Detection method of proteins in amyloid state, and set for detection of proteins in amyloid state |
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IPC classes for russian patent Detection method of proteins in amyloid state, and set for detection of proteins in amyloid state (RU 2509155):
       
 Method for hypocalcemia correction in children under age of two years old suffering convulsive disorder / 2508112
Invention relates to medicine, particularly to paediatrics. If ionised blood serum calcium is less than 0.9 mmole/l and the relation of ionised Ca/inorganic phosphorus is 0.3 or more, calcium gluconate 1.0 ml is introduced intramuscularly 7-10 times/day in a combination with the oral administration of 10% calcium chloride 5.0 ml 7-10 times/day and 0.5% alcohol solution of vitamin D2 8000 IU a day. If observing comorbid hypomagnesaemia (Mg below 0.66 mmole/l), 25% magnesium sulphate 1 ml is introduced intramuscularly once a day. If the calcium level is less than 0.9 mmole/l and the relation of ionised Ca/inorganic phosphorus is less than 0.3 instead of vitamin D20, Parathyreoidin 25-0.5 ml is administered intramuscularly 3 times a day. If ionised calcium is more than 1.1 mmole/l and magnesium is more than 0.66 mmole/l, 10% calcium chloride 5.0 ml is orally administered 7-10 times a day, and vitamin D2 8000 IU a day or substituting Parathyreoidin in a dose of 0.25-0.5 ml 3 times a day intramuscularly. After the calcium and magnesium levels are stably normalised, Parathyreoidin is replaced by 0.5% alcohol solution of vitamin D2.
 Moisture sensor / 2497130
Disclosed is a method of detecting urine, which involves: using a urine sensor having a matrix, the matrix containing a thermochromic substance distributed therein and a temperature varying means, where said temperature varying means is capable of either raising temperature or lowering temperature upon contact with urine so as to cause temperature change in the thermochromic substance, resulting in colour change; contacting the urine with the sensor matrix; and determining presence of urine based on colour change of the thermochromic substance. A sensor for determining presence or absence of urine is also disclosed.
Diagnostic technique for chronic generalised periodontitis / 2488115
Oral fluid is examined for the concentrations of mineral elements. A calcium/magnesium ratio is calculated, and if the value is 4.8-8.3, an increase of the saliva cadmium concentration and a propensity for developing chronic generalised periodontitis; the ratio of 8.4 and more, chronic generalised periodontitits is diagnosed if the saliva cadmium concentration exceeds 0.61 mcg/l.
Method for prediction of posthypoxic cardiopathy in newborns delivered by mothers suffering congenital cardiac failures / 2462716
Newborn's umbilical blood is examined for a level of endogenous nitrite, blood flow between aortic cusps, blood flow between pulmonary artery cusps, systolic blood pressure; diagnostic indexes F1 and F2 are calculated by formulae: F1=34.06*ABF+0.89*SBP+65.96*PBF-0.43*NOE-74.67 F2=26.29*ABF+0.73*SBP+54.74*PBF-0.15*NOE-51.45 wherein NOs is the level of endogenous nitrite in umbilical blood (mcmole/l); ABF is blood flow between aortic cusps (m/s); PBF is blood flow between pulmonary artery cusps (m/s); SBP is systolic blood pressure (mm Hg). If F1 is more than F2, posthypoxic cardiopathy is predicted, while F1 less than F2 shows the absence of risk of developing cardiopathy.
Early diagnostic technique for oral diseases in adolescents by microelement composition and lactic acid bacilli concentration in non-stimulated oral fluid / 2460076
Non-stimulated oral fluid is collected on an empty stomach and analysed for the concentrations of sodium, potassium, calcium, phosphorous and lactic acid bacilli. The relations Ca/P and Na/K are calculated. The values Ca/P 0.63, Na/K 1.2 and lactic acid bacilli count 103-104 enables diagnosing a risk of caries and inflammatory diseases of periodontium tissues; the values Ca/P 1.3, Na/K 0.9 and lactic acid bacilli count 105-106 enables diagnosing a risk of developing caries and no risk of inflammatory diseases of periodontium tissues; while the values Ca/P 1.1, Na/K 1.2 and lactic acid bacilli count 105-106 provides diagnosing a risk of caries and inflammatory diseases of periodontium tissues.
Method of laboratory diagnostics of hypertension and diabetes mellitus / 2407018
Content of sodium, potassium, calcium, ratio of sodium concentration to content of potassium in saliva of examined person are determined. If calcium content is more than 0.07 g/l and value of Na/K ratio is 0.12 and less -diabetes mellitus of 2-nd type is diagnosed.
 Method of pre-inoculation processing of samples collected from environment objects for release of microbacteria / 2402781
Milled samples of biological material are poured with physiological solution for 24 hours at room temperature, filtered through wadding, centrifuged for 20 min at 3-4 thousand revolutions per minute. After that 1.5% solution of lauryl sulphate per 1.5% solution of caustic soda is added to sediment, kept for 20 minutes, centrifuged for 20 minutes at 3-4 thousand revolutions per minute, sediment is washed with physiological solution 2-3 times and inoculated on nutrient medium.
 Method of predicting development of unilateral or bilateral nephrolithiasis / 2396913
Invention relates to medicine, in particular to urologic diagnostics and is intended for evaluation of prognosis of unilateral or bilateral stone formation development. Laboratory test of patient's urine is performed, by results PHU, MGU, R% and CH CAOX are obtained and linear classification discriminant functions are calculated by the following formulae: F1=-2076.27+22.89* PHU-9784.25* MGU+41.33* R%-2.76* CH CAOX, F2=-2068.17+23.82* PHU-9952.00* MGU+41.20* R%-2.98* CH CAOX, where: F1 - first classification discriminant function corresponds to group of patients with unilateral affection of kidneys, F2 - second discriminant function corresponds to group of patients with bilateral affection; PHU - pH of urine; MGU is concentration of urine magnesium, mole/l; R% is relative tubular reabsorption, %; CH CAOX is degree of urine saturation with calcium oxalate. After that obtained values of the first and second discriminant functions are compared, function whose value turns out to be higher, points to patient's belonging to corresponding group of patients.
Method for prediction of changing functional class of disease in patients with chronic cardiac failure / 2386973
Prediction of increasing the functional class of chronic cardiac failure (FC CCF) within the next year is ensured by determination of serum magnesium in blood serum (mmol/l) (XI), potassium of blood serum (mmol/l) (X2) and encephalic natriuretic peptide (pg/ml) (X3), observed diastolic dysfunction of left ventricle (LV) (1 - shown by cardiac ultrasound; 0 - not shown) (X4), high-grade ventricular premature beats (1 - shown by cardiac electrocardiogram; 0 - not shown) (X5) and cardiac fibrillation (1 - shown by cardiac ultrasound; 0 - not shown) (X6). The probability is calculated by formula Prob (event)=exp(eta)/(1+exp(eta)), where Prob (event) is probability of CCF progression, eta=-0.332674+2.12648*X1+6.34589*X2-0.00574387*X3+3.38776*X4=1-0.574371*X4+2-3.01702*X5=0; if Prob (event) is equal to 0, increasing the functional class is not expected; if Prob (event) is equal to 1, increasing the functional class by one unit is observed.
Laboratory saliva-composition diagnostic technique for oral diseases / 2367959
Invention refers to medicine, particularly to the laboratory diagnostic techniques for oral diseases based on the ultimate saliva composition. To implement the technique, a patient's saliva is analysed for the content of sodium, potassium, calcium and phosphorus. The ratios Ca/P and Na/K are calculated, and if Ca/P is less than 0.3, and Na/K is less than 0.2, dental caries is diagnosed. If the value Ca/P is less than 0.3, and Na/K exceeds 0.2, odontolith process is diagnosed. The values Ca/P exceeding 0.3 enables to consider oral health status as normal.
 Optical visualisation agents / 2484111
Visualisation agent contains a conjugate of formula (I) of benzopyrylium dye through a linker group with a 3-100-dimensional synthetic peptide which provides directed delivery to the biological target. Also disclosed is a pharmaceutical composition which contains said conjugate of formula (I), a set for preparing said pharmaceutical composition and methods for visualisation of a mammal body in vivo.
 Fast biosensor with reagent layer / 2482495
Detection system for detecting target molecules includes a sensor chip (1), having on its detecting surface (33) an immobilised target molecule or a capturing molecule for target molecules and a soluble reagent layer (5), having a labelled molecule for binding with the target. The group of inventions also relates to a sensor chip (1) and a method of detecting target molecules in a sample using said sensor chip.
 Identification of molecules modulating protein-protein interaction / 2476891
Group of inventions refers to methods and systems of analysis based on enzymatic degradation following protein-protein interaction for reporter modulation (activation or inactivation).
 Method of cell population discrimination and application thereof / 2397494
There is offered a method of discrimination and calculation of at least two populations of biological elements - carriers of specific signs, probably presented in a sample. The method provides the use of three different probes, each of which is specifically fixed with one of the populations of biological elements which are required to be detected. Each probe itself becomes detectable due to its proper marker, and two different markers specified have two emission spectra containing at least one common part (overlapping emission spectra), and the third one has the emission spectrum which essentially contain no common parts with two others (nonoverlapping spectrum).
 Device and method for detecting flourescent marked biological components / 2390024
Device comprises a measuring cavity for receiving and introducing a fluid sample. The measuring cavity has a set fixed thickness not exceeding 170 micrometres. The measuring cavity has a section fit for acquisition of its image. Within the measuring cavity, there is a dry reagent. The reagent contains as a component, a molecule conjugate with phosphor used for binding with biological components and with all other reacting components. The reacting components are soluble and/or suspended in the fluid sample. The method involves mixing of the reagent with the liquid sample to be introduced in the measuring cavity. A section of the sample in the measuring cavity is exposed to electromagnetic radiation of wavelength corresponding to wavelength of phosphor excitation. Phosphor marked biological components are detected through-thickness of the measuring cavity. Further, numerical analysis of the digital image follows to identify the biological components showing phosphor and to determine amounts of the biological components showing phosphor in the sample. The biological components are discernible on the digital image as fluorescing points emitting electromagnetic radiation of wavelength corresponding wavelength of phosphor emission.
 Method of multianalytic immune assay with using microparticles / 2379691
Invention refers to biology and medicine, namely to immunodiagnosis. There is offered method of multianalytic immune assay based on immunochemical, genetic and other types of reactions of biospecific binding analyte and ligands. There are mixed various categories of microparticles coated with biospecific reagents for binding of various required analytes and marked with one or more fluorochromes in various concentrations emitting a long-living fluorescence. The analysed sample and biospecific developing reagent marked with a detecting fluorochrome with a short-living fluorescence with its excitation area being outside that of fluorochromes with long-living fluorescence are added to the particle mixture. It is followed with reaction for biospecific complex formation. The prepared biospecific complexes are deposited on a solid-phase carrier. The fluorescence emission of all fluorochromes is excited with emitters in two spectral ranges herewith measuring an amount of long-living fluorescence in a time resolution mode to identify the microparticle and an amount of short-living fluorescence of detecting fluorochrome for measuring concentration of required analytes. Thus the concentration ratio of long fluorescing fluorochromes in microparticles for detecting the same type of analyte is constant, and for determining different types of analytes, the concentration ratio differs at least twice.
 Method of multyanalite immunoassay with use of microparticles / 2339953
On surface of porous membrane apply the reactionary admixture containing analyte, the first binding molecules bound to detecting substance and specific to analyte, the investigated sample and the particles, not capable to pass through the pores of a membrane covered with the second binding molecules, also specific to analyte, incubate an admixture for formation of a biospecific complex, wash an admixture from not bound reagents and register in a regimen of the time permission phosphorescence signals in spectral ranges of the detecting substances corresponding to a constant of time of attenuation of these substances. Determine the required analyte on a parity of measured phosphorescence signals, thus use on two kinds of the first and second binding molecules, each kind of the first binding molecule is bound to two detecting is long luminescing substances, for example chelate of europium and platinaporphyrine which parity of concentration in each first binding molecule is chosen in advance and corresponds to defined analyte.
Method for diagnosing estrogen- and progesteron-dependent genitalia abnormalities / 2312354
Method involves determining estradiol- and progesterone receptors concentration in mononuclear cells fraction of peripheral blood. The value being greater than 210 and 2050 receptors per cell, estrogen- and progesteron-dependent genitalia abnormalities are diagnosed, respectively.
 Method for determining substances transport intensity changes in prevailing directions between blood and non-mineralized organs / 2297001
Method involves introducing radioisotope to animals and further repeatedly determining radioisotope inclusions percent in blood and in and in non-mineralized organs in given time intervals, calculating relative radio-activity RRA for an animal examined at each time as ratio between radioisotope inclusions percent in non-mineralized organs to radioisotope inclusions percent in blood. Then transport intensity is determined in prevailing directions in each examination time by applying RRA difference factor (DFRRA), calculated as difference between the subsequent and previous RRA values. The received values of DFRRA factor changes are interpreted in terms of intensity time fluctuations and radioisotope transport direction for each organ.
Method for detecting toxic action of oral mucosal interferonotherapy / 2288474
One should daily introduce into oral cavity of inbred mouse BALB/c for about 3-5 d an olive-shaped melted edge of capron fish line of 0.5-2.0 mm diameter impregnated with glycerol-containing preparation of recombinant interferon-α, containing 104-106 IU/ml recombinant human interferon-α, then in a dead mouse it is necessary to determine against an intact mouse the morphofunctional state of hepatocytes, enterocytes, lymphoid tissue of regional lymph nodes to evaluate: the absence of toxic action of oral mucosal interferonotherapy in case of no degenerative alterations in hepatocytes, enterocytes, lymphoid tissue of regional lymph nodes in inbred mouse BALB/c subjected to oral mucosal interferonotherapy; the presence of toxic action of oral mucosal interferonotherapy at availability of degenerative alterations of hepatocytes and/or enterocytes, and/or lymphoid tissue of regional lymph nodes in inbred mouse BALB/c after oral mucosal interferonotherapy. The innovation increases information value of the method suggested.
 Method for prediction of deterioration probability of clinical course of psoriasis, progression into erythroderma considering immunological values, liver detoxification and metabolic functions / 2508903
Invention refers to medicine, namely to dermatology. To predict the deterioration probability of the clinical course of psoriasis, the progression into erythroderma, an integrated assessment of a pathoigenetic significance of the detected risk factors promoting the deterioration is made. What is determined is a prognostic coefficient (PC) for such criteria, as: sex; age; petrochemical workers; agricultural workers; construction workers; drivers; office staff; hyper insolation; adverse agricultural labour conditions (pesticide and fertiliser contact); petroleum contact; construction material contact; drug abuse; pockets of chronic bacterial and/or mycotic infections; frequent acute respiratory viral infections; chronic gastrointestinal and/or hepatic diseases; chronic pulmonary diseases; chronic respiratory infections; chronic otorhinolaryngological organs; disturbed liver detoxification; liver metabolic disturbance; circulating immune complex increase; immunoglobulin increase; immunoregulatory index decrease; stimulation index decrease; cytotoxic index decrease; antitissue autoantibody increase; blood skin antigen; leukocyte migration inhibition index increase. If a factor is absent, the related PC is considered to be zero. The derived PCs are summed up. If PC is 5.64-16.24, the low deterioration probability of the clinical course of psoriasis and the favourable prognosis; the PC value being 16.25-26.85 shows the medium deterioration probability and the relatively favourable prognosis; the value of 26.86-37.86 and more provides the high probability and the unfavourable prognosis, and the potential progression into erythroderma.
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FIELD: biotechnologies. SUBSTANCE: invention proposes a detection method of proteins in amyloid state, in which a specimen of lysate of yeast culture or tissue of a mammal is obtained, ionic detergent is added to the specimen, proteins are concentrated in an amyloid shape on a cellulose acetate membrane, and they are detected by means of aptomeres, their conjugates or antibodies specific to amyloid shape of proteins. Besides, a set for detection of proteins in amyloid state is proposed. EFFECT: invention can be used in medicine for diagnostics of amyloid diseases. 9 cl, 6 dwg, 7 ex
The technical field The present invention relates to molecular biology and medical diagnostics, in particular to a method of diagnosing amyloidosis, based on the detection of amyloids, including infectious variants of prions in biological fluids of animals and humans. Prior art Amyloids is insoluble protein aggregates with a fibrillar structure, and resistant to proteases. Amyloid fibrils are linear polymers consisting of ecovalence of connected protein molecules. These polymers are formed by the addition of protein monomers to their ends; however, the growth process is accompanied by enrichment of the joining of monomers of β-layers that are perpendicular to the axis of the polymers associated with conformational rearrangement with the formation of specific "cross-β structure. For amyloid is characterized by the interaction with some dyes, especially with Congo red and thioflavin T (Chiti f, Dobson C.M. 2006. Protein misfolding, functional amyloid, and human disease. Annu. Rev. Biochem. 75, 333-366). Classification of amyloidosis complex and ambiguous: allocate system amyloidosis (affecting many organs and organ-specific amyloidosis, primary amyloidosis (accumulation of amyloid causes disease) and secondary (accumulation of amyloid due to the network by some other disease). Finally, amyloidosis divided into non-infectious and infectious diseases (prion) (Sipe J.D., Cohen A.S. 2000. History of the amyloid fibril. J. Struct. Biol. 130, 88-98). Interest in the chemical and physical nature of amyloid caused, primarily, by their relation to a wider group of human and animal diseases. These diseases unite under a common name amyloidosis. Some of these diseases, due to the special form of amyloid, called prions, transmissive. Prion amyloids are described not only in mammals but also in lower eukaryotes, in which they act as achromosome inherited determinants. Infectious prion protein animal, called PrPScis a special conformationally altered the cellular isoform of the protein PrPCdifferent from it in its spatial structure. The infectious form of the protein PrPScgetting into the body from outside, or it spontaneously appearing, is able to stimulate the transformation of normal form of PrPCin pathogenic abnormal form of PrPScthrough protein-protein interactions. This ensures autocatalytic maintenance of prion protein isoforms (Prusmer, S.B., Scott, M.R., DeArmond,S.J., Cohen,F.E. (1998) Prion protein biology. Cell, 93, 337-348). Among the most famous animal diseases of this type include the so-called "mad cow disease". To : the most well-known human diseases, associated with the formation of amyloids, are amyloidosis Central nervous system (diseases of Creutzfeldt-Jakob, Alzheimer's disease, Parkinson's disease, Huntington's disease), and diabetes type 2 diabetes and some forms spinocerebellar ataxia, and cataract (Galkin A.P., Mironov, L.N., Zhuravleva GA, Inge-Vechtomov SG Genetics (2006) 42, No. 11, 1558-1570). Currently available methods for diagnosis of prion diseases is based on the study of the infectivity of tissue samples from diseased individuals or immunodetection pathogenic form of the protein PrP. The first method is potentially very sensitive, but its disadvantage is the long time between inoculation of infectious material in the experimental animal and the development of his disease. The possibility of detection of pathogenic form of the protein can be limited by its low content in the tissues and fluids (blood, urine, lymph, saliva). Received monoclinal antibodies that distinguish between pathogenic and normal form of the protein PrP (Korth, S., Stierii, C., Streit, P., Moser, M., Schaller, O., Fisher, R., Schultz-Schaeffer, W., Kretzschmar, H., Raeber, A., Braun, U., Ehrensperger, F., Homemann, S., Glockshuber, R., Riek, R., Billeter, M., Wuthrich, K. and Oesch, B. (1997) Prion PrPSc-specific an epitope defined by a monoclonal antibody. Nature, 390, 74-77; O Nuallain, B., Wetzel, R. (2002) Conformational Abs recognizing a generic amyloid fibril an epitope. Proc. Natl. Acad. Sci. USA, 99, 1485-1490). However, diagnostic methods practiced today is based on the research and topsony tissue samples of the brain. In recent time there have been attempts to develop new methods for diagnosis of prion diseases, based on different principles. One such approach is based on the ability of the prion protein to pass into the pathogenic form in vitro. In these experiments are mixed in vitro protein in the prion state, with its narional option and see the transformation of normal prion protein in the form. As we have noted above, this transformation is effective for protein Sup35 yeast, but not for protein PrP mammals. Developed a method of increasing the rate of conversion of PrP protein into the prion form. This method is based on periodic ultrasonic crushing prion aggregates (Saborio, G.P., Permanne, B. and Soto, C. (2001) Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature, 411, 810-813). A promising alternative to existing approaches for the diagnosis of prion diseases, as well as to create new drugs with potential therapeutic action is the procedure of designing and selective selection of short oligonucleotides (aptamers) DNA-type or RNA-type origin of a vast array of original mixture of chemically synthesized fragments with random sequence ("SELEX" - Systematic Evolution of Ligands by Exponential enrichment), Klug S.J., Famulok, M. (1994) All you wanted to know about SELEX. Molecular Biology Reports, 20, 97-107). So in the patent application PCT W02006138676 described group polynucleotides (aptamers)with the ability to bind with protein PrP, and PrP protein detection using these aptamers. However, so far there is no method of determining the concentration of pathogenic prion protein in biological fluids and tissues (except brain) sick animals due to its very small quantities. One of the main problems of creating a method of identifying the pathogenic prion protein is the concentration of the prion protein to detectable amounts. Description of the invention The aim of the present invention is to provide a fast, efficient and reliable method of detecting proteins in the amyloid state. As the result of careful study, the authors present invention has been developed a method of detecting proteins in the amyloid state, in which receive the sample lysate of yeast culture, tissue, or biological fluid of a mammal, concentrate the proteins in the amyloid form on cellulose acetate membrane and detects them using aptamers or antibodies. The pore size of the membrane is selected such that the delay of prion polymers, but ignore all other substances with lower molecular weight. Advantages and benefits of the present invention are the two who are simplicity, accessibility, efficiency, possibility of rapid and simultaneous analysis of a large number of samples and the high sensitivity of the method due to the unlimited possibilities of the concentration of proteins in the amyloid state by passing an unlimited number of test sample through the membrane. Thus, the present invention provides a method for non-invasive diagnosis of proteins in the amyloid state in biological fluids of a mammal, in particular humans, which currently is not. For detection of proteins in the amyloid state can be used aptamers and antibodies specific to proteins in the amyloid state. For example, earlier authors present invention through careful screening of selected aptamers with the ability to effectively and specifically bind to a protein Sup35 yeast S. cerevisiae in fibrillar amyloid condition and not to communicate with its Monomeric form, the possibility of using these aptamers for the detection of prion forms of proteins mammals, in particular PrP protein in infectious animal material, as well as polyglutamine proteins, for example Q70, which is the analog of the protein of huntingtin, and developed a way of detecting these prion forms of proteins using specified the aptamer (Russian patent application 2010125875). Thus, the aim of the present invention is the provision of a method of detecting proteins in the amyloid state, in which receive the sample lysate of yeast culture or tissue of a mammal or a sample of biological fluid of a mammal, add to the sample ionic detergent, concentrate the proteins in the amyloid form on cellulose acetate membrane and detects them using aptamers or antibodies. Also the aim of the present invention is the provision of the above-described method, in which the biological fluid is blood, lymph, saliva or urine of a mammal. Also the aim of the present invention is the provision of the above-described method in which the specified detergent is sodium lauroylsarcosine (sarkosyl). Also the aim of the present invention is the provision of the above-described method in which the concentration of proteins in the amyloid form on cellulose acetate membrane is performed by passing a sample of the lysate through a cellulose acetate membrane. Also the aim of the present invention is the provision of the above-described method, in which use cellulose acetate membrane with a pore size of 0.1-0.4 microns. Also the aim of the present invention is the provision of the above-described method in which the detection of proteins in AMI is odnom condition, adsorbed on cellulose acetate membrane, is performed with the use of aptamers, their conjugates or antibodies specific to proteins in the amyloid state. Also the aim of the present invention is the provision of the above-described method, wherein said conjugate contains a label, recognizable by an antibody. Another objective of the present invention is the provision of a kit for detection of proteins in the amyloid state, containing cellulose acetate membrane; ionic detergent; aptamers, its conjugate and/or antibody specific to the amyloid form of the protein; and instructions for use of the kit in accordance with the above-described method. Also the aim of the present invention is the provision described above, in which the cellulose acetate membrane has a pore size of 0.1-0.4 microns. Another objective of the present invention is the provision of the above set in which the specified detergent is sodium lauroylsarcosine (sarkosyl). More in detail, the present invention is described below. Detailed description of the present invention The concentration of proteins in the amyloid state in the method according to the present invention is carried out by transmission of the test sample through the membrane. The pore size of the membrane is selected such that the delay of the prion is aimery, but ignore all other substances with lower molecular weight. The authors of the present invention has been shown that it is preferable to use cellulose acetate membrane with a pore size of 0.1-0.4 μm, most preferably 0.2 μm. A sample of tissue culture mammal can be as raw cell lysate and the lysate, the pre-processed in order to clean it from proteins that do not form amyloids, to reduce nonspecific background binding of the aptamer with such proteins from the sample. However, the method according to the present invention is quite effective and has a high sensitivity due to the possibility of unlimited concentration in the absence of the necessary purification of protein, not forming amyloids, so that pre-treatment of the lysate before passing through the membrane is not required. The investigated samples can also be solutions of substances isolated from biological fluids or tissues of an animal or person, including those intended for the production of pharmaceuticals, biological additives or cosmetics. The investigated samples can also be a biological fluid of a mammal, in particular humans. Biological fluids that can be used in the method is according to the present invention, include blood, lymph, saliva, urine, but are not limited to. The possibility of transmission of an unlimited number of test samples through the membrane provides unlimited possibilities concentration of proteins in the amyloid state on the diaphragm that delivers high sensitivity of the method of detection according to the present invention and provides analysis of biological fluids with very small amounts of PrPSc. Thus, the present invention provides a method for non-invasive diagnosis of proteins in the amyloid state in biological fluids of a mammal, in particular humans, which currently is not. Testing the presence in the samples of the proteins in the amyloid state, adsorbed on the membrane, carried out with the use of aptamers or antibodies specific to proteins in the amyloid state. The term "aptamer"as used in the present invention, means a single-stranded oligonucleotide capable of binding with another molecule (molecule-target) and education with her relatively stable complex. When this binding occurs not at the expense of standard binding between base pairs according to the Watson-Crick due to hydrogen bonds, and at the expense of other types of non-covalent bonds. These types with the ides include non-covalent hydrogen bonds, electrostatic interaction, linking by van der Wals, hydrophobic interactions, or combinations thereof. This aptamer binds to the target molecule with a much higher affinato than with other molecules present in the sample. Methods design and characterization of the binding of the aptamer to the target molecule are well known in the prior art (see, for example, Lorsch and Szostak (1996) or U.S. patent No. 5,582,981, 5,595,877, 5,637,459). Themselves aptamers can be obtained by any known method, including chemical synthesis, methods of recombinant DNA, using standard cleaning methods. Also, the term "aptamer" refers to the secondary aptamers containing the so-called "consensus" sequence, obtained by analyzing two or more aptamers that bind the same target molecule. Typically, the length of the aptamer can vary from 10 to 40 or more nucleotides that are required for binding to the target molecule. Aptamers contain a nucleotide sequence that determines the binding specificity to the target molecule, but may also contain sites flanking this sequence, which is required for amplification of the entire aptamer PCR, or contain sites of restriction endonucleases for Legerova the Oia) in the plasmid cloning, etc. Such flanking sequences usually contain from 10 to 30 nucleotides, with some of these nucleotides may have a random structure. The aptamers according to the present invention can contain a variety of covalently linked, functional modifications and additional ligands required for detecting binding of the aptamer to the target molecule, for example fluorescent dyes, Biotin or various enzymes, the activity of which is easy to detect (peroxidase, luciferase, alkaline phosphatase, and the like). Such modified aptamers can be obtained by standard methods, well known to experts in the art, for example, during solid-phase synthesis of oligonucleotides or in solution using phosphotriesterase method. An example of aptamers that specifically bind to the proteins in the amyloid state, are aptamers S1-S4, containing a nucleotide sequence selected from the group consisting of sequences provided in the sequence Listing under the numbers SEQ ID NO:1-4, obtained by the authors of the present invention previously using procedures SELEX (Systematic Evolution ofLigands by Exponential enrichment) and is described in Russian patent application 2010125875. Aptamers S4 (SEQ ID NO:4) is preferred. These oligo is ucleotide have the ability to bind with proteins, forming amyloids, with high specificity and efficiency. Also, these oligonucleotides can be used to detect proteins form amyloids, in particular, the prion protein. For detection of binding oligonucleotides (aptamers) according to the present invention with proteins, forming amyloids can be used any methods known to experts in this field of technology. Can be applied to methods using radioactive isotopes, when binding with the target molecule used radioactively labeled oligonucleotides, such as32P-labeled oligonucleotides, and the degree of binding was determined after washing the sample for residual radioactivity. Can be used in quantitative PCR, such as real-time PCR, when determining the number of oligonucleotides, contacting the target molecule and the remaining part of the complex of the aptamer - target molecule, after denaturation of such a complex. Can also be used functional modification of oligonucleotides (aptamers) or more ligands required for detecting binding of the aptamer to the target molecule, for example fluorescent dyes, such as carboxyfluorescein (FAM), Biotin or various enzymes, the activity of which is easy to detect the AMB (peroxidase, luciferase, alkaline phosphatase, and the like). In case of using a biotinylated derivative of the oligonucleotide using a peroxidase, luciferase, and alkaline phosphatase fused with streptavidin or Avidya, with streptavidin and avidin binds to Biotin and peroxidase, alkaline phosphatase and luciferase are used to detect the binding. Can also be used in the conjugates of the aptamer is labeled, for example, FAM, and as a molecule that allows detection of binding of the specified aptamer with amyloid, using antibodies to the tag. One embodiment of the present invention provides a kit for detection of proteins in the amyloid state, containing cellulose acetate membrane; ionic detergent; aptamers, its conjugate and/or antibody specific to the amyloid form of the protein; and instructions for use of the kit in accordance with the method according to the present invention. Cellulose acetate membrane in the specified collection has a pore size of 0.1-0.4 μm, preferably 0.2 μm. The most preferred ionic detergent is sodium lauroylsarcosine (sarkosyl). Brief description of drawings Figure 1 shows a map of plasmid pL-prnP. Figure 2 shows a map of plasmid pL-moPnP. Figure 3 shows the results of the comparative analysis of the adsorption of protein lysates is of Rogga by passing the solution through a cellulose acetate or nitrocellulose membrane. Proteins were stained with nonspecific dye Ponco. Figure 4 shows the effect of detergents SDS and sarkosyl on the efficiency of adsorption of amyloid PrP on cellulose acetate membrane. The first point of the original lysate diluted 10 times in each of the next serial dilutions 4 more times. Figure 5 shows the results of the analysis of the binding specificity of the aptamer S4 with amyloid prion protein (PrP), educated his shortened (90-231) or full size (23-231) forms. The first point of the original lysate diluted 10 times in each of the next serial dilutions 4 more times. Figure 6 shows the effect of the presence of DNA and RNA to the binding specificity of aptamers with amyloid prion protein. "+" samples treated with enzymes Dnazol and RNase; "-" - samples not treated with these enzymes. The first point of the original lysate diluted 10 times in each of the next serial dilutions 4 more times. Examples The present invention will be described in more detail below with reference to the following not limiting the present invention to the Examples. Example 1. Construction of expression plasmids to obtain a full-sized protein of mouse PrP (23-231) and its shortened variant PrP (90-231). Mnogokupolnaya plasmid pL-PrnP containing the full sequence of the gene PrnP(23-231) mouse coding linakis is the notes from 23 to 231, under control of the CUP1 promoter was created on the basis of the vector pRS425 (Christianson T.W., Sikorski RS, Dante M., Shero J.H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors // Gene. 1992. 110, 119-122). A plasmid was obtained in two stages: the sequence of the CUP1 promoter from plasmid pRS316CG [Liu J.J., Lindquist S. Oligopeptide-repeat expansions modulate "protein-only inheritance in yeast // Nature. 1999. Vol.400. P.573-576] was embedded in the vector pRS425 sites XhoI-BamHI. Then, in the resulting structure on the sites BamHI and SacI was embedded sequence of a gene PrnP (23-231) mouse. The sequence encoding the protein fragment PrP mouse 23 to 231 amino acid amplified by PCR using as template the plasmid pcDNA3-l-3F4 (Narwa R & Harris DA. Prion proteins carrying pathogenic mutations are resistant to phospholipase cleavage of their glycolipid anchors // Biochemistry. 1999. 38(27) P: 8770-7), containing the gene LPR mouse, modified for detection of monoclonal antibodies 3F4 and primers PrnP (23-231) BamHI-F (SEQ ID NO:5) and PrnP SacI-R (SEQ ID NO:6). Used thermostable polymerase Pfu "Silex M" (Russia). Was used the following program for the amplification of fragments of the murine PrnP gene: 94°C for 2 min; 2 cycles of 94°C for 40 seconds, 49°C for 40 seconds; 72°C for 40 seconds; 24 cycle of 94°C for 40 seconds, 60°C for 40 seconds; 72°C for 40 seconds; and at the end of 72°C for 5 minutes. The correct sequence of nucleotides in the resulting plasmids were checked by sequencing with what ispolzovaniem standard primers 21M13F (SEQ ID NO: 7) and 29M13R (SEQ ID NO:8). Map of the resulting plasmid pL-PrnP shown in figure 1. Mnogokupolnaya plasmid pL-moPrP (90-231), containing the sequence of the PrnP gene (90-231) mouse, encoding amino acids 90 to 231, under control of the GPD promoter was created on the basis of the vector pRS425 (Christianson T.W., Sikorski RS, Dante M., Shero J.H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors // Gene. 1992. 110, 119-122). The plasmid was constructed by replacing the BamHI fragment-Sad containing PrP(90-231)-GFP (1,2 TPN), from the plasmid PGPD-PrP-GFP(LEU2) [rubel A.A., Saifutdinova A.F., Lada A.G., A. Nizhnikov, Inge-Vechtomov SG, Galkin A.P. Yeast chaperone Hsp104 regulates gene expression at the post transcriptional level // Mol. Biol. 2008. V.42. No. 1. P.123-130], a fragment of the SacII-SacI containing the sequence of PrP (90-231). The sequence encoding the protein fragment PrP mouse 90 to 231 amino acid amplified by PCR using as template the plasmid pcDNA3-1-3F4 (rubel A.A., Saifutdinova A.F., Lada A.G., A. Nizhnikov, Inge-Vechtomov SG, Galkin A.P. Yeast chaperone Hsp104 regulates gene expression at the post transcriptional level // Mol. Biol. - 2008. - V.42. No. 1. - P.123-130) and primers PmP (90-231)BamHI-F (SEQ ID NO:9) and PmP SacI-R (SEQ ID NO:6). Used the program for amplification described above. The correct sequence of nucleotides in the resulting plasmids were also determined as described above. Map of the resulting plasmid pL-moPrP shown in figure 2. Note the R 2. Transformation of Saccharomyces cerevisiae and getting lysates containing protein PrP. As the model used strain 74-D694ΔRNQ1 [psi-] MATa ade1-14 his3Δ200 leu3-112 trp1-289 ura3-52 rnq1::HIS3 yeast Saccharomyces cerevisiae (Salnikova, A.B., Kryndushkin D.S., Smimov V.N., Kushnirov V.V., Ter-Avanesyan MD Nonsense suppression in yeast cells overproducing Sup35 (eRF3) is caused by its non-heritable amyloids // J Biol Chem. 2005, 280, P.8808-8812). This strain of yeast expressed plasmid containing the sequence encoding PrP 23-231 or PrP 90-231, or as a negative control plasmid PRS425, on the basis of which were created by these plasmids. For transformation of yeast used a standard Protocol (Gietz, R.D., Schiesti, R.., Willems, .R., and Woods, R.A. (1995) Yeast 11, 355 to 360 above). Example 3. Getting lysates containing protein PrP Yeast were grown in 10 ml of minimal medium with appropriate supplements (adenine, uracil, essential amino acids and, if necessary, the copper sulfate as an inductor) to stationary phase. The cells were collected by centrifugation, resuspendable in 5 ml of distilled water was poured into 1 ml in a test tube "Eppendorf" and precipitated by centrifugation, resulting in each tube was 50-100 ál of cells. Next, the cells were frozen and kept at -70°C. To obtain the lysate was thawed one tube was added 3-fold volume of glass beads, 100 μl of Tris-saline buffer TBS (30 mm Tris-HCl pH 7,4,150 mm NaCl), soda is containing 10 mm EDTA, 10 mm phenylmethylsulfonyl of fluoride (PMSF), 1 mm dithiothreitol (DTT) and a cocktail of protease inhibitors (Complete®, Roche Applied Science), in the amount recommended by the manufacturer's Protocol. Cell debris was separated by centrifugation at 1500 g for 4 minutes and the clarified lysate containing polymers PrP protein, Packed up the tubes. Example 4. The concentration of proteins on membranes Cellulose acetate membrane (0.2 μm, Whatman GmbH OE66) moistened with distilled water and placed in a cell of the vacuum dot-blotter BioRad. Lysates of yeast cells containing protein PrP was diluted to the desired concentration in TBS buffer with an ionic detergent, making serial dilutions of these solutions in the same buffer and added into cell blotter 50 ál of the obtained solutions lysates. Using a vacuum (water) pump the solutions were pumped through the membrane. Then these cells were added in 100 μl of TBS buffer with an ionic detergent (SDS - sodium dodecyl sulphate or sarkosyl - laurylsarcosine sodium), incubated for 10 minutes, after which the buffer was removed and washed cells is 2 times 100 ál of the same buffer, then 2 times with 100 μl TBS buffer without detergent. After that, the membrane was stained with a dye solution Ponco-to estimate the amount of protein deposited on the membrane. For comparison lysates of yeast similarly passed through nitrate rulesnew membrane (0.45 μm, Macherey-Nagel, Germany), adsorbing proteins. Figure 3 shows that the majority of protein lysates of yeast well adsorbed on a nitrocellulose membrane and easily passes through acetylcellulose even at low dilutions of the lysates. Example 5. Study of the binding of aptamers and antibodies with protein PrP. The membrane was incubated for one hour in a solution of casein (4 mg/ml) in TBS buffer, pH 7.4 with 0.05% Tween-20. After that, the membrane was immersed in a solution of labeled aptamers or antibodies to the prion protein. Aptamers labeled with a fluorescent label FAM, 8 picograms were diluted in 0.5 ml of 5 mm phosphate buffer (PBS), pH 6.0, containing 0.05% Tween-20 and 200 mm urea. Aptamers were subjected to denaturation by boiling the resulting solution on a water bath for 5 minutes. Then the solution was cooled at room temperature for 5 minutes, kept for another 30 minutes at room temperature and then dissolved in 8 ml of the same phosphate buffer containing 0.4% casein. Antibodies were diluted in TBS buffer pH 7.4 with 0.05% Tween-20 and 4 mg/ml casein. After an hour incubation at room temperature the membrane was rinsed twice with distilled water and treated with the following solution. In the case of antibody - solution secondary antibodies conjugated with horseradish peroxidase in the same buffer. In the case of aptamers second solution served as antibodies to the FAM in the buffer is TBS. Then after an hour incubation with antibodies to FAM and a single washing of the membrane to the membrane was added a solution of secondary antibodies in TBS. After incubation with secondary antibodies conjugated with peroxidase, the membrane was washed 3 times for 15 minutes with TBS buffer with 0.05% solution of Tween-20 and then 2 times for 15 minutes with TBS buffer. The binding of aptamers or antibodies to the prion protein recorded on the device for the detection of chemiluminescence company Vilber Lourmat, using a mixture exhibiting solutions ECL West Dura system (Thermo Scientific). Example 6. Effect of detergents SDS and sarkosyl on the efficiency of adsorption of amyloid PrP on cellulose acetate membrane. Figure 4 presents the results of experiments on the concentration of amyloid PrP on cellulose acetate membrane in the presence of detergents SDS or sarkosyl (sodium lauroylsarcosine). From Figure 2 it is seen that the PrP amyloid dissolved in SDS and not adsorbed on cellulose acetate membrane. For concentration of PrP amyloid as a detergent should be used sarkosyl. Example 7. The proof of the binding specificity of the aptamers with the prion protein. To prove the specificity of binding of the aptamers with the prion protein, concentrated on cellulose acetate membrane, a similar experiment was performed with a solution of aptamers obtained for urokinase and have the same the size (70 nucleotides), as the aptamer to the prion protein (Skrypina NA, Savochkina LP, Beabealashvilli RSh. In vitro selection of single-stranded DNA aptamers that bind human pro-urokinase. Nucleosides Nucleotides Nucleic Acids 2004; 23:891-3). As can be seen from Figure 5, the aptamer to the urokinase is not associated with the full-size and truncated prion protein. It also shows that the sensitivity of the aptamer to the prion protein may be higher than monoclonal antibodies to the same protein. Despite the fact that used the aptamer shows a nonspecific binding control lysate, its interaction with prion protein not less than 10 times greater than background binding. To determine whether the membrane of the nonspecific binding of the aptamer with DNA and RNA present in the lysate of yeast, which can partially be delayed on the membrane, an experiment was conducted to remove DNA and RNA from the lysate by processing their Dnazol and RNase (at a concentration of 0.1 mg/ml each) for 30 minutes at room temperature. As can be seen from Fig.6, the presence or absence of DNA and RNA in the lysate did not affect the efficiency of binding of the aptamer with the prion protein. While this invention is described in detail with reference to the best mode for carrying out the invention, for a specialist in a specified field of technology it is obvious that can be done various changes and produced equiv the Lenten replacement and such modifications and substitutions are within the scope of the present invention. All cited documents are part of the description of the present invention and are incorporated in the present description in its entirety by reference. 1. Method of detecting proteins in the amyloid state, which includes the following stages: 2. The method according to claim 1, wherein the biological fluid is blood, lymph, saliva or urine of a mammal. 3. The method according to claim 1, characterized in that said detergent is sodium lauroylsarcosine (sarkosyl). 4. The method according to claim 1, characterized in that the concentration of proteins in the amyloid form on cellulose acetate membrane is performed by passing the sample through a cellulose acetate membrane. 5. The method according to claim 1, characterized in that use cellulose acetate membrane with a pore size of 0.1-0.4 microns. 6. The method according to claim 1, characterized in that pointed to by the second conjugate contains a label, recognizable by an antibody. 7. Kit for detection of proteins in the amyloid state, containing: 8. The kit according to claim 7, characterized in that the cellulose acetate membrane has a pore size of 0.1-0.4 microns. 10. The set of claim 8, characterized in that said detergent is sodium lauroylsarcosine (sarkosyl).
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