Preparations for serodiagnosis of diseases due to causative agents of ixode tick-borne Borreliosis (Lyme disease). Communication 2. Comparative study of recombinant enzyme immunoassay test-systems (rELISA) for serological diagnosis of ixode tick-borne Borreliosis]

One foreign and two Russian recombinant enzyme immunoassay test-systems were comparatively investigated under conditions of an encoded experiment. Sensitivity in the experiment with the Russian test-systems Borreliosis-ELISA-IgG and Lyme Best was 63.8 and 68.8% respectively. As for the test-system Borrelia IgG Recombinant, it was 47.5%. All the test-systems were highly specific (94.4 to 99.5%). The test-systems Lyme Best and Borrelia IgG Recombinant revealed partial cross reactions with sera from patients with systemic lupus erythematosus and leptospirosis.     

Preparations for serodiagnosis of diseases due to causative agents of ixode tick-borne borreliosis (Lyme disease). Communication 1. Comparative study of the 1st generation enzyme immunoassay test-systems for detection of antibodies to Borrelia burkdorferi sensu lato]

Four foreign and one Russian 1st generation test-systems for detecting class G antibodies or summary antibodies to Borrelia burkdorferi sensu lato were comparitively investigated with the use of the clinical material under conditions of an encoded experiment. Cross reactions with sera from patients with syphilis, Epstein-Barr infection, cytomegalovirus infection and systemic lupus erythematosus were observed. The best specificity and sensitivity parameters were provided by the Enzygnost Borreliosis test-system.     

Sensitivity of Escherichia coli O157:H7, a pathogen of hemorrhagic colitis, to antibacterial drugs]

Antibioticograms of enterohemorrhagic strains of serogroup O157 Escherichia coli isolated in the Russian Federation and Japan were comparatively studied. Strains with multiple drug resistance were detected. The main biochemical characteristics of the isolates were investigated. Significant differences in susceptibility spectra of the isolates and in their fermentative properties were revealed.     

The use of molecular biological methods for the individual characterization of Mycobacterium tuberculosis strains

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

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.     

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.     

Characterization of Russian universal kits for real-time PCR: Application to molecular oncodiagnosis

Russian real-time PCR (Q-PCR) kits and reference foreign kits were experimentally compared using serial dilutions (30–300,000 copies) of a plasmid containing a cDNA fragment of fusion PML-RARα (DNA standard). The Russian RialitiTM kit and ABI TaqMan PCR core reagent kit were similar in the efficiency of amplification (1.919 and 1.927, respectively) and in the coefficient of correlation obtained for the standard calibration plots (0.999 and 0.996, respectively). The RialitiTM kit was used to estimate the copy number of PML-RARα and ABL (internal control) in cDNA specimens obtained from peripheral blood and bone marrow of patients with acute promyelocytic leukemia and the chromosome translocation t(15;17), which generates chimeric PML-RARα. The PML-RARα copy number per μg total RNA ranged from 58,600 to 831,500 before therapy. After 3.5 months of chemotherapy, the patient showed no signs of the minimal residual disease: PML-RARα was undetectable by Q-PCR, which agreed with clinical data. Studies with the example of PML-RARα demonstrated the suitability of the RialitiTM kit for oncodiagnosis.     

Bartonella and bartonellosis--emerging and re-emerging infections. Taxonomy, bacteriology, pathogenesis and genetics

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.     

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.