COMPARATIVE STUDY OF MICROSCOPY AND PCR IN DETECTION OF MYCOBACTERIUM TUBERCULOSIS IN A TERTIARY CARE CENTRE

S. SANGEETHA¹, R. PRAKASH²*, S.A. LAKSHMINARAYANA³, K. ARCHANA RAO^4
1Rajarajeswari Medical College & Hospital, Kumbalgodu, Mysuru Road, Bengaluru, 560074, Karnataka, India
²Rajarajeswari Medical College & Hospital, Kumbalgodu, Mysuru Road, Bengaluru, 560074, Karnataka, India
³Rajarajeswari Medical College & Hospital, Kumbalgodu, Mysuru Road, Bengaluru, 560074, Karnataka, India
⁴Rajarajeswari Medical College & Hospital, Kumbalgodu, Mysuru Road, Bengaluru, 560074, Karnataka, India
* Corresponding Author : prakashssmc@gmail.com

Received : 25-11-2019     Accepted : 26-12-2019     Published : 30-12-2019
Volume : 11     Issue : 12       Pages : 1742 - 1744
Int J Microbiol Res 11.12 (2019):1742-1744

Keywords : Auramine O stain, IS6110, Microscopy, Mycobacterium tuberculosis, Polymerase Chain Reaction, Ziehl-Neelsen stain
Academic Editor : Kanagasanthosh K., Dey S., Mahajan V., Dhiraj B. Shekhawat, Maciej Cedzynski, Dr Sharada T. Rajan, Dr Sahin Direkel
Conflict of Interest : None declared
Acknowledgements/Funding : Authors are thankful to Rajarajeswari Medical College & Hospital, Kumbalgodu, Mysuru Road, Bengaluru, 560074, Karnataka, India. Authors are also thankful to Rajarajeswari Medical College & Hospital for supporting us in procuring the PCR kits and Mr. Prabhu and Mr. Basanth of Central Research Laboratory for helping us in running RT-PCR.
Author Contribution : All authors equally contributed.

Background & objectives: Tuberculosis (TB) continues to be the second the major cause of morbidity and mortality in India as well as worldwide. Early diagnosis is required to prevent transmission of TB and also to reach the WHO goal to end TB by 2035. TB is diagnosed in laboratories by smear microscopy and culture which either lacks sensitivity or delay in reporting. Hence, RT-PCR was compared with sputum microscopy to find an appropriate method to diagnose TB at the earliest. Methods: Forty-four sputum samples were collected from presumptive TB cases, sputum microscopy was done by Ziehl-Neelsen stain, Auramine O stain microscopy and RT-PCR was done by using primer IS6110 of Mycobacterium tuberculosis. Results: Of the forty-four sputum samples, ZN staining showed 48% positivity while Fluorescent stain showed 55% positivity in smear and PCR showed 71% positivity in the sputum samples. Interpretation & conclusions: PCR is a better method than staining techniques for detection of Mycobacterium tuberculosis. Fluorescent microscopy is more sensitive when compared to ZN stain but requires an expensive microscope.

1. Global Tuberculosis Report (2018) World Health Organization. France
2. Trent R.J. (2005) Molecular Medicine: An Introductory Text. 3 ed. Elsevier Academic Press.
3. Chan E.D., Heifets L., Iseman M.D. (2000) Tuber Lung Dis, 80, 131-140.
4. Steingart K.R., Henry M., Ng V., Hopewell P.C., Ramsay A., Cunningham J., et al. (2006) Lancet Infect Dis, 6, 570-81.
5. Eisenach K.D. (1998) In Mycobacterium: Molecular Biology and Virulence, pp. 161-179. Edited by C. Ratledge& J. Dale. Oxford: Blackwell Science.
6. Nagpal S., Chopra G.S., Oberoi A., Singh N., Varghese S.R. (2016) J Health Res, 3, 258-62.
7. Clarridge J.E. 3rd, Shawar R.M., Shinnick T.M., Plikaytis B.B. (1993) J Clin Microbiol, 31, 2049-56.
8. Ieven M., Goossens H. (1997) Clin Microbiol Rev, 10, 242-56.
9. Case finding and Diagnosis strategy in Technical and Operational Guidelines for TB Control in India (2016) Central Tuberculosis Division. Accessed on 09/02/2019
10. RNTCP (2017) National Strategic Plan for Tuberculosis Elimination 2017-2025. Accessed on 12/03/2019.
11. Kocagoz T., Yilmaz E., Ozkara S., Kocagoz S., Hayran M., Sachedeva M., et al. (1993) J Clin Microbiol, 31, 1435-1438.
12. Hooja S., Pal N., Malhotra B., Goyal S., Kumar V., Vyas L. (2011) Ind J Tuberculosis, 58, 51-53.
13. Pooja S., Asthana A.K., Madan M. (2014) JMID, 4, 141-4.
14. Sethi S., Yadav R., Mewara A., Dhatwalia S.K., Sharma M., Gupta D. (2012) Braz J Infect Dis, 16, 493-4.
15. Nimesh M., Joon D., Pathak A.K., Saluja D. (2013) J Infect, 67, 399-407.
16. Pinhata J.M.W., Cergole-Novella M.C., Carmo A.M.S., Silva R.R.F., Ferrazoli L., Sacchi C.T., et al. (2015) J Med Microbiol, 64, 1040-45.
17. Narayanan S., Parandaman V., Narayanan P.R., Venkatesan P., Girish C., Mahadevan S., et al. (2001) J Clin Microbiol, 39, 2006-8.
18. Radhakrishnan I., Manju Y.K., Kumar R.A., Mundayoor S. (2001) J Clin Microbiol, 39, 1683.
19. Broccolo F., Scarpellini P., Locatelli G., Zingale A., Brambilla A.M., Cichero P., et al. (2003) J Clin Microbiol, 41, 4565-4572.
20. Armand S., Vanhuls P., Delcroix G., Courcol R., Lemaiˆtre N. (2011) J Clin Microbiol, 49, 1772-1776.