ABSTRACT
The most widely discussed antibiotic-resistant tuberculosis strains ("W" and "B0/W148", "CAO") belong to L2/Beijing Lineage and are characterized by IS6110 insertion sequences at the NTF locus. We present a high-throughput, microbead-based method, called NTF-RINT for detection of IS in NTF and Rifampicin and Isoniazid Typing. This method provides tuberculosis diagnostic confirmation, screens for the so-called modern L2/Beijing sublineage and detects mutations involved in resistance to Rifampicin (RIF) and Isoniazid (INH).
Subject(s)
Bacteriological Techniques , DNA Transposable Elements , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , High-Throughput Nucleotide Sequencing , Mutation , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/diagnosis , DNA Mutational Analysis , Genotype , Humans , Kazakhstan/epidemiology , Molecular Epidemiology , Mycobacterium tuberculosis/pathogenicity , New York City/epidemiology , Phenotype , Polymerase Chain Reaction , Population Surveillance , Predictive Value of Tests , Reproducibility of Results , Tuberculosis, Multidrug-Resistant/epidemiology , Tuberculosis, Multidrug-Resistant/genetics , VirulenceABSTRACT
BACKGROUND: Kazakhstan remains a high-burden TB prevalence country with a concomitent high-burden of multi-drug resistant tuberculosis. For this reason, we performed an in depth genetic diversity and population structure characterization of Mycobacterium tuberculosis complex (MTC) genetic diversity in Kazakhstan with both patient and community benefit. METHODS: A convenience sample of 700 MTC DNA cultures extracts from 630 tuberculosis patients recruited from 12 out of 14 regions in Kazakhstan, between 2010 and 2015, was independently studied by high-throughput hybridization-based methods, TB-SPRINT (59-Plex, n = 700), TB-SNPID (50-Plex, n = 543). DNA from 391 clinical isolates was successfully typed by two methods. To resolve the population structure of drug-resistant clades in more detail two complementary assays were run on the L2 isolates: an IS6110-NTF insertion site typing assay and a SigE SNP polymorphism assay. RESULTS: Strains belonged to L2/Beijing and L4/Euro-American sublineages; L2/Beijing prevalence totaled almost 80%. 50% of all samples were resistant to RIF and to INH., Subtyping showed that: (1) all L2/Beijing were "modern" Beijing and (2) most of these belonged to the previously described 94-32 sublineage (Central Asian/Russian), (3) at least two populations of the Central Asian/Russian sublineages are circulating in Kazakhstan, with different evolutionary dynamics. CONCLUSIONS: For the first time, the global genetic diversity and population structure of M. tuberculosis genotypes circulating in Kazakhstan was obtained and compared to previous local studies. Results suggest a region-specific spread of a very limited number of L2/Beijing clonal complexes in Kazakhstan many strongly associated with an MDR phenotype.
Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/diagnosis , Adult , Aged , Antitubercular Agents/pharmacology , Antitubercular Agents/therapeutic use , DNA, Bacterial/isolation & purification , DNA, Bacterial/metabolism , Drug Resistance, Multiple, Bacterial/drug effects , Evolution, Molecular , Genetic Variation , Genotype , Humans , Kazakhstan/epidemiology , Male , Middle Aged , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/isolation & purification , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Hybridization/methods , Phenotype , Prevalence , Tuberculosis, Multidrug-Resistant/drug therapy , Tuberculosis, Multidrug-Resistant/epidemiology , Tuberculosis, Multidrug-Resistant/microbiology , Young AdultABSTRACT
In order to evaluate the prevalence of Chlamydia infection in asymptomatic patients, 568 materials were analyzed. Analysis were performed among women and men aged 16 - 45 years. The observed prevalence rate is 5.5% and is similar to the prevalence reported for other European countries. Our study demonstrates that the prevalence of asymptomatic Chlamydia infection in Sofia is low.
