中国畜牧兽医2016，Vol.43Issue(9)：2285-2290,6.DOI:10.16431/j.cnki.1671-7236.2016.09.010
猪瘟病毒巢式 RT-PCR 检测方法的建立与应用
Establishment and Application of Nested RT-PCR Assay for Detection of Classical Swine Fever Virus
摘要
Abstract
A nested RT-PCR for detection of classical swine fever virus (CSFV)was developed u-sing the total RNA of CSFV as a reverse transcription template,and two pairs of specific primers were designed based on the conserved region of CSFV E 2 gene.The specificity,sensitivity and repetition of CSFV nested RT-PCR were tested,and 35 samples taken from clinical suspicious CS-FV infected pigs were tested by the established nested RT-PCR assay.The results indicated that the nested RT-PCR assay was successfully established.The sensitivity and specificity of the nes-ted RT-PCR assay revealed that the minimum detectable virus content limit was 1 TCID50 ,and no products were amplified from the nucleic acid of ST normal cell or other 8 pathogenic viral or bac-terial microorganisms.The repetition test indicated the nested RT-PCR assay showed a good re-peatability.Meanwhile,1 9 positive samples were detected in 35 suspected samples,which was con-sistent with the results tested by sequencing.The study suggested that the established nested RT-PCR assay was specific,sensitive and suitable for early detection of CSFV.关键词
猪瘟病毒/巢式 RT-PCR/检测/建立/应用Key words
classical swine fever virus/nested RT-PCR/detection/establishment/application分类
农业科技引用本文复制引用
刘梅芬,王淑娟,闫若潜,王东方,曹伟伟,赵雪丽,李勤楠,李宁..猪瘟病毒巢式 RT-PCR 检测方法的建立与应用[J].中国畜牧兽医,2016,43(9):2285-2290,6.基金项目
河南省科技攻关项目“病死动物无害化处理技术及管理机制研究”(142102310224) （142102310224）
