中国兽医科学2017，Vol.47Issue(12)：1492-1496,5.DOI:10.16656/j.issn.1673-4696.2017.12.004
猪繁殖与呼吸综合征病毒GDr180疫苗株与其他毒株PCR-HRM鉴别检测方法的建立
Establishment of PCR-HRM detection method for differentiating porcine reproductive and respiratory syndrome virus vaccine strain GDr180 from other strains
摘要
Abstract
To establish a rapid method of PCR-HRMto differentiate porcine reproductive and respiratory syndrome virus(PRRSV)vaccine strain GDr180 and other strains,according to the alignment of a large number of PRRSV gene sequences published on NCBI,the differential basic sites of sequences between GDr180 strain and other strains were screened.PCR products were amplified by asymmetric PCR with the addition of the LC Green dye and the blocked probe of 3'end.The melting curves of PCR products were obtained and analyzed by HRMfunction mode.The results showed that the probes Tm value of the melting curves of GDr180 was 72.64±0.39℃ and the other strains was 65.63±0.69 ℃,so the difference between both ones was significant.The primers were unable to amplify five non-PRRSV virus such as porcine epidemic diarrhea virus (PEDV),porcine pseudorabies virus (PRY),classical swine fever virus (CSFV),porcine circovirus(PCV)and porcine rotavirus(PoRV),while the detectable sensitivity was 1 000 copies/μL.The study shows that combining PCR with HRM is a high throughput economical,simple and reliable method for differentiating GDrl80 from other strains of PRRSV.关键词
猪繁殖与呼吸综合征病毒/不对称PCR/HRMKey words
porcine reproductive and respiratory syndrome virus/asymmetrical PCR/HRM分类
农业科技引用本文复制引用
饶丹,郭鹏举,丛锋,黄碧洪,练月晓,肖丽,李秀珍,黄韧,张钰,陈梅丽..猪繁殖与呼吸综合征病毒GDr180疫苗株与其他毒株PCR-HRM鉴别检测方法的建立[J].中国兽医科学,2017,47(12):1492-1496,5.基金项目
广东省科技计划项目(2016A040403060,2015B070701003) （2016A040403060,2015B070701003）
