浙江农业学报

• 动物科学 • 上一篇    下一篇

一株江苏新型鸭肝炎病毒的鉴定

  

  1. (1 江苏省农业科学院, 国家兽用生物制品工程技术研究中心, 江苏 南京210014;2 江苏省出入境检验检疫局, 江苏 南京210001)
  • 出版日期:2014-11-25 发布日期:2014-12-02

Isolation and identification of a new serotype of duck hepatitis virus in Jiangsu

  1. (1 National Research Center of Veterinary Biological Engineering and Technology,Nanjing 210014, China;2 Lab of Animal Quarantine, APHIC, Jiangsu Entry\|Exit Inspection and Quarantine Bureau,Nanjing 210001, China)
  • Online:2014-11-25 Published:2014-12-02

摘要: 对江苏地区分离到的1株疑似鸭肝炎病毒毒株(JS株)进行传代繁殖、雏鸭回归试验和理化性能测定,结果表明,该毒株能在鸭胚上进行传代繁殖,E1~E6代鸭胚毒的ELD50为10438·02 mL-1 ~10617·02 mL-1;攻毒的雏鸭可产生与I型鸭肝炎病毒相同的症状和病变;3,7,14日龄易感雏鸭的LD50分别为1055·02 mL-1,10517·02 mL-1,10383·02 mL-1;病毒对氯仿、乙醚和胰酶不敏感,对酸(pH 30)稳定,对热不稳定。进一步进行交叉中和试验、交叉被动保护试验和VP1基因序列分析,结果显示,该毒株的抗原性与经典1型DHV差异显著;VP1序列与多株1型DHV的同源性在693%~933%;从进化关系来看,与韩国株的亲缘性最近。JS株属于我国流行的新型鸭肝炎病毒(命名为N\|DHV\|JS株)。

关键词: 新型鸭肝炎病毒, 鸭甲型肝炎病毒, 分离鉴定, 反转录PCR

Abstract: A suspected duck hepatitis virus(JS strain) was isolated from clinical dead ducklings with duck hepatitis symptoms in Jiangsu. The isolates were passed in duck embryos and the titers were 10-438 ELD50·02 mL-1 to 10-617 ELD50·02 mL-1. Animal regression test showed attack ducklings could reproduce original disease of clinical symptoms and pathological changes. The virus was not sensitive to chloroform, diethyl ether, trypsin; The virus was stable to acid\|resistance (pH 30) but not stable to heat\|resistance. The JS strain of antigen was not related to the classic strain of DHV\|1 as showed by in vitro cross\|neutralization assay and in vivo cross\|protection test. The strain was amplified by RT\|PCR using specific primers designed according to the published sequence of DHV. The sequence of JS strain consisted of about 720 bp shared the high identities of 693%-933% with VP1 gene of DHV\|1. Furthermore, based on the deduced gene sequences, the phylogenetic analysis indicated that the isolates were highly homologous with the DHV strains isolated in Korea. Therefore, JS strain represents a new serotype of DHV occurred in China (designated N\|DHV\|JS strain).

Key words: new serotype of DHV, duck hepatitis A virus, isolation and identification, RT\, PCR