Detection and differentiation of North American and European genotypes of porcine reproductive and respiratory syndrome virus in formalin-fixed, paraffin-embedded tissues by multiplex reverse transcription-nested polymerase chain reaction

Abstract

North American and European genotypes of porcine reproductive and respiratory syndrome virus (PRRSV) were distinguished in formalin-fixed, paraffin-embedded tissues from PRRSV-infected pigs by multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR). A method based on xylene deparaffinization followed by proteinase K digestion yielded RNA of quality for reliable and consistent RT-nPCR analyses. The PRRSV nucleic acid was detected in lung, mediastinal lymph node, tonsil, and liver samples from pigs inoculated with North American strain virus, European strain virus, or both North American and European strains. All 30 archival formalin-fixed, paraffin-embedded lung tissues from pigs naturally infected with PRRSV were positive for PRRSV by RT-nPCR amplication and gave a pattern that corresponded to the North American genotype. Multiplex RT-nPCR could be applied successfully to formalin-fixed, paraffin-embedded tissues for the detection and differentiation between North American and European genotypes of PRRSV.