Title: Comparison of Mycobacterium tuberculosis complex Yield and Contamination Rates using Lowenstein-Jensen with and without Antibiotics in Western Kenya

Authors: Okumu A, McCarthy K, Orwa J, Williamson J, Musau S, Alexander H, Cavanaugh S, Modi S, Cain K

 DOI:  https://dx.doi.org/10.18535/jmscr/v5i8.86

Abstract

Background: While molecular methods have been recently endorsed for diagnosis of tuberculosis (TB), mycobacterial culture remains the gold standard. Lowenstein-Jensen (LJ) is often used for the cultivation of Mycobacterium tuberculosis complex (MTBC); however contamination often renders a subset of cultures useless. We compared the MTBC yield and contamination rate of processed sputum inoculated on LJ with antibiotics (LJ PACT) to LJ without antibiotics (LJ).

Methodology: Sputum samples were obtained from people living with HIV enrolled in a TB screening study in western Kenya, processed using NALC/NaOH-Na citrate, then inoculated on LJ PACT and LJ media. Cultures were evaluated weekly with growth identified as acid-fast bacilli by Ziehl-Neelsen bright-field microscopy. MTBC and nontuberculous mycobacteria (NTM) were identified by immunochromatographic and line probe assays.

Results: A total of 700 sputum samples were cultured on both LJ PACT and LJ between March and June 2012. Of those cultured on LJ PACT, 29 (4.1%) grew MTBC, 613 (87.6%) were negative, 12 (1.7%) grew NTM, and 46 (6.6%) were contaminated; on LJ, 28 (4%) grew MTBC, 553 (79%) were negative, 9 (1.3%) grew NTM, and 110 (15.7%) were contaminated. The difference in contamination on LJ PACT and LJ was statistically significant (p‹0.0001), while the difference in MTBC growth was not (p=0.566).

Keywords: pulmonary tuberculosis, diagnosis, culture techniques, Nontuberculous mycobacteria, cross-sectional studies, Kenya, Lowenstein-Jensen.

References

  1. Tenover, F.C., et al., The resurgence of tuberculosis: is your laboratory ready? J Clin Microbiol, 1993. 31(4): p. 767-70.
  2. World Health Organization Global Tuberculosis Report 2015. http://apps.who.int/iris/bitstream/10665/191102/1/9789241565059_eng.pdf.
  3. Perkins, M.D., New diagnostic tools for tuberculosis. Int J Tuberc Lung Dis, 2000. 4(12 Suppl 2): p. S182-8.
  4. Wilson, M.L., Recent advances in the laboratory detection of Mycobacterium tuberculosis complex and drug resistance. Clin Infect Dis, 2011. 52(11): p. 1350-5.
  5. Parsons, L.M., et al., Laboratory diagnosis of tuberculosis in resource-poor countries: challenges and opportunities. Clin Microbiol Rev, 2011. 24(2): p. 314-50.
  6. Steingart, K.R., A. Ramsay, and M. Pai, Optimizing sputum smear microscopy for the diagnosis of pulmonary tuberculosis. Expert Rev Anti Infect Ther, 2007. 5(3): p. 327-31.
  7. Cavanaugh, J.S., et al., Survival among patients with HIV infection and smear-negative pulmonary tuberculosis - United States, 1993-2006. PLoS One, 2012. 7(10): p. e47855.
  8. Colebunders, R. and I. Bastian, A review of the diagnosis and treatment of smear-negative pulmonary tuberculosis. Int J Tuberc Lung Dis, 2000. 4(2): p. 97-107.
  9. Barnes, P.F., et al., Tuberculosis in patients with human immunodeficiency virus infection. N Engl J Med, 1991. 324(23): p. 1644-50.
  10. Getahun, H., et al., Diagnosis of smear-negative pulmonary tuberculosis in people with HIV infection or AIDS in resource-constrained settings: informing urgent policy changes. Lancet, 2007. 369(9578): p. 2042-9.
  11. Monkongdee, P., et al., Yield of Acid-fast Smear and Mycobacterial Culture for Tuberculosis Diagnosis in People with HIV. Am J Respir Crit Care Med, 2009.
  12. WHO, Laboratory services in tuberculosis control. Culture part III. 1998, Geneva, Switzerland: World Health Organization.
  13. Stop TB Department, W.H.O. Seventh meeting. Strategic and technical advisory group for tuberculosis (STAG-TB) report on conclusions and recommendations. 2007. Geneva, Switzerland: WHO.
  14. Alfred, N., et al., Optimising Mycobacterium tuberculosis detection in resource limited settings. BMJ Open, 2014. 4(3): p. e004093.
  15. Duque, A., et al., Evaluation of the BD Bactec MGIT 320 system for detection of mycobacteria and drug susceptibility testing of Mycobacterium tuberculosis. J Clin Microbiol, 2013. 51(10): p. 3403-5.
  16. Chihota, V.N., et al., Liquid vs. solid culture for tuberculosis: performance and cost in a resource-constrained setting. Int J Tuberc Lung Dis, 2010. 14(8): p. 1024-31.
  17. World Health Organization. Annex 4 TB burden, etsimates, notifications and treatment outcomes for individual countries and territories, WHO regions and the world. Global Tuberculosis Report 2016.
  18. Joloba, M.L., et al., What is the most reliable solid culture medium for tuberculosis treatment trials? Tuberculosis (Edinb), 2014. 94(3): p. 311-6.
  19. Heilig, C.M., et al., How we determined the most reliable solid medium for studying treatment of tuberculosis. Tuberculosis (Edinb), 2014. 94(3): p. 317-22.
  20. Kassaza, K., et al., Lowenstein-Jensen selective medium for reducing contamination in Mycobacterium tuberculosis culture. J Clin Microbiol, 2014. 52(7): p. 2671-3.
  21. Modi, S., et al., Performance of Clinical Screening Algorithms for Tuberculosis Intensified Case Finding among People Living with HIV in Western Kenya. PLoS One, 2016. 11(12): p. e0167685.
  22. Cavanaugh, J.S., et al., Comparative Yield of Different Diagnostic Tests for Tuberculosis among People Living with HIV in Western Kenya. PLoS One, 2016. 11(3): p. e0152364.
  23. Zeger, S.L. and K.Y. Liang, Longitudinal data analysis for discrete and continuous outcomes. Biometrics, 1986. 42(1): p. 121-30.
  24. Duwe, A.K., et al., In vitro cytotoxicity and antibiotic activity of polymyxin B nonapeptide. Antimicrob Agents Chemother, 1986. 30(2): p. 340-1.
  25. Zavascki, A.P., et al., Polymyxin B for the treatment of multidrug-resistant pathogens: a critical review. J Antimicrob Chemother, 2007. 60(6): p. 1206-15.
  26. Swenson, C.E., et al., In vitro and in vivo antifungal activity of amphotericin B lipid complex: are phospholipases important? Antimicrob Agents Chemother, 1998. 42(4): p. 767-71.
  27. Sainsbury, S., et al., Crystal structures of penicillin-binding protein 3 from Pseudomonas aeruginosa: comparison of native and antibiotic-bound forms. J Mol Biol, 2011. 405(1): p. 173-84.
  28. Karpanoja, P., et al., Connection between trimethoprim-sulfamethoxazole use and resistance in Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Antimicrob Agents Chemother, 2008. 52(7): p. 2480-5.
  29. Piersimoni, C., et al., Multicenter evaluation of the MB-Redox medium compared with radiometric BACTEC system, mycobacteria growth indicator tube (MGIT), and Lowenstein-Jensen medium for detection and recovery of acid-fast bacilli. Diagn Microbiol Infect Dis, 1999. 34(4): p. 293-9.
  30. Saddiqi, S.H., MGITTM Procedure Manual. For BACTECTM MGIT 960TM TB System. 2005. p. 81-84.
  31. Ssengooba, W., et al., An Early Morning Sputum Sample Is Necessary for the Diagnosis of Pulmonary Tuberculosis, Even with More Sensitive Techniques: A Prospective Cohort Study among Adolescent TB-Suspects in Uganda. Tuberc Res Treat, 2012. 2012: p. 970203.
  32. Islam, M.R., et al., Yield of two consecutive sputum specimens for the effective diagnosis of pulmonary tuberculosis. PLoS One, 2013. 8(7): p. e67678.
  33. World Health Organization. Automated real-time nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance: Xpert MTB/RIF assay for the diagnosis of pulmonary and extrapulmonary TB in adults and children: Policy update. 2013.
  34. Lima, D.M., et al., Identification of mycobacterium species in contaminated cultures by polymerase chain reaction. Chest, 2005. 127(4): p. 1283-8.
  35. Perkins, M.D. and J. Cunningham, Facing the crisis: improving the diagnosis of tuberculosis in the HIV era. J Infect Dis, 2007. 196 Suppl 1: p. S15-27.
  36. Rodrigues, C., et al., Evaluation of the bactec MGIT 960 TB system for recovery and identification of Mycobacterium tuberculosis complex in a high through put tertiary care centre. Indian J Med Microbiol, 2009. 27(3): p. 217-21.
  37. McCarthy, K.D., et al., Nontuberculous mycobacterial disease in patients with HIV in Southeast Asia. Am J Respir Crit Care Med, 2012. 185(9): p. 981-8.
  38. Oramasionwu, G.E., et al., The utility of stool cultures for diagnosing tuberculosis in people living with the human immunodeficiency virus. Int J Tuberc Lung Dis, 2013. 17(8): p. 1023-8.

Corresponding Author

Albert Okumu

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Kevin Cain

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