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Biomedical subjects

V I Golyshevskaia

Publications and source records attributed to V I Golyshevskaia.

At least 19 recordsLinked to original sources

[Preponderance of Mycobacterium tuberculosis strains of the family Beijing and risk factors of their transmission in the Samara Region].

Current molecular and epidemiological studies could reveal individual families of Mycobacterium tuberculosis strains. The purpose of the present study was to reveal the spread of Mycobacterium tuberculosis strains of the family Beijing in the Samara Region and to define risk factors for their transmission. This was a cross-sectional populational molecular epidemiological study that showed that the Beijing genotype prevailed among the obtained isolates (66.6%; 586/880) and it was encountered among convicts and young persons (RR 1.3; 95% CI 1.2-1.5 and RR 1.2; 95% CI 1.0-1.3, respectively), which is indicative of active and recent transmission. Multifactorial analysis indicated that male sex (OR 1.5; 95% CI 1.1-1.9), younger age (OR 1.3; 95% CI 1.1-1.7), homelessness (OR 5.6; 95% CI 1.1-6.3), and prior or current confinement (OR 2.0; 95 CI 1.5-2.7) were substantially associated with the risk of contamination with the strain of the Beijung family. Drug resistance, including multidrug resistance, was twice higher among the strains of this family.

Adult↗

[Evaluation of the efficiency of activity of the liposomal form of isoniazid against different types of mycobacteria in vitro].

M. avium and three M. tuberculosis strains were used as an example to study the bacteriostatic activity of the liposomal form of isoniazid in the liquid nutrient medium by serial dilutions. Unlike isoniazid in solution, its liposomal form was found to suppress the multi plication of all study mycobacterial samples and diminishes their viability, but at different concentrations. The liposomes that did not contain the drug exerted an inhibitory effect only on the M. tuberculosis strains H37Rv and Erdman. It is suggested that the change in isoniazid sensitivity results from the effect of liposomal phospholipids on some mycobacterial enzymes. Electron microscopy indicated that the liposomal form of isoniazid is able to penetrate into the alveolar macrophageal cytoplasm of guinea pigs just 30 min after incubation.

Antitubercular Agents↗

[The quality of bacteriological detection and determination of drug sensitivity of Mycobacterium tuberculosis by the participants of the External Clinical Laboratory Study Quality Assessment made by the Federal System in 2002-2003].

The paper presents the results of the assessment of the quality of cultural detection and determination of Mycobacterium tuberculosis (MBT) sensitivity at Russia's public health laboratories, which has been made by the Federal System for External Assessment of the Quality of Clinical Laboratory Studies in 2002-2003. The "Bacteriological Detection" Section involved 102 laboratories, of them 57 laboratories also participated in the section "Detection of Drug Resistance". Sixty-three laboratories operated as part of specialized tuberculosis-controlling service facilities and 39 laboratories worked as part of general medical care therapeutic-and-prophylactic institutions. A set of 5 lyophilized samples containing 2.5 x 10(6) M. tuberculosis H37Rv (n = 1); 2.5 x 10(4) M. tuberculosis H37Rv (n = 2); M. fortuitum (n = 1); E. coli in the mixture with autoclaving-killed M. tuberculosis H37Rv (n = 1). 77% of the laboratories could detect in the cultural sample with a high content of M. tuberculosis, by obtaining a correct result; 85% revealed MBT if only in every three MBT-containing samples; 15% of the laboratories could not find MBT in none of the three positive samples. 12% of the laboratories took the growth of microorganisms in the M. fortuitum-containing sample for that of M. tuberculosis. 58% of the laboratories obtained correct results if only in one sample containing M. tuberculosis H37Rv; 42% reported the detection of resistance to one or several antituberculous drugs in the samples containing the drug-sensitive strain H37Rv (a false drug resistance). All the results of determination of drug resistance (DR) in the M. fortium-containing sample were correct. The findings suggest that it is necessary to increase the professional competence of laboratory personnel to the level that ensures the adequate of fulfillment of the standard protocols for detecting MTB and determining DR, which have been introduced by the Ministry of Health of the Russian Federation through its order No. 109 on March 21, 2003.

Anti-Infective Agents↗

[Analysis of risk factors of the occurrence of drug resistance in patients with tuberculosis from civil and penitentiary sectors in the Samara Region].

The true prevalence rates of multidrug-resistant tuberculosis (MDRT) are unknown for most regions of Russia. This study was conducted in the Samara Region that differs from other regions in the rapid spread of HIV infection. The purpose of this study was to determine the primary and acquired resistance of Mycobacterium tuberculosis (MBT) to first-line antituberculous drugs in patients from civil and penitentiary sectors and to reveal risk factors of drug resistance of MBT. Six hundred patients (309 civilians and 291 prisoners who had been bacteriologically diagnosed as having tuberculosis. The authors have established the following:--in new cases, primary drug resistance is as follows: to isoniazid [38.9% (95% CI, 31.3-36.9%)], to rifampicin [25.9% (95% CI, 19.4-33.4%)] and to MDRT [23.0% (95% CI, 16.7-30.3%)];--in prisoners, the primary resistance of MBT was statistically more significant than in civilians;--male sex, in adequate prior or current treatment for tuberculosis for more than 4 weeks, the presence of fibrocavernous tuberculosis and previous prison stay are essential risk factors of the development of resistance of MBT to both any first-line drug and MDRT;--HIV infection is unassociated with resistance.

Adult↗

[Comparative characterization of molecular and microbiological methods for controlling chemotherapy in new cases of pulmonary tuberculosis].

The purpose of the study was to define a role of molecular genetic techniques in the evaluation of the efficacy of short-term chemotherapy in patients with pulmonary tuberculosis. A total of 203 patients with first detected tuberculosis patients who isolated Mycobacterium tuberculosis (MBT) were examined. Microbiological methods (bacterioscopy and seeding) and polymerase chain reaction (PCR) were used to detect MBT in new cases. During chemotherapy, there was a reduction in the number of positive results of both bacterioscopy and PCR, which serves as a sign of abacillation. Following 2 months of chemotherapy, 50% of the patients with negative bacterioscopy and seeding were found to have MBT DNA. In the patients with negative bacterioscopy and seeding, the results of PCR were also negative by month 6. In a group of patients without bacterial isolation, but with positive sputum tests for MBT DNA, the efficiency of chemotherapy was confirmed by the negative results of PCR. The latter is shown to enhance the sensitivity of bacteriological tests for MBT and to reduce the time of obtaining their results.

Antitubercular Agents↗

[Ways of enhancing the effectiveness of bacterioscopic diagnosis of tuberculosis in general health care facilities in case of pilot areas of Russia].

The experience of researchers of the Central Research Institute of Tuberculosis, Russian Academy of Medical Sciences, in the pilot areas of Russia has made it possible to perfect approaches to improving a bacteriological diagnosis of tuberculosis in general health care facilities. A decisive role in enhancing the effectiveness of detection of patients with tuberculosis by the Ziel-Nielsen test is played by the following factors: training of laboratory personnel; provision of laboratories with high-grade equipment and reagents; introduction of the methods and techniques unified and approved by the Ministry of Health of the Russian Federation; regular supervision of jurisdictional laboratories; outside control of the quality of studies. Proper organization of examination of patients, including an effectual selection of patients to be examined, the observance of the established frequency of their examination, and provision of the required quality of a taken sample greatly affect the efficiency of a microscopic diagnosis of bacillary patients. In addition, the centralization of performed microbiological tests for tuberculosis in the most trained, certified, and licensed large laboratories exercising a centralized control of the quality of microbiological studies is optimal.

Ambulatory Care Facilities↗

[Role of ultrasmall forms of Mycobacteria in the pathomorphology of tuberculosis].

Microbiological and histological studies made in patients with active tuberculosis revealed that 65.5% of cases had revertants from ultraminor forms (UMF) of Mycobacteria tuberculosis (MBT), which are typically bacterial in shape and referred to as representatives of the genus Mycobacterium tuberculosis. Cultivation of the diagnostic material containing MBT UMF the enriched liquid nutrient medium could detect rod-shaped bacterial forms of MBT in 18% of the patients with tuberculosis. Inoculation of the MBT UMF-containing material isolated from patients with tuberculosis to guinea-pigs revealed productive sarcoid-like changes in the organs of the experimental animals in 30.6% of cases. A postmortem study of the organs of experimental animals inoculated the material containing "visible, but not growing" mycobacteria detected paraspecific tissue reactions and infiltration of lung tissue by multiple cells of the macrophageal series.

Animals↗

[Comparison of nitrate reductase and automatic BACTEC MGIT 960 AST techniques for determining the drug sensitivity of mycobacteria tuberculosis].

The accelerated nitrate reductase method (NRM) developed at the Central Research Institute of Tuberculosis versus the automatic assay of drug sensitivity by means of a BACTEC 960 bacteriological analyzer was assessed. NRM was carried out, by using the Lówenstein-Jensen medium for 10 days. It is based on the detection of alive Mycobacteria tuberculosis, by recording their enzymatic activity. The study showed a good agreement of the results obtained by NRM with those obtained on a BACTEC 960 analyzer. Agreements were found for 52 isolates in 47 (90.4%) cases, the results disagreed in the testing of 5 (9.6%) cultures. The results of NRM were identical to those for 21 of the 22 cultures sensitive on a BACTEC 960 device; the coincidence was 95.5%. The sensitivity of NRM ranged from 88.2% (for rifampicin) to 96.3% (for isoniazid) and the specificity did from 96% (for isoniazid) to 100% (for streptomycin, rifampicin, and ethambutol). The positive prognostic value of NRM was 100% (for streptomycin, rifampicin, and ethambutol) and 96.3% (for isoniazid). The negative prognostic value of NRM ranged from 94.6 to 96.8% for individual drugs. The efficiency of NRM (a ratio of the number of correct results to the total number of results) was greater than 0.96, which suggests that this method and the BACTEC MGIT 960 AST technique may be regarded as rather comparable. The testing of NRM versus the automatic BACTEC MGIT 960 AST technique has indicated that the former may be successfully used to determine the sensitivity of Mycobacteria to the critical concentrations of first-line antituberculous agents.

Antitubercular Agents↗

[Microbiological blood studies in the diagnosis of tuberculosis in adolescents].

Luminescence bacterioscopy of capillary blood has been found to be an additional rapid laboratory method that verifies a clinical diagnosis in cases of negative sputum bacterioscopic tests in 48.8% of the adolescent patients. The Boy luminescence microscopy shows that in adolescent tuberculosis patients, Mycobacteria are significantly more frequently present in the capillary blood (72.4%) than in the sputum (30.9%) and bronchial washes (34.8%). The detection of Mycobacteria in the capillary blood has been evidenced by cultural tests of some venous blood samples in the BACTEC 460 TB system.

Adolescent↗

[Organizational and methodological approaches to improving the microbiological diagnosis of tuberculosis in Russia].

The practice of the researchers of the Central Research Institute of Tuberculosis in Russia's experimental areas yielded approaches to improving the microbiological diagnosis of tuberculosis in both specialized and general health care facilities. A decisive role in enhancing the efficiency of microbiological identification of patients with tuberculosis is played by the following factors: laboratory staff training, provision of laboratories with qualitative equipment and reagents, introduction of unified methods and those approved by the Ministry of Health of the Russian Federation, regular supervision of jurisdictional laboratories, and outside test quality control. The efficiency of microbiological diagnosis of bacillary patients primarily depends on the expediency of formation of groups of patients under study, the observance of the frequency of examinations, and the assurance of the quality of collected materials. With this, the centralization of microbiological tests for tuberculosis at the most trained, certified, and licensed large laboratories that exercise a centralized control over the quality of microbiological tests.

Bacteriological Techniques↗

[Analyzing the drug resistance of Mycobacterium tuberculosis in Russia's experimental areas introducing the WHO tuberculosis control programme].

The authentic data on the drug resistance of Mycobacterium tuberculosis can be obtained only provided that standard laboratory procedures and the unified standardized method for determining the resistance are used along with a compulsory outside quality control over the performance of this test. The supervisory activities of the researchers of the Central Research Institute of Tuberculosis (CRIT), Russian Academy of Medical Sciences, made in some regions of Russia included standardization of methods and their implementation quality control allowed them to reveal and to correct main errors in drug resistance testing. In this connection, this made it to record the true level of M. tuberculosis resistance to essential antituberculous agents in the areas investigated by the researchers of CRIT. Analyzing the data on the drug resistance of M. tuberculosis in the above areas leads to the conclusion that 2000 was marked by rather high rates of primary drug resistance; nearly a fourth of all new and prior cases of pulmonary tuberculosis studied for its drug resistance were carriers of a M. tuberculosis resistant to at least one antituberculous agent.

Bacteriological Techniques↗

[Determination of drug resistance of Mycobacterium tuberculosis by methods of proportions and absolute concentrations].

The studies have showed the frequency and spectrum of drug resistance, determined by the methods of proportions and absolute concentrations, to be virtually equal. This suggests that both methods are of equal diagnostic value. The method of absolute concentrations is rather accurate and similar to that of proportions in sensitivity, by ensuring comprehensive determination of the spectrum of drug resistance. Furthermore, the advantages of this method are its simplicity, accessibility, and cost-efficiency.

Antibiotics, Antitubercular↗

[Molecular characteristics of multiresistant clinical strains of Mycobacterium tuberculosis isolated in Russia].

The efficiency of tuberculosis control programs is largely determined by methods for rapid diagnosis of the agent. In comparison with the traditional methods, new molecular technologies for characterization of mycobacteria appear to be more promising, because the result can be obtained in almost no time. Sixty-five strains of M. tuberculosis isolated in various regions of Russia were investigated. Drug resistance and strain appurtenance of this sample were determined by classical (absolute concentrations method, IS6110-RFLP) and modern molecular genetic methods (detection of mutations in rpo B gene, DRE-PCR). The spectrum of mutations of the rpoB gene associated with rifampicin resistance was evaluated by direct sequencing. Mutations involving codons 531 (62.7%), 526 (18.6%), and 516 (10.2%) of rpoB gene predominated in the studied sample. The studied strains were discriminated into 52 individual strains by IS6110-RFLP and DRE-PCR typing. Analysis of the resultant genetic variants showed the predominance of M. tuberculosis family W. The efficiency of combined approach to screening for M. tuberculosis is discussed.

DNA-Directed RNA Polymerases↗

[Detection and characteristics of rpoB gene mutations in rifampicin-resistant clinical strains of M. tuberculosis].

The resistance of M. tuberculosis to rifampicin, one of the key agents used in the treatment of tuberculosis is due to point mutations in the rpoB gene encoding for the B-subunit of PNA polymerase. Based on the detection of such mutations, genotypic determinations of rifampicin resistance is a serous alternative to routine microbiological assays that take much more time. Nevertheless, the efficiency of genotypic methods largely depends how completely the resistance-associated mutations are studied and characterized. It is shown that the types and detection rates of certain rpoB mutations can greatly vary in the Mycobacterium strains spread in different geographical regions. By applying the approach based on the direct sequencing of PCR with rpoB gene fragments, the present paper analyzed 48 rifampicin-sensitive and 52 rifampicin-resistant clinical M. tuberculosis strains provided by Moscow tuberculous control facilities. Mutations responsible for rifampicin resistance were detected in 51 (98%) of the 52 resistant strains. The mutations involving codons 531 (46%), 526 (23%), and 516 (23%) of the rpoB gene were proved to be dominant. An unusual double mutation combining the replacement of F by L in codon 514 and previously uncharacterized methionine deletion in the position 515 was detected in the single investigated strain. The efficiency of the employed approach for rapid diagnosis of rifampicin-resistant M. tuberculosis strains is discussed.

Antibiotics, Antitubercular↗

[Biological properties of laboratory strains and clinical isolates of mycobacteria multiresistant to antitubercular preparations].

Multiresistant M. tuberculosis strains show varying drug resistance, virulence and growth rates. The count of cells of some multiresistant strains in the guinea-pig parenchymatous organs after intracardiac inoculation was comparable with that of after inoculation with clinical isolates. Furthermore, some multiresistant strains were not inferior to sensitive clinical isolates. The findings lead to the conclusion that there is a wide range in the virulence of multiresistant strains and hence they can present an epidemiological hazard.

Animals↗