Korean J Clin Microbiol.  2008 Apr;11(1):29-33. 10.5145/KJCM.2008.11.1.29.

Clinical Usefulness of Real-time PCR and Amplicor MTB PCR Assays for Diagnosis of Tuberculosis

• Jung CL
• Kim MK
• Seo DC
• Lee MA

• KMID: 2089805
• DOI: http://doi.org/10.5145/KJCM.2008.11.1.29

Abstract

BACKGROUND: PCR assay has provided a mean of more rapid and sensitive detection of Mycobacterium tuberculosis (MTB) complex than conventional acid- fast bacilli (AFB) smears and MTB cultures. Using the recently developed AdvanSure TB/NTM kit (LG Life Science Diagnostic Division, Korea), which could differentiate nontuberculous mycobacteria (NTM) from MTB, this study compared clinical usefulness of real-time PCR assay and Amplicor MTB PCR assay (Roche Molecular Systems, USA) for diagnosis of tuberculosis.
METHODS
A total of 213 specimens (148 respiratory and 65 nonrespiratory specimens) were tested by using real-time PCR, Amplicor MTB PCR, AFB smear, and MTB culture. The sensitivity and specificity of four methods were evaluated according to clinical diagnosis.
RESULTS
Of six NTM grown in culture, four (67%) were detected by real-time PCR. The overall agreement of real-time and Amplicor MTB PCR was 92% (191/207). The overall sensitivity and specificity were 91% and 87%, respectively, for real-time PCR, and 86% and 93% for Amplicor MTB PCR. In nonrespiratory specimens, the sensitivities of real-time PCR, Amplicor MTB PCR, AFB smear, and MTB culture were 67%, 60%, 13%, and 40%, respectively, and the specificity of the four methods were all 100%.
CONCLUSION
For diagnosis of tuberculosis, the sensitivity and specificity of the real-time PCR assay using AdvanSure TB/NTM kit and Amplicor MTB PCR were similar, and the former could differentiate NTM from MTB. The PCR assay can be considered as a more sensitive technique for the detection of MTB than the conventional AFB smear and culture.

Keyword

Mycobacterium tuberculosis; Nontuberculous mycobacteria; nonrespiratory; Real- time PCR

MeSH Terms

Biological Science Disciplines
Mycobacterium tuberculosis
Nontuberculous Mycobacteria
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Tuberculosis

Reference

References

1. Diagnostic Standards and Classification of Tuberculosis in Adults and Children. This official statement of the American Thoracic Society and the Centers for Disease Control and Prevention was adopted by the ATS Board of Directors, July 1999. This statement was endorsed by the Council of the Infectious Disease Society of America, September 1999. Am J Respir Crit Care Med. 2000; 161:1376–95.

2. Lew WJ. Tuberculosis situation in Korea. Tuberc Respir Dis. 1999; 46:301–10.

3. Lee JY, Choi HJ, Lee H, Joung EY, Huh JW, Oh YM, et al. Recovery rate and characteristics of nontuberculous mycobacterial isolates in a university hospital in Korea. Tuberc Respir Dis. 2005; 58:385–91.

4. Koh WJ, Kwon OJ, Yu CM, Jeon KM, Seo GY, Jeong MP, et al. Recovery rate of nontuberculous mycobacteria from acid-fast-bacilli smear-positive sputum specimens. Tuberc Respir Dis. 2003; 54:22–32.

5. Scientific committee in Korean academy of tuberculosis and respiratory disease. National survey of mycobacterial diseases other than tuberculosis in Korea. Tuberc Respir Dis. 1995; 42:277–94.

6. Cho SN and Brennan PJ. Tuberculosis: diagnostics. Tuberculosis. 2007; 87:S14–7.

7. Cho SN. Current issues on molecular and immunological diagnosis of tuberculosis. Yonsei Med J. 2007; 48:347–59.

8. Woods GL. Molecular techniques in mycobacterial detection. Arch Pathol Lab Med. 2001; 125:122–6.

9. Lee JS, Ji HS, Hong SB, Oh YM, Lim CM, Lee SD, et al. Clinical utility of polymerase chain reaction for the differentiation of nontuberculous mycobacteria in patients with acid-fast bacilli smear-positive specimens. Tuberc Respir Dis. 2005; 58:452–8.

10. Yu CM, Koh WJ, Rye YJ, Jeon K, Choi JC, Kang EH, et al. Usefulness of PCR test for M. tuberculosis for the differentiation of pulmonary tuberculosis and nontuberculous mycobacterial lung disease in patients with smear-positive sputum. Tuberc Respir Dis. 2004; 57:528–34.

11. Centers for Disease Control and Prevention (CDC). Update: Nucleic acid amplification tests for tuberculosis. MMWR Morb Mortal Wkly Rep. 2000; 49:593–4.

12. Michos AG, Daikos GL, Tzanetou K, Theodoridou M, Moschovi M, Nicolaidou P, et al. Detection of Mycobacterium tuberculosis DNA in respiratory and nonrespiratory specimens by the Amplicor MTB PCR. Diagn Microbiol Infect Dis. 2006; 54:121–6.

13. Choi WS, Shin SY, Kim JO, Kim MS, Lee HK. Usefulness of automated PCR test for the detection of Mycobacterium tuberculosis in fresh biopsy tissues. Tuberc Respir Dis. 2006; 61:54–9.

14. Shah S, Miller A, Mastellone A, Kim K, Colaninno P, Hochstein L, et al. Rapid diagnosis of tuberculosis in various biopsy and body fluid specimens by the AMPLICOR Mycobacterium tuberculosis polymerase chain reaction test. Chest. 1998; 113:1190–4.

15. Tortoli E, Tronci M, Tosi CP, Galli C, Lavinia F, Natili S, et al. Multicenter evaluation of two commercial amplification kits (Amplicor, Roche and LCx, Abbott) for direct detection of Mycobacterium tuberculosis in pulmonary and extrapulmonary specimens. Diagn Microbiol Infect Dis. 1999; 33:173–9.

16. Scarparo C, Piccoli P, Rigon A, Ruggiero G, Scagnelli M, Piersimoni C. Comparison of enhanced Mycobacterium tuberculosis amplified direct test with COBAS AMPLICOR Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis complex in respiratory and extrapulmonary specimens. J Clin Microbiol. 2000; 38:1559–62.

17. Levidiotou S, Vrioni G, Galanakis E, Gesouli E, Pappa C, Stefanou D. Four-year experience of use of the Cobas Amplicor system for rapid detection of Mycobacterium tuberculosis complex in respiratory and nonrespiratory specimens in Greece. Eur J Clin Microbiol Infect Dis. 2003; 22:349–56.

18. Rajalahti I, Vuorinen P, Nieminen MM, Miettinen A. Detection of Mycobacterium tuberculosis complex in sputum specimens by the automated Roche Cobas Amplicor Mycobacterium Tuberculosis Test. J Clin Microbiol. 1998; 36:975–8.

19. Brown TJ, Power EG, French GL. Evaluation of three commercial detection systems for Mycobacterium tuberculosis where clinical diagnosis is difficult. J Clin Pathol. 1999; 52:193–7.

20. D'Amato RF, Hochstein LH, Colaninno PM, Scardamaglia M, Kim K, Mastellone AJ, et al. Application of the Roche Amplicor Mycobacterium tuberculosis (PCR) test to specimens other than respiratory secretions. Diagn Microbiol Infect Dis. 1996; 24:15–7.

21. Reischl U, Lehn N, Wolf H, Naumann L. Clinical evaluation of the automated COBAS AMPLICOR MTB assay for testing respiratory and nonrespiratory specimens. J Clin Microbiol. 1998; 36:2853–60.

22. Lee MH. Evaluation of Amplicor PCR kit for direct detection of Mycobacterium tuberculosis from sputum specimens. Korean J Lab Med. 1996; 16:364–72.

23. Lee J, Kim Y, Park J, Ko W, Yang D, Kim S, et al. Clinical utility of bronchial washing PCR for IS6110 and Amplicor for the rapid diagnosis of active pulmonary tuberculosis in smear negative patients. Tuberc Respir Dis. 2001; 50:213–21.

24. Piersimoni C and Scarparo C. Relevance of commercial amplification methods for direct detection of Mycobacterium tuberculosis complex in clinical samples. J Clin Microbiol. 2003; 41:5355–65.