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中国一株变异猪流行性腹泻病毒的分离与遗传分析

Isolation and genetic analysis of a variant porcine epidemic diarrhea virus in China.

作者信息

Li Ren-feng, Tian Xiang-qin, Liu Yong, Xu Jing, Liu Dong-yang

出版信息

Pol J Vet Sci. 2016;19(1):65-73. doi: 10.1515/pjvs-2016-0009.

Abstract

Porcine epidemic diarrhea virus (PEDV) is having a severe effect on the pig breeding industry in central China. The mucosa and the content of the small intestine from newborn pre-weaned piglets with diarrhea were tested for the presence of PEDV by molecular and morphologic methods, and found to be positive. Negative-staining electron microscopy (EM) revealed the presence of coronavirus- like particles in the samples. The result of molecular detection by nested RT-PCR based on the amplification of the M gene was positive. Using a novel alternative method we successfully propagated the PEDV strain (CH/QX-2) in Vero cells, confirmed by ultrathin sections of the cells and Immunofluorescence assay (IFA). Phylogenetic analysis based on the partial S gene showed that the CH/QX-2 isolate was genetically closer to strains more commonly found in China, but differed genetically from two domestic strains (CH/S, 1986 and LZC, 2007), Korean strains (DR13, 2007), and the vaccine strain (CV777 vs) currently being used in China. CH/QX-2 formed a unique clade in the derived phylogenetic tree indicating that the CH/QX-2 strain currently circulating in central China is a new variant of PEDV. This study extends current knowledge on the diversity and epidemiology of PEDV.

摘要

猪流行性腹泻病毒(PEDV)正在对中国中部的养猪业产生严重影响。通过分子和形态学方法对腹泻的新生断奶前仔猪的小肠黏膜和内容物进行PEDV检测,结果呈阳性。负染电子显微镜(EM)显示样品中存在冠状病毒样颗粒。基于M基因扩增的巢式RT-PCR分子检测结果为阳性。我们使用一种新的替代方法成功地在Vero细胞中培养了PEDV毒株(CH/QX-2),通过细胞超薄切片和免疫荧光试验(IFA)得到证实。基于部分S基因的系统发育分析表明,CH/QX-2分离株在基因上与中国更常见的毒株更接近,但在基因上与两个国内毒株(CH/S,1986和LZ C,2007)、韩国毒株(DR13,2007)以及中国目前使用的疫苗毒株(CV777 vs)不同。CH/QX-2在推导的系统发育树中形成了一个独特的分支,表明目前在中国中部流行的CH/QX-2毒株是PEDV的一个新变种。本研究扩展了目前关于PEDV多样性和流行病学的知识。

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