A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Abstract

In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.

A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine

“…By simultaneously amplifying more than one target in the same reaction, multiplex PCR increasingly is used as a rapid and convenient assay in the clinical Y. Zhao, et al Journal of Virological Methods 269 (2019) 38-42 diagnosis (Henegariu et al, 1997;Hu et al, 2015;Liu et al, 2011;Wu et al, 2013). In this study, we developed a multiplex RT-PCR assay for simultaneously detecting four viral pathogens in clinical specimens.…”

“…Our results indicated that the mRT-PCR developed is a rapid and validated method for simultaneous detection of clinical specimens. Previous reports showed that co-infection of PRRSV/CSFV and procine teschovirus (PTV) (Liu et al, 2011) or PRRSV/CSFV and pseudorabies virus (PRV) (Hu et al, 2015) was detected by mRT-PCR in clinical specimens from commercial herds in China. In our experiments, five clinical specimens exhibited co-infection of PRRSV and CSFV and one specimen co-infected with PEDV and PRRSV based on mRT-PCR detection.…”

“…Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis coronavirus (TGEV), are currently the most important pathogens that have caused tremendous economic losses for swine industries worldwide. Under typical conditions of intensive swine production, it is ordinary for swine to be simultaneously infected with two or more viral pathogens (Hu et al, 2015;Liu et al, 2011). Multiple infections lead to atypical clinical signs.…”

“…Several investigators have previously developed PCR and real-time PCR based multiplex assays for the differential detection of related multiple pathogens or pathogen causing diseases with similar symptoms (Aguero et al, 2004;Hu et al, 2015;Liu et al, 2011;Settypalli et al, 2016;Thonur et al, 2012;Wu et al, 2013;Yue et al, 2009). To our knowledge, mRT-PCR for simultanous detection of CSFV, PRRSV, PEDV and TGEV infection of swine has not been reported.…”

Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples

“…; Hu et al . ) an enzyme‐linked immunosorbent assay (ELISA) (Depner et al . ) and the immunoperoxidase test (IPT) (Pan et al .…”