title: Classical against molecular-genetic methods for susceptibility testing of antituberculotics.
abstract: Tuberculosis currently belongs to rare respiratory diseases in Slovakia. However, the emergence and spread of multi-drug resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) are major challenges for global tuberculosis control, since the treatment of resistant forms creates both medical and financial problems. Cultivation methods of diagnosis are time-consuming, many times exceeding the time of the initial phase of tuberculosis treatment. Therefore, in the presented study we compared the standard procedures, based on the cultivation of mycobacteria and subsequent drug susceptibility testing to antituberculotics, with molecular-genetic methods using PCR diagnostic kits. The molecular-genetic testing enables to obtain direct and fast evidence of Mycobacterium tuberculosis, with genomic verification of resistance to the most important anti-tuberculosis drugs - isoniazid and rifampicin in MDR-TB, and ethambutol, aminoglycosides, and fluoroquinolones in XDR-TB. In 2012-2013, we confirmed 19 cases of drug-resistant tuberculosis in Slovakia. The resistance to rifampicin was confirmed in all strains with both methods. In two cases, the molecular-genetic testing did not show resistance to isoniazid, as confirmed by conventional cultivation. Furthermore, two strains demonstrating susceptibility in conventional microbiological testing to ethambutol and five strains to fluoroquinolones were verified as actually being resistant using a PCR method. Rapid diagnosis and identification of MDR-TB or XDR-TB strains using molecular-genetic testing is an essential tool for the timely and appropriate drug treatment and prevention of spread of drug resistant strains.
authors.affiliation: National Reference Laboratory for Mycobacteria, National Institute of Tuberculosis, Pulmonary Diseases and Thoracic Surgery, 1 Vysne Hagy St., 059 84, Vysoke Tatry, Slovakia, porvaznik@hagy.sk.
authors.forename: I
authors.initials: I
authors.lastname: Porvaznik
authors.forename: J
authors.initials: J
authors.lastname: Mokry
authors.forename: I
authors.initials: I
authors.lastname: Solovic
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology; D000995:Antitubercular Agents / Q000494:pharmacology*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004977:Ethambutol / Q000494:pharmacology; D054908:Extensively Drug-Resistant Tuberculosis / Q000175:diagnosis* / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology; D024841:Fluoroquinolones / Q000494:pharmacology; D005838:Genotype; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D016133:Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology; D018154:Slovakia / Q000453:epidemiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology
chemical_list: D000617:Aminoglycosides; D000995:Antitubercular Agents; D003470:Culture Media; D004269:DNA, Bacterial; D024841:Fluoroquinolones; D004977:Ethambutol; D007538:Isoniazid; D012293:Rifampin