Establishment of a PCR assay for detecting Bordetella bronchiseptica , Pasteurella multocida and Haemophilus parasuis

A PCR detecting Bb, Pm and HPS was developed based on published single PCRs for these pathogens. The PCR reagents, annealing temperature and the PCR procedure were optimized. A 237 bp fragment of Bb, a 457 bp of Pm and a 821 bp of HPS can be simultaneously amplified in one condition .The PCR can detect minimum 1.34×103cfu1/mL cells of Bb,2.6×102cfu/mL cells of Pm, 1.6×102cfu/mL cells of HPS. The sensitivity of the PCR is quite comparable with their reported uniplex PCRs.The assay is a potential tool for differentiation of the 3 pathogens and diagnosis of mixed infection by these bacteria.