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. 1993 Apr;31(4):1019–1021. doi: 10.1128/jcm.31.4.1019-1021.1993

Detection of Mycobacterium tuberculosis DNA in clinical samples by using a simple lysis method and polymerase chain reaction.

L Folgueira 1, R Delgado 1, E Palenque 1, A R Noriega 1
PMCID: PMC263610  PMID: 8463383

Abstract

We have evaluated the polymerase chain reaction for detection of Mycobacterium tuberculosis in clinical samples from patients with tuberculous infection. Two simple methods for mycobacterial DNA release have been compared: sonication and lysis with nonionic detergents and proteinase K. The more effective method was the enzymatic technique. By using this protocol with 75 specimens we detected M. tuberculosis DNA in all of the samples, whereas only 48 and 71 samples were positive by acid-fast staining and culture, respectively.

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Selected References

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  1. Brisson-Noel A., Aznar C., Chureau C., Nguyen S., Pierre C., Bartoli M., Bonete R., Pialoux G., Gicquel B., Garrigue G. Diagnosis of tuberculosis by DNA amplification in clinical practice evaluation. Lancet. 1991 Aug 10;338(8763):364–366. doi: 10.1016/0140-6736(91)90492-8. [DOI] [PubMed] [Google Scholar]
  2. Buck G. E., O'Hara L. C., Summersgill J. T. Rapid, simple method for treating clinical specimens containing Mycobacterium tuberculosis to remove DNA for polymerase chain reaction. J Clin Microbiol. 1992 May;30(5):1331–1334. doi: 10.1128/jcm.30.5.1331-1334.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cousins D. V., Wilton S. D., Francis B. R., Gow B. L. Use of polymerase chain reaction for rapid diagnosis of tuberculosis. J Clin Microbiol. 1992 Jan;30(1):255–258. doi: 10.1128/jcm.30.1.255-258.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Daniel T. M. Rapid diagnosis of tuberculosis: laboratory techniques applicable in developing countries. Rev Infect Dis. 1989 Mar-Apr;11 (Suppl 2):S471–S478. doi: 10.1093/clinids/11.supplement_2.s471. [DOI] [PubMed] [Google Scholar]
  5. Eisenach K. D., Cave M. D., Bates J. H., Crawford J. T. Polymerase chain reaction amplification of a repetitive DNA sequence specific for Mycobacterium tuberculosis. J Infect Dis. 1990 May;161(5):977–981. doi: 10.1093/infdis/161.5.977. [DOI] [PubMed] [Google Scholar]
  6. Eisenach K. D., Sifford M. D., Cave M. D., Bates J. H., Crawford J. T. Detection of Mycobacterium tuberculosis in sputum samples using a polymerase chain reaction. Am Rev Respir Dis. 1991 Nov;144(5):1160–1163. doi: 10.1164/ajrccm/144.5.1160. [DOI] [PubMed] [Google Scholar]
  7. KUBICA G. P., DYE W. E., COHN M. L., MIDDLEBROOK G. Sputum digestion and decontamination with N-acetyl-L-cysteine-sodium hydroxide for culture of mycobacteria. Am Rev Respir Dis. 1963 May;87:775–779. doi: 10.1164/arrd.1963.87.5.775. [DOI] [PubMed] [Google Scholar]
  8. Kadival G. V., Mazarelo T. B., Chaparas S. D. Sensitivity and specificity of enzyme-linked immunosorbent assay in the detection of antigen in tuberculous meningitis cerebrospinal fluids. J Clin Microbiol. 1986 May;23(5):901–904. doi: 10.1128/jcm.23.5.901-904.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pao C. C., Lin S. S., Wu S. Y., Juang W. M., Chang C. H., Lin J. Y. The detection of mycobacterial DNA sequences in uncultured clinical specimens with cloned Mycobacterium tuberculosis DNA as probes. Tubercle. 1988 Mar;69(1):27–36. doi: 10.1016/0041-3879(88)90037-2. [DOI] [PubMed] [Google Scholar]
  10. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Shoemaker S. A., Fisher J. H., Scoggin C. H. Techniques of DNA hybridization detect small numbers of mycobacteria with no cross-hybridization with non-mycobacterial respiratory organisms. Am Rev Respir Dis. 1985 May;131(5):760–763. doi: 10.1164/arrd.1985.131.5.760. [DOI] [PubMed] [Google Scholar]
  12. Sjöbring U., Mecklenburg M., Andersen A. B., Miörner H. Polymerase chain reaction for detection of Mycobacterium tuberculosis. J Clin Microbiol. 1990 Oct;28(10):2200–2204. doi: 10.1128/jcm.28.10.2200-2204.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Thierry D., Brisson-Noël A., Vincent-Lévy-Frébault V., Nguyen S., Guesdon J. L., Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol. 1990 Dec;28(12):2668–2673. doi: 10.1128/jcm.28.12.2668-2673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Victor T., du Toit R., van Helden P. D. Purification of sputum samples through sucrose improves detection of Mycobacterium tuberculosis by polymerase chain reaction. J Clin Microbiol. 1992 Jun;30(6):1514–1517. doi: 10.1128/jcm.30.6.1514-1517.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Yáez M. A., Coppola M. P., Russo D. A., Delaha E., Chaparas S. D., Yeager H., Jr Determination of mycobacterial antigens in sputum by enzyme immunoassay. J Clin Microbiol. 1986 May;23(5):822–825. doi: 10.1128/jcm.23.5.822-825.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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