title: Magnitude of gene mutations conferring drug resistance in mycobacterium tuberculosis isolates from lymph node aspirates in ethiopia.
abstract: OBJECTIVE
  Resistance to drugs is due to particular genomic mutations in the specific genes of Mycobacterium tuberculosis. Timely genetic characterization will allow identification of resistance mutations that will optimize an effective antibiotic treatment regimen. We determine the magnitude of gene mutations conferring resistance to isoniazid (INH), rifampicin (RMP) and ethambutol (EMB) among tuberculosis (TB) lymphadenitis patients.
  
  
  METHODS
  A cross sectional prospective study was conducted among 226 M.tuberculosis isolates from culture positive lymph node aspirates collected from TB lymphadenitis patients between April 2012 and May 2012. Detection of mutations conferring resistance to drugs was carried out using GenoType(®) MTBDRplus and GenoType® MTBDRsl assay.
  
  
  RESULTS
  Out of the 226 strains, mutations conferring resistance to INH, RMP, multidrug resistance tuberculosis (MDR-TB) and EMB were 8, 3, 2 and 2 isolates, respectively. There was no isolated strain that showed mutation in the inhA promoter region gene. All INH resistant strains had mutations in the katG gene at codon 315 with amino acid change of S315T1. Among rifampicin resistant strains, two isolates displayed mutations at codon 531 in the rpoB gene with amino acid change of S531L and one isolate was by omission of wild type probes at Q513L. According to mutations associated with ethambutol resistance, all of the isolates had mutations in the embB gene with aminoacid change of M306I. All isolates resistant to INH, RMP and MDR using BacT/AlerT 3D system were correctly identified by GenoType® MTBDRplus assay.
  
  
  CONCLUSIONS
  We observed mutations conferring resistance to INH at S315T1 of the katG gene, RMP at S531L and Q513L in the rpoB genes and EMB at M306I of the embB gene. In the absence of conventional drug susceptibility testing, the effort to develop easy, rapid and cost effective molecular assays for drug resistance TB monitoring is definitely desirable and the GenoType® MTBDRplus assay was found to be a useful method for diagnosis of resistance to INH, RMP and MDR from lymph node aspirates. Further molecular cluster analysis to determine transmission dynamics of mutated strain is required.
authors.affiliation: 1. College of Medicine and Health Sciences, Bahir Dar University, Bahir Dar, Ethiopia; ; 2. Institute of Medical Microbiology and Epidemiology of Infectious Diseases, University Hospital, University of Leipzig, Leipzig, Germany; ; 3. Institute of Clinical Immunology, University Hospital, University of Leipzig, Leipzig, Germany; ; 5. Translational Centre for Regenerative Medicine (TRM)-Leipzig, University Hospital, University of Leipzig, Leipzig, Germany.
authors.forename: Fantahun
authors.initials: F
authors.lastname: Biadglegne
authors.forename: Belay
authors.initials: B
authors.lastname: Tessema
authors.forename: Arne C
authors.initials: AC
authors.lastname: Rodloff
authors.forename: Ulrich
authors.initials: U
authors.lastname: Sack
mesh_terms: D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D003430:Cross-Sectional Studies; D005002:Ethiopia; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D008198:Lymph Nodes / Q000382:microbiology*; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D011446:Prospective Studies; D012293:Rifampin / Q000494:pharmacology; D018088:Tuberculosis, Multidrug-Resistant / Q000235:genetics
chemical_list: D000904:Antibiotics, Antitubercular; D007538:Isoniazid; D012293:Rifampin
keywords: Ethambutol; Ethiopia.; Isoniazid; M.tuberculosis; Mutation; Rifampicin; aspirates
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