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Quantification of Bordetella pertussis in clinical samples by colorimetric detection of competitive PCR products
Orebro Medical Center Hospital.
Orebro Medical Center Hospital.
KTH, Royal Institute of Technology, Stockholm.
KTH, Royal Institute of Technology, Stockholm.
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1998 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 106, no 7-12, 1041-1048 p.Article in journal (Refereed) Published
Abstract [en]

Quantification of microorganisms is an important part of the normal diagnostic work of a clinical microbiology laboratory. Traditionally the diagnosis of pertussis is subject to a yes or no approach with no quantitative dimension. This can, however, be of interest as a factor when judging the risk of a patient spreading the bacterium and as a research tool. The aim of the present study was to develop a PCR-based quantitative assay for Bordetella pertussis DNA in clinical nasopharyngeal aspirates by combining a quantitative PCR with a colorimetric detection principle, DIANA (detection of immobilised amplified nucleic acid). A competitor to the PCR target sequence in IS-481, containing a lac-operator, was constructed and calibrated, and a test protocol prepared. A total of 46 clinical nasopharyngeal aspirates, previously diagnosed using a standard nested PCR assay and quantified by culture, were analysed by the quantitative PCR. The method showed acceptable precision and accuracy considering that it estimates the total number of bacterial genomes while culture detects viable bacteria. Recognised advantages were the simple colorimetric detection, the inborn indication of a working PCR assay, and the possibility of obtaining results even when partial inhibition of the PCR assay was seen. In addition, the quantitative PCR result can be obtained within one day compared to 3–10 days for culture. The present results and the qualities of the quantitative PCR suggest that this assay will be a useful complement in routine diagnostics and in research.

Place, publisher, year, edition, pages
John Wiley & Sons, 1998. Vol. 106, no 7-12, 1041-1048 p.
Keyword [en]
Bordetella pertussis, whooping cough, quantitative PCR, nasopharyngeal samples
National Category
Medical and Health Sciences
Research subject
Biology
Identifiers
URN: urn:nbn:se:kau:diva-48377DOI: 10.1111/j.1699-0463.1998.tb00256.xOAI: oai:DiVA.org:kau-48377DiVA: diva2:1091444
Available from: 2017-04-26 Created: 2017-04-26 Last updated: 2017-05-03Bibliographically approved

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