First results of detection of PRRSV and CSFV RNA by SYBR Green I-based quantitative PCR.

Porcine reproductive and respiratory syndrome (PRRS) and classical swine fever (CSF) cause significant economic losses to the swine industry worldwide. As both diseases cause similar symptoms, rapid and reliable detection of these diseases is essential for disease surveillance. A quantitative SYBR Green I-based reverse transcription-polymerase chain reaction (RT-PCR) is described for simultaneous and differential diagnosis. The established RT-PCR for the quantitation of PRRSV and CSFV cDNA was found to provide a broad dynamic range, detecting from 10(3) to 10(11) and 10(2) to 10(11) copies of cDNA per reaction, respectively. Sensitivity and specificity of this method were compared with those of conventional RT-PCR and both were equal or superior to the reference method. Reproducibility was tested and the assay was proved very reliable. The assay is timesaving, easy to handle, and highly sensitive and specific. Therefore, it is a powerful tool for detecting PRRSV and CSFV simultaneously for routine outbreak investigation.