[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.

Application of water-soluble nanofilters for collection of airborne Mycobacterium tuberculosis DNA in hospital wards.

A simple inexpensive technique for collection of airborne biomarkers of nosocomial infections is described. Biomarkers were collected on water-soluble electrospun nanofilters attached to a household vacuum cleaner from 6-10m(3) of air in 10-15min within several wards of a tuberculosis clinic. Filters were then dissolved in water and tested for the presence of the IS6110 and regX3 genes of Mycobacterium tuberculosis (MTB) using real-time polymerase chain reaction.

[Development and use of a domestic test system for diagnosis of tuberculous infection on the basis of quantitative analysis of interferon-gamma induction in whole blood samples in vitro, by using specific recombinant antigens].

A test system was developed to detect tuberculous infection by qualitative analysis of interferon-gamma (IFN-gamma) in the plasma samples after 20-24-hour incubation of whole blood samples in the presence of Mycobacterium tuberculosis (MBT) antigens: tuberculin PPD and a mixture of the MBT-specific recombinant antigens ESAT-6 and CFP-10. The analysis used 3 test tubes each containing 1 ml of heparinized venous blood, one of which served as a control; the other two test tubes were employed to measure antigen-induced IFN-gamma production. Whether this test system might be used to determine primary tuberculous infection was studied in 277 children and adolescents.

[Polymerase chain reaction to determine and control drug-resistant Mycobacterium tuberculosis strains].

Real-time polymerase chain reaction was used to develop a one-stage procedure for molecular genetic analysis of Mycobacterium tuberculosis (MBT) DNA in order to determine mutations associated with drug resistance to the antituberculous agents: isoniazid and rifampicin. To analyze the spread of drug-resistance of the causative agent of tuberculosis in Russia, two thousand MBT strains were studied in 24 regions of all the federal districts. Testing 1406 MBT strains isolated by first detected and untreated patients revealed multidrug resistance (MDR) in 21.

[Use of polymerase chain reaction for the diagnosis and evaluation of chemotherapy for pulmonary tuberculosis].

Polymerase chain reaction (PCR) using a new procedure for preparing sputum samples for DNA isolation on the basis of immunomagnetic separation of mycobacteria was employed to examine sputum samples from 141 patients with first diagnosed pulmonary tuberculosis and 510 diagnostic materials from patients with nonspecific lung disease. In 47 patients with pulmonary tuberculosis, sputum bacterial isolation was followed up, by using PCR and conventional microbiological studies during regular, every 6-7-week, examination. The sensitivity of PCR employing the above procedure for preparing the samples averaged 66% versus 48% when a cultural study was applied.

[The effectiveness of Mycobacteria tuberculosis isolation by polymerase chain reaction: results of a randomized trial].

The results of testing sputum for Mycobacterium tuberculosis by different methods: polymerase chain reaction (PCR), luminescence microscopy, and culture tests were compared. Clinical sputum samples were studied in 62 patients with pulmonary tuberculosis and 72 patients with non-specific diseases of the lung. The specificity of PCR was 98.

[Direct genetic analysis of the rifampicin resistance of M. tuberculosis isolates in sputum samples].

The paper shows a rapid method for diagnosing the resistance of Mycobacterium tuberculosis to rifampicin in the testing of clinical sputum samples. The sputum samples from 12 patients ineffectively treated for pulmonary tuberculosis were treated by the immunomagnetic mycobacterial separation technique; polymerase chain reaction was used to perform the amplification and direct sequencing of the gene fragment rho poB by identifying the mutations responsible for mycobacterial rifampicin resistance. Other equal parts of the same sputum samples were cultured on liquid medium for 5 days and subsequently examined in the same manner and also cultured on the Löwenstein-Jensen solid medium, followed by the determination of rifampicin sensitivity by the routine procedure.

[Value of molecular biological methods in the diagnosis of urogenital tuberculosis].

Enzyme immunoassay (EIA) was used to study the diagnostic value of determination of serum Mycobacterium tuberculosis (MBT) antibodies in 41 patients with active urinary tract tuberculosis, 14 with inactive tuberculosis and 140 with nontuberculous diseases of the urinary system. The sensitivity of EIA was 73% with 88.6% sensitivity.

Detection of rifampicin-resistant Mycobacterium tuberculosis strains by hybridization and polymerase chain reaction on a specialized TB-microchip.

Two alternative methods for identification of rifampicin-resistant strains of Mycobacterium tuberculosis on biological microchips are developed. The methods are based on detection of point mutations and other rearrangements in the rpoB gene region determining rifampicin resistance. Hybridization on TB-microchip detects 30 mutant variants of DNA in rifampicin-resistant strains (about 95% of all resistant forms).

[Determination of antituberculous antibodies in the cerebrospinal fluid by an immunoenzyme assay method in patients with tuberculous meningitis].

Possibilities of using enzyme immunoassay to identify specific antibodies to M. tuberculosis in the cerebrospinal fluid of 44 patients with tuberculous meningitis and 81 cases with other diseases of the central nervous system were analysed. The assay sensitivity and specificity in an active period of tuberculous meningitis made up 100 and 97.

[Determination of antibodies to mycobacterial antigens in tuberculosis by erythrocyte immunoadsorption with a rosette marker].

A solid-phase enzyme immunoassay system for the determination of antibodies to mycobacterial antigens, based on the method of erythrocyte immunoadsorption in microchambers for immunological reactions, has been developed. To detect antibodies specifically bound with the solid-phase antigen, the affinity rosettes of Staphylococcus aureus strain Cowan I, carrying protein A, with erythrocytes conjugated with human gamma globulin have been used. The significant correlation of the titers of 34 sera, determined by means of erythrocyte immunoadsorption, with extinction values obtained in the solid-phase enzyme immunoassay of antibodies to Mycobacterium tuberculosis has been established.