Detection of rifampicin and izoniazid resistance in Mycobacterium tuberculosis strains from Samara Region (Central Russia)

Abstract

Recently high rates of tuberculosis incidence and prevalence are observed in civilian and prisons sectors in Russia. One of the main reasons for high morbidity levels and ineffectiveness of treatment is wide spreading of drug resistant Mycobacterium tuberculosis strains, but accurate and comprehensive information on levels of drug resistance among strains circulating in Central Russia is unavailable. Rifampicin and izoniazid resistance detection in TB isolates from Samara (Central Russia) civilian and prison TB hospitals and dispensaries in 2000–2002 by revealing mutations in rpoB, katG and inhA genes using Macroarray technique. Methods: A total of 342 M. tuberculosis isolates were tested using Macroarray method. It is based on multiplex amplification of rpoB, katG and inhA genes fragments (with three pairs of biotin labeled primers) following by dothybridization with normal and mutant oligonucleotide probes (fragments of rpoB, katG and inhA genes in which mutations occur) immobilized on nylon membrane strips. Mycobacterial DNA was extracted by heating of cell suspensions following by chloroform extraction. Streptavidin-alkaline phosphatase color development system was used for visualization of results.