Evaluation of a commercial rRNA amplification assay for direct detection ofMycobacterium tuberculosis in processed sputum

  • M. T. La Rocco
  • A. Wanger
  • H. Ocera
  • E. Macias
Article

Abstract

A commercial assay (AmplifiedMycobacterium tuberculosis Direct Test, Gen Probe) which combines transcription-mediated amplification of target rRNA with amplicon detection by a chemiluminescent DNA probe for the rapid detection ofMycobacterium tuberculosis in sputum was evaluated. The test was applied to consecutively collected, NALC/NaOH processed sputum sediments from two laboratories (H and L), each serving a different population of patients with pulmonary tuberculosis. Results were compared to those of fluorochrome staining and culture. A total of 760 specimens obtained from 246 patients were used for the study. The test was positive in 141 of 144 (98 %) specimens that were fluorochrome-positive and culture-positive forMycobacterium tuberculosis. Fifteen of 31 specimens that were fluorochromenegative, culture-positive were also assay-positive. A total of 312 specimens (100 patients) from laboratory H (prevalence = 10 %) and 448 specimens (146 patients) from laboratory L (prevalence = 34 %) were analyzed. Compared to culture, test sensitivity, specificity, positive predictive and negative predictive values were 65 %, 99 %, 94 % and 97 %, respectively, for laboratory H and 93 %, 99 %, 99 % and 97 %, respectively, for laboratory L. If the results were analyzed on the basis of at least one concordant result between the amplification assay and culture in three sputum samples per patient, then the sensitivity, specificity, positive and negative predictive values for identifying infected patients was 70 %, 99 %, 87 % and 97 %, respectively, for laboratory H, and 100 %, 98 %, 96 % and 100 %, respectively, for laboratory L.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Huebner RE, Good RC, Tokars JI: Current practices in mycobacteriology: results of a survey of state public health laboratories. Journal of Clinical Microbiology 1993, 31: 771–775.PubMedGoogle Scholar
  2. 2.
    Musial CE, Tice LS, Stockman L, Roberts GD: Identification of mycobacteria from culture by using the Gen-Probe rapid diagnostic system forMycobacterium avium complex andMycobacterium tuberculosis complex. Journal of Clinical Microbiology 1988, 26: 2120–2123.PubMedGoogle Scholar
  3. 3.
    Dewit D, Steyn L, Shoemaker S, Sogin M: Direct detection ofMycobacterium tuberculosis in clinical specimens by DNA amplification. Journal of Clinical Microbiology 1990, 28: 2437–2441.PubMedGoogle Scholar
  4. 4.
    Eisenach KD, Sifford MD, Cave MD, Bates JH, Crawford JT: Detection ofMycobacterium tuberculosis in sputum samples using a polymerase chain reaction. American Reviews of Respiratory Diseases 1990, 144: 1160–1163.Google Scholar
  5. 5.
    Forbes BA, Hicks KS: Direct detection ofMycobacterium tuberculosis in respiratory specimens in a clinical laboratory by polymerase chain reaction. Journal of Clinical Microbiology 1993, 31: 1688–1694.PubMedGoogle Scholar
  6. 6.
    Miyazaki Y, Hironobu K, Kohro S, Kaku M: Nested polymerase chain reaction for detection ofMycobacterium tuberculosis in clinical samples. Journal of Clinical Microbiology 1993, 31: 2228–2232.PubMedGoogle Scholar
  7. 7.
    Pierre C, Lecossier D, Boussougant Y, Bocart D, Joly V, Yeni P, Hance AJ: Use of a reamplification protocol improves sensitivity of detection ofMycobacterium tuberculosis in clinical samples by amplification of DNA. Journal of Clinical Microbiology 1991, 29: 712–717.PubMedGoogle Scholar
  8. 8.
    Jonas V, Alden MJ, Curry JI, Kamisango K, Knott CA, Lankford R, Wolfe JM, Moore DF: Detection and identification ofMycobacterium tuberculosis directly from sputum sediments by amplification of rRNA. Journal of Clinical Microbiology 1993, 31: 2410–2416.PubMedGoogle Scholar
  9. 9.
    Abe C, Hirano K, Wada M, Kazumi Y, Takahashi M, Fukasawa Y, Yoshimura T, Miyagi C, Goto S: Detection ofMycobacterium tuberculosis in clinical specimens by polymerase chain reaction and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test. Journal of Clinical Microbiology 1993, 31: 3270–3274.PubMedGoogle Scholar
  10. 10.
    Kent PT, Kubica GP: Public health mycobacteriology. A guide for the level III laboratory. Centers for Disease Control, Atlanta, 1985, p. 31.Google Scholar
  11. 11.
    Clarridge JE, Shawar RM, Shinnick TM, Plikaytis BB: Large-scale use of polymerase chain reaction for the detection ofMycobacterium tuberculosis in a routine mycobacteriology laboratory. Journal of Clinical Microbiology 1993, 31: 2049–2056.PubMedGoogle Scholar
  12. 12.
    Lipsley BA, Gates J, Tenover FC, Plorde JJ: Factors affecting the clinical value of microscopy for acid-fast bacilli. Reviews of Infectious Diseases 1984, 6: 214–222.PubMedGoogle Scholar
  13. 13.
    Snider DE, Hopewell PC, Mills J, Reichman LB: Mycobacterioses and the acquired immunodeficiency syndrome. American Reviews of Respiratory Diseases 1987, 136: 492–496.Google Scholar
  14. 14.
    Bass JB, Farer LS, Hopewell PC, Jacobs RF, Snider DE: Diagnostic standards and classification of tuberculosis. American Reviews of Respiratory Diseases 1990, 142: 725–735.Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1994

Authors and Affiliations

  • M. T. La Rocco
    • 1
  • A. Wanger
    • 2
  • H. Ocera
    • 1
  • E. Macias
    • 2
  1. 1.Department of PathologySt. Luke's Episcopal HospitalHoustonUSA
  2. 2.University of Texas Medical School at HoustonHoustonUSA

Personalised recommendations