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来自北美不同地理区域患断奶后多系统消耗综合征猪的2型猪圆环病毒(PCV - 2)的基因特征分析以及用于检测和区分PCV - 1与PCV - 2感染的差异聚合酶链反应-限制性片段长度多态性检测方法的建立

Genetic characterization of type 2 porcine circovirus (PCV-2) from pigs with postweaning multisystemic wasting syndrome in different geographic regions of North America and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infections with PCV-1 and PCV-2.

作者信息

Fenaux M, Halbur P G, Gill M, Toth T E, Meng X J

机构信息

Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342, USA.

出版信息

J Clin Microbiol. 2000 Jul;38(7):2494-503. doi: 10.1128/JCM.38.7.2494-2503.2000.

Abstract

Postweaning multisystemic wasting syndrome (PMWS) is an emerging disease in swine. Increasing evidence indicates that a variant strain of porcine circovirus (PCV), designated type 2 PCV (PCV-2), is responsible for PMWS. To determine the extent of genetic heterogeneity of PCV-2 isolates, the complete genomes of six PCV-2 isolates from different regions of North America were amplified by PCR and sequenced. Sequence and phylogenetic analyses confirmed that two distinct genotypes of PCV exist: nonpathogenic genotype PCV-1 and PMWS-associated genotype PCV-2. However, within the PCV-2 genotype, several minor branches that have been identified appear to be associated with geographic origins. The genomic sequences of two French PCV-2 isolates diverge the most from those of other PCV-2 isolates and form a distinct branch. Other minor but distinguishable branches have also been identified for a Taiwan PCV-2 isolate and two of the Canadian PCV-2 isolates. All the U.S. PCV-2 isolates are closely related, but the Canadian isolates vary, to some extent, in their genomic sequences. The data from this study indicate that although the genome of PCV-2 is generally stable among different isolates, PCV-2 isolates from different geographic regions vary in their genomic sequences. This variation may have important implications for PCV-2 diagnosis and research. On the basis of genetic analyses of available PCV strains, a universal PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed to detect and differentiate between infections with PCV-1 and PCV-2. This PCR-RFLP assay should be useful for studying the pathogenesis of PCV-2, for detecting PCV-2 infection in pigs from different geographic regions, and for screening donor pigs for use in xenotransplantation.

摘要

断奶后多系统消耗综合征(PMWS)是猪群中一种新出现的疾病。越来越多的证据表明,一种猪圆环病毒(PCV)变异株,即2型猪圆环病毒(PCV-2),是PMWS的致病原。为了确定PCV-2分离株的遗传异质性程度,通过聚合酶链反应(PCR)扩增并测序了来自北美不同地区的6株PCV-2分离株的全基因组。序列和系统发育分析证实存在两种不同的PCV基因型:非致病性基因型PCV-1和与PMWS相关的基因型PCV-2。然而,在PCV-2基因型内,已鉴定出的几个小分支似乎与地理来源有关。两株法国PCV-2分离株的基因组序列与其他PCV-2分离株的差异最大,并形成一个独特的分支。对于一株台湾PCV-2分离株和两株加拿大PCV-2分离株,也鉴定出了其他较小但可区分的分支。所有美国PCV-2分离株密切相关,但加拿大分离株的基因组序列在一定程度上有所不同。本研究的数据表明,尽管PCV-2的基因组在不同分离株中总体稳定,但来自不同地理区域的PCV-2分离株在基因组序列上存在差异。这种变异可能对PCV-2的诊断和研究具有重要意义。基于对现有PCV毒株的遗传分析,开发了一种通用的聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测方法,用于检测和区分PCV-1和PCV-2感染。这种PCR-RFLP检测方法对于研究PCV-2的发病机制、检测来自不同地理区域猪的PCV-2感染以及筛选用于异种移植的供体猪应该是有用的。

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