Volume 83, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Optimal tuberculosis testing usually involves sputum centrifugation followed by broth culture. However, centrifuges are biohazardous and scarce in the resource-limited settings where most tuberculosis occurs. To optimize tuberculosis testing for these settings, centrifugation of 111 decontaminated sputum samples was compared with syringe-aspiration through polycarbonate membrane-filters that were then cultured in broth. To reduce the workload of repeated microscopic screening of broth cultures for tuberculosis growth, the colorimetric redox indicator 2,3-diphenyl-5-(2-thienyl) tetrazolium chloride was added to the broth, which enabled naked-eye detection of culture positivity. This combination of filtration and colorimetric growth-detection gave similar results to sputum centrifugation followed by culture microscopy regarding mean colony counts (43 versus 48; = 0.6), contamination rates (0.9% versus 1.8%; = 0.3), and sensitivity (94% versus 95%; = 0.7), suggesting equivalency of the two methods. By obviating centrifugation and repeated microscopic screening of cultures, this approach may constitute a more appropriate technology for rapid and sensitive tuberculosis diagnosis in basic laboratories.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Kent PT, Kubica GP, , 1985. Public Health Mycobacteriology: A Guide for the Level III Laboratory. Atlanta, GA: Centers for Disease Control. [Google Scholar]
  2. Grandjean L, Martin L, Gilman RH, Valencia T, Herrera B, Quino W, Ramos E, Rivero M, Montoya R, Escombe AR, Coleman D, Mitchison D, Evans CA, , 2008. Tuberculosis diagnosis and multidrug resistance testing by direct sputum culture in selective broth without decontamination or centrifugation. J Clin Microbiol 46: 23392344.[Crossref] [Google Scholar]
  3. Mueller DH, Mwenge L, Muyoyeta M, Muvwimi MW, Tembwe R, McNerney R, Godfrey-Faussett P, Ayles HM, , 2008. Costs and cost-effectiveness of tuberculosis cultures using solid and liquid media in a developing country. Int J Tuberc Lung Dis 12: 11961202. [Google Scholar]
  4. Centers for Disease Control and Prevention, 2007. Biosafety in Microbiological and Biomedical Laboratories. Atlanta, GA: Centers for Disease Control and Prevention. [Google Scholar]
  5. Clark DE, , 2001. Safety and the Laboratory Centrifuge. Washington, DC: Division of Chemical Health and Safety, American Chemical Society. [Google Scholar]
  6. World Health Organization, 2002. WHO Global Burden of Disease (GBD) 2002 Estimates (Revised). Geneva: World Health Organization. [Google Scholar]
  7. World Health Organization, 2008. Global Tuberculosis C\control: Surveillance, Planning, Financing. Geneva: World Health Organization. [Google Scholar]
  8. Haley LD, Arch R, , 1957. Use of Millipore membrane filter in the diagnostic tuberculosis laboratory. Am J Clin Pathol 27: 117121. [Google Scholar]
  9. Smithwick RW, Stratigos CB, , 1981. Acid-fast microscopy on polycarbonate membrane filter sputum sediments. J Clin Microbiol 13: 11091113. [Google Scholar]
  10. McKinney RA, , 1958. The preparation of tuberculous sputum for membrane filter filtration. Am Rev Tuberc 77: 10191022. [Google Scholar]
  11. Tietz CJ, , 1950. Filtration as a method of concentrating tubercle bacilli for bacterioscopic examination [in German]. Zentralbl Bakteriol 155: 216221. [Google Scholar]
  12. Tortoli E, Mattei R, Savarino A, Bartolini L, Beer J, , 2000. Comparison of Mycobacterium tuberculosis susceptibility testing performed with BACTEC 460TB (Becton Dickinson) and MB/BacT (Organon Teknika) systems. Diagn Microbiol Infect Dis 38: 8386.[Crossref] [Google Scholar]
  13. Tovar M, Siedner M, Gilman R, Santillan C, Caviedes L, Valencia T, Jave O, Escombe A, Moore D, Evans C, , 2008. Improved diagnosis of pleural tuberculosis using the microscopic-observation drug-susceptibility technique. Clin Infect Dis 46: 909912.[Crossref] [Google Scholar]
  14. Oberhelman RA, Soto-Castellares G, Caviedes L, Castillo ME, Kissinger P, Moore DA, Evans C, Gilman RH, , 2006. Improved recovery of Mycobacterium tuberculosis from children using the microscopic observation drug susceptibility method. Pediatrics 118: e100e106.[Crossref] [Google Scholar]
  15. Moore DA, Evans CA, Gilman RH, Caviedes L, Coronel J, Vivar A, Sanchez E, Pinedo Y, Saravia JC, Salazar C, Oberhelman R, Hollm-Delgado MG, LaChira D, Escombe AR, Friedland JS, , 2006. Microscopic-observation drug-susceptibility assay for the diagnosis of TB. N Engl J Med 355: 15391550.[Crossref] [Google Scholar]
  16. Caviedes L, Lee TS, Gilman RH, Sheen P, Spellman E, Lee EH, Berg DE, Montenegro-James S, , 2000. Rapid, efficient detection and drug susceptibility testing of Mycobacterium tuberculosis in sputum by microscopic observation of broth cultures. The Tuberculosis Working Group in Peru. J Clin Microbiol 38: 12031208. [Google Scholar]
  17. Osato T, Shimizu H, , 1971. Studies on earlier detection of mycobacterial growth by application of STC (2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride). Kekkaku 46: 335340. [Google Scholar]
  18. Brady MF, Coronel J, Gilman RH, Moore DA, , 2008. The MODS Method for Diagnosis of Tuberculosis and Multidrug Resistant Tuberculosis. JoVE 18. Available at: http://www.jove.com/index/details.stp?id=845. [Google Scholar]
  19. Rutjes AW, Reitsma JB, Coomarasamy A, Khan KS, Bossuyt P, , 2007. Evaluation of diagnostic tests when there is no gold standard. A review of methods. Health Technol Assess 11: iii, ix51.[Crossref] [Google Scholar]
  20. Ratnam S, March SB, , 1986. Effect of relative centrifugal force and centrifugation time on sedimentation of mycobacteria in clinical specimens. J Clin Microbiol 23: 582585. [Google Scholar]

Data & Media loading...

  • Received : 18 Sep 2009
  • Accepted : 09 Feb 2010
  • Published online : 05 Oct 2010

Most Cited This Month

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