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Colorimetric Redox Indicator assay for rapid detection of multidrug-resistant Mycobacterium tuberculosis in South Gujarat region

Singh Shruti S., Desai Pratibha B.

Abstract


Tuberculosis (TB) has affected humans since antiquity. The burden of TB particularly with multi-drug-resistance (MDR) is increasing worldwide and has become a major public health concern. This study aims to perform Colorimetric Redox Indicator (CRI) assay for rapid detection of multidrug-resistant Mycobacterium tuberculosis in South Gujarat region. Total 234 samples were collected from South Gujarat region and were subjected for microscopy examination, culturing on Lowenstein-Jensen (LJ) medium and drug susceptibility testing by indirect CRI. Indirect CRI was performed on the culture isolates and the performance was evaluated and compared to that of the Conventional LJ Proportion Method. In our study, 4.95% and 14.85% samples were found to be RIF-Monoresistant and INH-Monoresistant respectively. 34.65% samples were detected as MDR. Majority of resistant strains had an MIC≥1 μg/ml for INH and MIC 1 μg/ml for RIF. While, most susceptible strains had an MIC≤0.031 μg/ml for INH and MIC≤0.062 μg/ml for RIF. As compared to Proportion method, the sensitivity and specificity by CRI for RIF were 62.5% and 100% respectively while for INH, the sensitivity and specificity were 100% and 93.47% respectively. In conclusion, the CRI assay appears to be simple, cheap and rapid method for detection of multidrug-resistant Mycobacterium tuberculosis, especially in poor-resource countries, with limited laboratory facilities. The technique may become a valid substitute to traditional time-consuming methods.
Keywords: M. tuberculosis, colorimetric redox indicator assay, minimal inhibitory concentration evaluation, multidrug-resistance

Cite this Article

Singh Shruti S., Desai Pratibha B. Colorimetric Redox Indicator assay for rapid detection of multidrug-resistant Mycobacterium tuberculosis in South Gujarat region. Research & Reviews: A Journal of Microbiology and Virology. 2018; 8(3): 65–70p.


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References


Global Tuberculosis Report 2013. Technical Report Series. WHO/HTM/TB/2013.15. World Health Organization, Geneva, Switzerland. 2014.

Chiang CY, Van Weezenbeek C, Mori T, Enarson D. Challenges to the global control of tuberculosis. Respirology. 2013; 18(4): 596-604.

Kent P.T. and Kubica G.P. Public health mycobacteriology. A guide for the level 111 laboratory, Centers for Disease Control and Prevention, Atlanta, USA. 1985.

RNTCP, Manual of Standard Operating Procedures (SOP) web address

http://www.tbcindia.nic.in/pdfs.

Palomino, J. C., Martin, A., Camacho, M. et al. Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis Antimicrobial Agents and Chemotherapy. 2002; 46(8):2720–2722.

Viera AJ, Garrett JM. Understanding interobserver agreement: the kappa statistic. Fam Med. 2005; 37(5):360-3.

Parikh R, Mathai A, Parikh S, Chandra Sekhar G, Thomas R. Understanding and using

sensitivity, specificity and predictive values. Indian Journal of Ophthalmology. 2008; 56(1): 45-50.

Iqbal S, Iqbal R, Khan MM, Hussain I, Akhtar A, Shabbir I. Comparison of two conventional techniques used for the diagnosis of tuberculosis cases. Int J. AgriBioi 2003; 5(4):545-7.

Vanisree R, Latha MK, Neelima A, Prasanti. Molecular detection of Rifampicin and Isoniazid resistance in culture isolates of newly diagnosed TB patients. International Journal of Medical Research & Health Sciences. 2014; 3(2):375-380.

Rishi, S., Sinha, P., Malhotra, B., & Pal, N. A comparative study for the detection of Mycobacteria by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination. Indian journal of medical microbiology. 2007; 25(4): 383.

Rajeswari R, Chandrasekaran V, Suhadev M, Sivasubramaniam S, Sudha G, Renu G. Factors associated with patient and health system delays in the diagnosis of tuberculosis in South India. The International Journal of Tuberculosis and Lung Disease. 2002; 6(9):789-95.

Dye C. Global epidemiology of tuberculosis. The Lancet. 2006; 367(9514): 938–40.

K.J. Seung, I.E. Gelmanova, G.G. Peremitin, V.T. Golubchikova, V.E. Pavlova, O.B. Sirotkina, G.V. Yanova, A.K. Strelis, The effect of initial drug resistance on treatment response and acquired drug resistance during standardized short-course chemotherapy for tuberculosis, Clin. Infect. Dis.2004; 39(9): 1321–8.

Takiff, H. E., L. Salazar, C. Guerrero, W. Philipp, W. M. Huang, B. Kreiswirth, S. T. Cole, W. R. Jacobs, Jr., and A. Telenti. Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. Antimicrob. Agents Chemother. 1994; 38(4):773–780.

Siddiqi, N., M. Shamim, S. Hussain, R. K. Choudhary, N. Ahmed, Prachee, S. Banerjee, G. R. Savithri, M. Alam, N. Pathak, A. Amin, M. Hanief, V. M. Katoch, S. K. Sharma, and S. E. Hasnain. Molecular characterization of multidrug-resistant isolates of Mycobacterium tuberculosis from patients in North India. Antimicrob. Agents Chemother. 2002; 46(2): 443–450.

Coban AY, Deveci A, Sunter AT, Palomino JC, Martin A. Resazurin microtiter assay for

isoniazid, rifampicin, ethambutol and streptomycin resistance detection in Mycobacterium tuberculosis: Updated meta-analysis. International Journal of Mycobacteriology. 2014 Dec; 3(4):230–41.

Rivoire N, Ravololonandriana P, Rasolonavalona T, Martin A, Portaels F, Ramarokoto H, Rasolofo Razanamparany V. Evaluation of the resazurin assay for the detection of multidrug-resistant Mycobacterium tuberculosis in Madagascar. The International Journal of Tuberculosis and Lung Disease. 2007 Jun 1; 11(6):683-8.

J.C. Palomino, A. Martin, M. Camacho, H. Guerra, J. Swings, F. Portaels, Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis, Antimicrob. Agents Chemother. 2002; 46(8): 2720–2722.

Khalifa RA, Nasser MS, Gomaa AA, Osman NM, Salem HM. Resazurin Microtiter assay plate method for detection of susceptibility of multidrug resistant Mycobacterium tuberculosis to second-line anti-tuberculous drugs. Egyptian Journal of Chest Diseases and Tuberculosis. 2013 Apr; 62(2):241–7.

Martin A, Camacho M, Portaels F, Palomino JC. Resazurin Microtiter assay plate testing

of Mycobacterium tuberculosis susceptibilities to Second-Line drugs: Rapid, simple, and inexpensive method. Antimicrobial Agents and Chemotherapy. 2003 Oct 23; 47(11):3616-3619




DOI: https://doi.org/10.37591/rrjomv.v8i3.343

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