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. 1995 Mar;33(3):596–601. doi: 10.1128/jcm.33.3.596-601.1995

Detection of Streptococcus pneumoniae in whole blood by PCR.

Y Zhang 1, D J Isaacman 1, R M Wadowsky 1, J Rydquist-White 1, J C Post 1, G D Ehrlich 1
PMCID: PMC227996  PMID: 7751363

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.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Barker R. H., Jr, Banchongaksorn T., Courval J. M., Suwonkerd W., Rimwungtragoon K., Wirth D. F. A simple method to detect Plasmodium falciparum directly from blood samples using the polymerase chain reaction. Am J Trop Med Hyg. 1992 Apr;46(4):416–426. doi: 10.4269/ajtmh.1992.46.416. [DOI] [PubMed] [Google Scholar]
  2. Bell L. M., Alpert G., Campos J. M., Plotkin S. A. Routine quantitative blood cultures in children with Haemophilus influenzae or Streptococcus pneumoniae bacteremia. Pediatrics. 1985 Dec;76(6):901–904. [PubMed] [Google Scholar]
  3. Davis T. E., Fuller D. D., Aeschleman E. C. Rapid, direct identification of Staphylococcus aureus and Streptococcus pneumoniae from blood cultures using commercial immunologic kits and modified conventional tests. Diagn Microbiol Infect Dis. 1992 May-Jun;15(4):295–300. doi: 10.1016/0732-8893(92)90014-k. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Dowson C. G., Hutchison A., Woodford N., Johnson A. P., George R. C., Spratt B. G. Penicillin-resistant viridans streptococci have obtained altered penicillin-binding protein genes from penicillin-resistant strains of Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5858–5862. doi: 10.1073/pnas.87.15.5858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Iralu J. V., Sritharan V. K., Pieciak W. S., Wirth D. F., Maguire J. H., Barker R. H., Jr Diagnosis of Mycobacterium avium bacteremia by polymerase chain reaction. J Clin Microbiol. 1993 Jul;31(7):1811–1814. doi: 10.1128/jcm.31.7.1811-1814.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Isaacman D. J., Karasic R. B. Lack of effect of changing needles on contamination of blood cultures. Pediatr Infect Dis J. 1990 Apr;9(4):274–278. doi: 10.1097/00006454-199004000-00010. [DOI] [PubMed] [Google Scholar]
  8. Isaacman D. J., Karasic R. B. Utility of collecting blood cultures through newly inserted intravenous catheters. Pediatr Infect Dis J. 1990 Nov;9(11):815–818. doi: 10.1097/00006454-199011000-00007. [DOI] [PubMed] [Google Scholar]
  9. Khan G., Kangro H. O., Coates P. J., Heath R. B. Inhibitory effects of urine on the polymerase chain reaction for cytomegalovirus DNA. J Clin Pathol. 1991 May;44(5):360–365. doi: 10.1136/jcp.44.5.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Klein J. O. Bacteremia in ambulatory children. Pediatr Infect Dis. 1984 May-Jun;3(3 Suppl):S5–S8. doi: 10.1097/00006454-198405001-00002. [DOI] [PubMed] [Google Scholar]
  11. Lipschik G. Y., Gill V. J., Lundgren J. D., Andrawis V. A., Nelson N. A., Nielsen J. O., Ognibene F. P., Kovacs J. A. Improved diagnosis of Pneumocystis carinii infection by polymerase chain reaction on induced sputum and blood. Lancet. 1992 Jul 25;340(8813):203–206. doi: 10.1016/0140-6736(92)90469-j. [DOI] [PubMed] [Google Scholar]
  12. McGuire B. S., Chandler F. W., Felz M. W., Huey L. O., Field R. S. Detection of Borrelia burgdorferi in human blood and urine using the polymerase chain reaction. Pathobiology. 1992;60(3):163–167. doi: 10.1159/000163717. [DOI] [PubMed] [Google Scholar]
  13. Ni H., Knight A. I., Cartwright K., Palmer W. H., McFadden J. Polymerase chain reaction for diagnosis of meningococcal meningitis. Lancet. 1992 Dec 12;340(8833):1432–1434. doi: 10.1016/0140-6736(92)92622-m. [DOI] [PubMed] [Google Scholar]
  14. Peter J. B. The polymerase chain reaction: amplifying our options. Rev Infect Dis. 1991 Jan-Feb;13(1):166–171. doi: 10.1093/clinids/12.5.166. [DOI] [PubMed] [Google Scholar]
  15. Pflugfelder S. C., Crouse C., Pereira I., Atherton S. Amplification of Epstein-Barr virus genomic sequences in blood cells, lacrimal glands, and tears from primary Sjögren's syndrome patients. Ophthalmology. 1990 Aug;97(8):976–984. doi: 10.1016/s0161-6420(90)32476-4. [DOI] [PubMed] [Google Scholar]
  16. Rappaport T., Sawyer K. P., Nachamkin I. Evaluation of several commercial biochemical and immunologic methods for rapid identification of gram-positive cocci directly from blood cultures. J Clin Microbiol. 1988 Jul;26(7):1335–1338. doi: 10.1128/jcm.26.7.1335-1338.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rowley A. H., Wald E. R. The incubation period necessary for detection of bacteremia in immunocompetent children with fever. Implications for the clinician. Clin Pediatr (Phila) 1986 Oct;25(10):485–489. doi: 10.1177/000992288602501001. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Teele D. W., Pelton S. I., Grant M. J., Herskowitz J., Rosen D. J., Allen C. E., Wimmer R. S., Klein J. O. Bacteremia in febrile children under 2 years of age: results of cultures of blood of 600 consecutive febrile children seen in a "walk-in" clinic. J Pediatr. 1975 Aug;87(2):227–230. doi: 10.1016/s0022-3476(75)80584-1. [DOI] [PubMed] [Google Scholar]
  20. Thiele D. The technique of polymerase chain reaction--a new diagnostic tool in microbiology and other scientific fields (review). Zentralbl Bakteriol. 1990 Sep;273(4):431–454. doi: 10.1016/s0934-8840(11)80451-2. [DOI] [PubMed] [Google Scholar]

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