Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis

Larisa N Ikryannikova; Maxim V Afanas'ev; Tat'yana A Akopian; Elena N Il'ina; Aleksey V Kuz'min; Elena E Larionova; Tat'yana G Smirnova; Larisa N Chernousova; Vadim M Govorun

doi:10.1016/j.mimet.2007.05.015

Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis

J Microbiol Methods. 2007 Sep;70(3):395-405. doi: 10.1016/j.mimet.2007.05.015. Epub 2007 May 31.

Authors

Larisa N Ikryannikova¹, Maxim V Afanas'ev, Tat'yana A Akopian, Elena N Il'ina, Aleksey V Kuz'min, Elena E Larionova, Tat'yana G Smirnova, Larisa N Chernousova, Vadim M Govorun

Affiliation

¹ Research Institute for Physical-Chemical Medicine of Ministry of Public Health of Russian Federation, 119992 Malaya Pirogovskaya 1a, Moscow, Russia. larisa@kge.msu.ru

PMID: 17602768
DOI: 10.1016/j.mimet.2007.05.015

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

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics
Catalase / genetics
DNA Probes / genetics
DNA-Directed RNA Polymerases
Databases, Nucleic Acid*
Drug Resistance, Multiple, Bacterial / genetics*
Genetic Markers / genetics
Humans
Isoniazid / pharmacology
Mycobacterium tuberculosis / chemistry
Mycobacterium tuberculosis / drug effects
Mycobacterium tuberculosis / genetics*
Point Mutation
Polymerase Chain Reaction / methods
Promoter Regions, Genetic
Rifampin / pharmacology
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Tuberculosis, Multidrug-Resistant / microbiology

Substances

Bacterial Proteins
DNA Probes
Genetic Markers
rpoB protein, Mycobacterium tuberculosis
Catalase
katG protein, Mycobacterium tuberculosis
DNA-Directed RNA Polymerases
Isoniazid
Rifampin