中国畜牧兽医2011，Vol.38Issue(1)：88-91,4.
山羊痘病毒和羊口疮病毒双重PCR检测方法的建立与初步应用
Establishment and Rudiment Application of Duplex PCR for Goatpox Virus and Orf Virus
摘要
Abstract
Two pairs of primers were designed according to the sequences of ITR gene of Goatpox virus (GPV) and H2L gene orf virus(ORFV) available in GenBank, respectively. Nucleic acids of GPV and ORFV were mixed as template, a duplex PCR assay was developed based on optimization of reaction conditions. Using the duplex PCR assay,the specific fragments 289 bp and 507 bp were amplified respectively from DNA samples of GPV and ORFV, but not amplified from foot-mouth disease virus,fowl pox virus,bluetongue virus and skin from normal healthy goat. The sensitivity of the duplex PCR reached 4 pg for GPV DNA and 0.4 pg for ORFV DNA,respectively. 12 clinical material were amplified by the duplex PCR assay,detection rate of GPV was 25％(3/12),detection rate of ORFV was 75％(9/12). The results showed that this duplex PCR assay was rapid, sensitive and specific for simultaneous diagnosis or differentiation of GPV and/or from ORFV infections.关键词
山羊痘病毒/羊口疮病毒/双重PCR/检测分类
畜牧业引用本文复制引用
向智龙,程振涛,卓建华,周碧君,欧德渊,鲜思美,刘芳,冉光鑫..山羊痘病毒和羊口疮病毒双重PCR检测方法的建立与初步应用[J].中国畜牧兽医,2011,38(1):88-91,4.基金项目
贵州省科学技术基金项目"贵州省羊口疮的病原学与检测技术研究"(黔科合J字[2010]2260) （黔科合J字[2010]2260）
贵州大学引进人才科研项目"贵州省羊口疮的诊断与防控技术研究"(贵大人基合字(2009)024号) （贵大人基合字(2009）
国家自然科学基金项目"基于山羊痘病毒P32基因的山羊粘膜免疫研究"(30940054) （30940054）
贵州大学2010年"SRT"项目"羊传染性脓疱病毒贵州株PCR鉴定及分子特征分析" （）
贵州大学2010年研究生创新基金创新项目"贵州省羊口疮的分子诊断技术及B2L表达研究". （）
