Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Dec;35(12):3082–3085. doi: 10.1128/jcm.35.12.3082-3085.1997

Diagnostic value of the strand displacement amplification method compared to those of Roche Amplicor PCR and culture for detecting mycobacteria in sputum samples.

S Ichiyama 1, Y Ito 1, F Sugiura 1, Y Iinuma 1, S Yamori 1, M Shimojima 1, Y Hasegawa 1, K Shimokata 1, N Nakashima 1
PMCID: PMC230126  PMID: 9399498

Abstract

We compared the ability of the semiautomated BDProbeTec-SDA system, which uses the strand displacement amplification (SDA) method, with that of the Roche Amplicor-PCR system and the Septi-Chek AFB culture system to directly detect Mycobacterium tuberculosis complex (MTB) and other mycobacteria in sputum samples. A total of 530 sputum samples from 299 patients were examined in this study. Of the 530 samples, 129 were culture positive for acid-fast bacilli with the Septi-Chek AFB system; 95 for MTB, 29 for M. avium-M. intracellulare complex (MAC), and 5 for other mycobacteria. The BDProbeTec-SDA system detected 90 of the 95 samples culture positive for MTB (sensitivity, 94.7%), and the Amplicor-PCR system detected 85 of the 95 samples culture positive for MTB (sensitivity, 89.5%). The specificity of each system, based on the clinical diagnosis, was 99.8% for SDA and 100% for PCR, respectively. Among the 29 samples culture positive for MAC, the BDProbeTec-SDA system detected MAC in 24 samples (sensitivity, 82.8%), whereas the Amplicor-PCR system detected MAC in 23 samples (sensitivity, 79.3%). The specificities of the systems were 98.3 and 100%, respectively. The high degrees of sensitivity and specificity of the BDProbeTec-SDA system suggest that it should be very useful in clinical laboratories for the rapid detection of mycobacteria in sputum samples.

Full Text

The Full Text of this article is available as a PDF (109.9 KB).

Selected References

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

  1. Abe C., Hirano K., Wada M., Kazumi Y., Takahashi M., Fukasawa Y., Yoshimura T., Miyagi C., Goto S. Detection of Mycobacterium tuberculosis in clinical specimens by polymerase chain reaction and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test. J Clin Microbiol. 1993 Dec;31(12):3270–3274. doi: 10.1128/jcm.31.12.3270-3274.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beavis K. G., Lichty M. B., Jungkind D. L., Giger O. Evaluation of Amplicor PCR for direct detection of Mycobacterium tuberculosis from sputum specimens. J Clin Microbiol. 1995 Oct;33(10):2582–2586. doi: 10.1128/jcm.33.10.2582-2586.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennedsen J., Thomsen V. O., Pfyffer G. E., Funke G., Feldmann K., Beneke A., Jenkins P. A., Hegginbothom M., Fahr A., Hengstler M. Utility of PCR in diagnosing pulmonary tuberculosis. J Clin Microbiol. 1996 Jun;34(6):1407–1411. doi: 10.1128/jcm.34.6.1407-1411.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bergmann J. S., Woods G. L. Clinical evaluation of the Roche AMPLICOR PCR Mycobacterium tuberculosis test for detection of M. tuberculosis in respiratory specimens. J Clin Microbiol. 1996 May;34(5):1083–1085. doi: 10.1128/jcm.34.5.1083-1085.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Carpentier E., Drouillard B., Dailloux M., Moinard D., Vallee E., Dutilh B., Maugein J., Bergogne-Berezin E., Carbonnelle B. Diagnosis of tuberculosis by Amplicor Mycobacterium tuberculosis test: a multicenter study. J Clin Microbiol. 1995 Dec;33(12):3106–3110. doi: 10.1128/jcm.33.12.3106-3110.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. D'Amato R. F., Wallman A. A., Hochstein L. H., Colaninno P. M., Scardamaglia M., Ardila E., Ghouri M., Kim K., Patel R. C., Miller A. Rapid diagnosis of pulmonary tuberculosis by using Roche AMPLICOR Mycobacterium tuberculosis PCR test. J Clin Microbiol. 1995 Jul;33(7):1832–1834. doi: 10.1128/jcm.33.7.1832-1834.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Devallois A., Legrand E., Rastogi N. Evaluation of Amplicor MTB test as adjunct to smears and culture for direct detection of Mycobacterium tuberculosis in the French Caribbean. J Clin Microbiol. 1996 May;34(5):1065–1068. doi: 10.1128/jcm.34.5.1065-1068.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doern G. V. Diagnostic mycobacteriology: where are we today? J Clin Microbiol. 1996 Aug;34(8):1873–1876. doi: 10.1128/jcm.34.8.1873-1876.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Down J. A., O'Connell M. A., Dey M. S., Walters A. H., Howard D. R., Little M. C., Keating W. E., Zwadyk P., Jr, Haaland P. D., McLaurin D. A., 3rd Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA. J Clin Microbiol. 1996 Apr;34(4):860–865. doi: 10.1128/jcm.34.4.860-865.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hellyer T. J., Fletcher T. W., Bates J. H., Stead W. W., Templeton G. L., Cave M. D., Eisenach K. D. Strand displacement amplification and the polymerase chain reaction for monitoring response to treatment in patients with pulmonary tuberculosis. J Infect Dis. 1996 Apr;173(4):934–941. doi: 10.1093/infdis/173.4.934. [DOI] [PubMed] [Google Scholar]
  13. Ichiyama S., Iinuma Y., Tawada Y., Yamori S., Hasegawa Y., Shimokata K., Nakashima N. Evaluation of Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test and Roche PCR-microwell plate hybridization method (AMPLICOR MYCOBACTERIUM) for direct detection of mycobacteria. J Clin Microbiol. 1996 Jan;34(1):130–133. doi: 10.1128/jcm.34.1.130-133.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iovannisci D. M., Winn-Deen E. S. Ligation amplification and fluorescence detection of Mycobacterium tuberculosis DNA. Mol Cell Probes. 1993 Feb;7(1):35–43. doi: 10.1006/mcpr.1993.1005. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Miller N., Hernandez S. G., Cleary T. J. Evaluation of Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test and PCR for direct detection of Mycobacterium tuberculosis in clinical specimens. J Clin Microbiol. 1994 Feb;32(2):393–397. doi: 10.1128/jcm.32.2.393-397.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moore D. F., Curry J. I. Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by Amplicor PCR. J Clin Microbiol. 1995 Oct;33(10):2686–2691. doi: 10.1128/jcm.33.10.2686-2691.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moore D. F., Curry J. I., Knott C. A., Jonas V. Amplification of rRNA for assessment of treatment response of pulmonary tuberculosis patients during antimicrobial therapy. J Clin Microbiol. 1996 Jul;34(7):1745–1749. doi: 10.1128/jcm.34.7.1745-1749.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Noordhoek G. T., van Embden J. D., Kolk A. H. Reliability of nucleic acid amplification for detection of Mycobacterium tuberculosis: an international collaborative quality control study among 30 laboratories. J Clin Microbiol. 1996 Oct;34(10):2522–2525. doi: 10.1128/jcm.34.10.2522-2525.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Schirm J., Oostendorp L. A., Mulder J. G. Comparison of Amplicor, in-house PCR, and conventional culture for detection of Mycobacterium tuberculosis in clinical samples. J Clin Microbiol. 1995 Dec;33(12):3221–3224. doi: 10.1128/jcm.33.12.3221-3224.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shah J. S., Liu J., Buxton D., Hendricks A., Robinson L., Radcliffe G., King W., Lane D., Olive D. M., Klinger J. D. Q-beta replicase-amplified assay for detection of Mycobacterium tuberculosis directly from clinical specimens. J Clin Microbiol. 1995 Jun;33(6):1435–1441. doi: 10.1128/jcm.33.6.1435-1441.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Spargo C. A., Haaland P. D., Jurgensen S. R., Shank D. D., Walker G. T. Chemiluminescent detection of strand displacement amplified DNA from species comprising the Mycobacterium tuberculosis complex. Mol Cell Probes. 1993 Oct;7(5):395–404. doi: 10.1006/mcpr.1993.1058. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Vlaspolder F., Singer P., Roggeveen C. Diagnostic value of an amplification method (Gen-Probe) compared with that of culture for diagnosis of tuberculosis. J Clin Microbiol. 1995 Oct;33(10):2699–2703. doi: 10.1128/jcm.33.10.2699-2703.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vuorinen P., Miettinen A., Vuento R., Hällström O. Direct detection of Mycobacterium tuberculosis complex in respiratory specimens by Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test and Roche Amplicor Mycobacterium Tuberculosis Test. J Clin Microbiol. 1995 Jul;33(7):1856–1859. doi: 10.1128/jcm.33.7.1856-1859.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Walker G. T., Nadeau J. G., Spears P. A., Schram J. L., Nycz C. M., Shank D. D. Multiplex strand displacement amplification (SDA) and detection of DNA sequences from Mycobacterium tuberculosis and other mycobacteria. Nucleic Acids Res. 1994 Jul 11;22(13):2670–2677. doi: 10.1093/nar/22.13.2670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wobeser W. L., Krajden M., Conly J., Simpson H., Yim B., D'costa M., Fuksa M., Hian-Cheong C., Patterson M., Phillips A. Evaluation of Roche Amplicor PCR assay for Mycobacterium tuberculosis. J Clin Microbiol. 1996 Jan;34(1):134–139. doi: 10.1128/jcm.34.1.134-139.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. van der Vliet G. M., Schukkink R. A., van Gemen B., Schepers P., Klatser P. R. Nucleic acid sequence-based amplification (NASBA) for the identification of mycobacteria. J Gen Microbiol. 1993 Oct;139(10):2423–2429. doi: 10.1099/00221287-139-10-2423. [DOI] [PubMed] [Google Scholar]

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

RESOURCES