畜牧兽医学报2012，Vol.43Issue(3)：416-423,8.
1株猪源牛病毒性腹泻病毒的分离与鉴定
Isolation and Identification of a Pig Bovine Viral Diarrhea Virus
摘要
Abstract
The aim of this study was to isolate virus from bovine viral diarrhea virus (BVDV) positive sample of the piglet and to study BVDV originating from pigs. BVDV positive samples from the piglet were inoculated in MDBK cells and a pig BVDV strain (SD0803) was obtained. In the study, cell culture, direct immunofluorescence assay, PCR amplification of 5'-UTR,electron microscopy and TCID50 were performed, moreover,bioinformatics software were used to analyze the characteristics of molecular evolution. The results showed that the virus strain hadn't caused cy-topathic effect (CPE) until the 13th passage on MDBK cell. SD0803 infected cells showed strong fluorescent signal. DNA fragment of 307 bp and 428 bp were amplified by PCR from 5'-UTR and Npro of SD0803. Under electron microscopy, virus particles were near round and enveloped with size of 50 nm in diameter. Virus titer of SD0803 was 10~65 TCID50 · 0. 2 mL~'. The homology comparison and phylogenetic analysis of 5'-UTR and Npro between SD0803 and representative BVDV isolates showed that SD0803 belongs to BVDV-1. However, SD0803 had an obvious dis-tance from the subtypes of BVDV-1 and formed a separate group according to the phylogenetic relationship and this showed that the isolate might be a new subtype of BVDV-1. So a pig originated BVDV SD0803 was successfully isolated and identified, and belongs to NCP BVDV-1 and maybe a probably new subtype of BVDV-1.关键词
牛病毒性腹泻病毒/进化分析/分离鉴定Key words
Bovine viral diarrhea virus/phylogenetic analysis/isolation and identification分类
农业科技引用本文复制引用
邓宇,文心田,郑浩,孙春清,张宏彪,蔺涛,张荣,龙进学,黄律,曹三杰,袁世山..1株猪源牛病毒性腹泻病毒的分离与鉴定[J].畜牧兽医学报,2012,43(3):416-423,8.基金项目
中央级公益性科研院所基本科研业务费专项资金项目(2010JB15) （2010JB15）
上海市技术标准专项项目(10DZ0503600) （10DZ0503600）
