title: Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis.
abstract: A MALDI TOF MS based minisequencing method has been developed and applied for the analysis of rifampin (RIF)- and isoniazid (INH)-resistant M. tuberculosis strains. Eight genetic markers of RIF resistance-nucleotide polymorphisms located in RRDR of rpoB gene, and three of INH resistance including codon 315 of katG gene and -8 and -15 positions of the promoter region of fabG1-inhA operon were worked out. Based on the analysis of 100 M. tuberculosis strains collected from the Moscow region in 1997-2005 we deduced that 91% of RIF-resistant and 94% of INH-resistant strains can be identified using the technique suggested. The approach is rapid, reliable and allows to reveal the drug resistance of M. tuberculosis strains within 12 h after sample isolation.
authors.affiliation: Research Institute for Physical-Chemical Medicine of Ministry of Public Health of Russian Federation, 119992 Malaya Pirogovskaya 1a, Moscow, Russia. larisa@kge.msu.ru
authors.forename: Larisa N
authors.initials: LN
authors.lastname: Ikryannikova
authors.forename: Maxim V
authors.initials: MV
authors.lastname: Afanas'ev
authors.forename: Tat'yana A
authors.initials: TA
authors.lastname: Akopian
authors.forename: Elena N
authors.initials: EN
authors.lastname: Il'ina
authors.forename: Aleksey V
authors.initials: AV
authors.lastname: Kuz'min
authors.forename: Elena E
authors.initials: EE
authors.lastname: Larionova
authors.forename: Tat'yana G
authors.initials: TG
authors.lastname: Smirnova
authors.forename: Larisa N
authors.initials: LN
authors.lastname: Chernousova
authors.forename: Vadim M
authors.initials: VM
authors.lastname: Govorun
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D002374:Catalase / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D012321:DNA-Directed RNA Polymerases; D030561:Databases, Nucleic Acid; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000187:drug effects / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D012293:Rifampin / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D018088:Tuberculosis, Multidrug-Resistant / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D015342:DNA Probes; D005819:Genetic Markers; C505805:rpoB protein, Mycobacterium tuberculosis; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases; D007538:Isoniazid; D012293:Rifampin