DEVELOPMENT OF PCR ASSAYS FOR SPECIES-SPECIFIC AND TYPE-SPECIFIC IDENTIFICATION OF PASTEURELLA-MULTOCIDA ISOLATES

Genomic subtractive hybridization of closely related Pasteurella multo cida isolates has generated clones useful in distinguishing hemorrhagi c septicemia-causing type B strains from other P. multocida serotypes. Oligonucleotide primers designed during the sequencing of these clone s have proved valuable in the development of PCR assays for rapid spec ies-and type-specific detection of P. multocida and of type B:2 in par ticular. This study demonstrated that the primer pair designed from th e sequence of the clone 6b (KTT72 and KTSP61) specifically amplified a DNA fragment from types B:2, B:5, and B:2,5 P. multocida and that the primers KMT1T7 and KMT1SP6 produced an amplification product unique t o all P. multocida isolates analyzed. It was also shown that PCR ampli fication performed directly on bacterial colonies or cultures represen ts an extremely rapid, sensitive method of P. multocida identification .