Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis.
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.
Ikryannikova
Larisa N
LN
Research Institute for Physical-Chemical Medicine of Ministry of Public Health of Russian Federation, 119992 Malaya Pirogovskaya 1a, Moscow, Russia. larisa@kge.msu.ru
Afanas'ev
Maxim V
MV
Akopian
Tat'yana A
TA
Il'ina
Elena N
EN
Kuz'min
Aleksey V
AV
Larionova
Elena E
EE
Smirnova
Tat'yana G
TG
Chernousova
Larisa N
LN
Govorun
Vadim M
VM
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
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