[The detection of antibodies to HCV F protein with immune enzyme analysis using synthetic peptide.]

Alternative reading frame encoding a single protein known as protein F or core + 1 / ARFP is located in the core region of the hepatitis C virus (HCV) genome. The presence of antibodies to the F protein of HCV in the serum of patients with chronic hepatitis C indicates the expression of this protein in vivo. In this study, to determine antibodies to the F protein of HCV in serum samples the methodology of the enzyme immunoassay (ELISA) was developed using the synthetic peptide F10 corresponding to the antigenic determinant of the F protein of the HCV subtype 1b. The immunogenicity and immunochemical specificity of synthetic F10 peptide has been demonstrated in laboratory animals (mice).

[The evaluation of detection rate of Mycoplasma hominis and Ureaplasma spp. using cultural technique and polymerase chain reaction].

In the clinical diagnostic laboratories of Chelyabinsk and St. Petersburg evaluation of detection rate of Mycoplasma hominis and Ureaplasma spp. was implemented using technique of polymerase chain reaction and cultural method. The reagents kits "Mycoplasma ACH-12" and "Ureaplasma ACH-12" were used to detect and determine antibiotics sensitivity of urogenital mycoplasma with determination of character of sensitivity of clinical isolates M.hominis and Ureaplasma spp. in sample of secretion of urethra and cervical channel. The results of study demonstrated that rate of detection of Ureaplasma spp. turned out significantly higher than M.hominis. The common coincidence of results in St. Petersburg between two techniques (polymerase chain reaction and cultural method) in case of detection of Ureaplasma spp. amounted to 97.5% and in case of detection of M.hominis - 93.5%.The common coincidence of the results in Chelyabinsk between two techniques (polymerase chain reaction and cultural method) in case of detection of Ureaplasma spp. amounted to 79.9% and in case of detection of M.hominis - 96.1%.

[Combined determination of spontaneous and antigen-induced production of cytokines for differential diagnostics of active tuberculosis of lungs and latent tuberculosis infection].

AIM: Choice of informative biomarkers for diagnostics of Mycobacterium tuberculosis infection and differential diagnostics of active tuberculosis (TB) of lungs and latent tuberculosis infection (LTBI). MATERIALS AND METHODS: 54 tuberculosis patients, 47 contact by TB, individuals and 43 healthy donors were examined. All the individuals included into the study had QuantiFERON-TB Gold In-Tube (Cellestis, Australia) carried out. Values of spontaneous (NIL) and antigen-induced (AG) production of 10 cytokines (EGF, MIP-1beta, VEGF, IL-2, IL-4, IL-6, IL-1alpha, IFN-alpha2, TGFalpha, TNFalpha) as well as sIL-2Ralpha and sCD40L using XMap technology were measured. IP-10 level was also determined in 48 individuals by using EIA. RESULTS: 6 out of 13 biomarkers distinguished active TB and LTBI. As a result of construction of a decision tree in JMP 9.0 program 3 most significant markers were selected. Use of combination of IFN(gammaAG-NIL), TGFalpha NIL and IL-6AG cytokines allowed to divide TB patients and contact individuals with sensitivity of 96.3% and specificity of 80.7% (AUC = 0.9). We also observed very high levels of IP-10 and IL-2 that correlated with IFN(gammaAG-NIL) (r = 0.71 and 0.79, respectively). CONCLUSION: IL-2 and IP-10 as well as IFNgamma may be used as helpful biomarkers as a first stage for diagnostics of (M.tb.) infection. At the second stage determination of IL-6, IFNgamma and TGFalpha for differential diagnostics of active TB and LTBI is proposed.

[The comparative value of quantiferonic test, neopterin and specific anti-tuberculosis antibodies in clinical diagnostic of tuberculosis of lungs].

The article deals with comparison of clinical diagnostic value of laboratory techniques of tuberculosis. The sample included 63 patients with tuberculosis of lungs. 49 persons of long-time contact with patients with tuberculosis and 28 healthy donors. The QuantiFERON-TB Gold In-Tube test was applied to total sampling. The content of neopterin and specific anti-tuberculosis antibodies in blood plasma was determined. According study data, the mentioned test is mostly informative for detection of tuberculosis contamination (sensitivity 64%, specificity 89%). However, the obvious shortcoming of this technique is the fact that it can't provide the differentiation between active and latent tuberculosis infection. As opposed to QuantiFERON-TB Gold In-Tube test, the detection of specific anti-tuberculosis antibodies (sensitivity 54%, specificity 94%) and neopterin (sensitivity 51%, specificity 92%) makes it possible to differentiate these two groups of patients.

[Development and use of test systems for the evaluation of sensitivity of Trichomonas vaginalisto preparations of the 5-nitroimidazole and 5-nitrofuran groups].

AIM: Development of test system for the evaluation of sensitivity of Trichomonas vaginalis to preparations of the 5-nitroimidazole and 5-nitrofuran groups. MATERIALS AND METHODS: Determination of minimal cidic concentration (MCC) of antiprotozoal preparations was carried out by cultivating laboratory T. vaginalis strains in wells of plates with nutrient medium containing varying concentrations of these preparations. Evaluation of vitality of the agent was determined by using trypan blue vital stain and bysubsequent growth ability in nutrient medium without antiprotozoal preparations. Construction of the test system was carried out by selecting conditions for the fixation of preparations in the plate wells and control of activity retention after the fixation. 109 isolates from patients with confirmed trichomoniasis diagnosis were used for the approbation of the test system. RESULTS: Cultivation of 10 strains showed that MCC of investigated preparations had the following values: metronidazole 5, tinidazole 1.25; secnidazole 2.5; nimorazole 1.25; ornidazole 2.5; clotrimazole 15; nifuratel 1.25 microg/ml. Studies of clinical material revealed single-type sensitivity of strains isolated during acute trichomoniasis, and varying--during chronic, while one strain had multidrug resistance. CONCLUSION: A simple test system available for routine laboratory work for the evaluation of sensitivity of T. vaginalis to preparations of the 5-nitroimidazole and 5-nitrofuran groups was developed. The efficacy of the test ensures high sensitivity, reproducibility and shorter procedure time as compared with classical method, thus allowing the selection of preparation for therapy with the highest probability.

[An immunoenzyme method for demonstrating Coxiella burnetii antigens]. / Immunofermentnyi metod indikatsii antigenov Coxiella burnetii.

Materials on the development of an enzyme immunoassay (EIA) system for the detection of the antigens of C. burnetii, the causative agent of Q rickettsiosis, are presented. The system is highly specific and effective with respect to both corpuscular antigens of phases 1 and 2 and soluble antigen (lipopolysaccharide). The sensitivity of this method varies within the range 5-100 ng/ml. The effectiveness of EIA as a quantitative (semiquantitative) control test used in the process of the production of Coxiella preparations has been demonstrated.

[A modification of an immunoenzyme test system for the serodiagnosis of opisthorchiasis]. / Modifikatsiia immunofermentnoi test-sistemy dlia serodiagnostiki opistorkhoza.

Based on the antigen derived from the whole Opisthorchis marita extraction in cats and A-peroxidase protein conjugate the authors modified the enzyme immunoassay system for the detection of IgG antibodies to the causative agent of opisthorchiasis. A vibroshaker used in all stages of study permitted them to reduce the time of the reaction and its analysis providing the visualized consideration of the results.

[Comparative evaluation of the immunoenzyme system used in the serodiagnosis of tuberculosis and non-tuberculous diseases]. / Sravnitel'naia otsenka immunofermentnoi test-sistemy pri ispol'zovanii dlia diagnostiki serodiagnostiki tuberkuleza i netuberkuleznykh zabolevanii.

A method for detection of antituberculous antibodies by enzyme immunoassay system developed by the authors, was tested on 317 blood serum samples taken from patients with tuberculosis and non-tuberculous diseases of the lungs, vertebral column, kidneys and genitalia. Depending on the process site, the detection rate of specific antibodies and their specificity made up 57-77% and 85-95%, respectively, which practically agrees with the data produced as a result of a simultaneously conduction of three routine serologic reactions.

[The use of commercial immunoenzyme and latex preparations for the demonstration of human rotavirus antigens]. / Ispol'zovanie kommercheskikh immunofermentnykh i lateksnykh preparatov dlia indikatsii antigenov rotaviusa cheloveka.

The results of using enzyme immunoassay and latex preparations for the diagnosis of rotavirus gastroenteritis are presented. High effectiveness of the enzyme immunoassay system developed in the USSR with latex diagnostic agents, such as Rotalex (Orion Diagnostica, Finland), Slidex Rota Kit (BioMérieux, France), The Wellcome Rotavirus Latex Kit (Wellcome Foundation Ltd., Great Britain), 48-63% and 21-41% respectively, has been noted. The results of the comparison of the system developed in the USSR with Wellcozyme Rotavirus, an enzyme immunoassay system manufactured by Wellcome Foundation Ltd. (Great Britain), are practically comparable. The results of the block test and the confirmation test used for control indicate that the Soviet preparation is specific. Materials on the practical evaluation of the assay system at health institutions are presented. Good prospects for the use of this system in the diagnosis of rotavirus gastroenteritis, as well as in the realization of epidemiological surveillance on this infection, are substantiated.

[Theophylline concentration in the blood of children with persistently recurring bronchial asthma]. / Issledovanie kontsentratsii teofillina v krovi u detei s uporno retsidiviruiushchei bronkhial'noi astmoi.

An antirecurrence effect of theophylline was studied in 32 children aged 3 to 12 years suffering from severe bronchial asthma. The drug dosage was determined by means of a stepwise increase of a daily dose in the range of 10-30 mg/kg/day under control of the patient's general condition and parameters of external respiratory function. Blood theophylline concentration was measured by an immunofluorescent method. The drug individual biotransformation was revealed. Euphylline proved to be a highly effective and safe antirecurrence agent in an individual selection of doses and administration regimens. Combination of theophylline with inhalation beta-adrenomimetics and corticosteroids potentiates its clinical effect without influencing biotransformation.

[Use of solid-phase immunoenzyme analysis for the serodiagnosis of pseudotuberculosis]. / Primenenie tverdofaznogo immunofermentnogo analiza dlia serodiagnostiki psevdotuberkuleza.

The diagnostic test system based on the solid-phase enzyme immunoassay (EIA) for the detection of antibodies to Yersinia pseudotuberculosis in the sera of patients with the use of Soviet-made preparations and reagents has been developed. The test has been performed in microchambers for immunological reactions, thus making it possible to decrease the consumption of reagents 10-20 times in comparison with the traditional technique with the use of plates. The results of the titration of 42 sera in EIA and in the passive hemagglutination test (PHAT) are indicative of the presence of positive correlation (r = 0.78; p less than 0.05) between antibody titers in EIA and PHAT. A fourfold or greater increase in antibody titers has been determined by means of EIA in 80% of cases and with the use of PHAT in 55% of cases. The minimum diagnostic titer yielded by EIA has been determined: 1:256.