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用于检测南部非洲地区口蹄疫病毒的新型多重引物逆转录聚合酶链反应的开发

Development of a New RT-PCR with Multiple Primers for Detecting Southern African Territories Foot-and-mouth Disease Viruses.

作者信息

Liu Ya-Li, Ding Yao-Zhong, Dai Jun-Fei, Ma Bing, He Ji-Jun, Ma Wei-Min, Lv Jian-Liang, Ma Xiao-Yuan, Ou Yun-Wen, Wang Jun, Liu Yong-Sheng, Chang Hui-Yun, Wang Yong-Lu, Zhang Qiang, Liu Xiang-Tao, Zhang Yong-Guang, Zhang Jie

机构信息

State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China.

Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, Jiangsu, China.

出版信息

J Vet Res. 2018 Dec 31;62(4):431-437. doi: 10.2478/jvetres-2018-0064. eCollection 2018 Dec.

Abstract

INTRODUCTION

The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information.

MATERIAL AND METHODS

A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay.

RESULTS

The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results.

CONCLUSIONS

A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.

摘要

引言

南非领土血清型口蹄疫病毒(FMDV),包括SAT1、SAT2和SAT3,具有极高的遗传变异性且新变种不断出现,因此有必要根据最新的基因组信息开发一种新的通用逆转录聚合酶链反应(RT-PCR)用于监测病毒。

材料与方法

基于SAT血清型病毒的1D2A2B基因,使用多重引物组建立了FMDV SAT-D8一步法RT-PCR。纳入FMDV A、O、C和亚洲1型血清型、其他水泡性疾病病毒、灭活的SAT病毒,以及从中国大陆采集的125份牛、羊、山羊和猪的组织样本用于评估该检测方法。

结果

新的RT-PCR被证明具有特异性,与欧亚FMDV、猪水泡病病毒(SVDV)、塞内卡山谷病毒(SVV)或牛羊猪的其他常见病毒病原体无交叉反应。检测灭活的SAT病毒时,清晰出现了约257 bp大小的扩增产物。然而,从中国大陆未发生SAT疫情的FMDV易感动物采集的所有125份样本结果均为阴性。

结论

FMDV SAT-D8一步法RT-PCR是一种有前景的用于口蹄疫病毒SAT血清型初步筛查的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/50dc/6364153/a9ed23e925de/jvetres-62-431-g001.jpg

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