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.
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.


RESULTS
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.
Maasstad Laboratory, Molecular Diagnostics Unit, Maasstad Hospital, Olympiaweg 350, 3078HT Rotterdam, The Netherlands. zeea@maasstadziekenhuis.nl.
Lieuwe
L
Roorda
Johannes
J
Buitenwerf
Jacobus M
JM
Ossewaarde
Anneke
A
van der Zee
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