[Characterization of the universal Russian reagent sets for real-time PCR and its application for molecular oncodiagnostic].

Universal Russian reagents for real time PCR were tested and compared with reference reagents provided by foreign companies. Testing was carried out on plasmids with cloning fragments (DNA-standards) of cDNA with chimeric (fusion) gene PML-RARalpha. Values of amplification efficiency of Russian and foreign reagents were measured on samples with serial dilutions (30-300000 copies) of cloned cDNA fragments of PML-RARalpha and internal control gene ABL. Amplification efficiencies of Russian and foreign reagents were found to be close one to another. Russian universal reagent kit RealityTM and ABI TaqMan Core Reagent Kit have amplification efficiencies 1.919 and 1.929, and correlation coefficients of copy numbers PML-RARalpha0.999 and 0.996, respectively. These values were determined by construction of a standard curve. To verify these results we studied also the samples of cDNA from blood and bone marrow of patients with acute promyelocytic leukemia. All samples posses translocation t(15;17), and appropriate chimeric gene PML-RARalpha. copy number in 1 microg of total RNA was in range 5.86 x 10(4)-8.315 x 10(5) before chemotherapy. No symptoms of minimal residual disease were found after 3.5 months since chemotherapy - fusion gene PML-RARalpha was not detected by real time PCR method. These results are in agreement with clinical data. Our investigations tend to show that application of RealityTM reagent set in real timePCR experiments gives correct results and may be used in molecular oncodiagnostics.

[RT-PCR-based evaluation of the activity of human papilloma virus infection in relapsing papillomatosis of the larynx].

RT-PCR-based examination of papilloma samples obtained from patients with relapsing papillomatosis of the larynx showed an incidence rate of human papilloma virus (HPV) amounting to 89%. The viral load level of the studied samples, when measured by concurrent RT-PCR HPV, differed by more than 130 times. It made, in the untreated patient, 1.2 x 10(9) hormonal equivalents/ml, i.e. 13-fold higher versus the patient who received pathogenetic therapy. Thus, the approach in question provides for a possibility to monitor the activity of papilloma viral infection and to evaluate the efficiency of different variations of pathogenetic therapy because the "classic" variant of PCR-detection is not informative in the discussed case.

[Fast detection of DNA of hepatitis B virus by real time PCR].

90 blood plasma samples from patients with suspected chronic viral hepatitis B (CVHB) were analyzed by real-time polymerized chain reaction (PCR). The findings were compared with the results obtained by 2 PCR electrophoresis-based test-systems; the sensitivity limit for the quantification of DNA of hepatitis B virus (HBV) was determined; in the present case, the limit corresponded to 30 replications of HBV DNA to reaction (600 GE/ml). The positive result of real-time PCR was registered in 53.3% of cases. The quantity of HBV DNA replications in blood plasma samples varied from 30 to 3.9 x 10(6) per reaction (600-7.8 x 10(7) GE/ml). Serological profiles were analyzed in 18 patients with the verified diagnosis of CVHB. HBV DNA was detected in blood of 65% of HBsAg(+)-patients. The markers of HBeAg replication were noted in 35.5% of patients; it is noteworthy, that HBeAg(+)-samples were characterized by a higher level of viral loads (> or = 10(6) GE/ml) versus HBeAg(-)-samples (> or = 6 x 10(3) GE/ml). An analysis of blood-plasma samples dynamically obtained from one patient with chronic renal insufficiency and CVHB showed a decreased level of viral load from 1.7 x 10(7) GE/ml to a negative finding of real-time PCR registered after a therapy course by zeffix. Hence, the automated and standardized real-time PCR, when used at a multi-field patient-care facility, ensures a better diagnosis of viral hepatitis B.

[Bartonella and bartonellosis--emerging and re-emerging infections: epidemiology, clinical course, diagnosis].

Information on the epidemiology, clinic and diagnostics of bartonellosis is updated. The importance of bartonellosis lies in the fact that its main risk group embraces patients with immunodeficiencies of different origin, the number of such patients constantly growing. The diagnostics of bartonellosis is insufficiently developed and the research on Bartonella organisms, except for the trunch (Volynia) fever causative agent, is insufficient in our country.

[Bartonella and bartonellosis--emerging and re-emerging infections. Taxonomy, bacteriology, pathogenesis and genetics]. / Bartonelly i bartonellezy--novye i vozvrashchaiushchiesia. Taksonomiia, bakteriologiia, patogenez i genetika.

The review updates knowledge on the taxonomy, bacteriology and genetics of Bartonella as well as pathogenesis of bartonellosis. The role of Bartonella in human pathology, formerly considered to be rather modest, causes now growing anxiety. In this connection Bartonella are now believed to be the causative agents of so-called emerging and re-emerging diseases, i.e. diseases, formerly unknown to man, and diseases, believed to be eradicated and playing at present no important role in human pathology. These microorganisms are a bright example of successes of molecular biology in the detection of microorganisms, as well as in their phylogenetics and systematics.

[The use of molecular biological methods for the individual characterization of Mycobacterium tuberculosis strains]. / Ispol'zovanie molekuliarno-biologicheskikh metodov dlia individual'noi kharakteristiki shtammov Mycobacterium tuberculosis.

The expediency of using molecular biological methods for the evaluation of M. tuberculosis clinical strains by individual genetic certification of circulating M. tuberculosis strains has been substantiated. Considerable genetic heterogeneity of M. tuberculosis strains isolated from patients in different regions of the Russian Federation has been established; this heterogeneity is due to the presence of differences in the number of copies (5-26) of element IS6110 in M. tuberculosis cells.

[Identification of nontuberculous mycobacteria by methods of molecular biology]. / Identifikatsiia netuberkuleznykh mikobakterii s pomoshch'iu molekuliarno-biologicheskikh metodov.

A number of clinical and laboratory strains of microorganisms belonging to different species of mycobacteria of the nontuberculous complex were tested with the use of the polymerase chain reaction and the restriction analysis. The unique restriction profiles of the following species of mycobacteria have been obtained: M. fortuitum VI, M. kansasii I, M. intracellulare, M. avium.

[Resistance of Pseudomonas mallei to tetracyclines: assessment of the feasibility of chemotherapy]. / Ustoichivost' sapnogo mikroba k tetratsiklinam: otsenka vozmozhnosti khimioterapii.

The spectrum of cross resistance in tetracycline resistant strains of Pseudomonas mallei was studied. Possible use of antibacterial drugs in the prevention and treatment of mallel due to such strains was investigated in the experiments on golden hamsters and monkeys. It was shown that the efficacy of the minocycline therapy depended on the level of the strain resistance to tetracycline antibiotics. The combination of biseptol and ofloxacin proved to be highly efficient in the treatment of mallei.

[Identification of the bacterium Pseudomonas mallei using Pseudomonas pseudomallei bacteriophages]. / Identifikatsiia bakterii Pseudomonas mallei s pomoshch'iu bakteriofagov Pseudomonas pseudomallei.

Phage production by Pseudomonas pseudomallei and Pseudomonas mallei strains has been studied. 32 P. pseudomallei bacteriophages have been isolated. Their spectrum of lytic action against P. pseudomallei, P. mallei and other Pseudomonas sp. has been defined. It has been shown that P. pseudomallei bacteriophages PP19, PP23, PP33 may be used for identification P. mallei among related Pseudomonas.

[Transduction of Pseudomonas mallei bacteria]. / Transduktsiia bakterii Pseudomonas mallei.

The transducing ability of 17 Pseudomonas pseudomallei bacteriophages has been studied. Five of them were found to be capable of transducing the transposon Tn7 markers TpR, SmR into the strain Pseudomonas mallei C-5. The transduced bacterial recipients became lysogenic and acquired resistance to trimethoprim and streptomycin. Transduction of transposon Tn7 has been confirmed by DNA-DNA hybridization.