Journal article
Multiplex PCR-based assay for detection of Bordetella pertussis in nasopharyngeal swab specimens
Journal of clinical microbiology, v 34(11), pp 2645-2649
Nov 1996
PMID: 8897157
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Abstract
A multiplex PCR-based assay was developed for the detection of Bordetella pertussis in nasopharyngeal swab specimens. The assay simultaneously amplified two separate DNA targets (153 and 203 bp) within a B. pertussis repetitive element and a 438-bp target within the beta-actin gene of human DNA (PCR amplification control). PCR products were detected by a sensitive and specific liquid hybridization gel retardation assay. A total of 496 paired nasopharyngeal swab specimens were tested by both the PCR-based assay and culture. Although 30 (6%) of the specimens inhibited the amplification of the beta-actin target, in all 29 specimens studied, the inhibition disappeared on repeat testing or was easily overcome with a 1:8 dilution or less of specimen digest. Of the 495 specimen pairs yielding a final evaluable result by the PCR-based assay, 19.0% were positive by the PCR-based assay, whereas 13.9% were positive by culture (P < 0.0001). After resolving the PCR-positive, culture-negative results by testing an additional aliquot from these specimens by the multiplex PCR-based assay, the PCR-based assay had a sensitivity and specificity of 98.9 and 99.7%, respectively, compared with values of 73.4 and 100%, respectively, for culture. In comparison with patients with culture-confirmed pertussis, those with PCR-positive, culture-negative results were older and more likely to have had prolonged cough, immunization with pertussis vaccine, or treatment with erythromycin. This multiplex PCR-based assay is substantially more sensitive than culture and identifies specimens that contain inhibitors of PCR.
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Details
- Title
- Multiplex PCR-based assay for detection of Bordetella pertussis in nasopharyngeal swab specimens
- Creators
- R M Wadowsky - Department of Otolaryngology, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USAR H Michaels - Department of Otolaryngology, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USAT Libert - Department of Otolaryngology, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USAL A Kingsley - Department of Otolaryngology, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USAG D Ehrlich - Department of Otolaryngology, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USA
- Publication Details
- Journal of clinical microbiology, v 34(11), pp 2645-2649
- Publisher
- American Society for Microbiology (ASM)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:A1996VM55600002
- Scopus ID
- 2-s2.0-0029998099
- Other Identifier
- 991014878431104721
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- Web of Science research areas
- Microbiology