›› 2008, Vol. 1 ›› Issue (2): 64-66.

• 生物技术 • 上一篇    下一篇

猪瘟病毒一步法RTPCR检测方法的建立

吴鑫,王军 ,张以芳   

  1. 云南农业大学动物科技学院,昆明 650201
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-02-20 发布日期:2008-02-20

Development of One Step RT-PCR Assay to Detect CSFV

WU Xin,WANG Jun,ZHANG Yifang
  

  1. College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-02-20 Published:2008-02-20

摘要: 本研究根据NCBI公布的猪瘟病毒石门毒株的E2基因序列,设计、合成了1对检测猪瘟病毒E2基因的RT-PCR引物。自疑似猪瘟病料及接猪瘟F114毒的PK-15细胞中提取总RNA,经一步法RT-PCR扩增E2基因。建立了猪瘟病毒E2基因RT-PCR检测方法,RT-PCR从26份疑似猪瘟病料中检出阳性病料24份,阳性检出率为92.3%;结果表明,建立的 RT-PCR方法具有良好的特异性和敏感性,可用于CSFV的分离鉴定、临床病料的检测和分子流行病学调查。

关键词: 猪瘟病毒; 一步法RTPCR; 检测

Abstract: Based on E2 gene sequences of CSFV Shimen published on NCBI, a pair of primers were designed and synthesized for detecting E2 gene. E2 gene was amplified by one step RT-PCR from total RNA, which had been extracted from tissues of pigs doubting infected by classical swine fever virus and PK-15 infected by CSFV F114. We compared IF method with one step RT-PCR which had been established. 24 samples of 26 samples doubting infected by CSFV were positive deteced by one step RT-PCR, the positive rate of one step RT-PCR was 92.3%. Results showed that the RT-PCR is specific and sensitive, and can be used as a method to identify CSFV isolates,detect samples of clinical cases and investigate molecular epidemiology of CSFV.

Key words: CSFV; one step RT-PCR; detection

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