title: Resistance to pyrazinamide in Russian Mycobacterium tuberculosis isolates: pncA sequencing versus Bactec MGIT 960.
abstract: Resistance to pyrazinamide (PZA) may impact clinical outcome of anti-tuberculosis chemotherapy. PZA susceptibility testing using MGIT 960 is not reliable and little information is available on the prevalence of PZA resistance in Russia. A collection of 64 clinical isolates of Mycobacterium tuberculosis, including 35 multidrug resistant and extensively drug-resistant (MDR/XDR), was analyzed for PZA resistance using MGIT 960, Wayne test, and sequencing of PZA resistance genes pncA, rpsA and panD. In addition, we analyzed 519 MDR-TB strains for susceptibility to PZA by MGIT 960. Sequencing of pncA revealed 17 of 25 (68%) MDR strains and all 10 XDR strains harboring pncA mutations. A correlation of φ = 0.81 between MGIT 960 and pncA sequencing was observed. Mutations in rpsA and panD not associated with PZA resistance as defined by MGIT 960 were identified. We found 1 PZA-resistant strain without mutations in known PZA resistance genes. Almost 73% of MDR-TB strains isolated in Moscow, Russia, were PZA-resistant by MGIT 960 testing of 519 MDR-TB clinical isolates. Further studies are needed to determine the role of rpsA and panD mutations in possible low-level PZA resistance and to identify the molecular basis of new PZA resistance in the isolate without known PZA resistance mutations.
authors.affiliation: Laboratory of Bacterial Genetics, Department of Genetics and Biotechnology, Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russia.
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Maslov
authors.affiliation: Laboratory of Bacterial Genetics, Department of Genetics and Biotechnology, Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Zaĭchikova
authors.affiliation: Department of Microbiology, Central TB Research Institute, Russian Academy of Medical Sciences, 2 Yauzskaya Alleya, 107564 Moscow, Russia.
authors.forename: Larisa N
authors.initials: LN
authors.lastname: Chernousova
authors.affiliation: Laboratory of Bacterial Genetics, Department of Genetics and Biotechnology, Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russia.
authors.forename: Kirill V
authors.initials: KV
authors.lastname: Shur
authors.affiliation: Laboratory of Bacterial Genetics, Department of Genetics and Biotechnology, Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russia.
authors.forename: Olga B
authors.initials: OB
authors.lastname: Bekker
authors.affiliation: Department of Microbiology, Central TB Research Institute, Russian Academy of Medical Sciences, 2 Yauzskaya Alleya, 107564 Moscow, Russia.
authors.forename: Tatiana G
authors.initials: TG
authors.lastname: Smirnova
authors.affiliation: Department of Microbiology, Central TB Research Institute, Russian Academy of Medical Sciences, 2 Yauzskaya Alleya, 107564 Moscow, Russia.
authors.forename: Elena E
authors.initials: EE
authors.lastname: Larionova
authors.affiliation: Department of Microbiology, Central TB Research Institute, Russian Academy of Medical Sciences, 2 Yauzskaya Alleya, 107564 Moscow, Russia.
authors.forename: Sofya N
authors.initials: SN
authors.lastname: Andreevskaya
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology & Immunology, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Laboratory of Bacterial Genetics, Department of Genetics and Biotechnology, Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russia. Electronic address: valerid@vigg.ru.
authors.forename: Valery N
authors.initials: VN
authors.lastname: Danilenko
mesh_terms: D000581:Amidohydrolases / Q000235:genetics*; D000995:Antitubercular Agents / Q000627:therapeutic use*; D001431:Bacteriological Techniques; D004252:DNA Mutational Analysis; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D054908:Extensively Drug-Resistant Tuberculosis / Q000175:diagnosis* / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D010641:Phenotype; D011237:Predictive Value of Tests; D015995:Prevalence; D011718:Pyrazinamide / Q000627:therapeutic use*; D012426:Russia / Q000453:epidemiology; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis* / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology
chemical_list: D000995:Antitubercular Agents; D011718:Pyrazinamide; D000581:Amidohydrolases; C487496:PncA protein, Mycobacterium tuberculosis
keywords: Drug susceptibility testing; MGIT960; Pyrazinamide; Resistance; Tuberculosis; pncA