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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Sep;30(9):2462–2466. doi: 10.1128/jcm.30.9.2462-2466.1992

Detection of Coxiella burnetti by DNA amplification using polymerase chain reaction.

A Stein 1, D Raoult 1
PMCID: PMC265524  PMID: 1401016

Abstract

The polymerase chain reaction (PCR) was used for the detection of Coxiella burnetti, an obligate intracellular bacterium and the etiologic agent of Q fever. A pair of primers derived from the C. burnetii superoxide dismutase gene served to amplify a targeted 257-bp fragment of genomic DNA. These primers were chosen on the basis of GenBank analysis, G + C ratio, and absence of secondary structure. This technique allowed the detection of as few as 10 C. burnetii organisms. C. burnetti was detected in tissue culture and in specimens from patients (heart valves). In all, 8 reference isolates and 22 new isolates of C. burnetii from France were successfully amplified. No amplification products were found when PCR was performed with 25 bacterial species that had been isolated in a clinical laboratory from patients with clinically similar infections. Amplification products of C. burnetii were confirmed by restriction enzyme digestion and dot blot hybridization. The method used here, a combination of PCR and restriction analysis, is a faster and more sensitive assay for C. burnetii than standard culture techniques.

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Selected References

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  1. Frazier M. E., Mallavia L. P., Samuel J. E., Baca O. G. DNA probes for the identification of Coxiella burnetti strains. Ann N Y Acad Sci. 1990;590:445–458. doi: 10.1111/j.1749-6632.1990.tb42253.x. [DOI] [PubMed] [Google Scholar]
  2. GIMENEZ D. F. STAINING RICKETTSIAE IN YOLK-SAC CULTURES. Stain Technol. 1964 May;39:135–140. doi: 10.3109/10520296409061219. [DOI] [PubMed] [Google Scholar]
  3. Heinzen R. A., Frazier M. E., Mallavia L. P. Nucleotide sequence of Coxiella burnetii superoxide dismutase. Nucleic Acids Res. 1990 Nov 11;18(21):6437–6437. doi: 10.1093/nar/18.21.6437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Heinzen R. A., Mallavia L. P. Cloning and functional expression of the Coxiella burnetii citrate synthase gene in Escherichia coli. Infect Immun. 1987 Apr;55(4):848–855. doi: 10.1128/iai.55.4.848-855.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Levy P. Y., Drancourt M., Etienne J., Auvergnat J. C., Beytout J., Sainty J. M., Goldstein F., Raoult D. Comparison of different antibiotic regimens for therapy of 32 cases of Q fever endocarditis. Antimicrob Agents Chemother. 1991 Mar;35(3):533–537. doi: 10.1128/aac.35.3.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Mallavia L. P., Whiting L. L., Minnick M. F., Heinzen R., Reschke D., Foreman M., Baca O. G., Frazier M. E. Strategy for detection and differentiation of Coxiella burnetii strains using the polymerase chain reaction. Ann N Y Acad Sci. 1990;590:572–581. doi: 10.1111/j.1749-6632.1990.tb42268.x. [DOI] [PubMed] [Google Scholar]
  7. Minnick M. F., Heinzen R. A., Douthart R., Mallavia L. P., Frazier M. E. Analysis of QpRS-specific sequences from Coxiella burnetii. Ann N Y Acad Sci. 1990;590:514–522. doi: 10.1111/j.1749-6632.1990.tb42261.x. [DOI] [PubMed] [Google Scholar]
  8. Minnick M. F., Heinzen R. A., Frazier M. E., Mallavia L. P. Characterization and expression of the cbbE' gene of Coxiella burnetii. J Gen Microbiol. 1990 Jun;136(6):1099–1107. doi: 10.1099/00221287-136-6-1099. [DOI] [PubMed] [Google Scholar]
  9. Minnick M. F., Small C. L., Frazier M. E., Mallavia L. P. Analysis of strain-specific plasmid sequences from Coxiella burnetii. Acta Virol. 1991 Nov;35(6):497–502. [PubMed] [Google Scholar]
  10. Raoult D., Bollini G., Gallais H. Osteoarticular infection due to Coxiella burnetii. J Infect Dis. 1989 Jun;159(6):1159–1160. doi: 10.1093/infdis/159.6.1159. [DOI] [PubMed] [Google Scholar]
  11. Raoult D., Piquet P., Gallais H., de Micco C., Drancourt M., Casanova P. Coxiella burnetii infection of a vascular prosthesis. N Engl J Med. 1986 Nov 20;315(21):1358–1359. [PubMed] [Google Scholar]
  12. Raoult D., Torres H., Drancourt M. Shell-vial assay: evaluation of a new technique for determining antibiotic susceptibility, tested in 13 isolates of Coxiella burnetii. Antimicrob Agents Chemother. 1991 Oct;35(10):2070–2077. doi: 10.1128/aac.35.10.2070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Raoult D., Vestris G., Enea M. Isolation of 16 strains of Coxiella burnetii from patients by using a sensitive centrifugation cell culture system and establishment of the strains in HEL cells. J Clin Microbiol. 1990 Nov;28(11):2482–2484. doi: 10.1128/jcm.28.11.2482-2484.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  15. Samuel J. E., Frazier M. E., Mallavia L. P. Correlation of plasmid type and disease caused by Coxiella burnetii. Infect Immun. 1985 Sep;49(3):775–779. doi: 10.1128/iai.49.3.775-779.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Samuel J. E., Frazier M. E., Mallavia L. P. Stability of plasmid sequences in an acute Q-fever strain of Coxiella burnetii. J Gen Microbiol. 1988 Jul;134(7):1795–1805. doi: 10.1099/00221287-134-7-1795. [DOI] [PubMed] [Google Scholar]
  17. Savinelli E. A., Mallavia L. P. Comparison of Coxiella burnetii plasmids to homologous chromosomal sequences present in a plasmidless endocarditis-causing isolate. Ann N Y Acad Sci. 1990;590:523–533. doi: 10.1111/j.1749-6632.1990.tb42262.x. [DOI] [PubMed] [Google Scholar]
  18. Stein A., Raoult D. Phenotypic and genotypic heterogeneity of 8 new human Coxiella burnetti isolates. Acta Virol. 1992 Jan;36(1):7–12. [PubMed] [Google Scholar]
  19. Vodkin M. H., Williams J. C. A heat shock operon in Coxiella burnetti produces a major antigen homologous to a protein in both mycobacteria and Escherichia coli. J Bacteriol. 1988 Mar;170(3):1227–1234. doi: 10.1128/jb.170.3.1227-1234.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weiss E. The biology of rickettsiae. Annu Rev Microbiol. 1982;36:345–370. doi: 10.1146/annurev.mi.36.100182.002021. [DOI] [PubMed] [Google Scholar]

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