54 results about "Heart Fatty Acid-Binding Protein" patented technology

The present invention relates to methods and compositions for symptom-based differential diagnosis, prognosis, and determination of treatment regimens in subjects. In particular, the invention relates to the use of biomarkers, either individually or in combinations with one another to rule in or out diseases of the aorta and its branches, most particularly aortic aneurysm and/or aortic dissection, and for risk stratification in such conditions. Preferred markers include one or more of creatine kinase-BB (CK-BB), creatine kinase-MB (CK-MB), acidic calponin, basic calponin, B-type natriuretic peptide (BNP), NT-proBNP, proBNP, BNP_79-108, BNP_3-108, caldesmon, caspase-3, D-dimer, soluble elastin fragments, endothelial cell-selective adhesion molecule (ESAM), fibrillin-1, heart-type fatty acid binding protein, MMP-9, myeloperoxidase, myoglobin, smooth muscle myosin, smooth muscle myosin heavy chain, TIMP-1, free cardiac troponin I, complexed cardiac troponin I, free and complexed cardiac troponin I, free cardiac troponin T, complexed cardiac troponin T, and free and complexed cardiac troponin T, and preferred assays are configured to detect these markers.

The invention discloses a fluorescence immunochromatographic assay method for quantitatively detecting hFABP (heart fatty acid binding protein) and a kit for quantitatively detecting the same. The fluorescence immunochromatographic assay method for quantitatively detecting the hFABP realizes quantitative fluorescence detection on the basis of optimizing components of a test strip by the aid of excellent fluorescent characteristics of quantum dots and by means of combining bicolor labeling technique and immunochromatographic assay. Compared with a conventional colloidal gold immunochromatographic assay method, the fluorescence immunochromatographic assay method has the advantages of fine labeling stability, low non-specificity, high sensitivity, wide linear range and accuracy in quantization. The kit is used for quantitatively detecting the hFABP, can be used for simultaneously detecting whole blood, blood serum and plasma samples, serves as a simple, accurate, specific and inexpensive detecting tool for early screening and prognosis evaluation of acute myocardial infarction, is applicable to hospitals at all levels, and is particularly beneficial to wide popularization in primary hospitals and clinics.

A detection kit of multiindex protein for diagnosing cardiovascular disease is prepared as having base plate and reaction holes on it set on reaction hole plate; including 12-200 sample holes and 5-8 standard piece holes in said reaction holes as each reaction hole bottom being set with solid phase carrier; enveloping micro lattice of eight antibody as anti-CRP, anti-Myoglobin, anti-cTnI, anti-cTnT, anti-NT-proBNP, anti-CK-MB, anti-H-FABP and anti-GPBB on said carrier for accurately diagnose said disease in multiple man-share.

The invention discloses a preparation method of joint-inspection test paper for cardiac troponin I (cTn I) and heart-type fatty acid binding protein (H-FABP). The preparation method comprises the following steps: using a membrane-spotting metal spraying device and a mixed liquid labeled by a cTn I labeled antibody and an H-FABP labeled antibody to spray metal to a pad, so as to obtain a gold pad; using a cTn I detection line coating buffer, an H-FABP detection line coating buffer and a control line coating buffer to respectively scribe on a polyvinyl chloride base plate adhered with nitrocellulose membrane, so as to obtain the polyvinyl chloride base plate spotted with the membrane; assembling a sample pad, the gold pad, the membrane spotted polyvinyl chloride base plate and absorbent paper together, and then cutting the assembled paper to obtain the detecting test paper. With adoption of the method, the detecting test paper obtained by the invention is based on the lateral flow immune chromatography technology, and can be used by manual operation and visible reading to judge whether the blood sample contains cardiac troponin I and heart-type fatty acid binding protein, so that the diagnosis is rapid and the result is accurate, and the body of the subject can be free of damage.

The invention discloses a TRF (time-resolved fluorescence) immunochromatography reagent for rapidly and quantitatively detecting H-FABP (heart fatty acid-binding protein) and a preparation method and belongs to the field of clinical medical diagnosis. The reagent comprises two parts including a test strip and a fluorescent liquid, wherein the test strip comprises a bottom plate, Fusion5, a nitrocellulose membrane and a water absorbent pad; the Fusion5, the nitrocellulose membrane and the water absorbent pad are horizontally and sequentially connected and fixed onto the bottom plate; the nitrocellulose membrane is coated with a detection line for H-FABP monoclonal antibodies 1 and a quality control line comprising rabbit IgG (immunoglobulin G) antibodies; the fluorescent liquid contains TRF microspheres labeled by H-FABP monoclonal antibodies 2 and TRF microspheres labeled by goat anti-rabbit antibodies. According to the reagent, the fluorescence intensity is improved by the aid of the TRF microspheres, background signals are reduced, meanwhile, the content of the H-FABP in whole blood, serum or plasma is quantitatively detected, and only 10-20 microliters of samples are required. The test strip is convenient, rapid, simple to operate, short in detection time, high in specialty, high in sensitivity, more accurate in detection result and applicable to rapid diagnosis for clinical POCT (point-of-care testing).

The invention discloses a myocardial infarction triple rapid detection kit and a preparation method for the myocardial infarction triple rapid detection kit. The kit is formed by taking novel heart markers, that is, human myeloperoxidase (MPO) and heart fatty acid binding protein (FABP3) as basis, and matching with any of myocardial infarction markers CRP (C reactive protein), cTnI and Lp-PLA2. The kit is characterized by being capable of warning myocardial infarction diseases much earlier compared with the existing gold-label kit, and capable of warning the risk of the whole process before and after the occurrence of myocardial infarction. The kit is composed of a test paper card and a colour comparison card or a colloidal gold quantitative reading meter, wherein an anti-MPO specific antibody, an anti-FABP3 specific antibody and a colloidal gold pad resisting to any of other myocardial infarction markers are labelled on the test paper card simultaneously; and novel myocardial infarction markers are detected simultaneously by a colloidal gold immunochromatography technology at the first time, and the kit has the characteristics of being capable of realizing early warning and whole-course monitoring, simple and convenient to operate, rapid, suitable for field detection, economical and practical, and the like.

The invention belongs to the immunology field in medicine and particularly relates to a B-cell epitope peptide of H-FABP and applications of an antibody thereof. The epitope peptide is shown in SEQ ID NO.5 and can be used for preparing hybridoma cell lines and secreting a corresponding monoclonal antibody. The monoclonal antibody is capable of being used for preparing a diagnostic reagent which is used for detecting the H-FABP and prepared on a large scale, provided with purity higher than 96%, good in specificity and the like; and a detection titer of a purified antibody through an indirect enzyme-linked immunosorbent assay (ELISA) reaches 1:256,000. Monoclonal antibodies and polyclonal antibodies prepared through the epitope peptide can be used for H-FABP detection in patients' blood, and a foundation is laid for early diagnosis of acute myocardial infarction (AMI).

The invention relates to a cardiac muscle control material, a preparation method therefor, a detection kit and a detection device for cardiac muscle. The cardiac muscle control material comprises quality control component and protection component, the quality control component comprises at least one of B-type natriuretic peptide, D-dimer, NT-proBNP, cardiac troponin I, myoglobin, creatine kinase isozyme and cardioid fatty acid binding protein and heart fatty acid binding protein. The protective component comprises 5 mM to 100 mM of a buffer agent, 0.5 % to 5% by weight of an excipient, 0.05 mMto 5 mM of an antioxidant, 0.05 mM to 15 mM of a protease inhibitor, 0.5% to 10% by weight of a stabilizer and 0.01% to 2% by weight of a surfactant. The cardiac muscle control material disclosed bythe invention is good in stability.

The invention relates to a heart-type fatty acid binding protein (H-FABP) detection kit and a making method thereof. The kit comprises a reagent R1, a reagent R2 and a standard substance; the reagent R1 comprises a solvent and solvates, the solvent is a buffer solution with the pH value of 7.0-7.6, and the solvates comprise a surfactant, a chelating agent, an accelerator, an antiseptic and purified water; the reagent R2 is an anti-human H-FABP antibody latex reagent; and the standard substance is a recombinant protein containing a quantitative amount of the H-FABP. The problems of bad anti-interference capability, low specificity, low reagent stability and narrow detection linearity range of latex enhanced immunoturbidimetery detection kits in the present market are solved. The kit disclosed in the invention has the characteristics of simple operation, high accuracy, good repeatability, high sensitivity and the like, can be used on a fully-automatic biochemical analyzer, a special protein analyzer or a spectrophotometer, and has a wide application prospect.

The purpose of this invention is to provide monoclonal antibodies which specific binding with heart-type fatty acid-binding protein (H-FABP), especially a pair of monoclonal antibodies specific binding with the heart-type fatty acid-binding protein (H-FABP), can develop specific, high accuracy kit in the early diagnosis of acute myocardial infarction.

The invention pertains to the field of the biotechnology. The invention relates to a recombinant protein. The recombinant protein includes two dominant antigenic epitopes of humanized heart type fatty acid binding protein, the amino acid sequence of the recombinant protein is converted into corresponding nucleotide sequence by adopting codon preferred by escherichia coli, chemical synthesis is performed on the nucleotide sequence and recombinant expression vector is constructed, so that protein expression level of the recombinant protein in the escherichia coli is increased. The invention also relates to the preparation of monoclonal antibody of the recombinant protein. Immunization, cell fusion and several rounds of screening are carried out to obtain hybridoma cell strain, the monoclonal antibody is purified and labeled with horse radish peroxidase (HRP), the best matching combination of the monoclonal antibody is determined by ELISA orthogonal experiment, and an early diagnosis application of myocardial infarction is achieved.

A heart-type fatty acid binding protein detection reagent kit, the kit body of the reagent kit comprises two reagents inside: reagent R1: buffer solution, polyethylene glycol, sodium azide, sodium ethylene diamine tetracetate, and bovine serum albumin; reagent R2: buffer solution, latex particles bound with heart-type fatty acid binding protein monoclonal antibody, Tween-20, sodium azide, sodium ethylene diamine tetracetate, and bovine serum albumin. Mixing a sample with the reagents in a specific volumetric ratio to conduct a series of reactions, placing the reactant under a semi/full automatic biochemical analyzer, and detecting the change speed of light absorbance at the position of 500nm dominant wavelength, thus the concentration of heart-type fatty acid binding protein can be figured out. The reagent kit has the advantages of accuracy, stability and convenience.

The invention discloses hybridoma cell strains 1-F10 (preservation number: C20122139 and preservation unit: Wuhan University) and 3-H5 (preservation number: C20122140 and preservation unit: Wuhan University), and a monoclonal antibody of an anti-heart-type fatty acid-binding protein (H-FABP) generated by the same. The hybridoma cell strains 1-F10 and 3-H5 are prepared through predicating epitopes in the H-FABP and corresponding antibodies for detection can be secreted; and the subtypes of the monoclonal antibodies of the anti-H-FABP, which are secreted by the hybridoma cell strains 1-F10 and 3-H5, are IgG2a and IgG2b. The antibodies generated by the cell strains 3-H5 and 1-F10 are respectively used as a capturing antibody and a detection antibody, so as to successfully detect the difference of the H-FABP in blood serums of healthy people and myocardial infarction patients, and develop a diagnostic reagent which has good specificity and high accuracy, and can diagnose acute myocardial infarction in early stage.

The invention relates to a determination reagent for heart-type fatty acid binding protein and a preparation method, and belongs to the technical field of methods for testing or analyzing materials by means of chemical or physical properties of determination materials. The determination reagent is prepared from a reagent R1 and a reagent R2, wherein the reagent R1 is a solution formed by dissolving bovine serum protein into purified water added with a buffer solution, and the reagent R2 is a solution formed by dissolving anti-human heart-type fatty acid binding protein monoclonal antibody (mouse) latex into purified water. When the determination reagent is applied to determination of the heart-type fatty acid binding protein, the determination reagent has the advantages of being rapid to determine, sensitive to determination, good in accuracy, high in specificity, good in stability and the like.

The invention relates to a diagnostic test paper for detecting an H-FABP. The test paper is characterized in that: the test paper is formed through adhering a sample pad, a cellulose nitrate membrane and an absorption pad to a base plate in sequence, arranging a gold-marking binding pad on the sample pad, and covering a PE label protective membrane on an upper layer; the gold-marking binding pad is coated with a labeled H-FABP mouse mono-antibody with colloidal gold; and the cellulose nitrate membrane is coated with a detection line which comprises the labeled H-FABP mouse mono-antibody and aquality control C line which comprises an H-FABP secondary antibody respectively. The diagnostic test paper has the advantages of strong specificity, high sensitivity and no need of any auxiliary instrument, and has a noninvasive advantage because a patient self can conveniently take a sample for the needed detection sample is urine; and simultaneously detection only needs 5 min, so the test paper provides a convenient and fast method for determining or eliminating myocardial infarction, thereby precious time is saved for early treatment.

The invention provides a rapid and quantitative detection device and a method for simultaneously detecting heart-type fatty acid-binding protein and cardiac troponin I. The device is characterized by being composed of detection test paper of the heart-type fatty acid-binding protein (FABP) and the cardiac troponin I (CTN1), a double-united clamping shell and an immunochromatography result judging and reading recorder which is internally provided with a corresponding standard curve and result judgment. The immunochromatography result judging and reading recorder is an optical detection system and is used for quantitatively judging a detection result; detection ranges to the FABP and the CTN1 are 5ng/mL-200ng/mL and 1ng/mL-100ng/mL. The detection is finished within 15-20 minutes. The rapid and quantitative detection device and the method have the advantages of united detection, simplicity and convenience in operation, rapid reaction, accurate results, suitableness for field detection and the like, and are suitable for accurate and simple clinical detection requirements.

The invention provides a kit for quantitatively detecting a heart-type fatty acid binding protein. The kit comprises a heart-type fatty acid binding protein calibrator, a magnetic particle coupled with an anti-biotin antibody, an acridine derivative-labeled heart-type fatty acid binding protein monoclonal antibody, a biotin-labeled heart-type fatty acid binding protein monoclonal antibody, a chemiluminescent substrate and a quality control substance. A preparation method of the kit comprises the following steps: preparing the calibrator from a pure heart-type fatty acid binding protein raw material; preparing the acridine derivative-labeled antibody; preparing a biotinylated antibody; coupling a magnetic particle with the anti-biotin antibody; sub-packaging the calibrator, a marker mixturesolution and the chemiluminescent substrate; and performing assembling to obtain the finished product. The kit has the advantages of high sensitivity, good specificity, high accuracy of a quantitative detection result, low use cost, and easiness in promotion and application.

The present invention relates to methods and compositions for monitoring, diagnosis, prognosis, and determination of treatment regimens in subjects suffering from or suspected of having a renal injury. In particular, the invention relates to using assays that detect one or more markers selected from the group consisting of Clusterin, Heart-type fatty acid binding protein, Hepatocyte growth factor, Interferon gamma, Interleukin-12 subunit beta, Interleukin-16, Interleukin-2, 72 kDa type IV collagenase, Matrix metalloproteinase-9, Midkine, and Serum amyloid P-component as diagnostic and prognostic biomarkers in renal injuries.

The invention especially relates to a fluorescence immunochromatographic method for quantitative detection of cardiac troponin I and heart-type fatty acid binding protein, belonging to the technical field of immunodetection and analysis. The fluorescence immunochromatographic method employs blood, an anti-cardiac troponin I antibody, an anti-heart-type fatty acid binding protein antibody, a fluorescence reagent and a detector, wherein the anti-cardiac troponin I antibody is one selected from a group consisting of or a combination of an anti-cardiac troponin I monoclonal antibody and an anti-cardiac troponin I polyclonal antibody; the anti-heart-type fatty acid binding protein antibody is one selected from a group consisting of or a combination of an anti-heart-type fatty acid binding protein monoclonal antibody and an anti-heart-type fatty acid binding protein polyclonal antibody; the fluorescence reagent is a liquid-phase and/or solid-phase material containing a fluorescent substance, and the fluorescent substance is one selected from a group consisting of or a combination of an organic fluorescence dye and a rare earth element fluorescence dye; and the detector is a fluorescence detector. According to the invention, human blood is used as a detection sample, and the method can effectively monitor cardiac troponin I and heart-type fatty acid binding protein in bood, has good specificity, good repeatability and high sensitivity, does not need special instrument and equipment and professional training, and is simple to operate, intelligible and distinct in results, easy to promote and applicable to on-site detection.

A method for ‘ruling out’ acute myocardial infarction (AMI) in a subject presenting with chest pain expected to be cardiac in nature. The method includes a) recording an Electrocardiography (ECG) reading from said subject, b) determining the amount of Heart-type Fatty acid binding protein (H-FABP) and cardiac Troponin T (cTnT) in a sample from said subject, c) comparing the results from b) to reference values for ruling out an acute Myocardial Infarction in a subject, and d) based on the results from steps a)-c) either ‘ruling out’ or ‘ruling in’ a diagnosis of acute Myocardial Infarction in said subject.

The invention discloses a magnetic bead time resolution immunofluorescence quantitative detection H-FABP (heart-type fatty acid binding protein) kit. The kit comprises an H-FABP monoclonal antibody-coated immunomagnetic bead, an H-FABP calibration product solution, a europium-marked H-FABP monoclonal antibody solution, cleaning liquid and an enhancement solution. The H-FABP monoclonal antibody-coated immunomagnetic bead is super paramagnetic bead with functional group modification and the diameter of 1 to 3 [mu]m and an H-FABP monoclonal antibody covalent conjugate. The kit is high in sensitivity, the sensitivity of H-FABP is 2 ng/mL, and a serum (plasma) sample does not need to be diluted; the detection time is short and a report can be output within 30 minutes; the demand quantity of samples is small and one-time sample loading only needs 50 [mu]L; the matched fully-automatic time resolution immunoanalyzer is simple in operation and labor-saving.

The invention discloses a heart-type fatty acid binding protein immunodetection reagent and a preparation and a detection method thereof, and concretely relates to heart-type fat binding protein immunodetection reagent and a preparation and a detection method thereof. The heart-type fatty acid binding protein immunodetection reagent comprises enzyme-labeled heart-type fat binding protein, and an indication reagent used for detecting a heart-type fat binding protein antibody-enzyme-labeled heart-type fat binding protein compound. The heart-type fatty acid binding protein immunodetection reagenthas the advantages that the above homogeneous immunodetection agent can conveniently, accurately and rapidly determine the heart-type fat binding protein content in a sample, and several samples canbe simultaneously determined on an automatic biochemical analyzer, high-flux rapid determination on heart-type fatty acid binding protein is realized, accuracy is high, specificity is strong, stability is good, and accuracy and detection efficiency are greatly increased.

The invention relates to a detection reagent for heart-type fatty acid binding protein and a preparation method of the detection reagent for the heart-type fatty acid binding protein. The detection reagent is prepared from a reagent I and a reagent II, wherein the volume ratio of the reagent I to the reagent II is 5 to 1; the reagent I comprises a 20-50mmol/L buffer solution, a 2%w/v coagulant, 2%w/v BSA and a 0.1%-0.5%w/v preservative; the reagent II comprises 0.1%-0.5%w/v anti-human H-FABP antibody-marked colloidal gold particles, a 2%-5%w/v stabilizer, the 20-50mmol/L buffer solution and the 0.1%-0.5%w/v preservative; the diameters of the anti-human H-FABP antibody-marked colloidal gold particles are 60nm-90nm. The detection reagent has the advantages of being high in detection sensitivity, large in linear detection range, easy and rapid in detection, good in operability and the like.

The invention relates to the expression and purification of human heart-type fatty acid-binding protein in escherichia coli DH5 alpha, which belongs to the biological technical field, the expression and purification comprises: firstly, preparing an oligonucleotide primer, secondly, amplifying with RT-PCR and determining a sequence, thirdly, constructing recombinant expression plasmids, fourthly, expressing heart-type fatty acid-binding protein through inducing, and fifthly, extracting and purifying the human heart-type fatty acid-binding protein. The expression and the purification of the invention have the beneficial effect that a PL promoter is enabled to enter a complete inhibitory state. H-FABP protein is expressed in PBV220 through utilizing the characteristic. We optimize the temperature and the time in the expression process, bacteria is increased under the temperature of 30DEG C, and high level expression is obtained through inducing for 5 hours under the temperature of 42 DEG C. The successful clone and expression of the human heart-type fatty acid-binding protein lay the foundation of further researching the significance of the heart-type fatty acid-binding protein in the diagnosis and therapeutic effect evaluating of diseases.

The invention provides a homogeneous immunometric fluorescent compound set for quickly and quantificationally detecting heart fatty acid binding-proteins (FABP) and a preparation method of the homogeneous immunometric fluorescent compound set. The homogeneous immunometric fluorescent compound set comprises a rare-earth element chelate marked anti-FABP monoclonal antibody, a near infrared fluorescent compound marked anti-FABP monoclonal antibody and FABP calibrators with series concentration. The homogeneous immunometric fluorescent compound set can be used for detecting the low-value FABP and the high-value FABP, particularly the low-value FABP, is low in cost, simple, quick and sensitive to operate and good in specificity, only needs to be matched with a special homogeneous fluoroimmunoassay detection instrument, and therefore, the homogeneous immunometric fluorescent compound set can be widely applied to medical examination places at different levels, particularly basic-level medical mechanisms including health clinics in towns and townships and has great significance on preventing heart and cerebral vessel events.

The invention relates to the technical field of medical detection, in particular to a kit for detecting heart-type fatty acid binding proteins by a latex enhanced immune turbidimetry method. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 contains a Tris buffer solution and polyethylene glycol p-isooctyl phenyl ether, and has the pH of 7.0 to 8.0; the reagent R2 contains mouse-anti-human heart-type fatty acid binding protein monoclonal antibody latex particles, a Tris buffer solution, albumin bovine serum, alkyl glycoside APG0810, mannitol and polyethylene glycol p-isooctyl phenyl ether, and has the pH of 7.0 to 8.0. According to the kit disclosed by the invention, the ratios of the latex particles with different particle sizes are strictly defined, so that the sensitivity and the specificity of detection are improved; therefore, a surfactant and stabilizing agent combination suitable for the system is obtained; on the premise of high measurement speed and sensitivity and high specificity, the whole system is high in stability and easy and convenient to operate, and is suitable for being matched with a clinically full-automatic or semi-automatic biochemical analyzer in use.