Detection of Mutations in folp1, rpoB and gyrA genes of M. leprae by PCR- direct sequencing – A rapid tool for screening drug resistance in leprosy

Sekar, B and Arunagiri, K and Narayanan, Sujatha and Menaka, K and Oommen, P K (2011) Detection of Mutations in folp1, rpoB and gyrA genes of M. leprae by PCR- direct sequencing – A rapid tool for screening drug resistance in leprosy. Leprosy Review, 82 (1). pp. 36-45. ISSN Print: 0305-7518 | Electronic: 2162-8807

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Abstract

Introduction: Conventional Mouse foot-pad (MFP) assay for screening drug resistance in M. leprae is cumbersome and time-consuming (approximately 6 to 12 months). Molecular targets for different anti-leprosy drugs have been well defined. Molecular tools for rapid detection of drug resistance in M. leprae have been standardised. A study to compare molecular methods with MFP assay in determining the drug susceptibility of M. leprae was carried out. Methods: Forty Bacteriological Index (BI) positive patients of leprosy with clinical features of relapse (25), new cases (11) and defaulters (4) were included in the study. A skin biopsy was done and the samples were processed using both MFP assay and Molecular method. PCR assays were carried out to amplify, 388 bp of folP1gene for dapsone resistance, 305 bp of rpoB gene for rifampicin resistance and 342 bp of gyrA gene for ofloxacin resistance, followed by direct DNA sequencing. Results: Significant growth in the MFP test was obtained in only 28 out of 40 biopsies processed (70%). Ten of these isolates were dapsone resistant; one isolate showed combined resistance against dapsone, rifampicin and clofazimine. Amplification for all three genes was successful in all the 40 (100%) samples. Among folP1 products sequenced, six isolates showed mutations at 53 (or) 55 amino acid positions. Those strains which showed high-level resistance with two log growth.

Affiliation:	ICMR-National Institute for Research in Tuberculosis
Item Type:	Article
Uncontrolled Keywords:	Detection, mutations, folp1, rpoB, gyrA genes, M. leprae, PCR- direct sequencing--a rapid tool, drug resistance, leprosy
Subjects:	Tuberculosis > Laboratory Research > Immunological
Divisions:	Basic Science Research > Immunology
Depositing User:	Dr. Rathinasabapati R
Date Deposited:	16 Jun 2022 07:35
Last Modified:	16 Jun 2022 07:35
URI:	http://eprints.nirt.res.in/id/eprint/1085

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