Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Nov;35(11):2733–2739. doi: 10.1128/jcm.35.11.2733-2739.1997

Molecular diagnosis of bacterial endocarditis by broad-range PCR amplification and direct sequencing.

D Goldenberger 1, A Künzli 1, P Vogt 1, R Zbinden 1, M Altwegg 1
PMCID: PMC230051  PMID: 9350723

Abstract

Broad-range PCR amplification of part of the 16S rRNA gene followed by single-strand sequencing was applied to samples of 18 resected heart valves from patients with infective endocarditis. The PCR results were compared with those of cultures of valves and with those of previous blood cultures. For two patients there was agreement with the cultures of the valves; for nine patients there was agreement with the previous blood cultures, which were positive, while the cultures of the valves were negative; a Streptococcus sp. and Tropheryma whippelii each were found in one patient with negative cultures (valve and blood); for two patients the cultures of the valves as well as the PCR results were negative but the blood cultures were positive; for one patient amplification was inhibited; and for two patients the PCR results were positive but the amplicons could not be sequenced. It is concluded that broad-range PCR is a promising tool for patients with culture-negative endocarditis and allows the detection of rare, noncultivable organisms.

Full Text

The Full Text of this article is available as a PDF (155.1 KB).

Selected References

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

  1. Ali A. S., Trivedi V., Lesch M. Culture-negative endocarditis--a historical review and 1990s update. Prog Cardiovasc Dis. 1994 Nov-Dec;37(3):149–160. doi: 10.1016/s0033-0620(05)80040-4. [DOI] [PubMed] [Google Scholar]
  2. Altwegg M., Fleisch-Marx A., Goldenberger D., Hailemariam S., Schaffner A., Kissling R. Spondylodiscitis caused by Tropheryma whippelii. Schweiz Med Wochenschr. 1996 Aug 31;126(35):1495–1499. [PubMed] [Google Scholar]
  3. Anderson B. E., Dawson J. E., Jones D. C., Wilson K. H. Ehrlichia chaffeensis, a new species associated with human ehrlichiosis. J Clin Microbiol. 1991 Dec;29(12):2838–2842. doi: 10.1128/jcm.29.12.2838-2842.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Böttger E. C., Teske A., Kirschner P., Bost S., Chang H. R., Beer V., Hirschel B. Disseminated "Mycobacterium genavense" infection in patients with AIDS. Lancet. 1992 Jul 11;340(8811):76–80. doi: 10.1016/0140-6736(92)90397-l. [DOI] [PubMed] [Google Scholar]
  5. Carbon P., Ehresmann C., Ehresmann B., Ebel J. P. The complete nucleotide sequence of the ribosomal 16-S RNA from Excherichia coli. Experimental details and cistron heterogeneities. Eur J Biochem. 1979 Oct 15;100(2):399–410. doi: 10.1111/j.1432-1033.1979.tb04183.x. [DOI] [PubMed] [Google Scholar]
  6. Chen K., Neimark H., Rumore P., Steinman C. R. Broad range DNA probes for detecting and amplifying eubacterial nucleic acids. FEMS Microbiol Lett. 1989 Jan 1;48(1):19–24. doi: 10.1016/0378-1097(89)90139-0. [DOI] [PubMed] [Google Scholar]
  7. Edwards U., Rogall T., Blöcker H., Emde M., Böttger E. C. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res. 1989 Oct 11;17(19):7843–7853. doi: 10.1093/nar/17.19.7843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fox G. E., Wisotzkey J. D., Jurtshuk P., Jr How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int J Syst Bacteriol. 1992 Jan;42(1):166–170. doi: 10.1099/00207713-42-1-166. [DOI] [PubMed] [Google Scholar]
  9. Frothingham R., Wilson K. H. Molecular phylogeny of the Mycobacterium avium complex demonstrates clinically meaningful divisions. J Infect Dis. 1994 Feb;169(2):305–312. doi: 10.1093/infdis/169.2.305. [DOI] [PubMed] [Google Scholar]
  10. Goldenberger D., Perschil I., Ritzler M., Altwegg M. A simple "universal" DNA extraction procedure using SDS and proteinase K is compatible with direct PCR amplification. PCR Methods Appl. 1995 Jun;4(6):368–370. doi: 10.1101/gr.4.6.368. [DOI] [PubMed] [Google Scholar]
  11. Goldenberger Daniel, Schmidheini Tobias, Altwegg Martin. Detection of Bartonella henselae and Bartonella quintana by a simple and rapid procedure using broad-range PCR amplification and direct single-strand sequencing of part of the 16S rRNA gene. Clin Microbiol Infect. 1997 Apr;3(2):240–245. doi: 10.1111/j.1469-0691.1997.tb00604.x. [DOI] [PubMed] [Google Scholar]
  12. Hadfield T. L., Warren R., Kass M., Brun E., Levy C. Endocarditis caused by Rochalimaea henselae. Hum Pathol. 1993 Oct;24(10):1140–1141. doi: 10.1016/0046-8177(93)90196-n. [DOI] [PubMed] [Google Scholar]
  13. Hamed K. A., Dormitzer P. R., Su C. K., Relman D. A. Haemophilus parainfluenzae endocarditis: application of a molecular approach for identification of pathogenic bacterial species. Clin Infect Dis. 1994 Oct;19(4):677–683. doi: 10.1093/clinids/19.4.677. [DOI] [PubMed] [Google Scholar]
  14. Hoen B., Selton-Suty C., Lacassin F., Etienne J., Briançon S., Leport C., Canton P. Infective endocarditis in patients with negative blood cultures: analysis of 88 cases from a one-year nationwide survey in France. Clin Infect Dis. 1995 Mar;20(3):501–506. doi: 10.1093/clinids/20.3.501. [DOI] [PubMed] [Google Scholar]
  15. Jalava J., Kotilainen P., Nikkari S., Skurnik M., Vänttinen E., Lehtonen O. P., Eerola E., Toivanen P. Use of the polymerase chain reaction and DNA sequencing for detection of Bartonella quintana in the aortic valve of a patient with culture-negative infective endocarditis. Clin Infect Dis. 1995 Oct;21(4):891–896. doi: 10.1093/clinids/21.4.891. [DOI] [PubMed] [Google Scholar]
  16. Kawamura Y., Hou X. G., Sultana F., Liu S., Yamamoto H., Ezaki T. Transfer of Streptococcus adjacens and Streptococcus defectivus to Abiotrophia gen. nov. as Abiotrophia adiacens comb. nov. and Abiotrophia defectiva comb. nov., respectively. Int J Syst Bacteriol. 1995 Oct;45(4):798–803. doi: 10.1099/00207713-45-4-798. [DOI] [PubMed] [Google Scholar]
  17. Kawamura Y., Hou X. G., Sultana F., Miura H., Ezaki T. Determination of 16S rRNA sequences of Streptococcus mitis and Streptococcus gordonii and phylogenetic relationships among members of the genus Streptococcus. Int J Syst Bacteriol. 1995 Apr;45(2):406–408. doi: 10.1099/00207713-45-2-406. [DOI] [PubMed] [Google Scholar]
  18. Raoult D., Fournier P. E., Drancourt M., Marrie T. J., Etienne J., Cosserat J., Cacoub P., Poinsignon Y., Leclercq P., Sefton A. M. Diagnosis of 22 new cases of Bartonella endocarditis. Ann Intern Med. 1996 Oct 15;125(8):646–652. doi: 10.7326/0003-4819-125-8-199610150-00004. [DOI] [PubMed] [Google Scholar]
  19. Regnery R. L., Anderson B. E., Clarridge J. E., 3rd, Rodriguez-Barradas M. C., Jones D. C., Carr J. H. Characterization of a novel Rochalimaea species, R. henselae sp. nov., isolated from blood of a febrile, human immunodeficiency virus-positive patient. J Clin Microbiol. 1992 Feb;30(2):265–274. doi: 10.1128/jcm.30.2.265-274.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Relman D. A., Loutit J. S., Schmidt T. M., Falkow S., Tompkins L. S. The agent of bacillary angiomatosis. An approach to the identification of uncultured pathogens. N Engl J Med. 1990 Dec 6;323(23):1573–1580. doi: 10.1056/NEJM199012063232301. [DOI] [PubMed] [Google Scholar]
  21. Relman D. A., Schmidt T. M., MacDermott R. P., Falkow S. Identification of the uncultured bacillus of Whipple's disease. N Engl J Med. 1992 Jul 30;327(5):293–301. doi: 10.1056/NEJM199207303270501. [DOI] [PubMed] [Google Scholar]
  22. Rickman L. S., Freeman W. R., Green W. R., Feldman S. T., Sullivan J., Russack V., Relman D. A. Brief report: uveitis caused by Tropheryma whippelii (Whipple's bacillus) N Engl J Med. 1995 Feb 9;332(6):363–366. doi: 10.1056/NEJM199502093320604. [DOI] [PubMed] [Google Scholar]
  23. Tunkel A. R., Kaye D. Endocarditis with negative blood cultures. N Engl J Med. 1992 Apr 30;326(18):1215–1217. doi: 10.1056/NEJM199204303261809. [DOI] [PubMed] [Google Scholar]
  24. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991 Jan;173(2):697–703. doi: 10.1128/jb.173.2.697-703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Welch D. F., Pickett D. A., Slater L. N., Steigerwalt A. G., Brenner D. J. Rochalimaea henselae sp. nov., a cause of septicemia, bacillary angiomatosis, and parenchymal bacillary peliosis. J Clin Microbiol. 1992 Feb;30(2):275–280. doi: 10.1128/jcm.30.2.275-280.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wendler D., Mendoza E., Schleiffer T., Zander M., Maier M. Tropheryma whippelii endocarditis confirmed by polymerase chain reaction. Eur Heart J. 1995 Mar;16(3):424–425. doi: 10.1093/oxfordjournals.eurheartj.a060928. [DOI] [PubMed] [Google Scholar]
  27. Wilson K. H., Blitchington R. B., Greene R. C. Amplification of bacterial 16S ribosomal DNA with polymerase chain reaction. J Clin Microbiol. 1990 Sep;28(9):1942–1946. doi: 10.1128/jcm.28.9.1942-1946.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wilson K. H., Blitchington R., Frothingham R., Wilson J. A. Phylogeny of the Whipple's-disease-associated bacterium. Lancet. 1991 Aug 24;338(8765):474–475. doi: 10.1016/0140-6736(91)90545-z. [DOI] [PubMed] [Google Scholar]
  29. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES