Differential diagnosis of DNA viruses related to reproductive disorder on sows by multiplex-PCR technique

Abstract

The newly emerged diseases caused by ASFV and PCV3 and their confirmed prevelance in Vietnam whereas most of available common commercial methods such as ELISA or realtime PCR designed for detecting single pathogen per reaction, highlighted a necessity for another diagnostic method to simultaneously detect and differentiate DNA viruses that are related to reproductive failures in sow herds including PCV2, PCV3, PPV, ASFV. In this communication, a diagnostic multiplex-PCR (mPCR) was established with pathogen-specific primers selected from previous studies and another set of primers designed for COX1 gene serving as an internal amplification control (IAC). The predicted products of PCV2, PCV3, PPV, ASFV and IAC were 702 bp, 223 bp, 380 bp, 278 bp and 463 bp, respectively. After optimization, the mPCR functioned specifically at 62°C. Results revealed the consistent detection limit at 100 copies/gene/reaction. In application, 185 serum samples from sows were used to examine the presence of the related pathogens. mPCR results showed that the mono-infection rate of PCV2, PCV3, PPV, and ASFV was 0% (0/185), 40% (74/185), 28.1% (52/185), and 48.1% (89/185), respectively. Regarding coinfection rate, the data indicated that coinfections of 2, 3 and 4 pathogens were 20%, 8.1% and 0% accordingly. In conclusion, the mPCR assay was successfully established and ready to serve for diagnosis of PCV2, PCV3, PPV and ASFV infection in reality with high specificity and sensitivity. It is a good contribution to a better understanding of the epidemiology of these diseases in swine.