重组酶介导扩增方法快速检测寨卡病毒  

Rapid detection of zika virus with recombinase aid amplification

作　　者：袁帅[1] 郑夔[1] 洪烨[1] 孙芳芳 黄吉城[1] 李小波[1] 戴俊[1] 师永霞[1] 康晓平[2] 李裕昌[2] YUAN Shuai;ZHENG Kui;Hong Ye;SUN Fang-fang;HUANG Ji-cheng;LI Xiao-bo;DAI Jun;SHI Yong-xia;KANG Xiao-ping;LI Yu-chang(Guangdong Inspection and Quarantine Technical Center,Guangzhou,Guangdong 510700,China)

机构地区：[1]广东检验检疫技术中心,广东广州510700 [2]军事医学科学院微生物流行病研究所

出　　处：《中国国境卫生检疫杂志》2018年第3期159-161,共3页Chinese Journal of Frontier Health and Quarantine

基　　金：国家"十三五"生物安全专项(20161202700);广东出入境检验检疫局科技计划项目(2016GDK46)

摘　　要：目的 建立寨卡病毒一步法重组酶聚合酶等温扩增检测方法(reverse transcription recombinase aid amplification,RT-RAA),为国境口岸提供现场快速检测方法。方法 根据寨卡病毒基因组保守片段NS2a设计引物和探针,建立寨卡病毒RT-RAA检测方法。用合成的含有寨卡病毒基因序列的质粒测试方法的重复性和灵敏度,用登革病毒、基孔肯雅病毒、黄热病毒、乙脑病毒核酸测试方法的特异性。结果 寨卡病毒一步法RT-RAA检测时间短(<20min),对寨卡病毒阳性质粒检测下限为10~3拷贝/μl,重复性好,与登革病毒、基孔肯雅病毒、黄热病毒、乙脑病毒无交叉反应。结论 建立的寨卡病毒一步法RT-RAA方法具有快速、特异以及灵敏的特点,适用于口岸现场快速检测。Objective To establish a one-step reverse transcriptase recombinase aid amplification（RT-RAA） assay for zika virus（ZIKV） detection for pathogen screening and rapid detection. Methods Accordding to the conserved genome sequences NS2a of ZIKV,the primers and probes were designed, and the RT-RAA method for ZIKV detection was established. The sensitivity,repeatability of the method were determined by synthetic plasmid containing ZIKV sequence, while the specificity was conducted by dengue virus,chikungunya virus,yellow fever virus,Japanese encephalitis virus detection. Results ZIKV was amplified with one-step RT-RAA performed with a short time（20 min）and low detecion limit 10~3 copies/μl of synthetic plasmid. It had no cross-reaction with dengue virus,chikungunya virus,yellow fever virus,Japanese encephalitis virus. Conclusion This RT-RAA assay was rapid,sensitive and specific. It can be used for the rapid detection of ZIKV at ports.

关 键 词：重组酶介导扩增 寨卡病毒 口岸 

分 类 号：R373.3[医药卫生—病原生物学]