摘要
为建立鉴别口蹄疫病毒O型、A型、AsiaⅠ型的RT-PCR快速检测方法,根据GenBank中已登录的口蹄疫病毒(FMDV)O型、A型、AsiaⅠ型高度保守的2 B基因序列设计了1条共用反转录引物,根据3个血清型的VP1特异性基因序列,设计了3对型特异性引物对;以FMDV样本总RNA反转录产物为模板,建立了鉴别FMDV O、A、AsiaⅠ型三重RT-PCR检测方法。该方法灵敏度高,最低检测限均为100拷贝/μL;重复性和稳定性好;特异性强,与其他7个对照病原核酸均呈现阴性反应;可检测出10份疑似临床样品中3个血清型的FMDV感染样品。表明本试验成功建立了鉴别FMDV O型、A型、AsiaⅠ型三重RT-PCR检测方法,可用于临床上FMDV O型、A型、AsiaⅠ型的分型鉴别检测。
In order to establish a rapid method for RT-PCR detection of foot-and-mouth disease virus types O,A and Asia I,a common reverse transcription primer was designed according to the highly conserved 2 B gene sequence of foot-and-mouth disease virus(FMDV)O,A,Asia I)registered in GenBank,and three pairs of type-specific primers were designed according to the VP 1 specific gene sequences of three serotypes.Using the total RNA reverse transcripts of FMDV samples as templates,a triple RT-PCR method was established to identify FMDV O,A and Asia I types.This method has high sensitivity,the lowest detection limit is 100 copies/μL,good repeatability and stability,strong specificity,negative reaction with other seven control pathogenic nucleic acids,and can detect FMDV infection samples of three serotypes in 10 suspected clinical samples.The results showed that this study successfully established a triple RT-PCR method for the identification of FMDV O,A and Asia I,which can be used for clinical typing and detection of FMDV O,A and Asia I types.
作者
马震原
闫若潜
赵雪丽
谢彩华
曹伟伟
王淑娟
王东方
王华俊
王翠
MA Zhen-yuan;YAN Ruo-qian;ZHAO Xue-li;XIE Cai-hua;CAO Wei-wei;WANG Shu-juan;WANG Dong-fang;WANG Hua-jun;WANG Cui(Henan Centre for Animal Diseases Control and Prevention,Zhengzhou 450008,China)
出处
《中国兽医杂志》
CAS
北大核心
2020年第8期32-35,共4页
Chinese Journal of Veterinary Medicine
基金
河南省科技攻关计划项目(142102310224)。
作者简介
马震原(1986-),女,兽医师,硕士,主要从事动物疫病防控技术研究,E-mail:mazhenyuan.1986@163.com;通讯作者:闫若潜,E-mail:yrq1688@126.com。
