Porcine reproductive and respiratory syndrome virus(PRRSV)infects alveolar macrophages,seriously damaging the immune system,leading to co-infection with other viruses,among which swine influenza virus(SIV)is also one of the pathogens.In order to rapidly detect and distinguish PRRSV genotypes and SIV subtypes,four pairs of specific primers were designed according to the conserved sequences of HA genes of H1 and H3 subtype SIV and N and M genes of the American and European PRRSVs.The primers were optimized for their concentrations and annealing temperatures and the specificity and sensitivity of this multiplex PCR method were also demonstrated for rapidly detecting these four pathogens.The detection limit of H1-SIV,H3-SIV,NA-PRRSV and EU-PRRSV was as low as 9.86× 100 copies/μL.A total of 155 clinical samples were then tested using this multiplex PCR method and single PCR methods.The results were consistent among these PCR methods,indicating that this multiplex PCR method could be used for rapid detection and type identification of these four viruses.
关键词
猪流感病毒/猪繁殖与呼吸综合征病毒/多重PCR
Key words
Swine influenza virus/Porcine reproductive and respiratory syndrome virus/multiplex PCR
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