Introduction. The use of an accelerated molecular genetic method for the diagnosis of tuberculosis and non-tuberculosis mycobacteria, which makes it possible to determine drug sensitivity to anti-tuberculosis drugs, reduces the time required for diagnosis and initiation of treatment. The purpose is to characterize the experience of using molecular genetic methods for the study of tuberculosis and non-tuberculosis mycobacteria in the Orenburg region. Materials and methods. The study was conducted on the basis of the Orenburg Regional Clinical Tuberculosis Dispensary among patients with tuberculosis, including those newly diagnosed, using molecular genetic research methods. Results. It is shown that over the period 2016-2023, the number of performed molecular genetic research methods among patients increased by 4.7%. At the same time, on average, 1.55 studies using this method were conducted for each patient during the study period. In most cases, respiratory material was analyzed using molecular genetic methods. However, over the past 8 years, the number of studies of patients' urine has increased 2.4 times. Among all patients, the most common problem was their resistance to isoniazid and rifampicin 52.6 [50.7;55.4] per 100 patients; A similar situation was found among newly diagnosed patients - 47.25[45.6;50.5] per 100 patients. Only one in three patients had sensitivity to isoniazid and rifampicin - 33.3[30.7;34.7] per 100 people among all patients and 37.6[36.1;39.1] per 100 newly diagnosed patients. In the period from 2016 to 2023, among patients examined by molecular genetic methods, the number of cases of mycobacteriosis increased 2.1 times among all patients and 3.3 times among newly diagnosed patients. Conclusion. The data obtained determine the need for widespread use of a molecular genetic method aimed at the rapid detection of tuberculosis and non-tuberculosis mycobacteria. Determining the sensitivity of anti-tuberculosis drugs of various chemotherapy regimens makes it possible to reduce the time required to identify patients with multidrug-resistant tuberculosis and prescribe an adequate chemotherapy regimen, which in turn shortens the duration of abacillation, improves the effectiveness of treatment and reduces the risk of the spread of drug-resistant strains of Mycobacterium tuberculosis.