Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1990 Nov;28(11):2437–2441. doi: 10.1128/jcm.28.11.2437-2441.1990

Direct detection of Mycobacterium tuberculosis in clinical specimens by DNA amplification.

D De Wit 1, L Steyn 1, S Shoemaker 1, M Sogin 1
PMCID: PMC268202  PMID: 2123884

Abstract

A polymerase chain reaction assay was developed for the amplification of a 336-bp repetitive fragment in the chromosome of Mycobacterium tuberculosis. The assay is specific for M. tuberculosis and can be used to detect the amount of DNA present in less than 10 organisms. It was used to demonstrate M. tuberculosis DNA in 14 of 26 clinical specimens (from cerebrospinal fluids, pleural fluid aspirates and biopsies, pericardial fluid aspirates, and an open lung biopsy), and it was shown to be at least as sensitive as conventional culture techniques with these specimens.

Full text

PDF
2437

Images in this article

Selected References

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

  1. 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]
  2. Daniel T. M. New approaches to the rapid diagnosis of tuberculous meningitis. J Infect Dis. 1987 Apr;155(4):599–602. doi: 10.1093/infdis/155.4.599. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Eisenach K. D., Crawford J. T., Bates J. H. Repetitive DNA sequences as probes for Mycobacterium tuberculosis. J Clin Microbiol. 1988 Nov;26(11):2240–2245. doi: 10.1128/jcm.26.11.2240-2245.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eisenstein B. I. The polymerase chain reaction. A new method of using molecular genetics for medical diagnosis. N Engl J Med. 1990 Jan 18;322(3):178–183. doi: 10.1056/NEJM199001183220307. [DOI] [PubMed] [Google Scholar]
  6. French G. L., Chan C. Y., Cheung S. W., Oo K. T. Diagnosis of pulmonary tuberculosis by detection of tuberculostearic acid in sputum by using gas chromatography-mass spectrometry with selected ion monitoring. J Infect Dis. 1987 Aug;156(2):356–362. doi: 10.1093/infdis/156.2.356. [DOI] [PubMed] [Google Scholar]
  7. French G. L., Teoh R., Chan C. Y., Humphries M. J., Cheung S. W., O'Mahony G. Diagnosis of tuberculous meningitis by detection of tuberculostearic acid in cerebrospinal fluid. Lancet. 1987 Jul 18;2(8551):117–119. doi: 10.1016/s0140-6736(87)92328-2. [DOI] [PubMed] [Google Scholar]
  8. Hobby G. L., Holman A. P., Iseman M. D., Jones J. M. Enumeration of tubercle bacilli in sputum of patients with pulmonary tuberculosis. Antimicrob Agents Chemother. 1973 Aug;4(2):94–104. doi: 10.1128/aac.4.2.94. [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. Patel R. J., Fries J. W., Piessens W. F., Wirth D. F. Sequence analysis and amplification by polymerase chain reaction of a cloned DNA fragment for identification of Mycobacterium tuberculosis. J Clin Microbiol. 1990 Mar;28(3):513–518. doi: 10.1128/jcm.28.3.513-518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Shankar P., Manjunath N., Lakshmi R., Aditi B., Seth P., Shriniwas Identification of Mycobacterium tuberculosis by polymerase chain reaction. Lancet. 1990 Feb 17;335(8686):423–423. doi: 10.1016/0140-6736(90)90268-a. [DOI] [PubMed] [Google Scholar]
  13. Yeager H., Jr, Lacy J., Smith L. R., LeMaistre C. A. Quantitative studies of mycobacterial populations in sputum and saliva. Am Rev Respir Dis. 1967 Jun;95(6):998–1004. doi: 10.1164/arrd.1967.95.6.998. [DOI] [PubMed] [Google Scholar]

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

RESOURCES