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United States Department of Agriculture

Agricultural Research Service

Title: Restriction Endonuclease Analysis Discrimnates Bordetella Bronchiseptica Isolates

Authors
item Sacco, Randy
item Register, Karen
item Nordholm, Gwen

Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 21, 2000
Publication Date: N/A

Interpretive Summary: Bordetella bronchiseptica causes respiratory disease in a number of animal species, including atrophic rhinitis in swine. In this study, we have developed a genetic method to easily distinguish between different isolates of B. bronchiseptica. Our approach is based on the use of genetic material from these organisms. The method that we have used is relatively easy to carry out compared to most previous methods for distinguishing among isolates of B. bronchiseptica. Further, our method may be used also as an identification system during outbreaks of atrophic rhinitis.

Technical Abstract: One hundred ninety-five Bordetella bronchiseptica isolates from 12 different host species worldwide were characterized by restriction enzyme analysis (REA). These isolates had previously been categorized into 21 PvuII ribotypes. Twenty restriction endonucleases were evaluated for use in REA. Digestion of chromosomal DNA with HinfI, followed by submarine electrophoresis in agarose gels and staining with ethidium bromide, produced DNA fragments in the 4.0- to 10-kb range which readily discriminated B. bronchiseptica isolates, resulting in 48 fingerprint patterns. Moreover, AluI digestion of chromosomal DNA produced 38 distinct fingerprint profiles with DNA fragments in the 6.0- to 20.0-kb range. While REA frequently provided more discriminatory power than ribotyping, there were examples where the use of ribotyping was more discriminatory than REA. Passage of selected isolates up to passage 25 did not change the eREA profile. Moreover, the Bvg phase did not alter the fingerprint profil of chromosomal DNA from B. bronchiseptica strains digested with HinfI or AluI. Based on the results presented herein, the combination of REA and ribotyping should provide valuable information in understanding the molecular epidemiology of B. bronchiseptica infections.

Last Modified: 10/23/2014
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