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Journal of Clinical Microbiology, March 2001, p. 924-929, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.924-929.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Organization of Pasteurella multocida cap Loci and Development of a Multiplex Capsular PCR Typing System†

Kirsty M. Townsend,1,* John D. Boyce,2 Jing Y. Chung,2 Alan J. Frost,1 and Ben Adler2

Veterinary Pathology and Anatomy, School of Veterinary Science and Animal Production, The University of Queensland, Brisbane, QLD 4072,1 and Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, VIC 3800,2 Australia

Received 11 September 2000/Returned for modification 27 November 2000/Accepted 21 November 2000

Current serotyping methods classify Pasteurella multocida into five capsular serogroups (serogroups A, B, D, E, and F) and 16 somatic serotypes (serotypes 1 to 16). In the present study, we have developed a multiplex PCR assay as a rapid alternative to the conventional capsular serotyping system. The serogroup-specific primers used in this assay were designed following identification, sequence determination, and analysis of the capsular biosynthetic loci of each capsular serogroup. The entire capsular biosynthetic loci of P. multocida A:1 (X-73) and B:2 (M1404) have been cloned and sequenced previously (J. Y. Chung, Y. M. Zhang, and B. Adler, FEMS Microbiol. Lett. 166:289-296, 1998; J. D. Boyce, J. Y. Chung, and B. Adler, Vet. Microbiol. 72:121-134, 2000). Nucleotide sequence analysis of the biosynthetic region (region 2) from each of the remaining three serogroups, serogroups D, E, and F, identified serogroup-specific regions and gave an indication of the capsular polysaccharide composition. The multiplex capsular PCR assay was highly specific, and its results, with the exception of those for some serogroup F strains, correlated well with conventional serotyping results. Sequence analysis of the strains that gave conflicting results confirmed the validity of the multiplex PCR and indicated that these strains were in fact capsular serogroup A. The multiplex PCR will clarify the distinction between closely related serogroups A and F and constitutes a rapid assay for the definitive classification of P. multocida capsular types.

* Corresponding author. Mailing address: Veterinary Pathology and Anatomy, School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia. Phone: 61 7 3365 3083. Fax: 61 7 3365 1355. E-mail: kirsty.townsend{at}mailbox.uq.edu.au.

dagger This paper is dedicated to the memory of our colleague, the late Rick Rimler, whose contribution to P. multocida research is gratefully acknowledged.

Journal of Clinical Microbiology, March 2001, p. 924-929, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.924-929.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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