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



Poor laboratory equipment and few human resources have made it difficult to implement microscopic diagnosis of pulmonary tuberculosis (TB) on a large scale basis worldwide. Three hundred sputum samples from patients in Cameroon were studied by using the CyScope, a new light-emitting, diode-based, fluorescence microscope, to compare auramine-rhodamine fluorescence with the conventional Ziehl-Neelsen staining method. Five fluorescence protocols were tested to reduce manipulation time. Smear positivity for acid-fast bacilli with the Ziehl-Neelsen staining method was 27.7% (83 of 300) compared with 33.3% (100 of 300) with the fluorescent method. Staining time with the modified fluorescence protocol could be reduced from 21 minutes to 10 minutes. This study confirmed that the fluorescence staining method is more sensitive than the Ziehl-Neelsen staining method. It is suggested that the training of laboratory technicians on fluorescence microscopy should be scaled up for increased disease control.


Article metrics loading...

Loading full text...

Full text loading...



  1. World Health Organization, 2009. Global Tuberculosis Control: Epidemiology, Strategy, Financing: WHO Report 2009. Geneva: World Health Organization, WHO.HTM/TB/2009.411.
  2. Sohn H, Minion J, Albert H, Dheda K, Pai M, , 2009. TB diagnostic tests: how do we figure out their costs? Expert Rev Anti Infect Ther 7: 723733.[Crossref]
  3. Harries AD, , 1998. An approach to the problems of diagnosing and treating adult smear-negative pulmonary tuberculosis in high-HIV-prevalence settings in sub-Saharan Africa. Bull World Health Organ 76: 651662.
  4. Getahun H, Harrington M, O'Brien R, Nunn P, , 2007. Diagnosis of smear-negative pulmonary tuberculosis in people with HIV infection or AIDS in resource-constrained settings: informing urgent policy changes. Lancet 369: 20422049.[Crossref]
  5. Göhde W, , 2008. A Fluorescence-Based Rapid Malaria Test for Resource-Constrained Areas. Clinical Laboratory International 32 (5). Available at: http://dev.cli-online.com/feature-articles/clinical-laboratory-articles/-6ec76ad800/index.html. Accessed July 30, 2010.
  6. Mizuno K, Chikamatsu K, Aono A, Azuma Y, Yamada H, Mitarai S, , 2009. Clinical evaluation of acid-fast smear examination with light emitting diode fluorescent microscopy. Kekkaku 84: 627629.
  7. World Health Organization, 1998. Laboratory Service in Tuberculosis Control. Part II. Microscopy. Geneva: World Health Organization.
  8. Angra P, Becx-Bleumink M, Gilpin C, Joloba M, Jost K, Kam KM, Kim SJ, Lumb R, Mitarai S, Ramsay A, Ridderhof J, Rieder HL, Selvakumar N, , 2007. Ziehl-Neelsen staining: strong red on weak blue or weak red under strong blue? Int J Tuberc Lung Dis 11: 11601161.
  9. Degommier J, , 1959. Fluorescence and relative acid-alcohol resistance of tubercule bacillus [in French]. Ann Inst Pasteur 96: 723733.
  10. Ait-Khaled N, Enarson D, , 1999. Tuberculose: Manuel pour les Étudiants en Médecine. Geneva: World Health Organization, WHO/CDS/TB/99.272.
  11. Githui W, Kitui F, Juma ES, Obwana DO, Mwai J, Kwamanga D, , 1993. A comparative study on the reliability of the fluorescence microscopy and Ziehl-Neelsen method in the diagnosis of pulmonary tuberculosis. East Afr Med J 70: 263265.
  12. Gebre N, Karisson U, Jonsson G, Macadem R, Wolde A, Assefa A, , 1995. Improved microscopical diagnosis of pulmonary tuberculosis in developping countries. Trans R Soc Trop Med Hyg 89: 191193.[Crossref]
  13. Noeske J, Dopico E, Torrea G, Wang H, Van Deun A, , 2009. Two vs. three sputum samples for microscopic detection of tuberculosis in high HIV prevalence population. Int J Tuberc Lung Dis 13: 842847.
  14. Ba F, Rieder HL, , 1999. A comparison of fluorescence microscopy with the Ziehl-Neelsen technique in the examination of sputum for acid-fast bacilli. Int J Tuberc Lung Dis 3: 11011105.
  15. Crampin C, Floyd SB, Mwaungulu F, Black G, Ndhlovu R, Mwaiyeghele E, Glynn JR, Warndorff DK, Fine PE, , 2001. Comparison of two versus three smears in identifying culture-positive tuberculosis patients in rural African setting with high HIV prevalence. Int J Tuberc Lung Dis 5: 994999.
  16. Kivihya-Ndugga LE, Van Cleeff MR, Githui WA, Nganga LW, Kibuga DK, Odhiambo JA, Klatser PR, , 2003. A comprehensive comparison of Ziehl-Neelsen and fluorescence microscopy for the diagnosis of tuberculosis in a resource-poor urban setting. Int J Tuberc Lung Dis 7: 11631171.
  17. Vamseedhar A, Mohan K, II Rekha BP, , 2009. Comparison of the modified fluorescent method and conventional Ziehl-Neelsen method in the detection of acid fast bacilli in lymphnode aspirates. Cytojournal 6: 13.[Crossref]
  18. Brändli O, , 2008. Comment diagnostiquer la tuberculose en 2008? Forum Med Suisse 8: 564568.
  19. Levy H, Sacho H, Kallenbach J, , 1969. A reevaluation of sputum microscopy and culture in the diagnosis of pulmonary tuberculosis. Chest 96: 11931197.
  20. Colebunders R, Bastian I, , 2000. A review of the diagnosis and treatment of smear-negative pulmonary tuberculosis. Int J Tuberc Lung Dis 4: 97107.
  21. Van Deun A, , 2005. Tuberculosis bacteriology—priorities and indications in high prevalence countries: position of the technical staff of the Tuberculosis Division of the International Union Against Tuberculosis and Lung Disease. Int J Tuberc Lung Dis 9: 355361.
  22. Rieder HL, Van Deun A, Kam KM, Kim SJ, Chonde TM, Trébucq A, Urbanczik R, , 2007. Priorities for Tuberculosis Bacteriology Services in Low-Income Countries. Second edition. Paris: International Union Against Tuberculosis and Lung Disease, 2829.
  23. Truant JP, Brett WA, Thomas W, Jr, 1962. Fluorescence microscopy of tubercle bacilli stained with auramine and rhodamine. Henry Ford Hosp Med Bull 10: 287296.
  24. Hänscheid T, , 2008. The future looks bright: low-cost fluorescent microscopes for detection of Mycobacterium tuberculosis and Coccidiae. Trans R Soc Trop Med Hyg 102: 520521.[Crossref]

Data & Media loading...

  • Received : 09 Mar 2010
  • Accepted : 19 Jul 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