title: Genetic variation of Mycobacterium tuberculosis circulating in Kharkiv Oblast, Ukraine.
abstract: BACKGROUND
  A persistent increase of tuberculosis cases has recently been noted in the Ukraine. The reported incidence of drug-resistant isolates of M. tuberculosis is growing steadily; however, data on the genetic variation of isolates of M. tuberculosis circulating in northern Ukraine and on the spectrum and frequency of occurrence of mutations determining resistance to the principal anti-tuberculosis drugs isoniazid and rifampicin have not yet been reported.
  
  
  METHODS
  Isolates of M. tuberculosis from 98 tuberculosis patients living in Kharkiv Oblast (Ukraine) were analyzed using VNTR- and RFLP-IS6110-typing methods. Mutations associated with resistance to rifampicin and isoniazid were detected by RFLP-PCR methods, and also confirmed by sequencing.
  
  
  RESULTS
  We identified 75 different genetic profiles. Thirty four (34%) isolates belonged to the Beijing genotype and 23 (23%) isolates belonged to the LAM family. A cluster of isolates belonging to the LAM family had significant genetic heterogeneity, indicating that this family had an ancient distribution and circulation in this geographical region. Moreover, we found a significant percentage of the isolates (36%) belonged to as yet unidentified families of M. tuberculosis or had individual non-clustering genotypes. Mutations conferring rifampicin and isoniazid resistance were detected in 49% and 54% isolates, respectively. Mutations in codon 531 of the rpoB gene and codon 315 of the katG gene were predominant among drug-resistant isolates. An association was found for belonging to the LAM strain family and having multiple drug resistance (R = 0.27, p = 0.0059) and also for the presence of a mutation in codon 531 of the rpoB gene and belonging to the Beijing strain family (R = 0.2, p = 0.04).
  
  
  CONCLUSIONS
  Transmission of drug-resistant isolates seems to contribute to the spread of resistant TB in this oblast. The Beijing genotype and LAM genotype should be seen as a major cause of drug resistant TB in this region.
authors: Institute of Chemical Biology and Basic Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russian Federation, Russia. maya.a.rot@gmail.com
authors: Maya A
authors: MA
authors: Dymova
authors: Oleksander O
authors: OO
authors: Liashenko
authors: Petro I
authors: PI
authors: Poteiko
authors: Valeriy S
authors: VS
authors: Krutko
authors: Eugeny A
authors: EA
authors: Khrapov
authors: Maxim L
authors: ML
authors: Filipenko
mesh_terms: D000328:Adult; D000995:Antitubercular Agents / Q000494:pharmacology; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D018598:Minisatellite Repeats; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D012150:Polymorphism, Restriction Fragment Length; D012293:Rifampin / Q000494:pharmacology; D014376:Tuberculosis / Q000453:epidemiology / Q000382:microbiology*; D014455:Ukraine / Q000453:epidemiology; D055815:Young Adult
chemical_list: D000995:Antitubercular Agents; D004269:DNA, Bacterial; D007538:Isoniazid; D012293:Rifampin