Development of a rapid multiplex PCR assay to genotype pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus

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DOI	Resolve DOI: https://doi.org/10.1128/JCM.02824-14
Author	Search for: Harper, Marina; Search for: John, Marietta; Search for: Turnic, Conny; Search for: Edmundsa, Mark; Search for: St. Michael, Frank¹; Search for: Adler, Ben; Search for: Blackall, P. J.; Search for: Cox, Andrew D.¹; Search for: Boyce, John D.
Name affiliation	National Research Council of Canada. Human Health Therapeutics
Format	Text, Article
Journal title	Journal of Clinical Microbiology
ISSN	0095-1137 1098-660X
Volume	53
Issue	2
Pages	477–485
Abstract	Pasteurella multocida is a Gram-negative bacterial pathogen that is the causative agent of a wide range of diseases in many animal species, including humans. A widely used method for differentiation of P. multocida strains involves the Heddleston serotyping scheme. This scheme was developed in the early 1970s and classifies P. multocida strains into 16 somatic or lipopolysaccharide (LPS) serovars using an agar gel diffusion precipitin test. However, this gel diffusion assay is problematic, with difficulties reported in accuracy, reproducibility, and the sourcing of quality serovar-specific antisera. Using our knowledge of the genetics of LPS biosynthesis in P. multocida, we have developed a multiplex PCR (mPCR) that is able to differentiate strains based on the genetic organization of the LPS outer core biosynthesis loci. The accuracy of the LPS-mPCR was compared with classical Heddleston serotyping using LPS compositional data as the “gold standard.” The LPS-mPCR correctly typed 57 of 58 isolates; Heddleston serotyping was able to correctly and unambiguously type only 20 of the 58 isolates. We conclude that our LPS-mPCR is a highly accurate LPS genotyping method that should replace the Heddleston serotyping scheme for the classification of P. multocida strains.
Publication date	2014-11-26
Publisher	American Society for Microbiology
Language	English
Peer reviewed	Yes
NPARC number	23000827
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Record identifier	12e0e9ba-b0d6-4739-9c04-1d41780309a1
Record created	2016-10-17
Record modified	2020-06-02

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Date modified:: 2021-04-22