中国兽医科学2017，Vol.47Issue(5)：569-576,8.DOI:10.16656/j.issn.1673-4696.2017.05.006
山羊痘病毒及口疮病毒ITR-059基因双重PCR检测方法的建立
Establishment of the duplex PCR assay based on ITR-059 gene for the detection of goatpox virus and orf virus
摘要
Abstract
To detect the goatpox virus(GTPV) and orf virus(ORFV) simultaneously and quickly,two pairs of primers for routine PCR assay,and two pairs of primers and relative probes for real-time PCR assay were designed based on the inverted terminal repeat(ITR) segment of GTPV and the 059 gene of ORFV,respectively.The probes were labeled with different fluorescent materials 5'JOE-Eclipse3'and 5'FAM-TAMRA3',respectively.The results showed that the developed GTPV ITR-L ORFV-059 duplex routine PCR assay was specific for GTPV and 0RFV with the detection sensitivity of 104copies/μL DNA.The established duplex ITR-059 probe real-time PCR assay was also proved specific for 0RFV and GTPV,and the fluorescent signal could be detected with a detection limit of 102.87copies/μL DNA and 102.52 copies/μL DNA for GTPV and ORFV,respectively.Finally,in the study the duplex routine PCR and the duplex real-time PCR assays for simultaneous detection of GTPV and ORFV were established preliminarily.关键词
山羊痘病毒/口疮病毒/二联/普通PCR/实时PCR/探针Key words
goatpox virus(GTPV)/orf virus(ORFV)/duplex/routine PCR/real-time PCR/probe分类
农业科技引用本文复制引用
赵文华,杨仕标,姚俊,李富祥..山羊痘病毒及口疮病毒ITR-059基因双重PCR检测方法的建立[J].中国兽医科学,2017,47(5):569-576,8.基金项目
云南省重点新产品开发计划项目(2014BB014) （2014BB014）
云南省现代农业奶牛产业技术体系建设项目[云财农(2009)171号] （2009）
