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Journal of Clinical Microbiology, December 2000, p. 4387-4393, Vol. 38, No. 12
0095-1137/00/$04.00+0

Restriction Endonuclease Analysis Discriminates Bordetella bronchiseptica Isolates

Randy E. Sacco,* Karen B. Register, and Gwen E. Nordholm

USDA/Agricultural Research Service, National Animal Disease Center, Ames, Iowa 50010

Received 10 April 2000/Returned for modification 16 August 2000/Accepted 21 September 2000

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 19 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 39 distinct fingerprint profiles with DNA fragments ranging from 6.0 to 20.0 kb. 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 REA profile. Moreover, the Bvg phase did not alter the fingerprint profile 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.

* Corresponding author. Mailing address: USDA/ARS, National Animal Disease Center, P.O. Box 70, 2300 Dayton Rd., Ames, Iowa. Phone: (515) 663-7354. Fax: (515) 663-7458. E-mail: rsacco{at}nadc.ars.usda.gov.

Journal of Clinical Microbiology, December 2000, p. 4387-4393, Vol. 38, No. 12
0095-1137/00/$04.00+0


This article has been cited by other articles:

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  • Register, K. B., Sacco, R. E., Nordholm, G. E. (2003). Comparison of Ribotyping and Restriction Enzyme Analysis for Inter- and Intraspecies Discrimination of Bordetella avium and Bordetella hinzii. J. Clin. Microbiol. 41: 1512-1519 [Abstract] [Full Text]  


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