Abstract:According to the gene sequences in GenBank of classical swine fever virus (CSFV), swine influenza virus (SIV), porcine circovirus type 2 (PCV2), porcine pseudorabies virus (PRV), porcine reproductive and respiratory comprehensive virus (PRRSV) gene sequences, designed five pairs of primers were used to amplify CSFV E2, SIV, PCV2 ORF2, PRV gB, PRRSV N gene. Reaction conditions were optimized by the establishment of a detection CSFV, SIV, PCV2, PRV and PRRSV multiple PCR (mPCR) diagnosis. Sensitivity and specificity results showed that the mPCR minimum of 5 of the virus nucleic acid detection volume were 106 gene copy number (about 10 pg). Against E. coli, staphylococcus, streptococcus, porcine parvovirus, infectious laryngotracheitis virus, Newcastle disease virus, porcine transmissible gastroenteritis virus, multiple PCR amplification results were negative. Clinical samples of multiple PCR results showed that the multiplex PCR method capable of SIV, CSFV, PCV2, PRV, PRRSV infection of single or mixed clinical samples for rapid diagnosis. Key words:CSFV;SIV;PCV2;PRV;PRRSV;multiple PCR