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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1990 Oct;28(10):2200–2204. doi: 10.1128/jcm.28.10.2200-2204.1990

Polymerase chain reaction for detection of Mycobacterium tuberculosis.

U Sjöbring 1, M Mecklenburg 1, A B Andersen 1, H Miörner 1
PMCID: PMC268147  PMID: 2121782

Abstract

A polymerase chain reaction for the specific detection of mycobacteria belonging to the Mycobacterium tuberculosis complex was developed. Using a single primer pair derived from the nucleotide sequence of protein antigen b of M. tuberculosis, we achieved specific amplification of a 419-base-pair DNA fragment in M. tuberculosis and M. bovis. After DNA was extracted from mycobacteria by using a simple, safe lysis procedure, we detected the 419-base-pair sequence in samples containing few mycobacteria. Preliminary data suggested that this technique could be applied to clinical specimens for early and specific diagnosis of tuberculosis.

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Selected References

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  1. Andersen A. B., Hansen E. B. Structure and mapping of antigenic domains of protein antigen b, a 38,000-molecular-weight protein of Mycobacterium tuberculosis. Infect Immun. 1989 Aug;57(8):2481–2488. doi: 10.1128/iai.57.8.2481-2488.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baess I. Isolation and purification of deoxyribonucleic acid from mycobacteria. Acta Pathol Microbiol Scand B Microbiol Immunol. 1974 Dec;82(6):780–784. doi: 10.1111/j.1699-0463.1974.tb02375.x. [DOI] [PubMed] [Google Scholar]
  3. Brisson-Noël A., Gicquel B., Lecossier D., Lévy-Frébault V., Nassif X., Hance A. J. Rapid diagnosis of tuberculosis by amplification of mycobacterial DNA in clinical samples. Lancet. 1989 Nov 4;2(8671):1069–1071. doi: 10.1016/s0140-6736(89)91082-9. [DOI] [PubMed] [Google Scholar]
  4. Daniel T. M., Debanne S. M. The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme-linked immunosorbent assay. Am Rev Respir Dis. 1987 May;135(5):1137–1151. doi: 10.1164/arrd.1987.135.5.1137. [DOI] [PubMed] [Google Scholar]
  5. Hance A. J., Grandchamp B., Lévy-Frébault V., Lecossier D., Rauzier J., Bocart D., Gicquel B. Detection and identification of mycobacteria by amplification of mycobacterial DNA. Mol Microbiol. 1989 Jul;3(7):843–849. doi: 10.1111/j.1365-2958.1989.tb00233.x. [DOI] [PubMed] [Google Scholar]
  6. Hartskeerl R. A., de Wit M. Y., Klatser P. R. Polymerase chain reaction for the detection of Mycobacterium leprae. J Gen Microbiol. 1989 Sep;135(9):2357–2364. doi: 10.1099/00221287-135-9-2357. [DOI] [PubMed] [Google Scholar]
  7. Kadival G. V., Chaparas S. D., Hussong D. Characterization of serologic and cell-mediated reactivity of a 38-kDa antigen isolated from Mycobacterium tuberculosis. J Immunol. 1987 Oct 1;139(7):2447–2451. [PubMed] [Google Scholar]
  8. Kennedy D. H., Fallon R. J. Tuberculous meningitis. JAMA. 1979 Jan 19;241(3):264–268. [PubMed] [Google Scholar]
  9. Larsson L., Odham G., Westerdahl G., Olsson B. Diagnosis of pulmonary tuberculosis by selected-ion monitoring: improved analysis of tuberculostearate in sputum using negative-ion mass spectrometry. J Clin Microbiol. 1987 May;25(5):893–896. doi: 10.1128/jcm.25.5.893-896.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Roberts M. C., McMillan C., Coyle M. B. Whole chromosomal DNA probes for rapid identification of Mycobacterium tuberculosis and Mycobacterium avium complex. J Clin Microbiol. 1987 Jul;25(7):1239–1243. doi: 10.1128/jcm.25.7.1239-1243.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  13. Shinnick T. M. The 65-kilodalton antigen of Mycobacterium tuberculosis. J Bacteriol. 1987 Mar;169(3):1080–1088. doi: 10.1128/jb.169.3.1080-1088.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  15. Sunderam G., McDonald R. J., Maniatis T., Oleske J., Kapila R., Reichman L. B. Tuberculosis as a manifestation of the acquired immunodeficiency syndrome (AIDS). JAMA. 1986 Jul 18;256(3):362–366. [PubMed] [Google Scholar]
  16. Thole J. E., Keulen W. J., De Bruyn J., Kolk A. H., Groothuis D. G., Berwald L. G., Tiesjema R. H., van Embden J. D. Characterization, sequence determination, and immunogenicity of a 64-kilodalton protein of Mycobacterium bovis BCG expressed in escherichia coli K-12. Infect Immun. 1987 Jun;55(6):1466–1475. doi: 10.1128/iai.55.6.1466-1475.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wolinsky E. Nontuberculous mycobacteria and associated diseases. Am Rev Respir Dis. 1979 Jan;119(1):107–159. doi: 10.1164/arrd.1979.119.1.107. [DOI] [PubMed] [Google Scholar]
  18. Young D., Kent L., Rees A., Lamb J., Ivanyi J. Immunological activity of a 38-kilodalton protein purified from Mycobacterium tuberculosis. Infect Immun. 1986 Oct;54(1):177–183. doi: 10.1128/iai.54.1.177-183.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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