摘要
目的建立检测多种蚊媒病毒的液相芯片方法。方法从NCBI数据库分别下载乙脑病毒(JEV)、登革病毒(DENV)、基孔肯雅病毒(CHIKV)、寨卡病毒(ZIKV)、黄热病毒(YFV)、裂谷热病毒(RVFV)基因组序列,比对分析后,在各病毒保守区域分别设计扩增引物和检测探针,以标准品质粒为模板,建立多重PCR扩增体系;将核酸探针偶联至荧光编码微球,建立液相芯片检测方法,并对方法进行评价。结果基于多重PCR和液相芯片技术的常见蚊媒病毒的检测方法能同时对其中任意1种、2种、3种、4种、5种和6种病毒进行检测,具有良好的灵敏性,JEV、DENV、CHIKV、RVFV的灵敏度为1×104拷贝/反应;ZIKV、YFV的灵敏度为1×105拷贝/反应。结论建立的方法能同时快速检测6种常见蚊媒病毒,为蚊媒传染病的预警和控制提供参考。
Objective To establish a liquichip method which can be used for detecting common mosquito-borne viruses. Methods The genomic sequence of Japanese encephalitis virus(JEV),dengue virus(DENV),chikungunya virus(CHIKV),Zika virus(ZIKV),yellow fever virus(YFV),and Rift Valley fever virus(RVFV) were all downloaded from the NCBI database. The amplification primers and detection probes in the conserved regions of each virus were designed after sequence alignment analysis,and a multiplex PCR amplification system using the standard plasmids of each virus as the template was established. The nucleic acid probes were coupled to fluorescent microspheres,and a liquichip detection system was established. The mosquito-borne viruses liquichip detection method was established and evaluated. Results The established method based on multiplex PCR and liquichip technology could detect any1,2,3,4,5 or 6 kind of JEV,DENV,CHIKV,ZIKV,YFV and RVFV simultaneously,showed high sensitivity and accuracy. The sensitivity for detecting JEV,DENV,CHIKV and RVFV was 1×104 copies/reaction,1×105 copies/reaction for ZIKV and YFV detection. Conclusion This method could be used for detecting 6 kinds of common mosquitoborne virus simultaneously and quickly,providing reference information for the early warning and control of mosquito-borne infectious diseases.
作者
周有良
赵亚峰
孙玉莲
葛斌鹏
杨国平
胡春凌
陈峰
杨庆贵
ZHOU You-liang;ZHAO Ya-feng;SUN Yu-lian;GE Bin-peng;YANG Guo-ping;HU Chun-ling;CHEN Feng;YANG Qing-gui(Suzhou Customs House,Suzhou,Jiangsu 215011,China;不详)
出处
《中国国境卫生检疫杂志》
CAS
2021年第4期233-236,240,共5页
Chinese Journal of Frontier Health and Quarantine
基金
南京海关科研项目(2020KJ05)。
