[The developoment of total complement activity assay based on complement-dependent immune liposome lysis].

BACKGROUND: The purpose of work was development of a fast and reproduced procedure for measurement of the total complement activity (TCA) in human or animal blood serum. MATERIALS AND METHODS: Steady at storage liposomes preparations, which surface sensitized 2,4-DNP haptens, and the internal volume contains calceine or sulforhodamine 101 are obtained. Complement-dependent immune lysis of liposomes at presence of the anti-2,4-DNP immunoglobulines and complement preparations from animals are investigated. RESULTS: It is shown that the degree of liposomes immune lysis depends on complement concentration in a wide range that can be used for definition of TCA level. Research of blood sera from patients has revealed correlation (r = 0.793) between data received with the help of liposome immunolytic systems, and the data of nephelometric analysis with application of suspension sheep erythrocytes. CONCLUSION: The method allows to define total complement activity in blood serum in 15 minutes without separation of reaction components. This might be useful for measurement TCA level at patients with various diseases and realization of scientific researches.

[Development of the quantitative immunochromatography tests for somatic deasease markers detection].

The purpose of work was creation of quantative immunochromatographic tests (ICT) for measuring concentrations of the marker substance associated with somatic diseases: immunoglobulin E (IgE), C-reactive protein (CRP) and fibrin D-dimer in blood serum (plasma), which is carried out with the help of videodigital analyzers of domestic development "Reflecom" and "Zondaj". ICT were designed in sandwich-format, using colloid gold and monoclonal antibodies. It is shown, that calibration curves received with the help of ICTand devices of videodigital registration, are well approximated by exponential dependence Y = Aexp(-x/B)+y0, where Y--device readings, x--analytes concentration, A, B, y0--constants. Samples of blood serum (plasma), under investigation received in conditions of a hospital from patients. Correlation with electrochemiluminescent immunoassay, enzyme linked immunoassay, latex-agglutinations assay and immunochromatographic method was observed. Sensitivity of the developed tests was 7,5 IU/ML for IgE, 5,7 ng/ml for CRP, 500 ng/ml for fibrin D-dimer. The developed analytical complex- videodigital analyzer "Zondaj" and ICT for quantitative measure of concentration IgE, fibrin D-dimer and CRP- can be successfully applied in laboratory practice and clinical laboratory researches alongside with actual immunochemistry methods.

[The use of fluorescent liposomal immunoanalysis for the diagnostics and indication of glanders and melioidosis].

The authors have developed a homogenous method for the detection of the cells, surface antigen, and Ag8 of glanders and melioidosis pathogens and antibodies to them, based on the compliment-dependent lysis of Ag8-sensitized liposomes. The method is highly specific; its sensitivity to antibodies to glanders and melioidosis pathogens is 24 to 440 times higher than that of RID: its sensitivity to Ag8 is 100 ng/ml corresponding to 1.25x10-10 M, and its sensitivity to cells is 10(5) to 10(6) cells/ml).

[The use of liposomes for detection of the surface lipopolysaccharide antigen, Vibrio cholerae cells, and antibodies against them].

A test system for determination of Vibrio cholerae cells, surface O-antigen, and antibodies against them was developed on the basis of complement-dependent lysis of liposomes sensitized by the lipopolysaccharide-dependent antigen from Vibrio cholerae 569B. The factors that affect the function of the liposomal reagent were studied, and the conditions for detecting antibodies and antigenic material were optimized. This system is highly specific and sensitive to be used for the determination of anticholeraic antibodies (30-50 times as effective as agglutination tests), lipopolysaccharide antigen (100 ng/ml, which corresponded to 3.0 ng of lipopolysaccharide in the sample studied), and Vibrio cholerae cells (3.3 x 10(7) m.b./ml, which corresponded to 10(6) m.b. in sample). It takes 30-40 min to detect the lipopolysaccharide antigen and 90 min to detect V. cholerae cells.

[Remote monitoring control techniques for biological air disperse systems]. / Distantsionnye metody kontrolia aérodispersnykh sistem biologicheskogo proiskhozhdeniia.

Described in the paper are the results of designing a combination-luminescence lidar for the automated express determination of the concentration and dynamic distribution of biological pollutants in the atmosphere. It was established as possible to detect simultaneously the signals of aerosols' luminescence and those of combined atmospheric nitrogen dispersion to ensure an effective monitoring of biological aerosols along the probing route.

[Efficiency criteria and basic parameters of powder inhalers]. / Kriterii éffektivnosti i osnovnye kharakteristiki poroshkovykh ingaliatorov.

A high therapeutic efficiency and ecological safety are typical of powder inhalers. However, the designs of devices, serially manufactured in many world countries, are different, which affects their technical parameters. There are also differences pertaining to the definition "efficiency criterion" of a powder inhaler. An attempt is made in the paper to substantiate the efficiency notion of the discussed medical equipment and to analyze the methodological approaches to define such efficiency in the laboratory setting.

[Change in the dynamics of a spin probe in complement-dependent lysis of a liposome]. / Izmenenie dinamiki spinovogo zonda pri komplementzavisimom immunnom lizise liposom.

The mobility of phospholipid chains in membranes of liposomes consisting of egg lecitin, cholesterol, dicetylphosphate, sensitized by the lipopolysaccharide antigen F. tularensis by the action of a homologous antiserum and a rabbit complement preparation was studied using 5- and 16-doxylstearate spin probes. It was shown that, during the immune lysis of liposome membranes, changes in the dynamics of spin probes occur, which correlate with the formation of transmembrane channels and exit of the fluorescent marker from the interior of liposomes. It was found that the ratio of the intensities I1/I2 of two low-field extrema in the ESR spectrum is most sensitive to changes in the liposome membrane that are induced by immune components.

[Distribution of biological particles in two-phase systems of water soluble polymers and salts]. / Raspedelenie biologicheskikh chastits v dvukhfaznykh sistemakh vodorastvorimykh polimerov i solei.

The paper discusses the problems of purification of Provachek's rickettsias used to prepare typhus vaccines and diagnostic kits. The currently available agents contain many admixtures of rickettsias products, more commonly yolk and chick embryo yolk sack cell detritus, which make it impossible to determine the true antigenic properties of rickettsias in the context of protection and diagnosis. By applying the well-known principle of distribution of biological particles in the two-phase systems of water soluble polymers, investigations were made to examine the distribution of rickettsias and their infected biological mass in the system: polyethylene glycol-6600, sodium-500 dextran sulfate, potassium phosphate, sodium chloride in order to prepare agents of pure rickettsias. It has been found that rickettsias are distributed in the polymer-enriched phase, their number and the degree of purity when isolated from the infected biological mass depends on the capacity of an interphase wherein tissue detritus sorption occurs.

[Development and design of a sampling kit for microbiological studies]. / Razrabotka i sozdnie komplekta otbora prod dlia mikrobiologicheskikh issledovanii.

The paper deals with the development and design of a kit for choosing samples from various objects for microbiological studies. The studies clarified the composition of the set and requirements for the complete set and its parts. The design of this set parts was made and the design documents were drawn up for manufacturing the sampling set. A programme and procedures for microbiological and pilot tests of the kit were developed and its trials were performed.

[Use of laser flow-type fluorescence aerosol particle counter to evaluate the concentration of microbes in the surface air under high dust content]. / Primenenie lazernogo protochno-fliuorestsentnogo schetchika chastits aérozolei dlia otsenki schetnoi i massovoi kontsentratsii mikrorganizmov v prizemnom sloe vozdukha v usloviiakh vysokoi zapylennosti.

The paper deals with the use of a laser flow-type fluorescence aerosol particle counter to evaluate the concentrations of microbes in the surface air under high dust content. Various circuits of flow-type optic aerosol recorders are analyzed. Flow spectral luminescence analysis of some particles flow while exciting the fourth harmonics of a pulse laser on yttrium-aluminium garnet with neodymium by ultraviolet radiation is shown to be the most optimum method for indication of individual aerosol particles. Experiments were conducted on the authors' model of a pilot plant based on this method. The model of a laser flow-type optic analyzer was developed for experimental studies that give a clear display of biological aerosols in complex aerosols. The laser flow-type analyzer-based unit developed may provide a fluorescence signal of aerosol particles in the flow of a sample and that light diffusion signal from them at an exciting light wavelength of 266 nm. Experiments with BVC aerosols and soil dust particles were conducted in different regions of Russia. They showed it possible to detect and to rapidly calculate soil microorganisms by laser flow-type fluorescence assay of individual particles when excited by ultraviolet radiation.

[Sulforodamine 101: novel effective marker in liposomal immunoassay]. / Sul'forodamin 101 - novyi éffektivnyi marker v liposomal'nom immunoanalize.

The new fluorescence marker Sulforodamine 101 that allows one to avoid a labour-consuming stage of purification of a marker substance (for example, calceine, other fluorescein derivatives) from hydrophobic admixtures that reduce the storage of a liposomal reagent was used in liposomal immunoassay. Sulforodamine 101 shows the maximum increase in a fluorescence signal when liposomes are destroyed with encapsulated marker, which is 45 times greater than does Sulforodamine B (8 times) and comparable with calceine (30 times). The comparative studies using liposomes containing calceine and Sulforodamine 101 have indicated that they are similar storage and sensitivity (50 ng/ml) of liposomal test system as determined by the concentration of F. tularensis polysaccharides in solutions.

[Biological safety problems on the brink of the 21st century]. / Problemy biologicheskoi bezopasnosti na poroge XXI veka.

The review deals with the prospects of development of means of bioindication, one of the key elements in the country's biological safety system. The level of their development is determined by the state-of-the-art of methodology and techniques for detection of ultralow concentrations of biomolecular markers and by the advances in bioengineering and molecular diagnosis. In the authors' opinion, great progress should be soon obvious in the design of specific indication means (biosensor devices and systems for multicomponent analysis due to the use of laser sources of stimulation and new long-luminescent markers (chelates of lanthanoids and metallic porphyrins) for the homogeneous and non-dissociative schemes of solid-phase analysis. This review is followed by the collected subject-matter papers of the scientists of the State Research Institute of Biological Instrument-Making Industry who have been actively engaged in the design of bioindication technical facilities.

[Membrane-filtration immunoassay: reagents, methods and the diagnostic and technical means for detection]. / Membranno-fil'tratsionnyi immunoanaliz: reagenty, metody, diagnosticheskie i tekhnicheskie sredstva indikatsii.

The comprehensive development of dot-EIA made at the State Research Institute of Biological Instrument-Making Industry has provided devices KIMF-02 and KIMF-03), a base of chemical reagents, immunoassays, test systems for detection of a wide range of causative agents of viral and bacterial infections, that of serodiagnosis of their related diseases. The KIMF-02 kit has undergone engineering and medical tests and recommended for the Ministry of Health of the Russian Federation to produce them in stock. The kit includes all required for analysis even in an ill-equipped laboratory, a set of attached agents ensures a valid visual recording of results. The developed procedures and test systems allow the immunoassay to be as sensitive as TIFA; however, they are laborious and much simpler in design. The simple and rapid procedures of dot-EIA are recommended for incorporation into the a package of laboratory methods for verification of the accumulation of virus-specific antigens in various biological substrata, environmental samples, for control of the activity of antigens and antibodies used in serological tests, for detection of specific antigens in the clinical samples, and for serodiagnosis of infections.

[Luminescent cytochemical methods of detecting microorganisms]. / Liuminestsentno-tsitokhimicheskie metody indikatsii mikroorganizmov.

The paper shows that the luminescence cytochemical technique can be used for identification of microorganisms and microbiological synthesis products. The method is based on the interaction of specific fluorescence probes (ANS, terbium ions, and beta-diketonate complexes of europium, as well as metal-containing porphyrines) with major microbial intracellular components and toxins. Unlike classical microbiological, immunochemical or biochemical methods of detection, the proposed method has a reasonable versatility, specificity, sensitivity, rapid action, and possible automation.

[The development and testing of test systems for the determination of herpes simplex virus and cytomegalovirus antigens by lanthanide immunofluorescence analysis]. / Razrabotka i aprobatsiia test-sistem dlia opredeleniia antigenov virusa prostogo gerpesa i tsitomegalovirusa metodom lantanidnogo immunofliuorestsentnogo analiza.

On the basis of the lanthanide immunofluorescent assay (LIFA) test systems for the determination of herpes simplex virus (HSV-LIFA) and cytomegalovirus (CMV-LIFA) antigens have been developed. The test system HSV-LIFA includes the sandwich of monoclonal antibodies (McAb) HSV A.3.3. or B-11 to type 2 HSV, strain BH; the test system CMV-LIFA includes the sandwich of rabbit McAb to CMV, strain AD 169. The approbation of the test systems has revealed that they insure the specific detection of HSV and CMV antigens in clinical specimens (urine, blood, liquor, saliva), the LIFA results well correlating with the data on the isolation of viruses in cell cultures, with the results obtained by other diagnostic methods and with clinical manifestations of diseases. LIFA has been shown to be more sensitive than the enzyme immunoassay.

[The detection of antibodies to the Venezuelan equine encephalomyelitis virus by immunoenzyme analysis using antigen purified in a water-polymer system]. / Vyiavlenie antitel k virusu venesuél'skogo éntsefalomielita loshadei metodom immunofermentnogo analiza s primeneniem ochishchennogo v vodno-polimernoi sisteme antigena.

Human and animal sera were tested for the presence of antibodies to Venezuelan equine encephalomyelitis (VEE) virus by direct enzyme immunoassay (EIA). For the test, the plates were sensitized with a VEE virus preparation purified in a two-phase system of water-soluble polymers. The proposed EIA variant was as specific as that with VEE antigen obtained by fractionation on sucrose cushion, but more sensitive. The high specificity of the assay allowed the antigen purified in the water-polymer system to be used for investigation of antigen relationships among viruses of the VEE complex.