摘要
本研究建立了一种简单、快速、准确的寨卡病毒可视化逆转录环介导等温扩增(RT-LAMP)检测方法。在线设计3套LAMP引物,利用实时浊度仪筛选最佳引物,加入羟基萘酚蓝,评估可视化RT-LAMP检测方法的灵敏度和特异性。建立的可视化RT-LAMP方法可检测的最低浓度为101copies/μL,高于普通PCR和荧光定量PCR 1~2个数量级;同时,该方法不与其他虫媒病毒产生交叉反应。该方法可为虫媒监测和临床诊断提供实验依据。
The study established a simple,rapid and accurate reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay for zika virus （ZIKV）.Three sets of LAMP primers were designed with online tools.The visual RT-LAMP assay was established with the optimized primers selected by real-time turbidimeter and Hydroxy naphthol blue （HNB）,which possessed high sensitivity and specificity.The detection limit of this method was 10 copies/reaction,which was 10-100 fold higher than the conventional PCR and quantitative real-time PCR.The method was high specificity without any cross-reaction with other arbovirus.This method provided an experimental tool for the arbovirus monitoring and clinical diagnosis.
出处
《寄生虫与医学昆虫学报》
CAS
北大核心
2017年第2期103-108,共6页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家重大传染病专项(2013ZX1004103-004)
国家自然科学基金重点项目(U1602223)
江苏省社会发展项目(BE2017620)
关键词
寨卡病毒
逆转录环介导等温扩增
可视化检测
羟基萘酚蓝
Zika virus
Reverse transcription loop mediated isothermal amplification
Visual detection
Hydroxy naphthol blue
