逆转录重组酶介导扩增技术快速检测基孔肯雅热病毒  

RAPID DETECTION OF CHIKUNGUNYA FEVER VIRUS WITH REVERSE TRANSCRIPTION RECOMBINASE AIDED AMPLIFICATION

作　　者：罗鹏[1] 姜梓凡 郑伟[1] 吴忠华[1] 吕沁风[1] 郭利川 应清界 王刚[1] LUO Peng;JIANG Zhi-Fan;ZHENG Wei;WU Zhong-Hua;LYU Qin-Feng;GUO Li-Chuan;YING Qing-Jie;WANG Gang(Zhejiang International Travel Healthcare Center, Hangzhou 310012, China;Zhejiang High School, Hangzhou 310000, China;Jiangsu Qitian Gene Bio-Sci＆Tech Co. , Ltd, Wuxi 214135, China)

机构地区：[1]浙江国际旅行卫生保健中心,杭州310012 [2]浙江省杭州高级中学,杭州310000 [3]江苏齐天基因生物科技有限公司,江苏无锡214135

出　　处：《寄生虫与医学昆虫学报》2017年第4期212-216,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家重点研发计划项目(2016YFF0203200);生物安全关键技术研发重点专项(2016YFC1202700);国家质检总局应急技术保障专项(2016IK278)

摘　　要：本研究通过使用逆转录酶,将基孔肯雅热病毒RNA首先逆转录为cDNA。选取基孔肯雅热病毒保守序列设计引物及探针,建立基孔肯雅热病毒重组酶介导一步法等温核酸扩增RT-RAA(Reverse transcription recombinase aided amplification,RT-RAA)快速检测法,并分析其灵敏度及特异性。结果显示,该方法整个过程在39℃进行,检测时间短(<20 min),检测下限可达100 copy,并与黄热病毒、日本乙型脑炎病毒、西尼罗病毒、登革病毒I型等蚊媒病毒不存在交叉反应。本文建立的基孔肯雅热病毒RT-RAA检测法,灵敏度高、特异性强、操作简便,适应于口岸基层基孔肯雅热病毒的快速检测。The Chikungunya fever virus （CHIKV） RNA was first transcribed into the cDNA by a reverse-transcriptase reaction, and the resulting product was then served as template for RT-RAA amplification. Universal primers and probe were designed according to the conserved genome sequence of the CHIKV to evaluate the specificity and sensitivity of the methods and develop a rapid one-step reverse transcription reeombinase aided amplification （RT-RAA） assay for （CHIKV） detection. The reaction was performed at a constant temperature of 39℃ with a short detection time （ 〈 20 minutes）. Our results indicated that the amplification is very sensitive which can detect 100 copies. The approach is also shown very specific as it had no cross reaction with other related viruses, such as Dengue virus, West Nile virus, Japanese encephalitis virus or Yellow fever virus. This RT-RAA assay was rapid, specific and sensitive, and might be utilized in rapid surveillance for CHIKV on ports.

关 键 词：重组酶介导扩增 基孔肯雅热病毒 分子检测 

分 类 号：R440[医药卫生—诊断学] R512.8[医药卫生—临床医学]