Differentiation between porcine reproductive and respiratory syndrome virus isolates by restriction fragment length polymorphism of their ORFs 6 and 7 genes.
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.
Gagnon
C A
CA
Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Laval.
Dea
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D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis; D002170:Canada; D019084:Fluorescent Antibody Technique, Indirect; D005814:Genes, Viral; D019590:Nucleocapsid Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D016366:Open Reading Frames; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D019318:Porcine Reproductive and Respiratory Syndrome / Q000276:immunology / Q000517:prevention & control / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D013552:Swine; D014481:United States; D014613:Vaccines, Attenuated; D014763:Viral Matrix Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D014613:Vaccines, Attenuated; D014763:Viral Matrix Proteins; D014765:Viral Vaccines