Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Jun;34(6):1407–1411. doi: 10.1128/jcm.34.6.1407-1411.1996

Utility of PCR in diagnosing pulmonary tuberculosis.

J Bennedsen 1, V O Thomsen 1, G E Pfyffer 1, G Funke 1, K Feldmann 1, A Beneke 1, P A Jenkins 1, M Hegginbothom 1, A Fahr 1, M Hengstler 1, G Cleator 1, P Klapper 1, E G Wilkins 1
PMCID: PMC229033  PMID: 8735089

Abstract

At present, the rapid diagnosis of pulmonary tuberculosis rests with microscopy. However, this technique is insensitive and many cases of pulmonary tuberculosis cannot be initially confirmed. Nucleic acid amplification techniques are extremely sensitive, but when they are applied to tuberculosis diagnosis, they have given variable results. Investigators at six centers in Europe compared a standardized PCR system (Amplicor; Roche) against conventional culture methods. Defined clinical information was collected. Discrepant samples were retested, and inhibition assays and backup amplification with a separate primer pair were performed. Mycobacterium tuberculosis complex organisms were recovered from 654 (9.1%) of 7,194 samples and 293 (7.8%) of 3,738 patients. Four hundred fifty-two of the M. tuberculosis isolates from 204 patients were smear positive and culture positive. Among the culture-positive specimens, PCR had a sensitivity of 91.4% for smear-positive specimens and 60.9% for smear-negative specimens, with a specificity of 96.1%. Analysis of 254 PCR-positive, culture-negative specimens with discrepant results revealed that 130 were from patients with recently diagnosed tuberculosis and 94 represented a presumed laboratory error. Similar analysis of 118 PCR-negative, culture-positive specimens demonstrated that 27 discrepancies were due to presumed uneven aliquot distribution and 11 were due to presumed laboratory error; PCR inhibitors were detected in 8 specimens. Amplicor enables laboratories with little previous experience with nucleic acid amplification to perform PCR. Disease in more than 60% of the patients with tuberculosis with smear-negative, culture-positive specimens can be diagnosed at the time of admission, and potentially all patients with smear-positive specimens can immediately be confirmed as being infected with M. tuberculosis, leading to improved clinical management.

Full Text

The Full Text of this article is available as a PDF (233.9 KB).

Selected References

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

  1. Böddinghaus B., Rogall T., Flohr T., Blöcker H., Böttger E. C. Detection and identification of mycobacteria by amplification of rRNA. J Clin Microbiol. 1990 Aug;28(8):1751–1759. doi: 10.1128/jcm.28.8.1751-1759.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clarridge J. E., 3rd, Shawar R. M., Shinnick T. M., Plikaytis B. B. Large-scale use of polymerase chain reaction for detection of Mycobacterium tuberculosis in a routine mycobacteriology laboratory. J Clin Microbiol. 1993 Aug;31(8):2049–2056. doi: 10.1128/jcm.31.8.2049-2056.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Wit D., Steyn L., Shoemaker S., Sogin M. Direct detection of Mycobacterium tuberculosis in clinical specimens by DNA amplification. J Clin Microbiol. 1990 Nov;28(11):2437–2441. doi: 10.1128/jcm.28.11.2437-2441.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Forbes B. A., Hicks K. E. Direct detection of Mycobacterium tuberculosis in respiratory specimens in a clinical laboratory by polymerase chain reaction. J Clin Microbiol. 1993 Jul;31(7):1688–1694. doi: 10.1128/jcm.31.7.1688-1694.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ichiyama S., Shimokata K., Takeuchi J. Comparative study of a biphasic culture system (Roche MB Check system) with a conventional egg medium for recovery of mycobacteria. Aichi Mycobacteriosis Research Group. Tuber Lung Dis. 1993 Oct;74(5):338–341. doi: 10.1016/0962-8479(93)90109-b. [DOI] [PubMed] [Google Scholar]
  7. Isenberg H. D., D'Amato R. F., Heifets L., Murray P. R., Scardamaglia M., Jacobs M. C., Alperstein P., Niles A. Collaborative feasibility study of a biphasic system (Roche Septi-Chek AFB) for rapid detection and isolation of mycobacteria. J Clin Microbiol. 1991 Aug;29(8):1719–1722. doi: 10.1128/jcm.29.8.1719-1722.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jonas V., Alden M. J., Curry J. I., Kamisango K., Knott C. A., Lankford R., Wolfe J. M., Moore D. F. Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by amplification of rRNA. J Clin Microbiol. 1993 Sep;31(9):2410–2416. doi: 10.1128/jcm.31.9.2410-2416.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mitchison D. A., Allen B. W., Manickavasagar D. Selective Kirchner medium in the culture of specimens other than sputum for mycobacteria. J Clin Pathol. 1983 Dec;36(12):1357–1361. doi: 10.1136/jcp.36.12.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mulder J., McKinney N., Christopherson C., Sninsky J., Greenfield L., Kwok S. Rapid and simple PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma: application to acute retroviral infection. J Clin Microbiol. 1994 Feb;32(2):292–300. doi: 10.1128/jcm.32.2.292-300.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nolte F. S., Metchock B., McGowan J. E., Jr, Edwards A., Okwumabua O., Thurmond C., Mitchell P. S., Plikaytis B., Shinnick T. Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization. J Clin Microbiol. 1993 Jul;31(7):1777–1782. doi: 10.1128/jcm.31.7.1777-1782.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Noordhoek G. T., Kolk A. H., Bjune G., Catty D., Dale J. W., Fine P. E., Godfrey-Faussett P., Cho S. N., Shinnick T., Svenson S. B. Sensitivity and specificity of PCR for detection of Mycobacterium tuberculosis: a blind comparison study among seven laboratories. J Clin Microbiol. 1994 Feb;32(2):277–284. doi: 10.1128/jcm.32.2.277-284.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Noordhoek G. T., van Embden J. D., Kolk A. H. Questionable reliability of the polymerase chain reaction in the detection of Mycobacterium tuberculosis. N Engl J Med. 1993 Dec 30;329(27):2036–2036. doi: 10.1056/NEJM199312303292713. [DOI] [PubMed] [Google Scholar]
  14. Pfyffer G. E., Kissling P., Wirth R., Weber R. Direct detection of Mycobacterium tuberculosis complex in respiratory specimens by a target-amplified test system. J Clin Microbiol. 1994 Apr;32(4):918–923. doi: 10.1128/jcm.32.4.918-923.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pierre C., Lecossier D., Boussougant Y., Bocart D., Joly V., Yeni P., Hance A. J. Use of a reamplification protocol improves sensitivity of detection of Mycobacterium tuberculosis in clinical samples by amplification of DNA. J Clin Microbiol. 1991 Apr;29(4):712–717. doi: 10.1128/jcm.29.4.712-717.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Small P. M., McClenny N. B., Singh S. P., Schoolnik G. K., Tompkins L. S., Mickelsen P. A. Molecular strain typing of Mycobacterium tuberculosis to confirm cross-contamination in the mycobacteriology laboratory and modification of procedures to minimize occurrence of false-positive cultures. J Clin Microbiol. 1993 Jul;31(7):1677–1682. doi: 10.1128/jcm.31.7.1677-1682.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Wolcott M. J. Advances in nucleic acid-based detection methods. Clin Microbiol Rev. 1992 Oct;5(4):370–386. doi: 10.1128/cmr.5.4.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zhang Y., Young D. Molecular genetics of drug resistance in Mycobacterium tuberculosis. J Antimicrob Chemother. 1994 Sep;34(3):313–319. doi: 10.1093/jac/34.3.313. [DOI] [PubMed] [Google Scholar]
  20. Zolg J. W., Philippi-Schulz S. The superoxide dismutase gene, a target for detection and identification of mycobacteria by PCR. J Clin Microbiol. 1994 Nov;32(11):2801–2812. doi: 10.1128/jcm.32.11.2801-2812.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Soolingen D., de Haas P. E., Hermans P. W., Groenen P. M., van Embden J. D. Comparison of various repetitive DNA elements as genetic markers for strain differentiation and epidemiology of Mycobacterium tuberculosis. J Clin Microbiol. 1993 Aug;31(8):1987–1995. doi: 10.1128/jcm.31.8.1987-1995.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES