚的发育不良,且GD21株可引起鸭胚死亡,死亡胚体出现全身性出血、充血等特征性病变;分离的2株DHAV-3均属于GⅠ基因型,且与之前在广东省流行的FS株、GD株和C-GY株等毒株亲缘关系较远,而与广东省外流行毒株SD株、JS株、CH株和EY株等亲缘关系较近。与参考株氨基酸序列比对,GD18株在第49位和第67位出现2个新的突变位点,而GD21株没有出现新的突变位点。说明在广东省内鸭群中可能存在由于引种或贸易过程而引入的DHAV-3新毒株的流行。
In order to understand the pathogens of infection in suspected cases of duck viral hepatitis occurring in two farms in Guangdong Province,this experiment was conducted to identify the virus using RT-PCR,virus isolation and sequence analysis of two diseased materials sent from two farms.The virus was isolated from 10-day-old SPF duck embryos and 9-day-old SPF chicken embryos by inoculation of the allantoic cavity after treatment of the disease material,and the whole genome of the virus and VP1 gene were amplified,and the homology alignment,amino acid sequence alignment and phylogenetic tree construction were carried out by using DNAStar 7.0 and MEGA Ⅹ according to the sequences of the isolates and related virus strains published in GenBank.The results showed that two strains of Duck hepatitis A virus type 3(DHAV-3)were successfully isolated and identified from the disease material,and were named GD18 and GD21.Both materials after challenge did not cause mortality and obvious lesions in chicken embryos,but caused stunting in duck embryos;and the isolate GD21 caused mortality in duck embryos,with characteristic lesions such as generalized hemorrhage and congestion in dead embryos.The two DHAV-3 isolates belonged to the GⅠgenotype and were distantly related to the previously prevalent strains FS,GD and C-GY in Guangdong Province,while they were more closely related to the out-of-province strains SD,JS,CH and EY.Aligned with the amino acid sequences of the reference strains,the isolate GD18 showed two new mutant sites at positions 49 and 67,while the isolate GD21 showed no new mutant site.The results suggested that there might be an epidemic of new strains of DHAV-3 introduced due to the introduction or trade process in duck populations in Guangdong Province.
作者
张玉倩
姚鑫炎
杨惠湖
庄园
袁生
黄淑坚
张雪莲
ZHANG Yuqian;YAO Xinyan;YANG Huihu;ZHUANG Yuan;YUAN Sheng;HUANG Shujian;ZHANG Xuelian(Foshan University,Foshan 528231,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2022年第24期69-75,137,共8页
Heilongjiang Animal Science And veterinary Medicine
基金
广东省自然科学基金面上项目(2022A1515012194)
广东省基础与应用基础研究区域联合基金-青年基金项目(2019A1515110157)
广东省教育厅普通高校重点领域专项(乡村振兴)(2020ZDZX1004)。
关键词
鸭甲肝病毒
分离
鉴定
进化分析
VP1基因
全基因组
Duck hepatitis A virus
isolation
identification
evolutionary analysis
VP1 gene
whole genome
