Sensitivity and specificity of direct and concentrated smear microscopy using culture and PCR based on IS6110 analysis for the detection of acid-fast Bacilli in suspected and having pulmonary tuberculosis

Paper Details

Research Paper 01/08/2012
Views (219) Download (2)

Sensitivity and specificity of direct and concentrated smear microscopy using culture and PCR based on IS6110 analysis for the detection of acid-fast Bacilli in suspected and having pulmonary tuberculosis

Md. Raihan Chowdhury, Kazi Saiful Islam, AGM Rakibuzzaman, Sadia Afrin, Md. Sarowar Jahan
Int. J. Biosci.2( 8), 67-75, August 2012.
Certificate: IJB 2012 [Generate Certificate]


Sensitivity and specificity of direct and concentrated AFB smear microscopy has been investigated by PCR analysis and culture methods to determine a rapid and cheap detection of pulmonary tuberculosis. In this study 305 patients were selected and 915 specimens were collected from suspected and hospital admitted patients. Patients were taken from Dhaka Central Jail Hospital and National TB Hospital, Dhaka, Bangladesh. All samples were smeared and Ziehl-Neelsen method and Lowenstein-Jensen (L-J) method were applied. PCR analysis and culture method was used to confirm the detection. Isolated DNA was used in PCR analysis. In this detection study PCR based IS6110 analysis was developed to identify the pulmonary tuberculosis rapidly and cheaply. A total of 915 samples were prepared for analysis and found 70.47% and 82.85% sensitivity in direct and concentrated smears respectively. Both type of smears (direct and concentrated) showed 100% sensitivity and specificity in culture and PCR IS6110 analysis. Since the direct and concentrated smears with the developed method of PCR IS6110 analysis showed significant detection, so that the proposed method was suggested for the detection analysis of Mycobacterium tuberculosis in pulmonary for the developing and poor countries.


Albert H. 2004. Economic analysis of the diagnosis of smear-negative pulmonary tuberculosis in South Africa: incorporation of a new rapid test, FASTPlaqueTB, into the diagnostic algorithm. Int J Tuberc Lung Dis. 8. 240-247.

Cattamanchi A, Dowdy DW, Davis JL, Worodria W, Yoo S, Joloba M, Matovu J, Hopewell PC and Huang L. 2009. Sensitivity of direct versus concentrated sputum smear microscopy in HIV-infected patients suspected of having pulmonary tuberculosis, BMC Infect Dis. 9, 53.

Contreras A. 1988. Pulmonary infection with NTM. Am Rev Resp Dis. 137, 142-152.

Eisenach, KD, Cave MD, Bates JH, and Crawford JT. 1990. Polymerase chain reaction amplification of a repetitive DNA sequence specific for Mycobacterium tuberculosis. J. Infect. Dis. 161, 977–981.

Jain Amita , Tiwari Vandana, R.S. Guleria, and R.K.Verma. 2002. Qualitative Evaluation of Mycobacterial DNA Extraction Protocols for Polymerase Chain Reaction. Molecular Biology Today 3, 43-50.

Hopewell PC. 1994. Overview of clinical tuberculosis. In: Bloom BR (ed) Tuberculosis: pathogenesis, protection, and control (1st ed). Washington, DC: American Society for Microbiology. pp.25–46.Chapter 3.

Marie Yvette C, Barez, MD, Myrna T, Mendoza MD, Regina S, Celada RMT, Heidi R and Santos RMT. 1995. Accuracy of AFB in Relation to TB Culture in Detection of Pulmonary Tuberculosis, Philippine Journal of Microbiology and Infectious Diseases 24(2), 33-36.

Peterson EM, Nakasone A, Platon-DeLeon JM, Jang Y, de la Maza LM, and Desmond E. 1999. Comparison of Direct and Concentrated Acid-Fast Smears To Identify Specimens Culture Positive for Mycobacterium spp. J Clin Microbiol. 37(11), 3564–3568.

Saceanu,C.A., N.C.Pfeiffer and T. McLean. 1993. Evaluation of sputum smears concentration by cytocentrifugation for detection of acid fast bacilli. J. Clin. Microbiology 31, 2371-2374.

Suárez PG, Floyd K, Portocarrero J, Alarcón E, Rapiti E, Ramos G, Bonilla C, Sabogal I, Aranda I, Dye C, Raviglione M and Espinal MA. 2002. Feasibility and cost-effectiveness of standardised second-line drug treatment for chronic tuberculosis patients: a national cohort study in Peru. Lancet. 359, 1980-1989.

The regional report. 2008. TB in South-East Asian Region (SEAR). World Health Organizaion, p.3.

V J Tevere, P L Hewitt, A Dare, P Hocknell, A Keen, J P Spadoro,and K K Young. 1996. Detection of M. tuberculosis by PCR amplification with pan-Mycobacterium primers and hybridization to an M. tuberculosis-specific probe. J Clin Microbiol. 34(4), 918-923.

Van Cleeff M, Kivihya-Ndugga L, Githui W, Ng’ang’a L, Kibuga D, Odhiambo J and Klatser P. 2005. Cost-effectiveness of polymerase chain reaction versus Ziehl-Neelsen smear microscopy for diagnosis of tuberculosis in Kenya. Int J Tuberc Lung Dis. 9, 877-883.

World Health Organization Report. 2010. Tuberculosis Fact Sheet.

World Health Organization. 2009. “Epidemiology”. Global tuberculosis control: epidemiology, strategy, financing. pp. 6–33.

World Health Organization. 2011. “The sixteenth global report on tuberculosis”.

World Health Organization. 2008. Global tuberculosis control: surveillance, planning, financing. WHO report, World Health Organization.WHO/HTM/TB/2008.393, WHO Report on the tuberculosis epidemic- Groups at risk, Klaudt K (ed) 1996,1-28.