The Journal of Medical Sciences

Register      Login

SEARCH WITHIN CONTENT

FIND ARTICLE

Volume / Issue

Online First

Archive
Volume  10 (2024)
Volume  9 (2023)
Volume  8 (2022)
Volume  7 (2021)
Volume  6 (2020)
Volume  5 (2019)
Volume  4 (2018)
Volume  3 (2017)
Volume  2 (2016)
Volume  1 (2015)
Related articles

VOLUME 10 , ISSUE 1--4 ( January-December, 2024 ) > List of Articles

RESEARCH ARTICLE

Comparison of Two Staining Methods (Ziehl–Neelsen Staining and Auramine O Staining) in Detecting Mycobacterium tuberculosis in Cytology Samples: A Cross-sectional Study

Debalina Roy, Sumitha Maniyan Prakash, Prabhakara Somanna, Jyothi Ananth Raj, Nikita Malave, Kantinedu Susairaj Jenefer Theresa

Keywords : Acid-fast bacilli, Auramine O stain, Cross-sectional study, Cytology, Fluorescent microscopy, Tuberculosis, Ziehl–Neelsen stain

DOI: 10.5005/jp-journals-10045-00305

License: CC BY-NC 4.0

Published Online: 15-11-2024

Copyright Statement:  Copyright © 2024; The Author(s).


Abstract

Aims and background: Tuberculosis is an important infectious disease of human beings from time immemorial. The conventional Ziehl–Neelsen (ZN) method for acid-fast bacilli (AFB) is an economical method for the diagnosis of tuberculosis. Fluorescent microscopy using Auramine O stain plays an important role in the detection of Mycobacterium because it is less time-consuming and lower magnifications can be used to examine smears. Our study aims to detect the accuracy of these techniques on various cytology samples received. Materials and methods: A total of 50 cytology samples collected from suspected cases of tuberculosis were included in the study. All samples were stained with both ZN stain and Auramine O stain and examined under a light microscope and fluorescent microscope respectively. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of both staining techniques were calculated and compared with the standard cartridge-based nucleic acid amplification test (CBNAAT) report. Results: Out of the total 50 samples, 35 (70.0%), 10 (20.0%), 4 (8.0%), and 1 (2.0%), are bronchoalveolar lavage (BAL) fluid, lymph node aspirates, pleural, and ascitic fluid respectively, of which, 39 (78%) were positive and 11 (22%) were negative by CBNAAT. In comparison with CBNAAT, the percentage of true positives was greater for the Auramine O stain than ZN stain (94.87 vs 74.4%). The PPV, NPV, and smear positivity of the Auramine O stain were higher compared to the ZN stain (97.4 vs 93.5, 83.3 vs 47.4, and 76 vs 62%, respectively). Conclusion: Auramine O staining with fluorescent microscopy has better accuracy compared to ZN staining with light microscopy for the diagnosis of tuberculosis in cytology specimens. Hence, fluorescent staining may be considered a superior investigation considering the importance of the early diagnosis and treatment of tuberculosis. Clinical significance: Auramine O staining with fluorescent microscopy can be utilized as a better routine investigation method for the diagnosis of tuberculosis bacilli in cytology samples.

PDF Share
  1. Bhandari R, Gaur DS, Kotwal A, et al. Comparison of Ziehl–Neelsen (ZN) staining and fluorescent (FL) staining in suspected cases of tuberculosis. Int J Pathol Clin Res 2021;7(1). DOI: 10.23937/2469-5807/1510122
  2. Ahmad F, Rehman A, Jadoon O. Histological comparison of two staining methods in identification of mycobacterium tuberculosis in granulomatous lymphadenitis. J Ayub Med Coll Abbottabad 2018;30(2):159–162. PMID: 29938410.
  3. Thakur B, Mehrotra R, Nigam JS. Correlation of various techniques in diagnosis of tuberculous lymphadenitis on fine needle aspiration cytology. Patholog Res Int 2013;2013:824620. DOI: 10.1155/2013/824620
  4. Kumar S, Gupta R, Kaushik N, et al. Role of autofluorescence technique in detection of mycobacterial bacilli on fine needle aspiration cytology in tubercular lymphadenitis in comparison to conventional methods. Int J Res Med Sci 2022;10(4):926–932. DOI: 10.18203/2320-6012.ijrms20220987
  5. Gulati HK, Mawlong M, Agarwal A, et al. Comparative evaluation of clinical, cytological and microbiological profile in abdominal vs. cervical lymph nodal tuberculosis with special emphasis on utility of Auramine-O staining. J Cytol 2021;38(4):191–197. DOI: 10.4103/JOC.JOC_61_20
  6. Murray SJ, Barrett A, Magee JG, et al. Optimisation of acid fast smears for the direct detection of mycobacteria in clinical samples. J Clin Pathol 2003;56(8):613–615. DOI: 10.1136/jcp.56.8.613
  7. Ulukanligil M, Aslan G, Tasçi S. A comparative study on the different staining methods and number of specimens for the detection of acid fast bacilli. Mem Inst Oswaldo Cruz 2000;95(6):855–858. DOI: 10.1590/s0074-02762000000600019
  8. Prasanthi K, Kumari AR. Efficacy of fluorochrome stain in the diagnosis of pulmonary tuberculosis co-infected with HIV. Indian J Med Microbiol 2005;23(3):179–181. DOI: 10.4103/0255-0857.16591
  9. Tansuphasiri U, Kladphuang B. Evaluation of sputum staining by modified cold method and comparison with Ziehl–Neelsen and fluorochrome methods for the primary diagnosis of tuberculosis. Southeast Asian J Trop Med Public Health 2002;33(1):128–135. PMID: 12118440.
  10. Lokeshwaran RS, Lavanya KB, Dhanalakshmi A, et al. Comparison of Auramine-O stain and Ziehl–Neelsen stain in the detection of tuberculous bacilli in fine needle aspiration of lymph nodes. J Evolution Med Dent Sci 2018;7(49):5268–5272. DOI: 10.14260/jemds/2018/1169
  11. Padmaja GV, Srujana K, Sadhana C. Comparison of Ziehl–Neelsen's stain, fluorescent stain with CBNAAT of sputum for the diagnosis of pulmonary tuberculosis. J Dr NTR Univ Health Sci 2019;8(4):238–243. DOI: 10.4103/JDRNTRUHS.JDRNTRUHS_46_19
  12. Dagar V, Heda S, Barsagade A, et al. Comparison of ZN staining and fluorescent microscopy in detection of acid fast bacilli in fine needle aspiration smears. IOSR J Dent Med Sci 2016;15(8):79–84. DOI: 10.9790/0853-1508047984
  13. Mistry Y, Rajdev S, Mullan S. Comparative study of ZN staining vs. flurochrome staining and impact of sample processing on diagnosis of tuberculosis from various clinical samples. Adv Microbiol 2016;6:953–958. DOI: 10.4236/aim.2016.613089
  14. Zaib-un-Nisa, Javed H, Zafar A, et al. Comparison of fluorescence microscopy and Ziehl–Neelsen technique in diagnosis of tuberculosis in paediatric patients. J Pak Med Assoc 2015;65(8):879–881. PMID: 26228336.
  15. Sharma M, Broor S, Maheshwari M, et al. Auramine-O staining vs Ziehl Neelsen staining: advantages and disadvantages. Indian J Chest Dis Allied Sci 2023;65(1):39–43. DOI: 10.5005/jp-journals-11007-0062
  16. Subramani P, Venkteswarao T, Malligere Lingaiah H, et al. Evaluation of multiple laboratory methods in the diagnosis of extrapulmonary tuberculosis. Int J Med Public Health 2014;4(4):482. DOI: 10.4103/2230-8598.144123
  17. Smithwick RW. Laboratory Manual for Acid-Fast Microscopy. 2nd ed. Atlanta: Center for Disease Control; 1976.
  18. Ramsay A, Cuevas LE, Mundy CJF, et al. New policies, new technologies: modelling the potential for improved smear microscopy services in Malawi. PLoS One 2009;4(11):e7760. DOI: 10.1371/journal.pone.0007760
  19. Cancer L. Some Aromatic Amines, Organic Dyes, and Related Exposures. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans Volume 99. Lyon: International Agency for Research on Cancer Press; 2010.
  20. 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):520–521. DOI: 10.1016/j.trstmh.2008.02.020
PDF Share
PDF Share

© Jaypee Brothers Medical Publishers (P) LTD.