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Journal of Clinical Microbiology, April 1998, p. 1096-1100, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Development of PCR Assays for Species- and Type-Specific Identification of Pasteurella multocida Isolates

Kirsty M. Townsend,1,* Alan J. Frost,1 Chiang W. Lee,1 John M. Papadimitriou,2 and Hugh J. S. Dawkins3

University of Western Australia Department of Pathology2 and Urological Research Centre,3 Queen Elizabeth II Medical Centre, Nedlands 6009 Western Australia, and Division of Veterinary Pathobiology, University of Queensland, Brisbane 4072 Queensland,1 Australia

Received 4 August 1997/Returned for modification 19 November 1997/Accepted 14 January 1998

Genomic subtractive hybridization of closely related Pasteurella multocida isolates has generated clones useful in distinguishing hemorrhagic septicemia-causing type B strains from other P. multocida serotypes. Oligonucleotide primers designed during the sequencing of these clones have proved valuable in the development of PCR assays for rapid species- and type-specific detection of P. multocida and of type B:2 in particular. This study demonstrated that the primer pair designed from the sequence of the clone 6b (KTT72 and KTSP61) specifically amplified a DNA fragment from types B:2, B:5, and B:2,5 P. multocida and that the primers KMT1T7 and KMT1SP6 produced an amplification product unique to all P. multocida isolates analyzed. It was also shown that PCR amplification performed directly on bacterial colonies or cultures represents an extremely rapid, sensitive method of P. multocida identification.

* Corresponding author. Mailing address: Division of Veterinary Pathobiology, School of Veterinary Science, The University of Queensland, Brisbane Qld 4072, Australia. Phone: 61 7 3365 2667. Fax: 61 7 3365 1355. E-mail: kirsty.townsend{at}mailbox.uq.edu.au.

Journal of Clinical Microbiology, April 1998, p. 1096-1100, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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