Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Jul;32(7):1721–1724. doi: 10.1128/jcm.32.7.1721-1724.1994

Detection of Streptococcus pneumoniae DNA in blood cultures by PCR.

M Hassan-King 1, I Baldeh 1, O Secka 1, A Falade 1, B Greenwood 1
PMCID: PMC263774  PMID: 7929764

Abstract

We have developed a PCR assay, with primers derived from the autolysin (lyt) gene, for the detection of Streptococcus pneumoniae DNA in blood cultures. The predicted fragment of 247 bp was detected in all strains of pneumococci, embracing 12 different serotypes that were tested. Although DNA extracted from four viridans streptococci spp. Streptococcus oralis, Streptococcus mitis, Streptococcus sanguis, and Streptococcus parasanguis) gave amplification products, these were quite different from the predicted fragment for S. pneumoniae. Application of the assay for diagnosis of septicemia caused by S. pneumoniae showed concordance between PCR and culture results. However, on four occasions PCR was positive in supernatants from both paired culture bottles while pneumococci were cultured from only one. Performing PCR on negative cultures in controlled studies such as vaccine trials may provide a sensitive tool for increasing case detection.

Full text

PDF
1723

Images in this article

1722Image
on p.1722
1723Image
on p.1723
1723Image
on p.1723

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Davis T. E., Fuller D. D. Direct identification of bacterial isolates in blood cultures by using a DNA probe. J Clin Microbiol. 1991 Oct;29(10):2193–2196. doi: 10.1128/jcm.29.10.2193-2196.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Denys G. A., Carey R. B. Identification of Streptococcus pneumoniae with a DNA probe. J Clin Microbiol. 1992 Oct;30(10):2725–2727. doi: 10.1128/jcm.30.10.2725-2727.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. García P., García J. L., García E., López R. Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli. Gene. 1986;43(3):265–272. doi: 10.1016/0378-1119(86)90215-5. [DOI] [PubMed] [Google Scholar]
  4. Guatelli J. C., Gingeras T. R., Richman D. D. Nucleic acid amplification in vitro: detection of sequences with low copy numbers and application to diagnosis of human immunodeficiency virus type 1 infection. Clin Microbiol Rev. 1989 Apr;2(2):217–226. doi: 10.1128/cmr.2.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Höltje J. V., Tomasz A. Specific recognition of choline residues in the cell wall teichoic acid by the N-acetylmuramyl-L-alanine amidase of Pneumococcus. J Biol Chem. 1975 Aug 10;250(15):6072–6076. [PubMed] [Google Scholar]
  6. Kalin M., Lindberg A. A. Diagnosis of pneumococcal pneumonia: a comparison between microscopic examination of expectorate, antigen detection and cultural procedures. Scand J Infect Dis. 1983;15(3):247–255. doi: 10.3109/inf.1983.15.issue-3.04. [DOI] [PubMed] [Google Scholar]
  7. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  8. Perlino C. A. Laboratory diagnosis of pneumonia due to Streptococcus pneumoniae. J Infect Dis. 1984 Jul;150(1):139–144. doi: 10.1093/infdis/150.1.139. [DOI] [PubMed] [Google Scholar]
  9. Pozzi G., Oggioni M. R., Tomasz A. DNA probe for identification of Streptococcus pneumoniae. J Clin Microbiol. 1989 Feb;27(2):370–372. doi: 10.1128/jcm.27.2.370-372.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rudolph K. M., Parkinson A. J., Black C. M., Mayer L. W. Evaluation of polymerase chain reaction for diagnosis of pneumococcal pneumonia. J Clin Microbiol. 1993 Oct;31(10):2661–2666. doi: 10.1128/jcm.31.10.2661-2666.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Tenover F. C. Diagnostic deoxyribonucleic acid probes for infectious diseases. Clin Microbiol Rev. 1988 Jan;1(1):82–101. doi: 10.1128/cmr.1.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tomasz A. Choline in the cell wall of a bacterium: novel type of polymer-linked choline in Pneumococcus. Science. 1967 Aug 11;157(3789):694–697. doi: 10.1126/science.157.3789.694. [DOI] [PubMed] [Google Scholar]
  13. Wolcott M. J. Advances in nucleic acid-based detection methods. Clin Microbiol Rev. 1992 Oct;5(4):370–386. doi: 10.1128/cmr.5.4.370. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES