Set of oligonucleotide primers and probes and method for quantitative determination of fetal dna in blood of pregnant woman based on analysis of hypermethylated fetal dna sections

FIELD: medicine.

SUBSTANCE: method is proposed for obtaining of DNA primers and probes to determine the presence and the relative amount of fetal DNA in a plasma sample of a pregnant woman. A method and a set are proposed to determine the presence and concentration of fetal DNA in a plasma sample of a woman.

EFFECT: effective detection of primer structures corresponding to the differentially methylated regions of the mother and fetus DNA.

17 cl, 1 dwg, 4 tbl, 1 ex



Same patents:

FIELD: medicine.

SUBSTANCE: content of calcium and protein in oral liquid is determined before and after physical load, as well as a day after physical load. Recovery of content of calcium ions and protein in oral liquid after physical load to initial values is considered to be a criterion of total recovery of athlete's-volleyball player's organism, with evaluating time interval, required for said process.

EFFECT: invention makes it possible to determine reserve abilities of organism and its adaptability in athletes-volleyball players to physical load.

2 dwg, 4 tbl

FIELD: chemistry.

SUBSTANCE: claimed is a portative analyser for the analysis of a biological fluid sample, which contains a body with a magazine, which has units for placing applied for the analysis diagnostic strips or test-strips, possessing a zone for a biological fluid, an analysing device with a slot-like receiver for the applied diagnostic strip or the test-strip, provided with electric contacts from one end, and an indicator device for the display of not fewer than one analysis result. The body from the back part side is made with a lowering, which forms a flat surface, on which along the body or transversely to it made are projections, dividing the flat surface of the lowered part into units for the placement of the diagnostic strips or test-strips and forming the magazine, located parallel in not fewer than one line. The units are closed with a demountable or an openable lid, which is a part of the body. 2 other versions of the portative analyser are also described.

EFFECT: extension of exploitation properties and increase of efficiency.

25 cl, 12 dwg

FIELD: medicine.

SUBSTANCE: spontaneous glow, an induction period and a fast flash amplitude over a period of 5 minutes of recording are measured, and an adaptation risk ratio (ARR) is calculated by formula: ARR=Cn-A/π, wherein: ARR is the adaptation risk ratio; Cn is the spontaneous glow; A is the fast flash amplitude; π is the induction period. If the ARR is 0.066-3.85 standard units, the adaptation is considered to be satisfactory, or compensated; the ARR falling within the range of 3.88-6.5 standard units stands for an adaptation tension; provided the ARR ranges 6.6-9.5 standard units, the adaptation is stated as unsatisfactory, whereas the ARR of 9.6 standard units and more shows an adaptation failure.

EFFECT: more accurate assessment ensured by determining adaptation-based grouping criteria.

1 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, and can be applicable for measuring catecholamines and their metabolites in complex array-based objects, including water-soluble, without additional sample preparation. A method is implemented by varying a principal analytical signal shaping and measurement network with the signal recorded in a sensitive biosensor layer; that is followed by a solid-phase fluorescence. The biosensor action is based on enzymatic derivatisation of catecholamines and their metabolites with organic amines (o-phenylene diamine, ethylene diamine) to form quinoxaline derivatives fluorescing in the range of 450-550 nm. The indicator test components are immobilised in the sensitive layer on the biosensor surface; that results in forming and recording the fluorescent signal on the solid surface directly in the reflection mode. The peak fluorescent signal intensity is generated when using a derivatizing agent presented by o-phenyldiamine and performing a process in the horseradish peroxidise concentration of 10-25 nM; in the hydrogen peroxide concentration of 250-500 mcM; in the o-phenylene diamine concentration of 50-100 mcM; in the concentration of catecholamines and their metabolites of 5-2000 nM. Buffer is phosphate buffer 5 mM of pH 9.5-10.0. The sensitive biosensor layer represents a double-layer film {chitosan - o-phenylene diamine/chitosan - peroxidase} applied as a smooth layer on the surface of a glass plate (14×40 mm).

EFFECT: invention provides the simple and sensitive measurement of catecholamines and their metabolites in the objects, which used to be difficult or impossible to analyse using optical detection method for instrumental reasons because the real object has a harmful influence, while the biosensors are insufficiently sensitive and reproducible.

4 cl, 4 dwg

FIELD: medicine.

SUBSTANCE: substance of the invention consists in simulating acute mesenteric ischemia in rats followed by measuring the lymphocyte count in the central venous blood, and decreasing the measured count by more than 80% from average normal values enables diagnosing intestinal necrosis.

EFFECT: higher accuracy and simplifying the diagnosis of intestinal necrosis accompanying mesenteric ischemia.

3 ex

FIELD: medicine.

SUBSTANCE: invention represents a method of differential diagnostics of chronic viral hepatitis and fatty liver disease in patients with dyslipidemia syndrome, consisting in the following: activity of the glutathione reductase antioxidant system enzyme (GLR), which is estimated by a spectrophotometric method, is determined in the patient's blood serum, and if GLR value is lower or equals 14.6 mcmol/l/min, the fatty liver disease is diagnosed, if GLR concentration is higher than 14.6 mcmol/l/min, chronic viral hepatitis is diagnosed.

EFFECT: reduction of trauma, increase of availability and simplicity of realisation with high sensitivity, specificity and effectiveness.

4 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to oncology. A combined scattering spectroscope is used to record a spectral series of the new growth and healthy tissue. A working portion of the spectroscope is placed directly above the examined region at 3-4 mm. Recording the spectra is followed by performing the statistical analysis of the results on a personal computer. Analysing involves identifying two phase signs that are a relation of peak intensities of the combined scattering in the bandwidths 1300-1340 cm-1 and 1640-1680 cm-1 to the bandwidth 1430-1470 cm-1. The combined scattering spectroscopic data obtained by each measurement are presented as a point on a phase plane. The point is found in one of three regions of the phase plane. According as which of the three regions of the plane comprises the point, either melanoma, or basal cell or squamous cell cancer, or no skin growths is diagnosed in the patient.

EFFECT: method enables obtaining objective data enabling differentiating various types of the skin growths, that provides high accuracy of the pre-operative diagnosis.

1 ex

FIELD: medicine.

SUBSTANCE: child's lachrymal fluid is examined to measure activity of γ-glutamyltransferase, and if the measured value is more than 11 Unit/l, the disease is diagnosed.

EFFECT: detecting myopia in the pre-school children when it is impossible to test the visual acuity according to the Orlova's chart.

4 ex

FIELD: biotechnologies.

SUBSTANCE: invention refers to a method of simultaneous determination of zinc cation concentration and zinc and copper cation ratio in blood serum, involving use of dithizone solution in carbon tetrachloride as the main reactant, incubation at room temperature in alkali medium, organic phase removal and colorimetry of colloid solutions of blood serum sample and standard solution at 535 nm, zinc cation concentration calculation, colorimetry of colloid solutions of blood serum sample and standard solution at 435 nm and molar ration calculation for zinc and copper cations.

EFFECT: determination of zinc cation concentration in blood serum together with zinc and copper cation ratio in the same sample without additional reactants.

4 ex

FIELD: chemistry.

SUBSTANCE: method is carried out as follows: crushing biological material containing O-(2,3-dihydro-2,2-dimethyl-7-benzofuranyl)-N-methyl carbamate, treating the material three times with a mixture of ethyl acetate and acetone taken in ratio of 1:1 by volume, each time for 30 minutes; merging the obtained extracts; evaporating the extractant; dissolving the residue in acetonitrile; diluting the obtained solution with water in ratio of 1:4 by volume; extracting with ethyl acetate twice with volume ratio of water to the organic phase of 1:1 at each extraction step; merging the ethyl acetate extracts; evaporating to a dry residue; dissolving the residue in acetonitrile; adding water to the solution to achieve volume ratio of acetonitrile to water of 4:6; adding the obtained solution to a Silasorb S18 sorbent column; carrying out a chromatographic process using a two-component mobile phase of acetonitrile-water in ratio of 4:6 by volume; merging eluate fractions containing O-(2,3-dihydro-2,2-dimethyl-7-benzofuranyl)-N-methyl carbamate; evaporating the eluent; dissolving the residue in methanol and performing determination with a physical-chemical method, which is chromatography-mass spectrometry, in a DB-5 MS EVIDEX capillary column with length of 25 m and inner diameter of 0.2 mm with a stationary phase which is 5% phenyl-95% methylpolysiloxane, with film thickness of the stationary phase of 0.33 mcm, using a helium carrier gas fed at a rate of 0.6 ml/min, and a mass-selective detector operating in electron impact mode, the initial thermostat temperature of the column being 70°C; maintaining said temperature for 3 min, then raising the temperature from 70°C to 290°C at a rate of 20°C/min; maintaining the final temperature of the column for 16 min; temperature of the injector is 250°C, temperature of the quadrupole is 150°C, temperature of the detector interface is 300°C; detecting strength of the signal resulting from charged particles formed when bombarding the analysed substance coming from the capillary column and falling into an ion source with an ionising electron beam with energy of 70 eV; recording the mass spectrum on the full ion current and calculating the amount of O-(2,3-dihydro-2,2-dimethyl-7-benzofuranyl)-N-methyl carbamate from the area of the chromatographic peak.

EFFECT: high sensitivity.

4 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to field of biotechnology, immunology and genetic engineering. Genetic modification contains deletion of endogenic low-affinity FcγR locus, where contain functional FcRγ-chain. Modified mice express up to five genes of low-affinity human FcγR on antigen-presenting cells of host immune system. Also described is method of screening substance, containing Fc site of human antibody by means of said mice. Invention can be used in the field of medicine.

EFFECT: presented are genetically modified mice, not related to man, and methods and compositions for their obtaining and application.

15 cl, 11 dwg, 6 tbl, 8 ex

FIELD: measurement equipment.

SUBSTANCE: group of inventions relates to the incubation of water samples. The incubator for water samples and the method for incubation of water samples is offered. The incubator is designed as an association of light- and heat-isolated cells. Each cell comprises a cover, a housing, a barrel with heat-isolated from the housing, a cell control unit, individual temperature stabilisation system and individual illumination stabilisation system. The individual illumination stabilisation system contains the light-emitting diode in the lower part of the barrel. The light-emitting diode provides the necessary level of light exposure of the sample. The incubation method is implemented in the offered incubator, temperature and constant illumination conditions are pre-set individually for each sample.

EFFECT: inventions allow to perform incubation of water samples in individual conditions of each sample.

4 cl, 2 dwg

FIELD: measurement equipment.

SUBSTANCE: group of inventions relates to the field of biochemistry. A biosensor system and test sensors (versions) are proposed to determine concentration of an analysed substance in a sample. The biosensor system includes a test sensor, comprising at least two electric conductors, a reaction agent containing a binding substance, including at least one water soluble polymer material, buffer salt, a water soluble medium for transfer of one or two electrons, containing at least 20% (wt/wt) of inorganic salt of non-transition metal, a fermentative system and a non-ionic surfactant. The biosensor system also comprises a measurement facility to measure speed of oxidation-restoration reaction of the analysed substance.

EFFECT: proposed group provides for accurate determination of dependence of an output signal of a test sensor, which contains compositions of reagent with low total concentration of salt, from concentration of analysed substance in samples of whole blood in a wide range of levels of hematocrit within the limits of not more than 7 seconds.

49 cl, 9 dwg, 7 tbl

FIELD: medicine.

SUBSTANCE: invention aims at a method for identifying microorganisms from a test blood culture sample. The method provides preparing a test sample, conducting a selective lysis and dissolving the cells other than microorganisms of the test sample, laminating the prepared lysate on a density buffer in a hermetically sealed container and centrifuging. The density buffer has a uniform density from approximately 1.025 g/ml to approximately 1.120 g/ml. The microorganisms passing through the above buffer are precipitated on a container bottom. The precipitate is analysed with the use of Raman spectroscopy that enables identifying a genus or a species of the microorganism.

EFFECT: invention enables identifying the microorganisms from the clinical samples over less than 120 min.

13 cl, 4 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to the field of biotechnology and cell technology. The claimed invention is aimed at the creation of pluripotent, multipotent and/or self-renewing cells, which are able to start differentiating in a culture into various types of cells and are capable of further differentiation in vivo. The claimed invention is also aimed at the creation of populations of the required differentiating cells, which can be transplanted to patients, genetic modification of endogenic cells and treatment of patients, suffering from diseases, intensity of which can be reduced by means of the said methods.

EFFECT: invention also claims methods of prevention, treatment or retardation of a disease, associated with an infection of immunodeficiency virus.

17 cl, 1 dwg, 13 ex

FIELD: biotechnology.

SUBSTANCE: biological sensor is described, as well as the method of creation of a biological sensor using thin films based on grapheme, graphene oxide or single-walled or multi-walled carbon nanotubes. The biological sensor comprises a substrate, a metal film on the surface of which the intermediate bonding layer is applied, made of a thin film of graphene or a thin film of graphene oxide or a thin film of carbon nanotubes. On the surface of the intermediate bonding layer the biospecific layer is adsorbed conformally and uniformly. The biospecific layer can be used as a layer of molecules of the binding partner of the analyte or a layer of a complex of biological molecules capable of interacting chemically with the molecules of the binding partner and forming a complex with them. Also the biospecific layer can be used as the hydrogel layer, on which the binding partner molecule and/or complex of binding partner molecules and biological molecules is deposited, which can form a chemical bond with the binding partner molecules. The described method of obtaining the biological sensor comprises the stages of applying a metal film, an intermediate bonding layer and a biospecific layer.

EFFECT: high sensitivity of the biosensor in combination with high biospecificity, expanding the range of application of the device, protection of the metallic film from the effects of the environment, the ability to detect large biological objects.

27 cl, 9 dwg

FIELD: chemistry.

SUBSTANCE: claimed invention relates to field of biophysics. Claimed are methods of determining space-time distribution of proteolytic enzyme activity in heterogeneous system, in accordance to which: provided is system in vitro, which contains sample of blood plasma, whole blood, water, lymph, colloidal solution, crystalloid solution or gel, and proteolytic enzyme or its precursor; fluorogenic, chromogenic or luminescent substrate for said enzyme id added; space distribution of signal of released substrate label is registered at specified time moments and space-time distribution of proteolytic enzyme activity is obtained by solving reverse problem of type "reaction-diffusion-convection" taking into account label binding with medium components. Also described is device for realisation of methods in accordance with claimed invention and method of diagnosing hemostasis disorders, based on their application.

EFFECT: invention can be further applied in the study of blood coagulation system and diagnostics of diseases associated with blood coagulation disorders.

22 cl, 6 dwg

FIELD: biotechnologies.

SUBSTANCE: invention relates to the field of gene engineering, specifically, to development of test systems based on fluorescent probes, which may be used as substrates for phospholipases A1 and A2. The probe with the name 1-[3-(2,4-dinitrophenylamino)propanoyl]-2-[8-(9-anthryl)-7E-octenoyl)]-sn-glycero-3-phosphocholine represents an analogue of phosphatidyl choline witth modified fatty acid chains, one of which carries the fluorescent group, and the other one - grouping-suppressor of fluorescence. The fluophor-suppressor pair is: 9-anthryl vinyl - 2,4-dinitrophenyl. The produced probe is included into the test system to determine activity of A2 phospholipase, group IIA, which also includes a micellar matrix, a buffer solution, diglyme and A2 phospholipase of apitoxin as standard.

EFFECT: invention makes it possible to determine activity of A2 phospholipase in blood serum in case of acute pathological processes, in tissue and cell extracts.

2 cl, 3 dwg, 5 ex

FIELD: biotechnologies.

SUBSTANCE: phospholipide fluorescent probe characterised by the following name 1-[13-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)tridecanoyl]-2-(10-{[(2-hydroxynaphthyl-1)azophenyl-4]azophenyl-4}decanoyl)-sn-glycero-3-phosphocholine, is used in the test system to determine activity of A2 phospholipase, group IIA (secFLA2(IIA)) in blood serum, which also contains vesicular and phospholipide matrix for inclusion of a probe, containing phosphatidyl choline, lisophosphatidyl choline and phosphatidyl glycerine, a buffer solution and A2 phospholipase of apitoxin as standard.

EFFECT: invention makes it possible to reliably detect activity of secFLA2 in human blood serum in clinical conditions.

2 cl, 5 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: enzyme immunoassay system for determining a probable time of the introduction of type 1 human immunodeficiency virus (HIV-1) infection, including group O HIV-1 in human serum (plasma). The system contains an immunosorbent of type 1 human immunodeficiency virus antigens (env HIV-1 and group O HIV-1), an assay diluent (PPC) and detecting reagents (conjugates, chromogen/substrate). The invention refers to a method for determining the probable time of the introduction of type 1 human immunodeficiency virus (HIV-1) infection, including group O HIV-1 in human serum (plasma) by analysing patient's blood serum with using the above enzyme immunoassay system.

EFFECT: invention enables the fast and simple determination of the probable time of the introduction of the HIV infection.

2 cl, 4 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: method includes treating a sample with a denaturing buffer solution, sorbing short RNA on a fibre glass sorbent in the presence of a chaotropic agent, followed by washing off non-bonded biopolymers and chemical reagents from the sorbent and eluting the end product. The biological fluid sample undergoes two-step denaturing, wherein at the first step two volumes of the denaturing buffer solution are added to the sample and at the second step two volumes of 96% ethanol and equal volume of chloroform are added to the obtained mixture, followed by stirring and separating the residue by centrifuging. Non-bonded biopolymers are washed off from the sorbent twice with the buffer solution and elution of the end product from the sorbent is carried out with the buffer solution.

EFFECT: simple method, short duration of the method and high output of short RNA.

3 cl, 3 tbl, 4 ex