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United States Department of Agriculture

Agricultural Research Service

Title: A Two-Color Hybridization Assay for Identification of Toxigenic Pasteurella Multocida and Bordetella Bronchiseptica - Hap Mtg.

Author
item Register, Karen

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: September 9, 1996
Publication Date: N/A

Technical Abstract: Toxigenic (tox+) P. multocida and B. bronchiseptica are etiologic agents of swine atrophic rhinitis. This report describes rapid, simultaneous identification of these organisms from mixed cultures. Plasmid pPMT, containing a P. multocida toxin gene fragment, was labeled using fluorescein-dUTP. This probe did not bind to nontoxigenic (tox-) strains of P. multocida isolated from geographically diverse locations. Plasmid pPMT and a previously described digoxigenin-labeled Bordetella-specific probe, pDLA5, were hybridized simultaneously to colony lifts from plates containing tox+ P. multocida, tox- P. multocida, and B. bronchiseptica. Bound probes were detected by sequential colorimetric development using a commercially available multicolor detection kit. Plasmid pPMT bound only to tox+ P. multocida and was visualized as a red reaction product. Plasmid pDLA5 hybridized only to B. bronchiseptica and was visualized as a blue reaction product. Tox- P. multocida gave no reaction product. This assay may be useful for identification of tox+ P. multocida and B. bronchiseptica in clinical specimens.

Last Modified: 4/19/2014
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