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An improved multiplex PCR for Actinobacillus pleuropneumoniae, Glaesserella australis and Pasteurella multocida

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Sun, X., Blackall, P. J., Daniel, P., Chandra, K., Jenkin, S. and Turni, C. (2021) An improved multiplex PCR for Actinobacillus pleuropneumoniae, Glaesserella australis and Pasteurella multocida. Journal of Microbiological Methods, 191 . p. 106360. ISSN 0167-7012

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Article Link: https://doi.org/10.1016/j.mimet.2021.106360

Publisher URL: https://www.sciencedirect.com/science/article/pii/S0167701221002281


Glaesserella australis, a newly described bacterial species, has been isolated from pig lungs that displayed lesions very similar to those caused by Actinobacillus pleuropneumoniae, prompting the need for a validated diagnostic tool. In this work, we have altered a multiplex PCR used for the identification of cultures of G. australis, A. pleuropneumoniae and Pasteurella multocida to be more sensitive and then evaluated the use of the altered diagnostic tool on cultures and directly on tissues. The altered multiplex PCR was validated using 47 related species, both type/reference strains and field isolates. The sensitivity was assessed by serial dilutions and used a mixture of target bacteria in different concentrations. Further, 166 lung samples from 54 farms from four Australian States were used to validate the ability of the multiplex PCR to detect bacteria in lung swabs. The multiplex PCR was specific for the three target species. The assay could detect a minimum of 40 colony forming units (CFU) of G. australis, 786 CFU of A. pleuropneumoniae and 238 CFU of P. multocida. The multiplex PCR yielded more positives than coventional bacteriological examination. From a total of 166 lung samples, 51.9%, 51.9% and 5.6% of farms were PCR positive for P. multocida, A. pleuropneumoniae and G. australis, respectively. The results suggested that the new multiplex PCR was specific, sensitive and out performed traditional culture. The prevalence of G. australis was not very high, but it was the dominant pathogen in infected pigs.

Item Type:Article
Business groups:Animal Science
Keywords:Multiplex PCR Prevalence
Subjects:Science > Microbiology
Science > Microbiology > Microorganisms in the animal body
Science > Microbiology > Bacteria
Animal culture
Animal culture > Swine
Live Archive:08 Nov 2021 02:23
Last Modified:08 Nov 2021 02:23

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