Development of a 23S rRNA-based PCR assay for the identification of Pasteurella multocida

Aims: The aim of this work was to develop a rapid diagnostic test for Paste urella multocida. Methods and Results: A polymerase chain reaction (PCR) assay using primers derived from the 23S rRNA gene sequence of Past. multocida was developed. T he PCR assay correctly identified all 144 isolates of Past. multocida teste d, including type strains of the three subspecies as well as the reference strains for the Heddleston and Carter typing schemes. Of 20 closely related bacteria from the family Pasteurellaceae tested, only the type strains of Past. canis biovar 2 and Past. avium biovar 2 were positive. These two bact eria, formerly known as Bisgaard Taxon 13, are the closest phylogenetic rel atives of Past. multocida based on 16S ribosomal rRNA. All phylogenetically unrelated avian and porcine organisms tested were negative. Conclusions: This PCR enables rapid identification of Past. multocida colon ies from avian or porcine origin. Significance and Impact of the Study: Veterinary diagnostic laboratories ca n use this PCR to rapidly and accurately diagnose fowl cholera and porcine pasteurellosis.