RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A–D from Clinical Specimens

作     者：CHEN Wei WENG Yu Wei HE Wen Xiang ZHU Ying YU Ting Ting XIE Jian Feng ZHENG Kui Cheng YAN Yan Sheng ZHANG Yong Jun ZHANG Wen Chang CHEN Wei;WENG Yu Wei;HE Wen Xiang;ZHU Ying;YU Ting Ting;XIE Jian Feng;ZHENG Kui Cheng;YAN Yan Sheng;ZHANG Yong Jun;ZHANG Wen Chang

作者机构：The School of Public HealthFujian Medical UniversityFuzhou 350122FujianChina Fujian Center for Disease Control and PreventionFujian Provincial Key Laboratory of Zoonosis ResearchFuzhou 350001FujianChina 

基　　金：National Science and Technology Major Projects[No.2017ZX10104001 and No.2017ZX10103008] Fujian Provincial Natural Science Foundation[No.2016J01350] 

出 版 物：《Biomedical and Environmental Sciences》 (生物医学与环境科学（英文版）)

年 卷 期：2020年第33卷第11期

页      码：829-838页

摘      要：Objective To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses(HEVs)from clinical samples and to contribute to etiological surveillance of HEV-related *** A panel of RT-nPCR assays,consisting of published combined primer pairs for VP1 genes of HEV A–C and in-house designed primers for HEV-D,was established in this *** sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID50 perμL and copies perμL,and the newly established methods were tested in clinical specimens collected in recent *** The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID50 perμL and 10 virus copies perμL,and for the complete VP1 gene was 1 CCID50 perμL and 100 virus copies perμL,using serially-diluted virus stocks of five *** a proof-of-concept,25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance *** This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A–D,providing rapid,sensitive,and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.

主 题 词：Clinical specimens Human enterovirus A–D VP1 gene Polymerase chain reaction 

学科分类：1004[医学-公共卫生预防医学类] 100401[100401] 10[医学] 

核心收录：

D　O　I：10.3967/bes2020.113

馆 藏 号：203995649...