中国畜牧兽医2017，Vol.44Issue(7)：2112-2118,7.DOI:10.16431/j.cnki.1671-7236.2017.07.029
猪瘟病毒实时荧光定量PCR检测方法的建立与应用
Establishment and Application of Real-time Quantitative PCR Assay for Detection of Classical Swine Fever Virus
摘要
Abstract
A Real-time quantitative PCR assay for detection of classical swine fever virus (CSFV) was developed using the specific probe and primers designed basing on the E2 gene of CSFV.The Real-time quantitative PCR assay was established using the total RNA of CSFV as template.The specificity, sensitivity and repeatability of the assay were tested, and samples taken from clinic suspicious CSFV infected pigs had been testified by the established assay.The results indicated that the Real-time quantitative PCR assay was successfully established, and showed a good linear relationship at a template range of 101 to 106 copies/μL with a coefficient correlation of 0.999;The specificity of the assay revealed that amplifications were showed on CSFV samples, but other pathogens had no amplifications;The sensitivity of the assay was 10 copies/μL nucleic acid and 1 TCID50 /mL virus;Meanwhile,19 positive samples were detected, which were consistent with results of CSFV detected by Nested RT-PCR, cloning and sequencing.The eatablished Real-time quantitative PCR assay was specific, sensitive rapid and suitable for early detection and epidemiological study of CSFV.关键词
猪瘟病毒/实时荧光定量PCR/特异性/重复性/敏感性/应用Key words
classical swine fever virus/Real-time quantitative PCR/specificity/repeatability/sensitivity/application分类
农业科技引用本文复制引用
王淑娟,刘梅芬,闫若潜,班付国,赵雪丽,马震原,王华俊,王翠,赵明军..猪瘟病毒实时荧光定量PCR检测方法的建立与应用[J].中国畜牧兽医,2017,44(7):2112-2118,7.基金项目
河南省科技创新人才计划项目"猪重要疫病快速检测集成技术研究"(174200510003) （174200510003）
