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通过猪繁殖与呼吸综合征病毒ORF6和ORF7基因的限制性片段长度多态性对病毒分离株进行鉴别

Differentiation between porcine reproductive and respiratory syndrome virus isolates by restriction fragment length polymorphism of their ORFs 6 and 7 genes.

作者信息

Gagnon C A, Dea S

机构信息

Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Laval.

出版信息

Can J Vet Res. 1998 Apr;62(2):110-6.

Abstract

Three distinct antigenic profiles were identified by comparing the reactivities of 15 Canadian field isolates, the attenuated U.S. vaccine (Ingelvac MLV) strain and 2 European reference strains (Lelystad and Weybridge) of the porcine reproductive and respiratory syndrome virus (PRRSV) by indirect immunofluorescence with a set of 4 monoclonal antibodies to the nucleocapsid (N) protein and 2 other to the matrix (M) protein. In the present study, 9 Canadian isolates for which the sequences were determined appeared closely related to 2 U.S. reference strains (ATCC VR-2332 and ATCC VR-2385) with amino acid identities varying between 90 to 98% for the M and N proteins; substitutions in the nucleotide sequences were distributed randomly throughout the ORFs 6 and 7 genes, and most were 3rd base silent mutations. In comparison, more than 30% divergence was demonstrated with the Lelystad virus. Furthermore, differentiation between North American and European isolates, and between field isolates and the MLV strain could be achieved by cutting PCR-amplified products encompassing both ORFs 6 and 7 genes with 4 restriction endonucleases. When taken individually, BsaJI and AluI were the more appropriate restriction enzymes for distinguishing the vaccine strain from field isolates. The results obtained suggest that the restriction fragment length polymorphism of the genomic region covering the ORFs 6 and 7 genes may be a valuable tool to differentiate among PRRSV isolates.

摘要

通过间接免疫荧光法,用一组4种针对核衣壳(N)蛋白的单克隆抗体和另外2种针对基质(M)蛋白的单克隆抗体,比较15株加拿大田间分离株、美国减毒疫苗(Ingelvac MLV)株以及猪繁殖与呼吸综合征病毒(PRRSV)的2株欧洲参考株(莱利斯塔德株和韦布里奇株)的反应性,确定了三种不同的抗原谱。在本研究中,已测定序列的9株加拿大分离株似乎与2株美国参考株(ATCC VR - 2332和ATCC VR - 2385)密切相关,M蛋白和N蛋白的氨基酸同一性在90%至98%之间;核苷酸序列中的替换随机分布在开放阅读框6和7基因中,且大多数是第三位碱基沉默突变。相比之下,与莱利斯塔德病毒的差异超过30%。此外,通过用4种限制性内切酶切割包含开放阅读框6和7基因的PCR扩增产物,可以实现北美分离株与欧洲分离株之间以及田间分离株与MLV株之间的区分。单独来看,BsaJI和AluI是区分疫苗株与田间分离株的更合适的限制性内切酶。所获得的结果表明,覆盖开放阅读框6和7基因的基因组区域的限制性片段长度多态性可能是区分PRRSV分离株的一种有价值的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d010/1189456/6d922d70cee6/cjvetres00014-0034-a.jpg

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