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

Harper, Marina, John, Marietta, Turni, Conny, Edmunds, Mark, St. Michael, Frank, Adler, Ben, Blackall, P. J., Cox, Andrew D. and Boyce, John D. (2015) Development of a rapid multiplex PCR assay to genotype pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus. Journal of Clinical Microbiology, 53 2: 477-485. doi:10.1128/JCM.02824-14

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ350968_OA.pdf Full text (open access) application/pdf 644.03KB 0

Author Harper, Marina
John, Marietta
Turni, Conny
Edmunds, Mark
St. Michael, Frank
Adler, Ben
Blackall, P. J.
Cox, Andrew D.
Boyce, John D.
Title Development of a rapid multiplex PCR assay to genotype pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
ISSN 0095-1137
1098-660X
Publication date 2015-02-01
Year available 2015
Sub-type Article (original research)
DOI 10.1128/JCM.02824-14
Open Access Status File (Publisher version)
Volume 53
Issue 2
Start page 477
End page 485
Total pages 9
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
Subject 2726 Microbiology (medical)
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.
Formatted 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.
Keyword Microbiology
Microbiology
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Alliance for Agriculture and Food Innovation
Official 2016 Collection
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 14 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 15 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 10 Feb 2015, 10:42:25 EST by System User on behalf of Qld Alliance for Agriculture and Food Innovation