中国人兽共患病学报2017，Vol.33Issue(11)：991-995,1001,6.DOI:10.3969/j.issn.1002-2694.2017.11.008
多重PCR结合核酸侵入反应及纳米金显色技术检测病毒性脑炎脑膜炎病原体
Application of multiplex PCR combined with invasive reaction and chromogenic reaction catalyzed by gold nanoparticles in detection of encephalitis and meningitis virus
摘要
Abstract
We developed a method for detecting encephalitis and meningitis virus by using multiplex PCR combined with invasive reaction and a chromogenic reaction catalyzed by gold nanoparticles.Primers were designed based on the conservative regions of encephalitis and meningitis virus (Eastern equine encephalitis virus,EEEV;Western equine encephalomyelitis virus,WEEV;West Nile virus,WNV;Nipah virus,NiPA;Japanese encephalitis virus,JEV).Multiplex PCR system,invasive reaction and a chromogenic reaction catalyzed by gold nanoparticles were established to detect different encephalitis and meningitis virus in one reaction.Tick-borne encephalitis virus (TBEV),St Louis encephalitis virus (StLEV),Chikungunya virus (CHIKV) and Dengue virus(DV) were used to test its specificity.Quantitative RNA transcribed in vitro and PCR fragments were used to assess its sensitivity.Clinical specimens collected from JEV patients were detected by this method.A method for detecting encephalitis and meningitis virus by using multiplex PCR,invasive reaction and a chromogenic reaction catalyzed by gold nanoparticles were successfully established.This method can detect targeted pathogens specifically,and it has no cross reaction with TBEV,StLEV,CHIKV and DV.The detecting limitation for different targets was 103 copies/μL.Clinical samples were positive for JEV nucleic acids for above assay.The method presented here has characteristic of high specificity,sensitivity and throughput.The results can be observed by visual inspection.This method has broad application prospects in pathogen detection.关键词
病毒性脑炎脑膜炎/多重PCR/核酸侵入反应/纳米金显色Key words
encephalitis and meningitis virus/multiplex PCR/invasive reaction/chromogenic reaction catalyzed by gold nanoparticles分类
医药卫生引用本文复制引用
樊欢,戚宇华,朱政,崔仑标,葛以跃,赵康程,吴涛,史智扬..多重PCR结合核酸侵入反应及纳米金显色技术检测病毒性脑炎脑膜炎病原体[J].中国人兽共患病学报,2017,33(11):991-995,1001,6.基金项目
艾滋病和病毒性肝炎等重大传染病防治科技重大专项(No.2013ZX10004103)资助 Supported by Grand Science and Technology Special Program "AIDS and Viral Hepatitis and Other Major Infectious Diseases Prevention and Control" (No.2013ZX10004103) （No.2013ZX10004103）
