1887

Abstract

The role of the polymerase chain reaction (PCR) in the diagnosis of tuberculosis in clinical practice remains to be defined: most results have been based on sputum samples. This study systematically compared the relative sensitivity and specificity of a single simplified method for different clinical samples. A wide range of clinical samples, including sputum, bronchoalveolar lavage fluid, cerebrospinal fluid, pleural fluid, gastric aspirate, pus and tissues (both fresh and paraffin-embedded) was tested. This method did not require routine DNA extraction before PCR, and consisted of an optimised single tube PCR amplification designed with different sets of time and temperature profiles. A total of 398 samples from 293 patients was studied. The sensitivity was 100% for all types of specimens, while the specificity ranged from 95% for sputum to 88% for bronchoalveolar lavage fluid and pleural fluid and to 85% for non-pulmonary specimens. This study showed that it was possible to employ a single simplified method with minor modifications for a wide range of specimens in clinical practice without loss of sensitivity and specificity.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-46-2-164
1997-02-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jmm/46/2/medmicro-46-2-164.html?itemId=/content/journal/jmm/10.1099/00222615-46-2-164&mimeType=html&fmt=ahah

References

  1. 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; 161:977–981
    [Google Scholar]
  2. Pao C. C., Yen T. S. B., You J.-B., Maa J.-S., Fiss E. H., Chang C.-H. Detection and identification of Mycobacterium tuberculosis by DNA amplification. J Clin Microbiol 1990; 28:1877–1880
    [Google Scholar]
  3. Böddinghaus B., Rogall T., Flohr T., Blöcker H., Böttger E. C. Detection and identification of Mycobacteria by amplification of rRNA. J Clin Microbiol 1990; 28:1751–1759
    [Google Scholar]
  4. Manjunath N., Shankar P., Rajan L., Bhargava A., Saluja S. Shriniwas. Evaluation of a polymerase chain reaction for the diagnosis of tuberculosis. Tubercle 1991; 72:21–27
    [Google Scholar]
  5. 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; 30:255–258
    [Google Scholar]
  6. Kolk A. H. J., Schuitema A. R. J., Kuijper S. Detection of Mycobacterium tuberculosis in clinical samples by using polymerase chain reaction and a nonradioactive detection system. J Clin Microbiol 1992; 30:2567–2575
    [Google Scholar]
  7. Soini H., Skumik M., Liippo K., Tala E., Viljanen M. K. Detection and identification of Mycobacteria by amplification of a segment of the gene coding for the 32-kilodalton protein. J Clin Microbiol 1992; 30:2025–2028
    [Google Scholar]
  8. Miyazaki Y., Koga H., Kohno S., Kaku M. Nested polymerase chain reaction for detection of Mycobacterium tuberculosis in clinical samples. J Clin Microbiol 1993; 31:2228–2232
    [Google Scholar]
  9. Pierre C., Lecossier D., Boussougant Y. Use of a reamplification protocol improves sensitivity of detection of Mycobacterium tuberculosis in clinical samples by amplification of DNA. J Clin Microbiol 1991; 29:712–717
    [Google Scholar]
  10. 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; 30:1331–1334
    [Google Scholar]
  11. Liedtke W., Opalka B., Zimmermann C. W., Schmid E. Different methods of sample preparation influence sensitivity of Mycobacterium tuberculosis and Borrelia burgdorferi PCR. PCR Methods Applic 1994; 3:301–304
    [Google Scholar]
  12. Wilson S. M., McNerney R., Nye P. M., Godfrey-Faussett P. D., Stoker N. G., Voller A. Progress toward a simplified polymerase chain reaction and its applification to diagnosis of tuberculosis. J Clin Microbiol 1993; 31:776–782
    [Google Scholar]
  13. Pierre C., Olivier C., Lecossier D., Boussougant Y., Yeni P., Hance A. J. Diagnosis of primary tuberculosis in children by amplification and detection of mycobacterial DNA. Am Rev Respir Dis 1993; 147:420–424
    [Google Scholar]
  14. Schluger N. W., Kinney D., Harkin T. J., Rom W. N. Clinical utility of the polymerase chain reaction in the diagnosis of infections due to Mycobacterium tuberculosis . Chest 1994; 105:1116–1121
    [Google Scholar]
  15. 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; 30:1514–1517
    [Google Scholar]
  16. de Lassence A., Lecossier D., Pierre C., Cadranel J., Stern M., Hance A. J. Detection of mycobacterial DNA in pleural fluid from patients with tuberculous pleurisy by means of the polymerase chain reaction: comparison of two protocols. Thorax 1992; 47:265–269
    [Google Scholar]
  17. Shawar R. M., El-Zaatari F. A. K., Nataraj A., Clarridge J. E. Detection of Mycobacterium tuberculosis in clinical samples by two-step polymerase chain reaction and nonisotopic hybridization methods. J Clin Microbiol 1993; 31:61–65
    [Google Scholar]
  18. Sritharan V., Barker R. H. A simple method for diagnosing M. tuberculosis infection in clinical samples using PCR. Mol Cell Probes 1991; 5:385–395
    [Google Scholar]
  19. Clarridge J. E., 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; 31:2049–2056
    [Google Scholar]
  20. Noordhoek G. T., Kolk A. H., Bjune G. Sensitivity and specificity of PCR for detection of Mycobacterium tuberculosis: a blind comparison study among seven laboratories. J Clin Microbiol 1994; 32:277–284
    [Google Scholar]
  21. Beige J., Lokies J., Schaberg T. Clinical evaluation of a Mycobacterium tuberculosis PCR assay. J Clin Microbiol 1995; 33:90–95
    [Google Scholar]
  22. Yuen K. Y., Chan K. S., Chan C. M. Use of PCR in routine diagnosis of treated and untreated pulmonary tuberculosis. J Clin Pathol 1993; 46:318–322
    [Google Scholar]
  23. Thierry D., Cave M. D., Eisenach K. D. IS6110, an IS-like element of Mycobacterium tuberculosis complex . Nucleic Acids Res 1990; 18:188
    [Google Scholar]
  24. 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; 144:1160–1163
    [Google Scholar]
  25. Higuchi R. Simple and rapid preparation of samples for PCR. In Erlich H. A. (ed) PCR technology – principles and applications for DNA amplification New York: Stockton Press; 198931–38
    [Google Scholar]
  26. Ng W. C., Guan R., Tan M. F. Hepatitis C virus genotypes in Singapore and Indonesia. J Viral Hepat 1995; 2:203–209
    [Google Scholar]
  27. Yap E. P. H., McGee J. O’D. Short PCR product yields improved by lower denaturation temperatures. Nucleic Acids Res 1991; 19:1713
    [Google Scholar]
  28. Ford N., Nolan C., Ferguson M. Analysis and cloning of eukaryotic genomic DNA. In Sambrook J., Fritsch E. F., Maniatis T. (eds) Molecular cloning: a laboratory manual 2nd edn Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 198945–46
    [Google Scholar]
  29. Kolk A. H. J., Noordhoek G. T., de Leeuw O., Kuijper S., van Embden J. D. A. Mycobacterium smegmatis strain for detection of Mycobacterium tuberculosis by PCR used as internal control for inhibition of amplification and for quantification of bacteria. J Clin Microbiol 1994; 32:1354–1356
    [Google Scholar]
  30. de Wit D., Wootton M., Allan B., Steyn L. Simple method for production of internal control DNA for Mycobacterium tuberculosis polymerase chain reaction assays. J Clin Microbiol 1993; 31:2204–2207
    [Google Scholar]
  31. Roberts G. D., Thompson G. R. Bacteriology and bacteriologic diagnosis of tuberculosis. In David Schlossberg. (ed) Tuberculosis New York: Springer-Verlag; 199355
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-46-2-164
Loading
/content/journal/jmm/10.1099/00222615-46-2-164
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error