5种猪病多重PCR检测方法的建立

Establishment of the Multiplex PCR for Detection of 5 Kinds of Swine Disease

To establish a method for simultaneous detection of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV), a multiplex PCR was developed with a set of specific primers designed based on the conserved sequences of CSFV, PRRSV, PRV, PCV2 and PPV. Under the optimized conditions of multiplex PCR，five special fragments of 167 (CSFV)，433 (PRRSV)，305 (PRV), 559 (PCV2) and 882 bp (PPV) were amplified with a detection limit of 220, 1.6, 72, 400 and 370 pg, respectively. But the multiplex PCR amplification results of swine influenza virus （SIV）, Japanese encephalitis virus （JEV）, Streptococcus suis （SS） and porcine epidemic diarrhea virus （PEDV） were negative．The results showed that the multiplex PCR method was capable of CSFV, PRV, PRRSV， PCV2, PPV infection of single or mixed clinical samples for rapid diagnosis.