This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Register, K. B.
Right arrow Articles by Yersin, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Register, K. B.
Right arrow Articles by Yersin, A. G.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 2005, p. 5567-5573, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5567-5573.2005

Analytical Verification of a PCR Assay for Identification of Bordetella avium

Karen B. Register* and Andrew G. Yersin{dagger}

Respiratory Diseases of Livestock Research Unit, USDA, Agricultural Research Service, National Animal Disease Center, P.O. Box 70, Ames, Iowa 50010

Received 3 March 2005/ Returned for modification 1 August 2005/ Accepted 15 August 2005

Bordetella avium is the etiologic agent of turkey coryza or bordetellosis, a respiratory disease responsible for substantial economic losses to the turkey industry. At present, identification of this bacterium relies on isolation and biochemical testing. Although a PCR for the detection of B. avium was proposed a number of years ago (P. H. Savelkoul, L. E. de Groot, C. Boersma, I. Livey, C. J. Duggleby, B. A. van der Zeijst, and W. Gaastra, Microb. Pathog. 15:207-215, 1993), lack of analytical verification precludes its use as a diagnostic tool. Furthermore, a number of details pertaining to the reaction conditions used are missing or unclear. In the present study we have identified an optimal set of PCR conditions for use with the previously described primer pair and determined the limit of detection under these conditions to be approximately 20 pg. Assay sensitivity is 100%, based on an analysis of 72 B. avium isolates from diverse geographic locations and covering a time span of at least 25 years. Evaluation of a separate group of 87 bacterial isolates from poultry, comprising both gram-positive and gram-negative commensals and pathogens representing 11 genera, demonstrated an assay specificity of 98.8%. Reproducibility is 100% using either purified genomic DNA or boiled cell lysates less than 3 days old. Sequence analysis of the B. avium PCR amplicons identified only three occasional sequence polymorphisms. These data indicate the B. avium PCR assay can provide clinically significant results.

* Corresponding author. Mailing address: Respiratory Diseases of Livestock Research Unit, USDA/ARS/National Animal Disease Center, P.O. Box 70, 2300 Dayton Road, Ames, Iowa 50010. Phone: (515) 663-7700. Fax: (515) 663-7458. E-mail: kregiste{at}nadc.ars.usda.gov.

{dagger} Present address: Kemin Industries, Inc., 2100 Maury St., Des Moines, IA 50317.

Journal of Clinical Microbiology, November 2005, p. 5567-5573, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5567-5573.2005




This article has been cited by other articles:

  • Register, K. B., Sanden, G. N. (2006). Prevalence and Sequence Variants of IS481 in Bordetella bronchiseptica: Implications for IS481-Based Detection of Bordetella pertussis. J. Clin. Microbiol. 44: 4577-4583 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»