[Genomic polymorphism in Mycobacterium tuberculosis strains]. / Issledovanie genomnogo polimorfizma shtammov Mycobacterium tuberculosis.

Different genomic fingerprinting techniques (universal probes, such as rRNA genes, phage M13 DNA, IS 6110 probe) have been used to investigate the genomic polymorphism of Mycobacterium tuberculosis strains isolated in different geographical regions of Russia and in some CIS countries. As shown with the use of these techniques and a specially developed PCR-mediated system for genetic typing, M.tuberculosis strains are genotypically heterogeneous in regions with a sporadic level of tuberculosis morbidity and genotypically homogeneous in regions with elevated morbidity and mortality levels. The evaluation of the effectiveness of the genetic typing of M.tuberculosis with the use of different genomic fingerprinting techniques has made it possible to propose the optimum 3-stage scheme for the differentiation of M.tuberculosis strains: (1) the typing of all isolated strains of the PCR-mediated test system; (2) the typing of several selected M.tuberculosis strains with the use of 1S 6110 probe (2-3 strains of each detected PCR-RFLP [correction of PLRF] genotypes); (3) the typing of M.tuberculosis strains, containing 1 copy of 1S 6110 or not containing such sequence, with the use of probes (phage M13 DNA) detecting hypervariable sequences in M.tuberculosis genomes.

[New microbiological Techniques in diagnosis of tuberculosis]. / Primenenie novykh mikrobiologicheskikh tekhnologii v diagnostike tuberkuleza.

Microbial spectrum in patients with disorders of the upper respiratory tract consisted mainly of streptococci, staphylococci and pneumococci in 1991-1993, 1994 and later, respectively. Most specific for detection of nontuberculous flora was brushing with protecting agar plugs in combination with transport medium and anaerobic culturing. Direct immunofluorescence allows detection of antigens M. chlamydia in 29.4%, M. pneumoniae in 18.5% of cases. In diagnosis of tuberculosis the following new technologies were introduced: bacterioscopy with previous culturing of the material on liquid culture media followed by biological test and polymerase-chain reaction; determination of drug resistance based on nitrate reductase activity of M. tuberculosis indicating resistance of M. tuberculosis on liquid media in 4-7 days, dense egg media in 8-12 days, usage of Popesku medium enables correction of tuberculosis chemotherapy one month after the test initiation; M. tuberculosis identification using Western blot with monoclonal antibodies in some cases for 7 days.

[Use of microscopy and inoculation for bacteriologic diagnosis in tuberculosis patients by the materials of Ivanovo region]. / Ispol'zovanie mikroskpii i poseva dlia diagnostiki bacteriovydeleniia u bol'nykh tuberkulezom po materialam Ivanovskoi oblasti.

The paper describes guidelines for isolation of Mycobacterium tuberculosis by bacterioscopy. It shows high detection rates for Mycobacteria in patients included into the programme. The detection rates of patients by bacterioscopy are not different in two quarters.

[The detection of mycobacteria in children and adolescents by using the polymerase chain reaction]. / Vyiavlenie mikobakterii s pomoshch'iu polimerazno-tsepnoi reaktsii u detei i podrostkov.

A polymerase chain reaction (PCR) was used for determination of mycobacterial DNA in clinical samples from children and adolescents. The results were positive in 23 and 53% of cases, respectively, while standard microbiological methods failed to show presence of any bacteria in the samples. Case histories contained information on high incidence of positive Mantoux test. Microbiologically the diagnosis was confirmed only in 7 adolescents. In children mycobacteria were not found.

[S and R forms of Mycobacterium tuberculosis as a problem of producing cloned and phenotypically competent strains]. / S- i R-formy Mycobacterium tuberculosis kak problema polucheniia klonirovannykh i fenotipicheski polnotsennykh shtammov.

The authors suggest that animals and humans carry M. tuberculosis in S form. These mycobacteria are virulent, do not form conglomerates and represent biologically most potent form. The state of R-form M. tuberculosis results from their culturing on artificial culture media which fail to secure compatible conditions for cells. M. tuberculosis S-form is convenient for microbiological and molecular-genetic research. The authors offer a method of biological production of single M. tuberculosis in S form.

[Biological properties of Mycobacterium tuberculosis in relation to sensitivity to antibacterial preparations]. / Biologicheskie svoistva mikobakterii tuberkuleza v zavisimosti ot chuvstvitel'nosti k antibakterial'nym preparatam.

Biological properties of M. tuberculosis were investigated in 265 patients with tuberculosis of respiratory organs. Slow but more massive growth in culture medium is characteristic for drug-resistant M.tuberculosis. 32 patients discharging drug-resistant M.tuberculosis had unstable drug resistance to isoniazid, kanamycin, ethambutol and streptomycin. It is suggested that drug-resistant M.tuberculosis by virulence are similar to sensitive varieties.

[Determination of drug sensitivity of Mycobacterium tuberculosis on solid and liquid culture media]. / K voprosu ob opredelenii lekarstvennoi chuvstvitel'nosti mikobakterii tuberkuleza na plotnykh i zhidkikh pitatel'nykh sredakh.

The paper gives a comparative characterization of some unfavourable factors influencing antituberculous agents which are a part of dense egg and liquid media in order to determine the drug resistance of Mycobacterium tuberculosis. The liquid media are more preferable primarily, mainly by preserving the activity of antituberculous agents to set accelerated methods for the detection of the spectrum of and the resistance of clinical mycobacterial isolates.

[Identification of tuberculosis pathogens in clinical material using the polymerase chain reaction]. / Identifikatsiia vozbuditelia tuberkuleza v klinicheskom materiale s pomoshch'iu metoda polimeraznoi tsepnoi reaktsii.

The authors examined the possibility of detecting M. tuberculosis cells in various types of diagnostic material (sputum, blood, bone marrow, bronchoalveolar lavage fluid) from tuberculosis patients using polymerase chain reaction (PCR). The developed PCR-based test systems helped detect M. tuberculosis in 48 (90.6%) out of 53 tuberculosis patients, in contrast to much slower microbiological methods which permitted detection of Mycobacteria in only 21 (39.6%) patients. High specificity and virtually no false-positive results of PCR were demonstrated in testing diagnostic material from patients with chronic nonspecific pulmonary diseases and from children with lympholeukemia and anemia.

[Biological characteristics of M. tuberculosis and difficulties in microbiological diagnosis of tuberculosis]. / Biologicheskie osobennosti M. tuberculosis i trudnosti mikrobiologicheskoi diagnostiki tuberkuleza.

The paper outlines difficulties in tuberculosis diagnosis and effectiveness of various methods of mycobacteria identification. The latter along with direct and indirect determination of mycobacterial drug resistance require perfection. There is a need in investigation of diagnostic samples from tuberculous and respiratory sarcoidosis patients for mycobacterial granular forms. A multimodality diagnostic approach to examination of clinical material from tuberculous patients is advocated.

[Is sarcoidosis a chronic persistent infection?]. / Iavliaetsia li sarkoidoz khronicheskoi persistiruiushchei infektsiei?

The study carried out with the use of microbiological diagnostic methods has revealed that in 67% of cases specimens obtained from sarcoidosis patients for analysis contain different forms of mycobacteria (typical Mycobacterium tuberculosis and granular forms of mycobacteria). The content of typical and granular forms of mycobacteria detected in diagnostic specimens has been shown to differ, depending on the clinical form of sarcoidosis: as a rule, in cases of the sluggish course of sarcoidosis granular forms of mycobacteria are detected, while during the exacerbation of the disease and in cases of the acute course of newly diagnosed sarcoidosis the proportion of typical M.tuberculosis increases. To verify M.tuberculosis with greater certainty, two highly sensitive and specific amplification test systems have been developed on the basis of polymerase chain reaction. In this article the goals of microbiological and molecular genetic investigations which may jointly give direct proofs of the etiological importance of mycobacteria in sarcoidosis are considered and discussed; sarcoidosis may probably be regarded as chronic persistence infection.

[Test-systems of polymerase chain reaction for determining the tuberculosis pathogen]. / Test-sistemy na osnove polimeraznoi tsepnoi reaktsii dla identifikatsii vozbuditelia tuberkulëza.

Two polymerase chain reaction-based test systems were used to identify Mycobacteria tuberculosis by using a clinical material for practical health purposes. The former test system strictly specifically enables M. tuberculosis to be detected. The latter also allows one to differentiate a BCG vaccine strain from the M. tuberculosis. The test systems were tested by using 11 clinical sputum samples, three of them were taken from patients with fibrocavernous tuberculosis, two from those with focal tuberculosis and one from that with infiltrative one, 5 from those with non-specific pulmonary diseases (chronic bronchitis and asthma). In addition, one serum sample from a patient with fibrocavernous tuberculosis was examined. All tests obtained from patients with tuberculosis were positive, whereas those from patients with non-specific pulmonary diseases were negative. This suggests that differentiation of BCG vaccine strain from M. tuberculosis can be used both in model experiments and in the study of clinical material lysates.

[The use of DNA probes for the diagnosis of pulmonary tuberculosis]. / Primenenie DNK-zondov dlia diagnostiki legochnogo tuberkuleza.

Mycobacterium-specific nonradioactive DNA probes have been tested able to detect mycobacteria in clinical samples (sputum, bronchoalveolar lavage) for 48 hours. The probes sensitivity makes up 1000-10,000 cells. Molecular hybridization confirms the results of DNA-probe diagnostic conclusion of tuberculosis more frequently than the results of standard microbiological tests.

[The multidimensional character of the niches in Gause's law applicable to prokaryotes and the methods for the spatial separation of competing associates]. / Mnogomernost' nish v zakone Gauze primenitel'no k prokariotam i metody prostranstvennogo razdeleniia konkuriruiushchikh assotsiantov.

The multidimensional niche in Gause's law includes, with respect to prokaryotes, the following factors: the alimentary factor, the symbiosis-antagonism complex, the respiratory function, the concentration of associants, mobility. Cultivation in U-shaped tubes is the optimum method for the detection of spatial separation on account of such factors as respiration and mobility.