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Development and application of Taq Man-MGB fluorescence quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus
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2008
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Diagnostic VirologyMolecular Diagnostic TechniquesQuantitative Taq ManViral DiagnosticsEmerging Infectious DiseasesMedicineVirus LoadPathogenesisImmunologyPathologyVirologyPorcine DiseaseSwine VirusRespiratory Syndrome VirusVirus TransmissionMolecular DiagnosticsFifth Passage PrrsvAnimal Virus
A quantitative Taq Man reverse transcription-polymerase chain reaction(RT-PCR) for detecting PRRSV was deve-loped using specific primers and fluorogenetic probe based on the ORF7 gene of porcine reproductive and respiratory syndrome virus(PRRSV).The established method was highly specific,sensitive and reproducible.It had a linear dynamic range of the 101 copies/μL to 107 copies/μL and could detect the least 10 copies of target DNA and 1 TCID50 virus.The assay was succesffuly used to test on serum and tissue samples collected from experimentally infected pigs at various times following infection.Pigs infected with the fifth passage PRRSV exhibitedd a rapid increase in virus load in serum on day 3 and reached a peak of 1010 copies/mL on day 7,while pigs infected with the sixty-fifth passage virus had a lower virus load which peaked at 106 copies/mL on day 21.The virus concentrations in the lung were 109 copies/g,which was much higher than that in other tissues.