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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins / genetics
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Catalase / genetics
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DNA Probes / genetics
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DNA-Directed RNA Polymerases
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Databases, Nucleic Acid*
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Drug Resistance, Multiple, Bacterial / genetics*
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Genetic Markers / genetics
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Humans
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Isoniazid / pharmacology
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Mycobacterium tuberculosis / chemistry
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Mycobacterium tuberculosis / drug effects
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Mycobacterium tuberculosis / genetics*
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Point Mutation
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Polymerase Chain Reaction / methods
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Promoter Regions, Genetic
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Rifampin / pharmacology
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
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Tuberculosis, Multidrug-Resistant / microbiology
Substances
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Bacterial Proteins
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DNA Probes
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Genetic Markers
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rpoB protein, Mycobacterium tuberculosis
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Catalase
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katG protein, Mycobacterium tuberculosis
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DNA-Directed RNA Polymerases
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Isoniazid
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Rifampin