A comparative study of diagnostic yield of Ziehl-Neelsen staining and fluorescent staining in diagnosis of primary pulmonary tuberculosis in a resource limited country like India

Maryam Faridi; Nazish Fatima; Sumit Varshney; Mohammad Shameem; Indu Shukla

doi:10.18203/2394-6040.ijcmph20242163

Authors

Maryam Faridi Department of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
Nazish Fatima Department of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
Sumit Varshney Department of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
Mohammad Shameem Department of Tuberculosis and Other Respiratory Diseases, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
Indu Shukla Department of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India

DOI:

https://doi.org/10.18203/2394-6040.ijcmph20242163

Keywords:

TB, Mycobacterium tuberculosis, ZN staining, Fluorescent microscopy

Abstract

Background: Tuberculosis (TB) is a major public health disease. Early diagnosis and treatment are essential for effective TB control program. In developing countries, diagnosis of TB is mainly by microscopy because of the simplicity of the procedure and cost-effectiveness. In the present study the two methods were compared to derive a conclusion to adopt the better method to provide more effective services for early diagnosis of TB.

Methods: This study was conducted to compare the Ziehl-Neelsen (ZN) staining with Fluorescent staining in the diagnosis of primary pulmonary TB patients. A total of 514 pulmonary tuberculosis suspected patients were included for a period of 2 years. Sputum samples were taken and subjected to both ZN staining and fluorescent staining and results were compared. All samples were subjected to solid culture and taking it as gold standard sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of both the staining methods were defined.

Results: Out of total 514 samples, 265 (51.56%) were ZN positive and 326 (63.43%) samples were positive for FM. Positive samples detected by ZN staining were compared with positive samples by FM. Sensitivity, specificity, PPV, NPV for ZN microscopy were 80.76%, 97.27%, 98.05% and 74.79% respectively. Sensitivity, specificity, PPV, NPV for FM were 98.71%, 95.63%, 97.47% and 97.76% respectively.

Conclusions: In developing country like India where there are large number of cases and gross financial constraints microscopy can be adopted as the main method of diagnosis of pulmonary TB and preferably FM because of higher sensitivity and lesser time taking character of the method.

Metrics

Metrics Loading ...

References

Ba F, Rieder HL. A comparison of fluorescence microscopy with the Ziehl-Neelsen technique in the examination of sputum for acid-fast bacilli. Int J Tuberc Lung Dis. 1999;3(12):1101-5.

Deshmukh SR, Mantri SB, Kendre PB, Nagoba BS. A comparison of sputum examination for acid fast bacilli by modified Schaeffer & Fulton stain, Ziehl-Neelsen stain & cold stain. Indian J Med Res. 1996;103:294-5.

Goyal R, Kumar A. A comparison of Ziehl-Neelsen staining and fluorescent microscopy for diagnosis of pulmonary tuberculosis. IOSR J Dent Med Sci. 2013;8(5):5-8.

World Health Organization. Global tuberculosis report 2023. Available at: https://www.who.int/ teams/global-tuberculosis-programme/tb-reports/ global-tuberculosis-report-2023#:~:text=The%2 0WHO%20Global%20Tuberculosis%20Report,TB%20commitments%2C%20strategies%20and%20targets. Accessed on 05 May 2024.

Ryu YJ. Diagnosis of pulmonary tuberculosis: recent advances and diagnostic algorithms. Tuberc Respir Dis (Seoul). 2015;78(2):64-71.

Steingart KR, Ramsay A, Pai M. Optimizing sputum smear microscopy for the diagnosis of pulmonary tuberculosis. Expert Rev Anti Infect Ther. 2007;5(3):327-31.

Cohen RA, Muzaffar S, Schwartz D, Bashir S, Luke S, McGartland LP, et al. Diagnosis of pulmonary tuberculosis using PCR assays on sputum collected within 24 hours of hospital admission. Am J Respir Crit Care Med. 1998;157(1):156-61.

Mathew P, Kuo YH, Vazirani B, Eng RH, Weinstein MP. Are three sputum acid-fast bacillus smears necessary for discontinuing tuberculosis isolation? J Clin Microbiol. 2002;40(9):3482-4.

Hänscheid T. The future looks bright: low-cost fluorescent microscopes for detection of Mycobacterium tuberculosis and Coccidiae. Trans R Soc Trop Med Hyg. 2008;102(6):520-1.

Kivihya-Ndugga LE, Van Cleeff MR, Githui WA, Nganga LW, Kibuga DK, Odhiambo JA, Klatser PR. 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. 2003;7(12):1163-71.

Bell WJ, Brown PP. Fluorescence microscopy in the laboratory diagnosis and assessment of pulmonary tuberculosis. Central Afr J Med. 1962;8(1):4-9.

Tripathy S, Kumar R, Singh SD. Prevalence of multidrug resistant pulmonary tuberculosis in North Bihar. J Clin Diagnost Res. 2015;9(11):LC09.

Mukherjee A, Saha I, Sarkar A, Chowdhury R. Gender differences in notification rates, clinical forms and treatment outcome of tuberculosis patients under the RNTCP. Lung India. 2012;29(2):120.

Sahoo UC, Singh SK, Basa S, Mohan U. A study of the epidemiological pattern of pulmonary tuberculosis patients undergoing treatment at tuberculosis units of Lucknow. Asian J Biomed Pharm Sci. 2015;20.

Myneedu VP, Singhal R, Khayyam KU, Sharma PP, Bhalla M, Behera D, et al. First and second line drug resistance among treatment naïve pulmonary tuberculosis patients in a district under Revised National Tuberculosis Control Programme (RNTCP) in New Delhi. J Epidemiol Global Health. 2015;5(4):365-73.

Thapa B, Reza LW, Kumar AM, Pandey A, Satyanarayana S, Chadha S. Light Emitting Diode Fluorescence Microscopy increased the detection of smear-positives during follow-up of Tuberculosis patients in India: program implications. BMC Res Notes. 2015;8:1-4.

Hooja S, Pal N, Malhotra B, Goyal S, Kumar V, Vyas L. Comparison of Ziehl Neelsen & Auramine O staining methods on direct and concentrated smears in clinical specimens. Indian J Tuberculosis. 2011;58(2):72-6.

Kelamane S, Mispah P. Sensitivity and Specificity of Conventional Fluorescent Microscopy to diagnose Pulmonary Tuberculosis among Pulmonary tuberculosis symptomatics in Karimnagar. Sch J App Med Sci. 2017;5:391-5.

Singh BK, Sharma SK, Sharma R, Sreenivas V, Myneedu VP, Kohli M, et al. Diagnostic utility of a line probe assay for multidrug resistant-TB in smear-negative pulmonary tuberculosis. PLoS One. 2017;12(8):e0182988.

Laifangbam S, Singh HL, Singh NB, Devi KM, Singh NT. A comparative study of fluorescent microscopy with Ziehl-Neelsen staining and culture for the diagnosis of pulmonary tuberculosis. Kathmandu University Med J. 2009;7(27):226-30.

Laifangbam S, Huidrom LS, Khumanthem SD. Detection of Mycobacterium tuberculosis by three methods and their correlation to chest X-ray findings and CD4 T-lymphocyte counts in human immunodeficiency virus-pulmonary tuberculosis coinfection. J Med Soc. 2013;27(3):203-7.

Bhalla M, Sidiq Z, Sharma PP, Singhal R, Myneedu VP, Sarin R. Performance of light-emitting diode fluorescence microscope for diagnosis of tuberculosis. Int J Mycobacteriol. 2013;2(3):174-8.

Gupta S, Shenoy VP, Bairy I, Muralidharan S. Diagnostic efficacy of Ziehl-Neelsen method against fluorescent microscopy in detection of acid fast bacilli. Asian Pacific J Trop Med. 2010;3(4):328-9.

Bansal R, Sharma PK, Jaryal SC, Gupta PK, Kumar D. Comparison of sensitivity and specificity of ZN and fluorescent stain microscopy with culture as gold standard. J Tuberc Res. 2017;5(02):118.

Timalsina B, Kutu B, Pradhan R, Maharjan B. A Comparative Study on Ziehl-Neelsen Staining (Light Microscopy), Auramine O Staining (Iled-Fluorescent Microscopy) and Culture on LJ Media of Sputum Samples for the Diagnosis of Pulmonary Tuberculosis. Int J Med Biomed Sci. 2015;1(1):4-14.

Suhasini S, Mamilla R. Sensitivity of FM staining versus ZN staining in diagnosing sputum smear positive PTB. IOSR-JDMS. 2015;14(3):29-33.

Goyal R, Kumar A. A comparison of Ziehl-Neelsen staining and fluorescent microscopy for diagnosis of pulmonary tuberculosis. IOSR J Dent Med Sci. 2013;8(5):5-8.

Golia S, Hittinahalli V, Nirmala AR, Sangeetha KT, Asha SK. A comparative study of auramine staining using led fluorescent microscopy with Ziehl-Neelsen staining in the diagnosis of pulmonary tuberculosis. J Evol Med Dent Sci. 2013;2(20):3450-7.