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Open Access Technical Note

A real-time PCR assay with improved specificity for detection and discrimination of all clinically relevant Bordetella species by the presence and distribution of three Insertion Sequence elements

Lieuwe Roorda1, Johannes Buitenwerf1, Jacobus M Ossewaarde12 and Anneke van der Zee1*

Author Affiliations

1 Maasstad Laboratory, Molecular Diagnostics Unit, Maasstad Hospital, Olympiaweg 350, 3078HT Rotterdam, The Netherlands

2 Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands

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BMC Research Notes 2011, 4:11  doi:10.1186/1756-0500-4-11

Published: 21 January 2011

Abstract

Background

In Dutch laboratories molecular detection of B. pertussis and B. parapertussis is commonly based on insertion sequences IS481 and IS1001, respectively. Both IS elements are more widely spread among Bordetella species. Both Bordetella holmesii, and B. bronchiseptica can harbour IS481. Also, IS1001 is found among B. bronchiseptica. IS481, and IS1001 based PCR thus lacks specificity when used for detection of specific Bordetella spp.

Findings

We designed a PCR based on IS1002, another IS element that is present among Bordetella species, and exploited it as a template in combination with PCR for IS481, and IS1001. In combining the PCRs for IS481, IS1001, and IS1002, and including an inhibition control, we were able to detect and discriminate all clinically relevant Bordetella species.

Conclusions

We developed an improved PCR method for specific detection of B. pertussis, B. parapertussis, B. holmesii, and B. bronchiseptica.