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Evaluation of an extended diagnostic PCR assay for detection and verification of the common causes of bacterial meningitis in CSF and other biological samples
1999 (English)In: Molecular and Cellular Probes, ISSN 0890-8508, E-ISSN 1096-1194, Vol. 13, no 1, 49-60 p.Article in journal (Refereed)
Abstract [en]

A seminested polymerase chain reaction (PCR)-based diagnostic assay was evaluated for detection and verification of Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, Steptococcus agalactiae and Listeria monocytogenes in cerebrospinal fluid (CSF) and other biological samples. A general bacterial amplicon from the 16S rRNA gene was amplified in a first step, and species-specific regions in a second. The detection level was 4 fg DNA/reaction, corresponding to about one bacterial genome per reaction tube. Sample preparations (Dynabeads DNA DIRECT kit) were assayed from 140 bacterial strains suspended in saline. In CSF the detection level for bacteria was 103 CFU ml-1 for N. meningitidis, H. influenzae and S. pneumoniae, 104 CFU ml-1 for Escherichia coli and 105 CFU ml-1 for S. agalactiae and L. monocytogenes. The detection levels for these bacteria were the same in the other tested biological samples, like blood with or without culture media. Clinical CSF samples were evaluated from 71 patients with proven bacterial meningitis, as were 61 CSF samples from individuals without bacterial meningitis. The diagnostic sensitivity of the assay in detecting bacteria in general was 0.97, and for the specific species in the clinical CSF samples 0.87-0.94. The specificity was 1.0 for detecting bacteria in general. Some cross-reactions were noted within the streptococcus group. The PCR results were verified by banding patterns of HaeIII digested PCR products.

Place, publisher, year, edition, pages
1999. Vol. 13, no 1, 49-60 p.
Keyword [en]
Food Engineering
Keyword [sv]
Livsmedelsteknik
National Category
Food Science
Identifiers
URN: urn:nbn:se:ri:diva-9292DOI: 10.1006/mcpr.1998.0218PubMedID: 10024433OAI: oai:DiVA.org:ri-9292DiVA: diva2:967166
Available from: 2016-09-08 Created: 2016-09-08Bibliographically approved

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CiteExportLink to record
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