Establishment of a multiplex RT-PCR assay for identification of atmospheric virus contamination in pig farms

Spread of pathogens in pig farms not only causes transfection of diseases to other pigs or even farmers working in the farms, but also induces pollution to the living atmospheric environment of the residents around the farm. Therefore, it is necessary to establish a rapid and simple monitoring method. In this study, full genome sequences of common viruses were analyzed in pig farms, in combination with the design of primers, optimization of the reaction parameters, so as to establish a multiplex RT-PCR assay for the identification of classical swine fever virus (CSFV), Japanese encephalitis virus (JEV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus Type 2 (PCV-2), porcine pseudorabies virus (PRV) and porcine parvovirus virus (PPV), which are common in pig farms. This method has a minimal detectable concentration of 10−3 ng/μL, which is highly specific. Furthermore, multiplex RT-PCR was applied to examine air samples from 4 pig farms located in different cities of China. The results were in line with those obtained by single PCR. Therefore, this study can be expected to provide essential technique support for the early warning mechanism as well as disease prevention and control system against the major viruses.