摘要
本研究根据非洲马瘟病毒(AHSV)S7基因保守序列设计特异性引物和探针,建立了检测AHSV的实时荧光反转录重组酶聚合酶扩增(RT-RPA)方法。该方法特异性强,仅AHSV检测为阳性,蓝舌病病毒(BTV)、马流感病毒(EIV)、马动脉炎病毒(EAV)等病原检测均为阴性;灵敏性高,最低检出限为2.36×10^(1)copies/μL;反应快速,检测时间为20 min,同时具有良好的重复性。利用该方法和实时荧光RT-PCR平行检测60份临床样品,符合率为100%。结果表明,本研究建立的AHSV RT-RPA检测方法特异、敏感、快速且结果可靠,适用于AHSV的高通量、快速检测。
A real-time fluorescent reverse transcription-recombinase polymerase amplification(RT-RPA)assay was developed to detect African horse sickness virus(AHSV)using specific primers and probes that were designed based on conservative sequence of S7 gene of the virus.With strong specificity,the assay could detect positive AHSV,and negative bluetongue virus(BTV),equine influenza virus(EIV),equine arteritis virus(EAV)and other pathogens;with high sensitivity,the minimum detection limit was 2.36×10^(1)copies/μL;with rapid reaction,the detection time was 20 min,and good repeatability was also observed.60 clinical samples were parallelly detected by the assay and real-time fluorescent RT-PCR,with 100%coincidence rate.It was concluded that the above established method was specific,sensitive,rapid and reliable,and appropriate for high-throughput and rapid detection of AHSV.
作者
史卫军
林彦星
黄超华
曹琛福
曾少灵
吴江
刘建利
陈兵
阮周曦
花群义
Shi Weijun;Lin Yanxing;Huang Chaohua;Cao Chenfu;Zeng Shaoling;Wu Jiang;Liu Jianli;Chen Bing;Ruan Zhouxi;Hua Qunyi(Animal and Plant Inspection and Quarantine Center,Shenzhen Customs,Shenzhen 518045,China)
出处
《中国动物检疫》
CAS
2022年第7期119-123,共5页
China Animal Health Inspection
基金
深圳海关科研项目(2020SZHG001)。
