Journal article
Detection of Streptococcus pneumoniae in whole blood by PCR
Journal of clinical microbiology, v 33(3), pp 596-601
Mar 1995
PMID: 7751363
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Streptococcus pneumoniae is a major cause of bacteremia in both children and adults. Currently, the diagnosis of pneumococcal bacteremia relies on the isolation and identification of the bacteria from blood cultures. We have developed a sensitive assay for the detection of S. pneumoniae in whole blood by the PCR. A specific primer-probe set (JM201 and JM202 primers with JM204 probe) designed from the penicillin-binding protein 2B gene was demonstrated to reproducibly detect between 10 and 100 fg of input purified S. pneumoniae DNA. This assay system was shown to be inclusive for all strains of S. pneumoniae evaluated, including 15 different serotypes and a battery of penicillin-resistant and -sensitive strains. The specificity of this PCR-based assay was demonstrated by its inability to support amplification from a series of human, bacterial, and yeast genomic DNAs. A general specimen preparation method which should be suitable for the purification of DNA from any pathogens in whole blood was developed. With this protocol it was possible to detect S. pneumoniae-specific DNA from whole blood specimens inoculated with as little as 4 CFU/ml. Copurified human blood DNA, ranging from 0 to 4.5 micrograms per PCR, did not affect the sensitivity of S. pneumoniae detection by PCR. A blinded clinical trial was used to compare the PCR-based assay with standard microbiological blood culture for the detection of S. pneumoniae bacteremia in 36 specimens obtained from pediatric patients seen in the emergency room of Children's Hospital of Pittsburgh.
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Details
- Title
- Detection of Streptococcus pneumoniae in whole blood by PCR
- Creators
- Y Zhang - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USAD J Isaacman - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USAR M Wadowsky - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USAJ Rydquist-White - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USAJ C Post - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USAG D Ehrlich - Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania, USA
- Publication Details
- Journal of clinical microbiology, v 33(3), pp 596-601
- Publisher
- American Society for Microbiology (ASM)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:A1995QG83000015
- Scopus ID
- 2-s2.0-0028837072
- Other Identifier
- 991014877985704721
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- Web of Science research areas
- Microbiology