The Development of a Multiplex Reverse Transcription Polymerase Chain Reaction for Detection of Porcine Reproductive and Respiratory

Abstract

A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). A set of three pairs of primer were designed based on the sequence of high conservative region of PRRSV. The diagnostic accuracy of the multiplex RT-PCR assay was evaluated using 25 field clinical samples, 10 reference strains and 6 true-negative samples. Simultaneously, the specificity and sensitivity of this method were evaluated. The result indicated that this assay could reliably differentiate between PRRSV and other swine viral disease, such as classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV). Meanwhile, this assay was shown to be 10-fold more sensitive than the conventional single RT-PCR (conventional sRT-PCR) method. The results indicated that the multiplex RT-PCR developed in this study has high sensitivity, strong specificity and easy operation,it is not only suitable for diagnosis of samples of insignifiance content,but also it has important applied value in getting rid of animals with recessive virus and veterinary quarantine .The method may provide a new avenue to the rapid detection of this important pathogen in one reaction.