COMPARATIVE ANALYSIS OF TUBERCULOSIS BACTERIA IN SALIVA AND SPUTUM USING ZIEHL-NEELSEN, PAPANICOLAOU AND BLEACH MODIFIED PAPANICOLAOU INDUCED FLUORESCENT MICROSCOPY
Background: This research contrasted the identification of Mycobacterium tuberculosis in saliva and sputum samples using Ziehl-Neelsen, Papanicolaou, and Bleach Modified Papanicolaou fluorescent microscopy.
Methodology: Sputum and saliva samples were collected in a clean sterile, leak-proof, wide-mouth containers early in the morning. Each saliva and sputum sample was further divided into 3 groups, Group A: sputum stained by Ziehl-Neelsen staining method, Group B: sputum smears stained by Papanicolaou staining method, Group C: sputum smears stained by bleach modified Papanicolaou staining method. saliva and sputum infected by Papanicolaou and observed under a Fluorescent Microscope for tubercle bacilli..
Results: The results obtained from saliva stained smears showed 63.3% of cases positive in both BMP and Pap stained saliva samples and 46.6% of cases positive in ZN stained saliva samples. Further grading of the individual staining method was carried out. Scant bacilli; 20% in Pap and 10% in BMP, 1+; 30% in Pap and 33.3% in BMP, 2+;10% in Pap and 6% in BMP and 3+-3.3% in Pap and 13% in BMP. ZN stained saliva samples scant bacilli; 56.6%, 1+; 13%, 2+; 10% and 3+; nil. Sputum samples stained by BMP, Pap, and Zn showed 100% positive results (30/30). But the grading differed among individual staining techniques.
Conclusion: In our study sputum samples showed 100% positivity in comparison to saliva samples with 63.33%. Though the results of the saliva sample were lower than that of the sputum samples, the BMP method used in our study is first of its kind to the best of our knowledge. It is a simpler, convenient, and economical method that can be used in routine diagnosis of TB.
2. Nelson KE, Williams CF, Masters Williams C. Early history of infectious disease: epidemiology and control of infectious diseases. Infectious Disease Epidemiology. 2000:3-16.
3. Chadha VK. Tuberculosis epidemiology in India: a review. The international journal of tuberculosis and lung disease. 2005 Oct 1;9(10):1072-82.
4. Khatri GR, Frieden TR. Controlling tuberculosis in India. New England Journal of Medicine. 2002 Oct 31;347(18):1420-5.
5. Sreeramareddy CT, Qin ZZ, Satyanarayana S, Subbaraman R, Pai M. Delays in diagnosis and treatment of pulmonary tuberculosis in India: a systematic review. The International Journal of Tuberculosis and Lung Disease. 2014 Mar 1;18(3):255-66.
6. Brewer TF. Preventing tuberculosis with bacillus Calmette-Guerin vaccine: a meta-analysis of the literature. Clinical Infectious Diseases. 2000 Sep 1;31(Supplement_3):S64-7.
7. Ahmad Z, Amin SS. Role of tuberculin test, FNAC and ELISA in the diagnosis of tuberculous lymphadenitis. J Indian AcadClin Med. 2003 Oct;4:292-5.
8. Bhatt CP, Timalsina B, Kutu B, Pradhan R, Maharjan B, Shrestha B. A Comparision of Laboratory Diagnostic Methods of Tuberculosis and Aetiology of Suspected Cases of Pulmonary Tuberculosis. SAARC Journal of Tuberculosis, Lung Diseases and HIV/AIDS. 2015 Apr 11;11(2):1-6.
9. Suitters, B. T., &Brøgger, S. A. (1967). Some aspects of laboratory investigations in a mass campaign against tuberculosis. Bulletin of the World Health Organization, 36(5), 837–845.
10. Dewan PK, Lal SS, Lonnroth K, Wares F, Uplekar M, Sahu S, Granich R, Chauhan LS. Improving tuberculosis control through public-private collaboration in India: literature review. Bmj. 2006 Mar 9;332(7541):574-8.
11. Harries AD, Maher D, Nunn P. Practical and affordable measures for the protection of health care workers from tuberculosis in low-income countries. Bulletin of the World Health Organization. 1997;75(5):477.
12. Selvakumar N, Gomathi M, Rehman F, Narayanan P. Evaluation of a two-reagent cold staining method for detection of acid-fast bacilli. The International Journal of Tuberculosis and Lung Disease. 2002 Aug 1;6(8):728-31.
13. Pollock HM, Wieman EJ. Smear results in the diagnosis of mycobacterioses using blue light fluorescence microscopy. Journal of clinical microbiology. 1977 Mar 1;5(3):329-31.
14. Selvakumar N, Ravikumar D, Sivagamasundari S, Gopi PG, Narayanan PR. A novel method of staining acid-fast bacilli in sputum containers. Indian Journal of Medical Research. 2006 Jun 1;123(6):776.
15. Gilpin C, Kim SJ, Lumb R, Rieder HL, Van Deun A, Working Group on Sputum Smear Microscopy. Critical appraisal of current recommendations and practices for tuberculosis sputum smear microscopy [Workshop Report]. The International Journal of Tuberculosis and Lung Disease. 2007 Sep 1;11(9):946-52.
16. Chen P, Shi M, Feng GD, Liu JY, Wang BJ, Shi XD, Ma L, Liu XD, Yang YN, Dai W, Liu TT. A highly efficient Ziehl-Neelsen stain: identifying de novo intracellular Mycobacterium tuberculosis and improving detection of extracellular M. tuberculosis in cerebrospinal fluid. Journal of clinical microbiology. 2012 Apr 1;50(4):1166-70.
17. Uddin MK, Chowdhury MR, Ahmed S, Rahman MT, Khatun R, van Leth F, Banu S. Comparison of direct versus concentrated smear microscopy in detection of pulmonary tuberculosis. BMC research notes. 2013 Jul 25;6(1):291
18. Cattamanchi A, Davis JL, Pai M, Huang L, Hopewell PC, Steingart KR. Does bleach processing increase the accuracy of sputum smear microscopy for diagnosing pulmonary tuberculosis. Journal of clinical microbiology. 2010 Jul 1;48(7):2433-9.
19. Peerapur BV, Bahadur AK, Divakar M. Sodium Hypochlorite Sedimentation Technique: A Simple Way to Enhance Sensitivity of Ziehl-Neelsen Stain in Diagnosis of Smear Negative Pulmonary Tuberculosis. Journal of Krishna Institute of Medical Sciences (JKIMSU). 2014 Jul 1;3(2).
20. Annam V, Karigoudar MH, Yelikar BR. Improved microscopical detection of acid-fast bacilli by the modified bleach method in lymphnode aspirates. Indian Journal of Pathology and Microbiology. 2009 Jul 1;52(3):349.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright © Author(s) retain the copyright of this article.