动物医学进展Issue(12)：11-16,6.
非洲马瘟病毒实时荧光定量 RT-PCR 检测方法的建立
Development of Real-time Fluorescent Quantitative RT-PCR for Detection of AHSV Based on SYBR GreenⅠ
摘要
Abstract
African horse sickness virus (AHSV)is a double-stranded ribonucleic acid (dsRNA)virus which can in-fect all the equid animals.Based on the VP7-ORF sequences from GenBank and results of sequencing for nine sero-types of AHSV previously completed by our own team,a pair of specific primers located on the segment S7 were designed for SYBR GreenⅠquantitative detection,in order to develop one kind of quick method for detecting the AHSV.Results showed that the primer pair was identified to be specific for each of the 9 serotypes of AHSV only and specific fluorescent signal could be detected,but not for the related viruses including BTV,EHDV and normal lymphoid tissue of horse without specific fluorescent signal.The built positive recombinant plasmid (AHSV9-pMD-18T-VP7)in our lab was used for a positive quantitative template to establish a standard curve.Finally,the method of pre-denaturalized template based on “three-step”reaction was established in the study.It was demon-strated that the SYBR GreenⅠfluorescent quantitative RT-PCR for detection of AHSV could be used for the rapid diagnosis and quantitative detection with detection limit of 102 copies/μL cDNA.关键词
非洲马瘟病毒/血清型/SYBR Green I/实时荧光定量 RT-PCRKey words
AHSV/serotype/SYBR GreenⅠ/real-time fluorescent quantitative RT-PCR分类
农业科技引用本文复制引用
赵文华,杨仕标,李富祥..非洲马瘟病毒实时荧光定量 RT-PCR 检测方法的建立[J].动物医学进展,2013,(12):11-16,6.基金项目
公益性行业（农业）科研专项 （）
