PCR Detection and Identification of Avian Pasteurella Multocida in Clinical Samples Based on the KMT, Sequence

Abstract

Polymerase Chain Reaction (PCR) was used for the detection and identification of Pasteurella multocida (PM)in poultry clinical samples with the KMT,SP/KMT,T, primer pair. A total of 54 clinical samples (liver, spleen, cloacae swabs)were assayed and finger prints of 14 samples corresponded with the 460bp expected band size. Results were unambiguous as finger prints were single and distinct. The sensitivity test of the KMT,SP/ KMT,T, primer pair gave 10⁻⁵ dilution (0.03025µg) as the limit of detection using ethidium bromide staining of amplified DNA in agarose gel electrophoresis. The PM-PCR was specific and reproduceble. Southern hybridization results confirmed the PM-PCR results and have therefore been shown to be a sensitive and reproducible confirmatory assay. The KMT, sequence based primers (KMT,SP/KMT,T,) proved sensitive and reliable to be used for the detection and identification of PM in clinical samples.