Application of multiplex PCR combined with invasive reaction and chromogenic reaction catalyzed by gold nanoparticles in detection of encephalitis and meningitis virus
10.3969/j.issn.1002-2694.2017.11.008
- VernacularTitle: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
- Author:
Huan FAN
1
;
hua Yu QI
;
Zheng ZHU
;
biao Lun CUI
;
yue Yi GE
;
cheng Kang ZHAO
;
Tao WU
;
yang Zhi SHI
Author Information
1. 卫生部肠道病原微生物重点实验室
- Keywords:
encephalitis and meningitis virus;
multiplex PCR;
invasive reaction;
chromogenic reaction catalyzed by gold nanoparticles
- From:
Chinese Journal of Zoonoses
2017;33(11):991-995,1001
- CountryChina
- Language:Chinese
-
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.
