Pyrosequencing for rapid detection of tuberculosis resistance to Rifampicin and Isoniazid in Syrian and Lebanese clinical isolates

M Hamze; M B Ismail; A K Rahmo; F Dabboussi

doi:10.1016/j.ijmyco.2015.05.007

Pyrosequencing for rapid detection of tuberculosis resistance to Rifampicin and Isoniazid in Syrian and Lebanese clinical isolates

Int J Mycobacteriol. 2015 Sep;4(3):228-32. doi: 10.1016/j.ijmyco.2015.05.007. Epub 2015 Jun 3.

Authors

M Hamze¹, M B Ismail¹, A K Rahmo², F Dabboussi³

Affiliations

¹ Health and Environment Microbiology Laboratory, Azm Center for Research in Biotechnology and its Applications, Doctoral School of Sciences and Technology, Lebanese University, Lebanon; Department of Medical Laboratory Sciences, Faculty of Public Health, Lebanese University, Lebanon.
² National Commission for Biotechnology, Damascus, Syria.
³ Health and Environment Microbiology Laboratory, Azm Center for Research in Biotechnology and its Applications, Doctoral School of Sciences and Technology, Lebanese University, Lebanon; Department of Medical Laboratory Sciences, Faculty of Public Health, Lebanese University, Lebanon. Electronic address: fdabboussi@ul.edu.lb.

PMID: 27649870
DOI: 10.1016/j.ijmyco.2015.05.007

Abstract

Background: Rapid and accurate techniques are always welcomed for the detection of resistant strains of Mycobacterium tuberculosis MTB.

Objectives: The objective of this study is to evaluate the pyrosequencing technology for the detection of MTB resistance to Rifampicin (RIF) and Isoniazid (INH) in Syrian and Lebanese clinical strains; 66 strains resistant to INH, among them 56 resistant also to RIF, were tested.

Methods: Four pyrosequencing assays were optimized and applied to the following loci: rpoBrpoB RIF resistance-determining region, katG, the promoter regions of inhA and ahpC-oxyR intergenic region.

Results: The prevalence of mutations on codon 315 of the katG gene, inhA and ahpc-oxyR were 42.4%, 21.2% and 9.0%, respectively, which make an overall sensitivity of 72.6% for INH resistance. All RIF-resistant strains contained at least one non-synonymous codon change in the sequenced rpoB region (507-533) relative to the ATCC reference strain. The RIF drug resistance region (RRDR) sequencing identified 96 modified codons representing 34 different mutations.

Conclusions: The high sensitivity and the short turnaround time combined with multilocus sequencing of several isolates in parallel make pyrosequencing an attractive method for drug resistance screening for MTB.

Keywords: Isoniazid; Lebanon; M. tuberculosis; Pyrosequencing; Rifampicin; Syria.

Publication types

Evaluation Study

MeSH terms

Antitubercular Agents / pharmacology*
Codon
DNA Mutational Analysis
Drug Resistance, Bacterial / drug effects*
Drug Resistance, Bacterial / genetics*
Genes, Bacterial
Humans
Isoniazid / pharmacology*
Lebanon / epidemiology
Microbial Sensitivity Tests
Mutation, Missense
Mycobacterium tuberculosis / drug effects*
Mycobacterium tuberculosis / genetics
Public Health
Refugees
Rifampin / pharmacology*
Sensitivity and Specificity
Syria / epidemiology
Tuberculosis, Multidrug-Resistant / drug therapy
Tuberculosis, Multidrug-Resistant / epidemiology
Tuberculosis, Multidrug-Resistant / microbiology*

Substances

Antitubercular Agents
Codon
Isoniazid
Rifampin