摘要
为实现对韦塞尔斯布朗病病毒(Wesselsbron virus,WSLV)的快速检测,根据WSLV NS5基因保守序列,设计特异性扩增引物和探针,建立WSLV的TaqMan荧光定量RT-PCR和RT-PCR检测方法,并验证方法的敏感性、特异性及临床可行性。结果显示:所建立的两种方法的核酸检测下限分别为10、100 copies/μL,对日本脑炎病毒、西尼罗病毒和坦布苏病毒核酸均无交叉反应;分别利用建立的两种检测方法对100份入境马匹样品进行检测,未检出WSLV阳性,与南非参考实验室提供的双重荧光RT-PCR检测方法结果一致。结果说明,所建的两种方法敏感性高、特异性强,可用于临床样本检测,为韦塞尔斯布朗病的出入境检验检疫、流行病学调查及防控提供了技术支撑。
In order to rapidly detect Wesselsbron virus(WSLV),specific primers and probe were designed based on conserved sequence of NS5 gene of WSLV to establish TaqMan-based fluorescent RT-PCR assay and RT-PCR assay,and their sensitivity,specificity and clinical feasibility were verified.The results showed that the established methods could detect nucleic acids at the lowest limits of 10 and 100 copies/μL,respectively,and failed to crossly react with the nucleic acids of Japanese encephalitis virus,West Nile virus and Tambusu virus;100 samples collected from entry horses were tested by the two methods,respectively,and no WSLV-positive samples were detected,which was consistent with the results by dual fluorescence RT-PCR assay provided by the reference laboratory in South Africa.In conclusion,the established methods were with high sensitivity,strong specificity and good stability,and could be used to detect clinical samples,supporting entry-exit inspection and quarantine,epidemiological investigation,prevention and control of Wesselsbron disease.
作者
王奕婷
张风荣
冯之航
郁蕾
薛俊欣
陈翔
王艳
赵光伟
Wang Yiting;Zhang Fengrong;Feng Zhihang;Yu Lei;Xue Junxin;Chen Xiang;Wang Yan;Zhao Guangwei(Technical Center for Animal&Plant&Food Inspection and Quarantine of Shanghai Customs,Shanghai 200135,China;Shandong Vocational Animal Science and Veterinary College,Weifang 261061,Shandong,China;Shanghai Customs,Shanghai 200135,China;College of Veterinary Medicine,Southwest University,Chongqing 402460,China)
出处
《中国动物检疫》
CAS
2024年第10期96-101,共6页
China Animal Health Inspection
基金
上海市科委技术标准专项(21DZ2207700)。