Detection of Pasteurella multocida in blood by Real-Time PCR

摘要

Haemorrhagic septicaemia (HS) is one of the most important bacterial diseases of cattle and buffalo caused by P. multocida serotype B:2 or E:2. The present study evaluated the efficacy of a real time PCR in the diagnosis of HS. Blood was collected at36 hr post-infection both from control and P. multocida infected animals in EDTA and DNA was extracted using pure link genomic DNA isolation kit. The extracted DNA was subjected to real-time PCR using P. midtocida specific PCR primers KMT1T7 and KMT1SP6.The assay was able to detect the P. multocidaDNA in case of infected animal, whereas no amplification was observed in case of control animal. The dissociation curve obtained in the assay confirmed the specificity of the amplified products indicating theprecision of the PCR.