Immunochromatographic test system for pathogenic helicobacter pylori strains detection
SUBSTANCE: immunochromatographic test system for detection of pathogenic H. pylori strains based on cagA protein is a multimembrane composite consisting of a nitrocellulose membrane with a membrane impregnated with a monoclonal antibody conjugate, a HP-387 clone at a concentration of 20 mcg⋅cm-3 with nano-particles of colloidal gold with an average diameter of 30 nm, a sample membrane and an adsorbing membrane. Monoclonal antibodies, HP-1811 clone at a concentration of 12 mcg⋅cm-3 are applied to the analytical membrane in the test zone, and in the control zone - goat anti-species antibodies against mouse Ig are applied at a concentration of 10 mcg⋅cm-3.
EFFECT: invention provides determination of the said protein in various biological materials.
1 dwg, 1 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: presented group of inventions refers to medicine. Presented are methods and devices for measuring processed polypeptide neurotoxin in a solution by subtracting the amount of partially processed and unprocessed neurotoxin from an amount of total neurotoxin. Presented are kits that comprise a system of first capture antibody, second capture antibody and detection antibody, media for computing an amount of mature polypeptide neurotoxin and related instructions.
EFFECT: presented group of inventions enables measuring processed neurotoxin in a preparation effectively.
24 cl, 4 dwg, 3 tbl
SUBSTANCE: differential pre-operative diagnosis of the fast growth patterns of leiomyoma is ensured by measuring the relative content of CD3+CD56+CD158i lymphocytes in the peripheral venous blood of a woman with the fast-growing leiomyoma. If the derived value is equal to 0.8% or less, the true growth of leiomyoma is diagnosed, while the value of more than 0.8% shows the false growth of myoma.
EFFECT: using the given technique enables the preoperative diagnosis of the fast growth patterns of leiomyoma in the females of reproductive age that enables developing the optimum management of the patient in good time and selecting the conservative and surgical methods of treating.
1 tbl, 3 ex
SUBSTANCE: invention refers to immunological methods of analysis and can be used for mass screening of congenital diseases in newborn children. That is ensured by immobilising in a microplate well first immune-specific components to thyrotropin, immunoreactive trypsin, thyroxin and 17α-OH-progesterone in the form of discrete microregions, placing in the well a paper disk of a dry blood sample, extracting the detectable markers from dry blood stains by adding into the well a buffer containing danazol and 8-anilinonaphthalene-1-sulfonic acid and 17α-OH-progesteron antibodies, which are bound to extracted 17α-OH-progesterone to form an immune complex with anti-species antibodies in the respective microregion simultaneously, then adding into the microplate well a reaction solution of second immune-specific components, containing a mixture of biotin-conjugated antibodies to thyrotropin, immunoreactive trypsin and thyroxin, and a 17α-OH-progesteron-protein-biotin conjugate for preparing the immune biotin-labelled complexes in the discrete microregions, removing the paper disk of the dry blood sample, adding a streptavidin and Pt-coproporphyrin conjugate to prepare the phosphorescent biotin-streptavidin complexes in the microregions on the bottom of the microplate well and detecting the label phosphoresce emission, by scanning the discrete microregions by a focused laser beam sequentially.
EFFECT: using the given method enables detecting biomarkers specifically and qualitatively in the newborn's capillary blood stain sample that enables diagnosing three newborn's congenital conditions.
10 cl, 7 dwg, 10 tbl
SUBSTANCE: invention refers to medicine, namely to child neurology. A method includes detecting the clinical signs of a disease, measuring erythrocytes with micronuclei in a child's peripheral blood. The novel in the method for determining a degree of severity of the disease in the children with ICP is that the clinical signs of the disease are assessed by the Rivermead Mobility scale, and then an enzyme immunoassay is used to determine the qualitative content of the tumour necrosis factor (TNF-α) in the ICP children's saliva; the measured and determined values are compared to the clinical signs, and if the measured erythrocytes with micronuclei make 0.55±0.14%, saliva TNF-α increases to 13.23±5.2 pg/ml, a moderate severity is stated, while erythrocytes with micronuclei making 1.27±0.87% and more, and saliva TNF-α being 25.41±5.42 pg/ml and more show the severe disease.
EFFECT: invention enables providing the simple, non-invasive, accessible instant method for determining a degree of severity of the disease and prescribing the timely treatment.
5 dwg, 8 tbl
SUBSTANCE: invention represents an instant diagnostic technique for acute intestinal infections (AIIs), involving detecting indication markers of the AII aetiology with the use of laboratory immunology tests, differing by the fact that the AII aetiology is stated in children of an early age category, preferentially in the newborn children; that is accompanied by measuring the concentration of cytokine, interleukin IL-10 in coprofiltrate and diagnosing chronic placental insufficiency (CPI); a probability (P) of the bacterial AII aetiology is calculated; the value P of more than 50% testifies to the bacterial AII aetiology, while the value P being less than 50% shows the absence of the bacterial AII aetiology, and enables considering the diagnostics second stage to be necessary, which implies measuring the concentration of cytokine, interleukin IL-4 in coprofiltrate; the time of latching the newborn child to the breast is established with considering the type of feeding; that is combined with calculating a probability (P) of the viral or viral-bacterial AII aetiology, with the value P of more than 50% testifying to the viral AII aetiology, while the value being less than 50% makes it possible to state the viral-bacterial AII aetiology.
EFFECT: more accurate diagnosing of the aetiology of acute intestinal infection and simplifying the diagnostic procedure.
SUBSTANCE: venous blood and oral fluid are sampled from a patient in any sequence. An oral fluid biomarker is presented by carcino-embryonal antigen (CEA). A blood plasma biomarker is presented by neuron-specific enolase. If the oral fluid CEA is found in an amount of 196.8-318.4 ng/ml, while an amount of plasma neuron-specific enolase is 26.3-34.2 ng/ml, sarcoma of jaw bones is diagnosed. If the oral fluid CEA is found in an amount of 1228.2-1762 ng/ml with an amount of plasma neuron-specific enolase being 38.67-57.67 ng/ml, squamous cell carcinoma of jaw bones is diagnosed. The findings are used to predict a therapeutic approach to the patient.
EFFECT: invention provides high-sensitive and accurate differential diagnostic technique of maxillary and mandibular malignant new growths on the day of visit to a doctor.
2 cl, 6 ex
SUBSTANCE: group of inventions relates to medicine and deals with a method of determining in an individual's sample of a protein of neutrophilic origin, gelatinase-associated lipocalin (NGAL), for determination of acute kidney injury in the individual, where the predominating amount of monomer and/or heterodimer forms of NGAL protein, in comparison with the dimer form of NGAL protein, shows that NGAL protein originates from the individual's kidneys and that the individual has acute kidney injury, whereas equal or predominating amount of the dimer form of NGAL protein, in comparison with the monomer or heterodimer form of NGAL protein shows that NGAL protein originates from the individual's neutrophils and that the said individual does not have acute kidney injury; a set for determination of relative amounts of monomer, dimer or heterodimer forms of NGAL; application of the device in the said method.
EFFECT: group of inventions ensures more accurate diagnosis and therefore contributes to better directed treatment.
17 cl, 2 ex, 10 dwg, 1 tbl
SUBSTANCE: invention is a method of determining the nonspecific resistance of pathogenic microorganisms to antibiotics and the fact of the presence of bacterial biofilms on the basis of measurement of catalytic activity of phosphodiesterases cleaving the cyclic diguanosine monophosphate, with a threshold sensitivity of 50 pg/ml, comprising: 1) isolating the target-phosphodiesterase from lysed bacterial cells; 2) binding of phosphodiesterase with biotin-conjugated antibodies specific for non-catalytic domains of phosphodiesterase; 3) affinity purification of complexes formed by target-phosphodiesterase and biotin-conjugated antibody using paramagnetic particles containing neutravidin or its analogs that bind biotin; 4) interacting of the complexes of phosphodiesterase/biotin-conjugated antibody, immobilised on paramagnetic particles with complexes containing a-di-GMP in the form of G-quadruplex systems with intercalate dye, which is accompanied by decrease in the intensity while destruction of complexes of intercalate dye with c-di-GMP; 5) measurement of decrease of fluorescence upon hydrolysis with c-di-GMP and destruction of complex of c-di-GMP with intercalate dye, followed by quantitative estimation of the phosphodiesterase activity based on calibration curves made using known amounts of the recombinant enzyme of phosphodiesterase identical to the test target; 6) identification of increased level of phosphodiesterase activity detected by the test antibiotic-resistant bacterial strains capable of biofilm formation, as compared with the level of phosphodiesterase activity that can be detected for the control strains of bacteria of the same species not having the antibiotic resistance and the ability to form biofilms.
EFFECT: method enables to determine the nonspecific resistance of pathogenic microorganisms to antibiotics and to establish the fact of the presence of bacterial biofilms.
4 dwg, 5 ex
SUBSTANCE: invention relates to the field of immunology, namely to enzyme-immunoassay, in particular to a method of detecting forms of vascular endothelial growth factor (VEGF) with a size more than 110 amino acids in a biological sample. The method includes the following stages: contact and incubation of the biological sample with an uptake reagent, immobilised on a solid substrate, where the uptake reagent contains a monoclonal antibody, which recognises and specifically binds with residues, in quantity more than 110, from human VEGF; separation of the biological sample from the immobilised uptake reagents; contact of the immobilised molecular complex of the reagent of the uptake-target with detected antibody, which binds with VEGF domains, responsible for binding with KDR and/or FLT1 receptor, or which binds with an epitope in VEGF1-110; measurement of the level of VEGF110+, bound with reagents of the uptake, with application of means of detection for the detected antibody. Set of immune assay reagents for detection of VEGF110+ forms in the biological sample. An antibody 5C3, obtained from hybridoma 5C3.1.1 with a depositary number PTA-7737, with the said antibody 5C3 binding VEGF110+ forms, including VEGF121+. Hybridoma 5C3.1.1, deposited in ATCC with the depositary number PTA-7737, to obtain the monoclonal antibody 5C3.
EFFECT: application of the claimed invention makes it possible to increase accuracy of detecting VEGF isoforms, which must not include isoform VEGF110 and must obligatory include isoform VEGF121.
25 cl, 3 dwg, 2 tbl, 1 ex
SUBSTANCE: invention represents an immunoassay reagent which contains an agent binding to an analyte in a diluent, and glycosaminoglycan in an amount sufficient to decrease non-specific binding in an analyte sample. In the presented immunoassay reagent, the analyte is troponin I binding to the analyte; the agent is a biotin-modified anti-troponin I antibody, and glycosaminoglycan is chondroitin sulphate. Also, the invention provides a composition containing the troponin I binding agent, and chondroitin sulphate in an amount sufficient to decrease non-specific binding in the troponin I sample. What is also provided is a method of detecting the analyte in the sample wherein non-specific binding is decreased by the use of glycosaminoglycan.
EFFECT: method improvement.
5 ex, 5 dwg
SUBSTANCE: invention relates to investigation and analysis of high-molecular weight materials via infrared spectroscopy when determining the composition of polyacrylate and polyacrylonitrile copolymers to control the quality of carbon fibre. The method includes measuring the infrared absorption spectrum of films of test samples of polyacrylonitrile fibre using Fourier transform infrared spectroscopy in the 3000-800 cm-1 region and determining the content of acrylonitrile and methyl acrylate from normalised spectra based on characteristic peaks thereof. During sample preparation, the number of sample preparation steps is maximally simplified and reduced and dimethyl sulphoxide which does not absorb in the operating infrared region is used. When processing the obtained infrared spectra, the method employs adjustment of the entire base line, smoothing of the shape of the peaks of the investigated compounds and decomposition of the composite peak at 1733±3 cm-1 into components.
EFFECT: invention provides a method for reproducible, precision and sensitive determination of basic components of polyacrylonitrile.
2 cl, 3 tbl, 1 dwg
SUBSTANCE: invention relates to genetic engineering and biotechnology. Disclosed is a method of evaluating bioactivity of chemical compounds, where the first step includes transient transfection of cell line HEK 293 with plasmid vector pX-Y-neo (X is any eukaryote transcription factor, Y is a proteotypic peptide corresponding to said transcription factor), which contains a minimal human adenovirus type 5 promoter; a green fluorescent protein gene; a nucleotide sequence which codes the binding site of the transcription factor; a nucleotide sequence which codes the proteotypic peptide; a neomycin resistance gene; the second step includes determining the activity of the transcription factor via fluorescent analysis and chromatographic-mass spectrometer measurement of the content of the proteotypic peptide in the transfected cell culture in the presence of the test substance compared to a transfected intact cell culture.
EFFECT: invention provides fast and highly sensitive evaluation of bioactivity of chemical compounds.
2 dwg, 1 ex
SUBSTANCE: device for research of physical-mechanical properties of tuberous roots comprises a frame (1) with an electric motor (2) attached to it, on a shaft of which a removable disc (3) is mounted with the surface under study, and the guide (4) on which a movable trolley (5) is mounted. The movable trolley (5) is connected on one side to the screw mechanism (7) through the spring (6), and on the other side to a load (8) through the block (9). The device is provided with a frequency converter (13) which enables to adjust smoothly the speed of rotation of the removable disc (3), and also a screw mechanism (15) with a guide by which the gap between the trolley (5) and the removable disc (3) is implemented.
EFFECT: increase in the accuracy of the results of research on friction process of rest and movement of tuberous roots on different surfaces.
FIELD: measurement equipment.
SUBSTANCE: invention relates to the field of assessment of atmospheric air pollution degree and may be used in monitoring of atmospheric air in background and urbanised territory. The method provides for identification of a test site with size of 25×25 m, detection of external criteria of lichen on the test site, detection of available indicator types of lichen and frequency of their occurrence. On the basis of the produced data the lichenoindex is calculated, according to the provided classification of lichenoindex, they determine degree of atmospheric air pollution.
EFFECT: invention makes it possible to detect degree of atmospheric air pollution by lichen.
4 tbl, 1 ex
SUBSTANCE: object is positioned on porous substrate, fixed to the substrate surface and scanned by probe microscopy method. Substrate with through pores of smaller size than the diameter of a study object is used, and an object is fixated by laminar flow of liquid or gas supplied to the substrate from the side of scanning, with clamping force exerted by the flow on an object within 10-12-10-3 N range.
EFFECT: possible study of structures and mechanical properties of organic and inorganic objects, enhanced information content of nano and micro object studies by probe microscopy.
SUBSTANCE: invention relates to ecology. The invention provides a method of determining the quality of the environment by EPR-spectroscopy of lichens, including the collection of samples of lichen thalli from the trunks of trees growing in an industrial and background area, which is not contaminated by anthropogenic emissions into the environment, cleaning, drying, grinding, which is characterised by the fact, that drying is carried out at a temperature of 85-95°C to constant weight and ground, the EPR spectra are removed, from which the concentration of paramagnetic centres is determined, in the excess of the concentration of the paramagnetic centres in the lichen samples collected in the industrial zone, over the concentration of the paramagnetic centres of the lichen samples from the background area the low quality of the environment in the industrial zone is detected, and in case of equal concentrations of the paramagnetic centres - the acceptable quality of the environment is detected, and in the studies the samples of the same species of lichen are used.
EFFECT: invention provides the improvement of the method of lichenoindication, improvement of the quality of evaluation of the test objects, obtaining objective result.
2 ex, 2 tbl, 3 dwg
SUBSTANCE: method of determining ammonium compounds in the atmosphere of livestock complexes comprises collecting samples of lichen from trees growing in the background zone, which has no emissions of pollutants into the atmosphere. The data for the samples of lichen collected in the zone of pollutants emission to the atmosphere is compared with data for laboratory standards by IR spectroscopy method. For obtaining the standards under laboratory conditions the interaction process of lichen of the background zone with emission of pollutants contributing to formation of ammonium sulphate is simulated. Lichen Parmelia sulcata is used as bioindicator.
EFFECT: invention enables to determine the level of ammonium compounds in the atmosphere of livestock complexes.
2 tbl, 1 dwg
SUBSTANCE: invention relates to ferrous metallurgy and can be used to determine chemical composition of materials containing lump metal and used as raw material in cast iron production. The method involves separation of material into metal and slag fractions, measurement of metal fraction weight, grinding of slag fraction down to the fineness of 5 mm at most and determination of weight ratio of total ferrum and of necessary components in it by complete acid digestion, calculation of weight ratio of total ferrum and of components in the material, after grinding a sample is taken with the fineness of from 0.16 mm to 5 mm at most and chemical analysis is performed.
EFFECT: improved information value and reliability of analysis.
SUBSTANCE: invention refers to medicine and can be used for the prediction of the early stage of lymphocyte apoptosis. That is ensured by isolating cells, incubating for 48 hours at temperature 37°C with 5% CO2 with adding apoptosis inductor, dexamethasone in the concentration of 10-4 mole/ml. A lymphocyte viability is quantified by trypan blue inclusion, the recovered and oxidised glutathione concentrations are measured in lymphocyte lysate after the 30-minute pre-incubation with 2-vinylpyridine 10 mM. The early stage of lymphocyte apoptosis is stated, if observing an integrated decrease of the recovered glutathione concentration by 17% and more and an increase of the oxidised glutathione concentration by 19% and more as compared to the reference.
EFFECT: using the presented method in medical practice enables predicting the antioxidant state of the patient's body accompanying various diseases as shown by the early stage of lymphocyte apoptosis evaluated.
SUBSTANCE: method includes the selection and preparation of samples to be analysed, selection of specified volumes of solutions of a test system components, placement of the samples to be analysed and the test system components into a cuvette, registration of chemiluminiscence with further quantitative estimation of its value with taking into account the background signal of chemiluminiscence. The weight of porting of the sample of the material to be analysed is taken such that corresponds to the value of a specific surface 0.20±0.05 m2/g, and in case when it is not possible to determine the value of the specific surface of the sample to be tested, the weight of the taken portion is 0.010±0.005 g. The portion of the sample of the material to be analysed is placed into a cuvette with the further successive addition of the test system components: 0.01M solution of luminal in 0.5 NaOH solution and a solution of hydrogen peroxide of a 20-30% concentration to fill the working space in the cuvette, keeping the ratio luminal:hydrogen peroxide equal to 2:5. After that, values of chemiluminiscence are registered for 125 minutes and the total value of chemiluminiscence is calculated.
EFFECT: identification of the free-radical activity of solid materials by the method of the chemiluminiscence registration by means of the system of chemical reagents without the application of biological substrates in the test system.
3 tbl, 3 ex
SUBSTANCE: method is based on contacting a membrane test strip with an analysed fluid sample and initiating thereby a motion along the test strip membranes of reagents being parts of the sample or coating the membrane, and forming the immune complexes to be detected in the course of reactions in the membrane pores or on the surface thereof. A distinguishing feature of the presented method for antigen detection is that the test strip is coated additionally within the test sample contact area with a certain amount of specific antibodies, which react to the detected antigen expected to be found in the sample, when a fluid front moves and block a certain number of binding sites. The number of the coating free antibodies is specified so that the low content thereof in the analysed sample being of no diagnostic importance ensures blocking the binding sites completely that prevents the antigen from binding in the analysed area of the test strip and from developing a destructive staining in the analysed area.
EFFECT: presented approach enables reliable diagnosing based on the detection results of the antigens of gastrointestinal disorders, avoiding the achievement of positive test results for the low-antigen samples, which testifies to no development of the disease in an individual.
1 tbl, 2 ex