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Journal of Clinical Microbiology, October 2008, p. 3228-3231, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00386-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification and Evaluation of New Target Sequences for Specific Detection of Bordetella pertussis by Real-Time PCR{triangledown}

William S. Probert,1* Janet Ely,1 Kimmi Schrader,1 Jessica Atwell,1 Angela Nossoff,2 and Stanley Kwan3

Microbial Diseases Laboratory, California Department of Public Health, Richmond, California 94804,1 Alameda County Public Health Department Laboratory, Oakland, California 94607,2 Yolo County Health Department Laboratory, Woodland, California 956953

Received 26 February 2008/ Accepted 15 August 2008

A comparative analysis of the Bordetella pertussis, B. bronchiseptica, and B. parapertussis genome assemblies permitted the identification of regions with significant sequence divergence and the design of two new real-time PCR assays, BP283 and BP485, for the specific detection of B. pertussis. The performance characteristics of these two assays were evaluated and compared to those of culture and an existing real-time PCR assay targeting the repetitive element IS481. The testing of 324 nasopharyngeal specimens indicated that, compared to culture, the BP283 assay had a sensitivity and specificity of 100 and 96.8% and the BP485 assay had a sensitivity and specificity of 92.3 and 97.1%. Notably, B. holmesii was isolated from two specimens that were positive by the IS481 assay but negative by the BP283 and BP485 assays. These two assays represent an improvement in specificity over those of PCR assays targeting only IS481 and may be duplexed or used in conjunction with existing PCR assays to improve the molecular detection of B. pertussis.

* Corresponding author. Mailing address: Microbial Diseases Laboratory, California Department of Public Health, Richmond, CA 94804. Phone: (510) 412-3754. Fax: (510) 412-3706. E-mail: Will.Probert{at}cdph.ca.gov

{triangledown} Published ahead of print on 27 August 2008.

Journal of Clinical Microbiology, October 2008, p. 3228-3231, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00386-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Fry, N. K., Duncan, J., Wagner, K., Tzivra, O., Doshi, N., Litt, D. J., Crowcroft, N., Miller, E., George, R. C., Harrison, T. G. (2009). Role of PCR in the diagnosis of pertussis infection in infants: 5 years' experience of provision of a same-day real-time PCR service in England and Wales from 2002 to 2007. J Med Microbiol 58: 1023-1029 [Abstract] [Full Text]  


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