Journal article
Development of a simplex polymerase chain reaction-based assay for the detection of Neisseria meningitidis
Molecular diagnosis, v 4(3), pp 233-239
01 Sep 1999
PMID: 10553023
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Background: This study was undertaken to develop a sensitive and specific polymerase chain reaction (PCR)-based assay for Neisseria meningitidis and Neisseria gonorrhoeae that could ultimately be incorporated into a multiplex assay designed to screen cerebrospinal fluid (CSF) for a panel of pyogenic bacterial species associated with bacterial meningitis.
Methods: N. meningitidis-specific primers were designed from porA gene sequences with the aid of a commercial software program and were used to develop and validate a clinical assay using a liquid hybridization-gel retardation detection system. The analytic sensitivity of the assay was determined using CSF spiked with N. meningitidis and comparing the PCR-based results with culture.
Results: Analytic sensitivity experiments showed the assay's limit of detection to be 100 fg purified input target DNA and 0.0125 colony-forming units of N. meningitidis spiked into CSF Specificity experiments showed the assay could detect all strains of N. meningitidis and N .gonorrhoeae tested, but did not support amplification of the commensal neisserial species or a panel of other human bacterial pathogens.
Conclusions: This PCR-based assay for pathogenic neisserial species is sensitive and specific and suitable for incorporation into a multiplex assay for the clinical differentiation of aseptic and septic meningitis.
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Details
- Title
- Development of a simplex polymerase chain reaction-based assay for the detection of Neisseria meningitidis
- Creators
- J Z Glustein - Children's Hospital of PittsburghY Z Zhang - Center for Genomic Sciences, Department of Human Genetics, MCP-Hahnemann University, Pittsburgh, Pennsylvania, USAR M Wadowsky - University of PittsburghG D Ehrlich - Genomic (Brazil)
- Publication Details
- Molecular diagnosis, v 4(3), pp 233-239
- Publisher
- Elsevier
- Number of pages
- 7
- Grant note
- R01DC002148 / NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute on Deafness & Other Communication Disorders (NIDCD) DC02148 / NIDCD NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute on Deafness & Other Communication Disorders (NIDCD)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000083409200007
- Scopus ID
- 2-s2.0-0032701058
- Other Identifier
- 991019168391504721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biotechnology & Applied Microbiology
- Medical Laboratory Technology
- Medicine, Research & Experimental