Rapid detection of multidrug resistant tuberculosis in respiratory specimens at a tertiary care centre in south coastal Karnataka using Genotype MTBDR plus assay

  • Vishnu Prasad Shenoy Department of Microbiology, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India
  • Ajay Kumar Department of Microbiology, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India
  • Kiran Chawla Department of Microbiology, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India
Keywords: Genes, GenoType MTBDR plus, Mutations, Resistance, Tuberculosis


Background and Objectives: Despite high prevalence of MDR-TB in India very limited information about MDR-TB and mutation patterns in rpoB, inhA and katG genes among MDR-isolates of Mycobacterium tuberculosis in south coastal Karnataka region is available; thus present study is an attempt to explore the extent of MDR-TB and mutation patterns prevalent among clinical isolates in this region using GenoType MTBDR plus assay. Materials and Methods: A total of 256 sputum samples from Pulmonary TB patients suspected of MDR-TB were tested by GenoType MTBDR plus as per manufacturer’s guidelines for detection of mutations conferring resistance to rifampicin and isoniazid. The results of GenoType MTBDR plus were recorded and analysed using SPSS version 22. For all analyses, a p value <0.05 was considered statistically significant. Results: Fifty (19.53%) isolates were found MDR, 32 (12.50%) isolates were found mono-resistant to isoniazid and 15 (5.86%) isolates were found mono-resistant to rifampicin. Eleven isolates (4.3%) were found NTM. Mutation in codon S531L, S315T1 and C15T were most common in rpoB, katG and inhA genes respectively. Unknown mutations were found in 50.77% (33/65), 3.66% (3/82) and 26.83% (22/82) isolates for rpoB, katG and inhA genes respectively. Hetero-resistance in MDR, rifampicin monoresistant and isoniazid monoresistant isolates was found to be 26% (13/50), 20% (3/15) and 34.37% (11/32) respectively. Conclusion: Mutation in codon S531L, S315T1 and C15T were most common mutations associated with MDR-TB. Further high number of isolates showed mutations in unknown regions and hetero-resistance thus more elaborate studies based on sequencing are desirable in this region.


World Health Organization. Global tuberculosis report 2016. Available from: http:// www. who.int/tb/publications/global_report/en/

TB statics for India. Available from : http://www.tbfacts.org/tb-statistics-india/

Chiang CY, Yew WW. Multidrug-resistant and extensively drug resistant tuberculosis. Int J Tuberc Lung Dis. 2009; 13:304–311.

Wang JY, Lee LN, Hsu HL, Hsueh PR, Luh KT. Performance assessment of the DR. MTBC Screen assay and the BD ProbeTec ET system for direct detection of Mycobacterium tuberculosis in respiratory specimens. J Clin Microbiol 2006; 44:716-719.

Piersimoni C, Olivieri A, Benacchio L, Scarparo C. Current perspectives on drug susceptibility testing of Mycobacterium tuberculosis complex: the automated nonradiometric systems J Clin Microbiol 2006; 44:20-28.

Udwadia ZF. Totally drug resistant-tuberculosis in India: The bad just got worse. J Assoc Chest Physicians 2016; 4:41-42.

Boehme CC, Nabeta P, Hillemann D, Nicol MP, Shenai S, Krapp F, et al. Rapid molecular detection of tuberculosis and rifampin resistance. N Engl J Med 2010; 363:1005–1015.

Pang Y, Xia H, Zhang Z, Li J, Dong Y, Li Q, et al. Multicenter evaluation of gene chip for detection of multidrug-resistant Mycobacterium tuberculosis. J Clin Microbiol 2013; 51:1707–1713.

Hillemann D, Weizenegger M, Kubica T, Richter E, Niemann S. Use of the genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis complex isolates. J Clin Microbiol 2005; 43:3699–3703.

Bai Y, Wang Y, Shao C, Hao Y, Jin Y .GenoType MTBDRplus Assay for Rapid Detection of Multidrug Resistance in Mycobacterium tuberculosis: A Meta-Analysis. PLoS ONE 2016; 11: e0150321.doi:10.1371/journal.pone.0150321

Drobniewski F, Cooke M, Jordan J, Casali N, Mugwagwa T, Broda A, et al. Systematic review, metaanalysis and economic modelling of molecular diagnostic tests for antibiotic resistance in tuberculosis. Health Technol Assess 2015; 19:1–188.

Central TB Division. Manual of sputum smear fluorescence microscopy. Available from: http://tbcindia.nic.in/WriteReadData/l892s/7890638455Flourescence_Microscopy%20Manual.pdf

Buyankhishig B, Oyuntuya T, Tserelmaa B, Sarantuya J, Lucero MG, et al. Rapid molecular testing for multi-resistant tuberculosis in mongoliya: A diagnostic accuracy study. Int J Mycobacteriol 2012: 40–44.

GenoType MTBDR plus Instructions for use. Available from: https://www.ghdonline. org/uploads/MTBDRplusV2_0212_304A-02-02.pdf.

Meaza A, Kebede A, Yaregal Z, Dagne Z, Moga S, Yenew B, et al. Evaluation of genotype MTBDR plus VER 2.0 line probe assay for the detection of MDR-TB in smear positive and negative sputum samples. BMC Infect Dis 2017; 17:280.

Omer ZB, Mekonnen Y, Worku A, Zewde A, Medhin G, Mohammed T, et al. Evaluation of the GenoType MTBDRplus assay for detection of rifampicin-and isoniazid-resistant Mycobacterium tuberculosis isolates in central Ethiopia. Int J Mycobacteriol 2016; 5:475-481.

Rao P, Chawla K, Shenoy VP, Mukhopadhyay C, Brahmavar V, Kamath A, et al. Study of drug resistance in pulmonary tuberculosis cases in south coastal Karnataka. J Epidemiol Glob Health 2015; 5:275-281.

Shivaswamy U, Neelambike SM. Drug resistance pattern of mycobacterial isolates in HIV and non-HIV population in South India. Lung India 2016; 33:27-31.

Munje R, Deshmukh R, Tumane K. Multidrug-resistant TB among previously treated TB cases: A retrospective study in Nagpur, India. Indian J Tuberc 2015; 62: 207-210.

Maurya AK, Singh AK, Kant S, Umrao J, Kumar M, Kushwaha RA, et al. Use of GenoType® MTBDRplus assay to assess drug resistance and mutation patterns of multidrug-resistant tuberculosis isolates in northern India. Indian J Med Microbiol 2013; 31:230-236.

Kumar P, Balooni V, Singh S. Genetic mutations associated with rifampicin and isoniazid resistance in MDR-TB patients in North-West India. Int J Tuberc Lung Dis 2015; 19:434-439.

Yadav RN, Singh BK, Sharma SK, Sharma R, Soneja M, Sreenivas V, et al. Comparative Evaluation of GenoType MTBDR plus Line Probe Assay with Solid Culture Method in Early Diagnosis of Multidrug Resistant Tuberculosis (MDR-TB) at a Tertiary Care Centre in India. PLoS ONE 2013;8(9):e72036

Barnard M, Albert H, Coetzee G, O'brien R, Bosman ME. Rapid molecular screening for multidrug-resistant tuberculosis in a high-volume public health laboratory in South Africa. Am J Respir Crit Care Med 2008; 177:787-792.

Albert H, Bwanga F, Mukkada S, Nyesiga B, Ademun JP, Lukyamuzi G, et al. Rapid screening of MDR-TB using molecular Line Probe Assay is feasible in Uganda. BMC Infect Dis 2010; 10:41.

Huyen MN, Tiemersma EW, Lan NT, Cobelens FG, Dung NH, Sy DN, et al. Validation of the GenoType® MTBDRplus assay for diagnosis of multidrug resistant tuberculosis in South Vietnam. BMC Infect Dis 2010; 10:149.

Singhal R, Myneedu VP, Arora J, Singh N, Sah GC, Sarin R. Detection of multi-drug resistance & characterization of mutations in Mycobacterium tuberculosis isolates from North-Eastern States of India using GenoType MTBDRplus assay. Indian J Med Res 2014; 140:501-506.

Mokrousov I, Narvskaya O, Otten T, Limeschenko E, Steklova L, Vyshnevskiy B. High prevalence of KatG Ser315Thr substitution among isoniazid-resistant Mycobacterium tuberculosis clinical isolates from north western Russia, 1996 to 2001. Antimicrob Agents Chemother 2002; 46:1417-1424.

Tolani MP, D'souza DT, Mistry NF. Drug resistance mutations and heteroresistance detected using the GenoType MTBDR plus assay and their implication for treatment outcomes in patients from Mumbai, India. BMC Infect Dis 2012; 12:9.

How to Cite
Shenoy V, Kumar A, Chawla K. Rapid detection of multidrug resistant tuberculosis in respiratory specimens at a tertiary care centre in south coastal Karnataka using Genotype MTBDR plus assay. IJM. 10(5):275-80.
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