[The application of lytic micro-bacteriophage В29 for accelerated phenotype detection of sensitivity mycobacteria of tuberculosis to anti-tuberculosis medications.].

In conditions of prevalence of medicine-resistant strains of mycobacteria of tuberculosis necessity in accelerated, including phenotype techniques of detection of sensitivity of mycobacteria to anti-microbial chemotherapeutic medications in clinical samples is an actual issue. The results of application of accelerated phenotype techniques of detection of sensitivity of clinical strains of mycobacteria of tuberculosis to anti-microbial chemotherapeutic medications on the basis application of lytic mycobacteriophage D29 are presented. The principle of technique is in evaluation of reproduction of mycobacteriophage in cells of mycobacteria of tuberculosis in presence of sensitive to them anti-bacterial medications. The reproduction of mycobacteriophage is evaluated by quantitative analysis of phage DNA in polymerase chain reaction in real-time. The study used 102 clinical strains of mycobacteria of tuberculosis obtained after primary cultivation or re-cultivation in tubes of MGIT system (Bactec). After positive results of growth of mycobacteria of tuberculosis were obtained, the samples were incubated during 48 hours in CO2 incubator in the presence of critical concentrations of 10 widely applied in case of treatment tuberculosis medicinal substances in liquid nutrient medium Middlebrook 7H9 enriched with components OADC, in format of 24 well cultural plate with volume of nutrient medium 1 ml per well. Whereupon, in plate wells deposited 2x103 plaque-forming units of mycobacteriophage D29. After 24 hours a qualitative detection of phage DNA was implemented with polymerase chain reaction in real-time using reagents phage D29 ("Syntol", Russia). The increasing of threshold level of fluorescence of Ct more than to 2 cycles in samples with antibiotic as compared with control testifies sensitivity of the analyzed strain of mycobacteria of tuberculosis to antibiotic. The level of coincidence made up to 91% in comparative study with inoculation in Lowenstein-Jensen nutrient medium. The level of coincidence made up to 96% in comparative study with Bactec test-system of limited number of strains with establishment of sensitivity for 10 medications. The data was confirmed concerning inverse relationship of value ∆Ct and minimal inhibiting concentration of medication. The supposed high efficiency of possible reagents' set on the basis of presented technique on cost/quality criterion.