Differentiation of North American and European porcine reproductive and respiratory syndrome virus genotypes by in situ hybridization.
Non-radioactive probes that can detect specifically North American and European isolates of porcine reproductive and respiratory syndrome virus (PRRSV) in formalin-fixed paraffin-embedded tissues by in situ hybridization were developed. These probes allow the differentiation between North American and European genotypes of the PRRS virus as well as the detection of both genotypes. Two amplified cDNA products generated by polymerase chain reaction (PCR), one from the cDNA of the Canadian PRRSV LHVA-93-3 isolate and the second one from the European Lelystad isolate, and labelled with digoxigenin were utilized as probes. The LHVA-93-3 derived probe was found to detect Canadian and USA PRRSV isolates in infected cells, while the Lelystad derived probe hybridized only with European isolates. The specificity of both probes was also demonstrated on formalin-fixed tissues collected from PRRSV infected pigs. Furthermore, by combining the LHVA-93-3 (North American) probe and the Lelystad (European) probe, successful detection of both PRRSV genotypes in fixed tissues could be achieved.
Larochelle
R
R
Laboratoire d'hygiène vétérinaire et alimentaire, Agriculture et Agroalimentaire Canada, Saint-Hyacinthe, Québec. larocheller@em.agr.ca
Magar
R
R
D000818:Animals; D002170:Canada; D015342:DNA Probes; D005060:Europe; D005557:Formaldehyde; D005838:Genotype; D017403:In Situ Hybridization / Q000379:methods*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000473:pathology / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D013552:Swine; D016707:Tissue Fixation; D014481:United States
D015342:DNA Probes; D005557:Formaldehyde