Development of a reverse transcription quantitative polymerase chain reaction-based assay for broad coverage detection of African and Asian Zika virus lineages

查看全文 作　　者：Yang [1,2]Yang;Gary [1,2]Wong;Baoguo [3]Ye;Shihua [2]Li;Shanqin [1]Li;Haixia [1]Zheng;Qiang [1,2]Wang;Mifang [4]Liang;George F [1,2,5,6]Gao;Lei [1]Liu;Yingxia [1]Liu;Yuhai [1,2]Bi

机构地区：[1]Shenzhen Key Laboratory of Pathogen and Immunity, State Key Discipline of Infectious Disease,Shenzhen Third People’s Hospital;[2]CAS Key Laboratory of Pathogenic Microbiology and Immunology, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, Institute of Microbiology, Center for Influenza Research and Early-warning (CASCIRE), Chinese Academy of Sciences;[3]China-Japan Union Hospital of Jilin University;[4]Key Laboratory for Medical Virology, NHFPC, National Institute for Viral Disease Control and Prevention, China CDC;[5]Office of Director-General, Chinese Center for Disease Control and Prevention;[6]Savaid Medical School, University of Chinese Academy of Sciences

出　　处：《Virologica Sinica》索引2017年第32卷第3期,共8页

基　　金：supported by the National Science and Technology Major Project(2016ZX10004222);the Sanming Project of Medicine in Shenzhen(ZDSYS201504301534057);the Key specialized fund for infectious diseases in Shenzhen City(No.201161);the intramural special grant for influenza virus research from the Chinese Academy of Sciences(KJZD-EW-L09 and KJZD-EWL15);the Shenzhen Science and Technology Research and Development Project(JCYJ20160427151920801 and JCYJ20160427153238750);G.F.G.is a leading principal investigator of the National Natural Science Foundation of China(NSFC)Innovative Research Group(81621091);supported by the Youth Innovation Promotion Association of Chinese Academy of Sciences(CAS)(2017122);G.W.is the recipient of a Banting Postdoctoral Fellowship from the Canadian Institutes of Health Research(CIHR);the President’s International Fellowship Initiative from the CAS 摘　　要：The Zika virus(ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome(GBS) and microcephaly in humans. The ZIKV is genetically diverse and can be separated into Asian and African lineages. A rapid, sensitive, and specific assay is needed for the detection of ZIKV across various pandemic regions. So far, the available primers and probes do not cover the genetic diversity and geographic distribution of all ZIKV strains. To this end, we have developed a one-step quantitative reverse transcription polymerase chain reaction(qRT-PCR) assay based on conserved sequences in the ZIKV envelope(E) gene. The detection limit of the assay was determined to be five RNA transcript copies and 2.94 × 10^(–3) 50% tissue culture infectious doses(TCID50) of live ZIKV per reaction. The assay was highly specific and able to detect five different ZIKV strains covering the Asian and African lineages without nonspecific amplification, when tested against other flaviviruses. The assay was also successful in testing for ZIKV in clinical samples. Our assay represents an improvement over the current methods available for the detection ZIKV and would be valuable as a diagnostic tool in various pandemic regions. 关键词：逆转录聚合酶链反应病毒检测覆盖血统亚洲非洲反转录聚合酶链反应特异性扩增

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Development of a reverse transcription quantitative polymerase chain reaction-based assay for broad coverage detection of African and Asian Zika virus lineages

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