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作者

Wang, M. Ren, Q. Zhang, Z. Zhang, L. Carr, M.J. Li, J. Zhou, H. Shi, W.

作者单位

National Virus Reference Laboratory, School of Medicine, University College Dublin, Belfield, Dublin 4, Ireland Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, 001-0020, Japan a Key Laboratory of Etiology and Epidemiology of Emerging Infectious Diseases in Universities of Shandong, Taishan Medical College, Taian, Shandong 271000, China Department of Children's Medical Laboratory Diagnosis Center, Qilu Chidren's Hospital of Shandong University, Jinan, Shandong 250022, China

刊名

Journal of Virological Methods

年份

2018

卷号

Vol.258

页码

7-12

ISSN

0166-0934

关键词

CVA10 CVA16 CVA6 Enterovirus EVA71 HFMD Multiplex PCR

摘要

Hand, foot and mouth disease is a pediatric disease associated with infection by enterovirus genotypes. The major HFMD EV pathogens are enterovirus A71 and coxsackievirus A16 ; however, recently, coxsackievirus A6 and coxsackievirus A10 have also emerged. EV genotypes cannot be distinguished on clinical grounds and a new methodology for the rapid detection of the four major HFMD EV genotypes is urgently required. In the present study, a multiplex real-time PCR assay was established for the ...更多

Hand, foot and mouth disease is a pediatric disease associated with infection by enterovirus genotypes. The major HFMD EV pathogens are enterovirus A71 and coxsackievirus A16 ; however, recently, coxsackievirus A6 and coxsackievirus A10 have also emerged. EV genotypes cannot be distinguished on clinical grounds and a new methodology for the rapid detection of the four major HFMD EV genotypes is urgently required. In the present study, a multiplex real-time PCR assay was established for the simultaneous detection of CVA6, CVA10, CVA16 and EVA71. The specificity and sensitivity of the assay was determined on a validation panel of clinical samples, comprising cerebrospinal fluid , blood , feces and throat swabs . The results showed that the multiplex real-time PCR exhibited high specificity, no cross-reactivity with other EV genotypes, lower limits of detection for CVA6, CVA10, CVA16 and EVA71 were 4 × 10 3 , 4 × 10 2 , 5 × 10 2 , and 3 × 10 3 copies/μL, respectively and had comparable sensitivity to singleplex assays testing clinical samples. The multiplex real-time PCR methodology established in this study can be employed for the rapid detection of the four most prevalent HFMD-associated EVs, for epidemiologic surveillance of circulating EV genotypes and for assessing treatment responses and vaccine studies.收起

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