Title: A Comparative Study of Auramine O Staining Using LED Fluorescent Microscopy with Ziehl –Neelsen Staining in the Examination of Sputum For Acid Fast Bacilli
Authors: Okonkwo, R. C, Anyabolu, A. E, Onwunzo M. C, Ifeanyichukwu, M. O*, Chukwuka, C. P1, Enemuo, E, Ngwu A. M
DOI: http://dx.doi.org/10.18535/jmscr/v3i8.14
DIDS : 08.2015-82156638
Abstract
Background: Despite all interventions, tuberculosis (TB) remains a public health issue in Nigeria. Effective TB treatment and control demand early case detection. The ZN staining technique, which is the method of choice in the DOTS program can have problem of low sensitivity. Hence, the need for more sensitive alternative methods. Use of culture is the gold standard but it is not feasible in resource limited settings. The fluorescent Light Emitting Diode (LED) microscopy (FM) method using auramine O stain offers a viable alternative.
Aims and Objectives: To study the efficacy of the fluorescent LED microscopy in the diagnosis of pulmonary tuberculosis (PTB) in comparison with the Z N method and to determine which of the methods is faster in the detection of Acid Fast Bacilli (AFB).
Methodology: Early morning sputum samples were collected from 630 pulmonary tuberculosis suspects (PTB) attending the TB (DOTS) centre of Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria. Two smears, measuring 1x2 cm in diameter, made from each specimen received were stained, one with Ziehl Neelsen staining method and the other with auramine O fluorescent staining method.
Results:Microscopy results obtained from both methods were recorded according to the IUALTD/WHO guideline as adopted by the National Tuberculosis programme (NTP). The results showed comparable sensitivities. The positivity rate was 51/630(8.1%) for ZN and 55/630(8.7%) for FM. However, FM was found to be faster than ZN technique.
Conclusions: The faster reading and the ease of use of the LED fluorescent microscope together with its general acceptability actually support its use in the DOTS program, not as an alternative but as an adjuvant, especially during the introductory phase which should be monitored.
Keywords:Tuberculosis, DOTS, LED, ZN.
References
1. United States Agency for International Development (USAID) Nigeria (2012) HIV/AIDS and Tuberculosis.
2. WHO (2011) Sixteenth global report on Tuberculosis: Executive Summary.
3. United States Embassy Nigeria (2011) Nigeria HIV Fact Sheet, September 2011. http://nigeria.usembassy.org.
4. Federal Ministry of Health (FMOH)(2010) Department of Public Health: National Tuberculosis and Leprosy Control Programme (NTBLCP),Worker’s Manual Revised 5th Edition.
5. Steingart K R, Megan H, Virenne N G. Sputum processing method to improve the sensitivity of smear microscopy for tuberculosis : A Systematic Review. The Lancet Infect. Dis 2009: 6 (10): 664-6774.
6. Mark D P, Giorgio R, Alimuddin Z. Progress towards improved tuberculosis diagnostics for developing countries. Lancet 2006: 367: 942-43.
7. Allen J. A modified Ziehl - Neelse stain for mycobacteria. Medical Laboratory Sciences 1992: 49: 99-1
8. Infectious Disease Survey of America (IDSA). The Forum for Collaborative HIV Research. HIV/TB Co infection: Basic Facts, 2007.
9. International Union Against Tuberculosis and Lung Diseases (IUATLD) technical guide : sputum examination for tuberculosis by direct microscopy in low income countries. 5th ed. 2000.
10. Cambanis A, Ramsay A, Wirkom V, Tata E, Cuevas L E. Investing time in microscopy : an opportunity to optimize smear based case detection of tuberculosis of tuberculosis. Int. J. Tuberc. Lung Dis 2007: 11: 40-45.
11. Cuevas L E, Vassin M A, Al-Sonboli, Lawson N, BAhader J. A multi-country non –inferioriy cluster randomize trial of front –loaded smear microscopy for the diagnosis of pulmonary tuberculosis. PLOS Med 2011: 8 (7): el000403.
12. Saroj H, Nital P, Bharti M, Sumit G, Vipin K, Leela V. Comparison of Ziehl- Neelsen and Auramine staining methods on direct and concentrated smears in clinical specimens. Indian Journal of Tuberculosis 2011: 58:72-76.
13. Reider H L. (1999) A Comparison of Fluorescence Microscopy with Ziehl – Neelsen technique in the examination of Sputum for Acid Fast Bacilli. International Journal of Tuberculosis and Lung Disease 1999: 3 (12): 1101-1105.
14. Steingart K R, Ramsay A, Pai M. Optimizing sputum smear microscopy for the diagnosis of pulmonary tuberculosis. Expert Rev Anti Infect Ther 2007: 5 (3): 327-31.
15. Prasnthi K, Kumari A R. Efficacy of fluorochrome stain in the diagnosis of pulmonary tuberculosis patients coinfected with HIV. Indian J Med Microboil 2005: 23:179-85.
16. Ritu K, Anita P, Ashish K, Asthana, Agrawal R K. (2007). Comparison between ZN and Auramine O staining for detection of Acid Fast Bacilli. Asian Journal of Microbiology, Biotechnology and Environmental Sciences Paper 2007: 9 (4): 1035-1036.
17. Saroj G, Vivek H, Nirmala A R, Sangeetha K T, Asha S, Kamath B. A comparative study of auramine staining using LED fluorescent microscopy with ZN staining in the diagnosis of pulmonary tuberculosis. Journal of Evolution of Medical and Dental Sciences 2013: 2 (20): 3450-6.
18. Bonnet M, Gagnidzel L, Githui W, Guerin P J, Bonte L, Varaine F, Ramnsay A. Performance of LED-Based fluorescence Microscopy to diagnose tuberculosis in a peripheral health centre in Nairobi. PLOS One Journal 2011: 6(2): e17214.