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Development of a 23S rRNA-based PCR assay for the identification of Pasteurella multocida
1Queensland Department of Primary Industries, Agency for Food and Fibre Sciences, Animal Research Institute, Moorooka, Queensland 4105, Australia. miflinj@dpi.qld.gov.au
Aims:
The aim of this work was to develop a rapid diagnostic test for Pasteurella multocida.
Methods And Results:
A polymerase chain reaction (PCR) assay using primers derived from the 23S rRNA gene sequence of Past. multocida was developed. The PCR assay correctly identified all 144 isolates of Past. multocida tested, 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 bacteria, formerly known as Bisgaard Taxon 13, are the closest phylogenetic relatives of Past. multocida based on 16S ribosomal rRNA. All phylogenetically unrelated avian and porcine organisms tested were negative.
Conclusion:
This PCR enables rapid identification of Past. multocida colonies from avian or porcine origin.
Significance And Impact Of The Study:
Veterinary diagnostic laboratories can use this PCR to rapidly and accurately diagnose fowl cholera and porcine pasteurellosis.
Insights
A new polymerase chain reaction (PCR) assay rapidly identifies Pasteurella multocida, a key cause of fowl cholera and porcine pasteurellosis. This diagnostic tool enhances accuracy for veterinary laboratories.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
Background:
- Pasteurella multocida is a significant pathogen causing diseases in avian and porcine species.
- Accurate and rapid identification of P. multocida is crucial for effective disease management.
Purpose of the Study:
- To develop a rapid and specific diagnostic test for Pasteurella multocida.
Main Methods:
- Development of a polymerase chain reaction (PCR) assay using primers targeting the 23S rRNA gene of P. multocida.
Main Results:
- The PCR assay demonstrated high specificity, correctly identifying 144 P. multocida isolates.
- Cross-reactivity was minimal, with only closely related Pasteurellaceae strains (Past. canis biovar 2 and Past. avium biovar 2) showing positive results.
- All phylogenetically unrelated avian and porcine organisms tested were negative.
Conclusions:
- The developed PCR assay provides a rapid and accurate method for identifying P. multocida colonies.
- This test is valuable for veterinary diagnostic laboratories for diagnosing fowl cholera and porcine pasteurellosis.