Comparison of Culture, Direct Microscopy, and Polymerase Chain Reaction Results for Detection of Mycobacterium Tuberculosis Complex in Clinical Specimens
Abstract
Aim: Tuberculosis is a chronic, necrotizing disease known since prehistoric times. The most important stage of the tuberculosis control program is the microbiological identification, typing and detection of drug resistance of the Mycobacterium tuberculosis complex for the detection of active cases. The aim of this study is to compare the results of the methods used in the detection of M.tuberculosis complex in clinical samples.
Materials and Methods: The presence of M. tuberculosis complex in various clinical specimens sent to the Medical Microbiology Laboratory from patients with a pre-diagnosis of tuberculosis between January 2016 and January 2022 was investigated retrospectively. In microbiological diagnosis, Ehrlich-Ziehl-Neelsen (EZN) staining method, culture (BACTEC MGIT 320 automated system and Löwenstein Jensen (LJ) medium, and polymerase chain reaction (PCR) tests were used.
Results: A total of 1706 samples were evaluated. The mean age of the patients was 46.49±25.77 years and 1025 (60.1%) were male. EZN method was positive in 32 (1.9%), culture in 48 (2.8%), and PCR in 40 (2.3%) samples. Samples were mostly sent from the departments of chest diseases (52.8%) and pediatrics (24.2%).
Conclusion: Culture, which is the gold standard method, was found to be the most reliable method for demonstrating the presence of M. tuberculosis complex. In the rapid diagnosis of tuberculosis, PCR method gives more reliable results than EZN method. As a result, since the culture method, gives late results, extensive routine use of molecular tests such as PCR is needed for accurate diagnosis of tuberculosis in a short time and to detect drug resistance.
References
- 1. World Health Organization. Global tuberculosis report https://www.who.int/tb/publications/global_report/en/. Accessed 15 October 2019.
- 2. Tomioka H, Namba K. Development of antituberculous drugs. current status and future prospects. Kekkaku 2006;81(12):753-74.
- 3. Baylan O. tensively drug resistant and extremely drug resistant tuberculosis forms after multi-drug resistant tuberculosis: new faces of the old disease. Mikrobiyol Bul 2011;45(1):181-95.
- 4. Colonel A. Mycobacterium: General Characteristics, Laboratory Diagnosis and Staining Procedures. In: Başustaoğlu A. Clinical Microbiology. 1st ed.Atlas Bookstore, Ankara. 2009:543-72.
- 5. Yaman G, Parlak M, Demirol M, Güdücüoğlu H, Berktaş M. Evaluation of EZN Staining and Tuberculosis Culture Results of Various Clinical Samples. Ortadogu Med J 2012;4(1):19-22.
- 6. Ozkara SV. Tuberculosis Laboratory Diagnostic Methods. V Tuberculosis Symposium and VI. Tuberculosis Laboratory Diagnostic Methods Applied Course; 18-21 October 2007; Ankara, Turkey.
- 7. Ceyhan İ, Şimşek H, Tarhan G. Comparison and evaluation of Lowenstein-Jensen medium and 2% Ogawa medium for the diagnosis of tuberculosis. Mikrobiyol Bul 2012;46(1):33-8.
- 8. Pfyffer G, Brown-Elliott BA, Wallace RJ. Mycobacterium: General characteristic, isolation, and staining procedures. Murray PR, Baron EJ, Jorgensen JH (Eds). Manuel of Clinical Microbiology. 8th ed. Washington: ASM Pres, 2003; 532-60.
- 9. Bulut Y. Molecular diagnostic Methods and Innovations. Trabzon mycobacteria days; 2009; Trabzon, Turkey.
- 10. Moore DF, Curry JI. Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by ligase chain reaction. J Clin Microbiol 1998; 36:1028-31.
- 11. Cheng VC, Yew WW, Yuen KY. Molecular diagnostics in tuberculosis. Eur J Clin Microbiol Infect Dis 2005; 24:711-20.
- 12. Alp A. Current status in laboratory diagnosis of tuberculosis. Hacettepe Med J 2011; 42:28-33.
- 13. Centers for Disease Control and Prevention. Updated guidelines for the use of nucleic acid amplification tests in the diagnosis of tuberculosis. Morb Mortal Wkly Rep 2009;58: 7-10.
- 14. Zeka AN, Tasbakan S, Cavusoglu C. Evaluation of the GeneXpert MTB/RIF assay for rapid diagnosis of tuberculosis and detection of rifampin resistance in pulmonary and extrapulmonary specimens. J Clin Microbiol 2011; 49: 4138-41
- 15. Köksalan OK. Tuberculosis diagnosis and its weaknesses today. ANKEM 2010;24:61-3.
- 16. Nyendak MR, Lewinsohn DA, Lewinsohn DM.New diagnostic methods for tuberculosis. Curr Opin Infect Dis 2009;22:174-82.
- 17. Malbruny B, Le Marrec G, Courageux K, Leclercq R, Cattoir V. Rapid and efficient detection of Mycobacterium tuberculosis in respiratory and non-respiratory samples. Int J Tuberc Lung Dis 2011;15:553–55.
- 18. Greco S, Girardi E, Navarra A, Saltini C. Current evidence on diagnostic accuracy of commercially based nucleic acid amplifi cation tests for the diagnosis of pulmonary tuberculosis. Thorax 2006; 61:783–90.
- 19. Kivihya-Ndugga L, van Cleeff M, Juma E, Kimwomi J, Githui W, Oskam L, and Klatser P. Comparison of PCR with the routine procedure for diagnosis of tuberculosis in a population with high prevalences of tuberculosis and human immunodeficiency virus. J Clin Microbiol, 2004;42(3):1012-15.
Comparison of Culture, Direct Microscopy, and Polymerase Chain Reaction Results for Detection of Mycobacterium Tuberculosis Complex in Clinical Specimens
Abstract
Aim: Tuberculosis is a chronic, necrotizing disease known since prehistoric times. The most important stage of the tuberculosis control program is the microbiological identification, typing and detection of drug resistance of the Mycobacterium tuberculosis complex for the detection of active cases. The aim of this study is to compare the results of the methods used in the detection of M.tuberculosis complex in clinical samples.
Materials and Methods: The presence of M. tuberculosis complex in various clinical specimens sent to the Medical Microbiology Laboratory from patients with a pre-diagnosis of tuberculosis between January 2016 and January 2022 was investigated retrospectively. In microbiological diagnosis, Ehrlich-Ziehl-Neelsen (EZN) staining method, culture (BACTEC MGIT 320 automated system and Löwenstein Jensen (LJ) medium, and polymerase chain reaction (PCR) tests were used.
Results: A total of 1706 samples were evaluated. The mean age of the patients was 46.49±25.77 years and 1025 (60.1%) were male. EZN method was positive in 32 (1.9%), culture in 48 (2.8%), and PCR in 40 (2.3%) samples. Samples were mostly sent from the departments of chest diseases (52.8%) and pediatrics (24.2%).
Conclusion: Culture, which is the gold standard method, was found to be the most reliable method for demonstrating the presence of M. tuberculosis complex. In the rapid diagnosis of tuberculosis, PCR method gives more reliable results than EZN method. As a result, since the culture method, gives late results, extensive routine use of molecular tests such as PCR is needed for accurate diagnosis of tuberculosis in a short time and to detect drug resistance.
References
- 1. World Health Organization. Global tuberculosis report https://www.who.int/tb/publications/global_report/en/. Accessed 15 October 2019.
- 2. Tomioka H, Namba K. Development of antituberculous drugs. current status and future prospects. Kekkaku 2006;81(12):753-74.
- 3. Baylan O. tensively drug resistant and extremely drug resistant tuberculosis forms after multi-drug resistant tuberculosis: new faces of the old disease. Mikrobiyol Bul 2011;45(1):181-95.
- 4. Colonel A. Mycobacterium: General Characteristics, Laboratory Diagnosis and Staining Procedures. In: Başustaoğlu A. Clinical Microbiology. 1st ed.Atlas Bookstore, Ankara. 2009:543-72.
- 5. Yaman G, Parlak M, Demirol M, Güdücüoğlu H, Berktaş M. Evaluation of EZN Staining and Tuberculosis Culture Results of Various Clinical Samples. Ortadogu Med J 2012;4(1):19-22.
- 6. Ozkara SV. Tuberculosis Laboratory Diagnostic Methods. V Tuberculosis Symposium and VI. Tuberculosis Laboratory Diagnostic Methods Applied Course; 18-21 October 2007; Ankara, Turkey.
- 7. Ceyhan İ, Şimşek H, Tarhan G. Comparison and evaluation of Lowenstein-Jensen medium and 2% Ogawa medium for the diagnosis of tuberculosis. Mikrobiyol Bul 2012;46(1):33-8.
- 8. Pfyffer G, Brown-Elliott BA, Wallace RJ. Mycobacterium: General characteristic, isolation, and staining procedures. Murray PR, Baron EJ, Jorgensen JH (Eds). Manuel of Clinical Microbiology. 8th ed. Washington: ASM Pres, 2003; 532-60.
- 9. Bulut Y. Molecular diagnostic Methods and Innovations. Trabzon mycobacteria days; 2009; Trabzon, Turkey.
- 10. Moore DF, Curry JI. Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by ligase chain reaction. J Clin Microbiol 1998; 36:1028-31.
- 11. Cheng VC, Yew WW, Yuen KY. Molecular diagnostics in tuberculosis. Eur J Clin Microbiol Infect Dis 2005; 24:711-20.
- 12. Alp A. Current status in laboratory diagnosis of tuberculosis. Hacettepe Med J 2011; 42:28-33.
- 13. Centers for Disease Control and Prevention. Updated guidelines for the use of nucleic acid amplification tests in the diagnosis of tuberculosis. Morb Mortal Wkly Rep 2009;58: 7-10.
- 14. Zeka AN, Tasbakan S, Cavusoglu C. Evaluation of the GeneXpert MTB/RIF assay for rapid diagnosis of tuberculosis and detection of rifampin resistance in pulmonary and extrapulmonary specimens. J Clin Microbiol 2011; 49: 4138-41
- 15. Köksalan OK. Tuberculosis diagnosis and its weaknesses today. ANKEM 2010;24:61-3.
- 16. Nyendak MR, Lewinsohn DA, Lewinsohn DM.New diagnostic methods for tuberculosis. Curr Opin Infect Dis 2009;22:174-82.
- 17. Malbruny B, Le Marrec G, Courageux K, Leclercq R, Cattoir V. Rapid and efficient detection of Mycobacterium tuberculosis in respiratory and non-respiratory samples. Int J Tuberc Lung Dis 2011;15:553–55.
- 18. Greco S, Girardi E, Navarra A, Saltini C. Current evidence on diagnostic accuracy of commercially based nucleic acid amplifi cation tests for the diagnosis of pulmonary tuberculosis. Thorax 2006; 61:783–90.
- 19. Kivihya-Ndugga L, van Cleeff M, Juma E, Kimwomi J, Githui W, Oskam L, and Klatser P. Comparison of PCR with the routine procedure for diagnosis of tuberculosis in a population with high prevalences of tuberculosis and human immunodeficiency virus. J Clin Microbiol, 2004;42(3):1012-15.
Details
| Primary Language | English |
|---|---|
| Subjects | Clinical Sciences |
| Journal Section | Original Article |
| Authors | |
| Early Pub Date | October 24, 2022 |
| Publication Date | October 29, 2022 |
| Submission Date | July 9, 2022 |
| Published in Issue | Year 2022 Volume: 32 Issue: 5 |
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