ISSN : 0975-5276
EISSN : 0975-9174
N. ANISH KUMARAN1, P. KENNEDY KUMAR2*, K.S. SRIDHARAN3
1Department of Microbiology, Sriramachandra Institute of Higher Education & Research, Chennai, 600116, Tamil Nadu, India
2Department of Microbiology, Sriramachandra Institute of Higher Education & Research, Chennai, 600116, Tamil Nadu, India
3Department of Microbiology, Sriramachandra Institute of Higher Education & Research, Chennai, 600116, Tamil Nadu, India
* Corresponding Author : kennychennai1973@gmail.com
Received : 01-07-2020 Accepted : 28-07-2020 Published : 30-07-2020
Volume : 12 Issue : 7 Pages : 1893 - 1897
Int J Microbiol Res 12.7 (2020):1893-1897
Keywords : Cartridge Based Nucleic Acid Amplification Test, Matrix-assisted laser desorption ionization - time-of-flight, Fluorescent microscopy, Bleach concentration
Academic Editor : Dr Carlos Aa Chagas, Tandel D. H.
Conflict of Interest : None declared
Acknowledgements/Funding : Authors are thankful to Department of Microbiology, Sriramachandra Institute of Higher Education & Research, Chennai, 600116, Tamil Nadu, India
Author Contribution : All authors equally contributed
Background: Mycobacterium infection results in chronic granulomatous lesion requiring drug treatment for a prolonged period. We compared different methods for diagnosis of Mycobacterium infection in respiratory samples in this study and speciated the positive samples using MALDI-TOF MS. Aim: To evaluate different methods of smear microscopy, comparing Cartridge based nucleic acid amplification test with convention culture methods and speciation of the isolates by MALDI-TOF MS. Settings and Design: A cross sectional study was done in SRMC&RI. Methods & Material: Respiratory samples both before and after concentrations (Modified Petroff’s & Bleach method) were stained by Kinyoun’s modification. Fluorescent staining performed on unconcerated samples. The smears were interpreted as per National Tuberculosis Elimination Program. Smear positive samples were cultivated in Lowenstein-Jensen slope and incubated at 37oC for 8 weeks. LJ growths were subjected to MALD-TOF MS for speciation. Results: Of the 399 samples 7.3% was positive by Kinyoun’s modification without concentration. Modified Petroff’s and Bleach concentration methods enhanced the positivity to 8% and 8.3% respectively. Fluorescent staining showed 10.8% sample positivity. Of these 44 positive samples, 41 grew in conventional LJ medium. However, CB-NAAT identified them within 2 hours of inoculation. On speciation of these isolates by MALDI-TOF MS, Mycobacterium tuberculosis complex were predominant followed by Mycobacterium avium spp., Mycobacterium arupense and Mycobacterium szulgai. Statistical analysis showed 100% specificity in all microscopy methods, however, sensitivity varied as follows: Unconcentrated Kinyoun’s-53.7%; Modified Petroff’s-61.1%; Bleach concentration-59.3%; Fluorescent staining -79.6%. Conclusion: Fluorescent microscopy is a good technique for initial screening as It is imperative to include CBNAAT for early detection and RIF resistance. Species identification is also essential for accurate treatment.
1. Park K. (2011) Epidemiology of communicable disease; 21st ed. Jabalpur, India, M/s Banarasidas Bhanot Publishers; 9th ed. 2011, 164-165.
2. World Health Organisation. (2019) Global Tuberculosis Report 2019. 1-2.
3. Haldar S, Bose M, Chakrabarti P, Daginawala HF, Harinath BC, Kashyap RS, et al. (2011) Tuberculosis., 91(5),414-26.
4. Ates Guler S, Bozkus F, Inci MF, Kokoglu OF, Ucmak H, Ozden S, et al. (2015) Med Princ Pract., 24(1),75-9.
5. Cambau E, Drancourt M. (2014) Clin Microbiol Infect., 20(3),196-201.
6. Cudahy P, Shenoi S V. (2016) Postgrad Med J., 92(1086),187-93.
7. Singhal R, Myneedu VP. (2015) Int J Mycobacteriology., 4(1),1-6.
8. World Health Organization. (2011) Fluorescent light-emitting diode (LED) microscopy for diagnosis of tuberculosis Policy statement. 1-12.
9. Evans CA. (2011) PLoS Med., 8(7),e1001064.
10. Sebastian G, Nagaraja SB, Vishwanatha T, Voderhobli M, Vijayalakshmi N, Kumar P. (2018) J Appl Microbiol., 124(1),267-73.
11. Neuschlova M, Vladarova M, Kompanikova J, Sadlonova V, Novakova E. (2017) Identification of Mycobacterium Species by MALDI-TOF Mass Spectrometry. In 2017. p. 37-42.
12. Revised National Tuberculosis Control Programme. (2016) Technical and Operational Guidelines for TB Control in India 2016. Central Tuberculosis Division.
13. Kulkarni HK, Pinto M, Wiseman J, Jayaprakash T. (2015) Int J Curr Microbiol App Sci., 4(12),536-40.
14. Firdaus S, Kaur IR, Kashyap B, Avasthi R, Singh NP. (2017) J Clin Tuberc Other Mycobact Dis., 8,6-12.
15. Gebre-Selassie S. (2003) Trop Doct., 33(3),160-2.
16. Uddin MKM, Chowdhury MR, Ahmed S, Rahman MT, Khatun R, Van Leth F, et al. (2013) BMC Res Notes., 6(1).
17. Rahman F, Munshi SK, Kamal SM, Rahman AM, Rahman MM, Noor R. (1970) Stamford J Microbiol., 1(1),46-50.
18. Bansal R, Sharma PK, Jaryal SC, Gupta PK, Kumar D. (2017) J Tuberc Res., 05(02),118-28.
19. Reed SL, Mamo G, Gossa E, Jasura M, Getahun M, Lemma E, et al. (2011) Int Health., 3(3),160-4.
20. Baruwa P, Bawri S, Ali S, Phukan C, Tayal B. (2008) Lung India., 25(3),118.
21. Neelu Sree P, Terin J, Dutta H, Kalyani M. (2018) Asian J Pharm Clin Res., 11(4),110-3.
22. Ängeby KAK, Hoffner SE, Diwant VK. (2004) Int J Tuberc Lung Dis., 8(7),806-15.
23. Tripathi K, Tripathi PC, Nema S, Shrivastava AK, Dwiwedi K, Dhanvijay AK. (2014) Int J Recent Trends Sci Technol., 10(3),461-4.
24. Workineh M, Maru M, Seman I, Bezu Z, Negash M, Melku M, et al. (2017) Ethiop J Health Sci., 27(5),459-64.
25. Chinedum OK, Emwiomwan A, Emmanuel Ifeanyi O, Babayi A. (2017) Ann Clin Lab Res., 05(04).
26. Moraa Orina G, Adoka Ong’wen S, Stephen Amolo A, Thomas Orindi O. (2017) Mycobact Dis., 07(03).
27. Bruchfeld J. (2000) Transactions of the Royal Society of Tropical Medicine and Hygiene, 94(6), pp.677-680. 2000;
28. Steingart KR, Ng V, Henry M, Hopewell PC, Ramsay A, Cunningham J, et al. (2006) Lancet Infect Dis., 6(10),664-74.
29. Cattamanchi A, Davis JL, Pai M, Huang L, Hopewell PC, Steingart KR (2010) J Clin Microbiol., 48(7),2433-9.