33 results about "Hook effect" patented technology

The invention discloses a urine microalbumin immunoturbidimetry kit and a detection method, and belongs to the technical field of biology. The invention relates to a urine microalbumin immunoturbidimetry kit, which comprises a reagent R1, a reagent R2, a reagent R3 and a reagent R4, wherein the reagent R1 comprises trihydroxymethyl aminomethane, a coagulant, a blocking agent, sodium chloride, bovine serum albumin, trehalose, a preservative and ultrapure water; the reagent R2 is prepared from hydroxyethyl piperazine ethanesulfonic acid, sodium chloride, bovine serum albumin, glycerol, trehalose, a preservative, a mALB monoclonal antibody and ultrapure water; the mALB monoclonal antibody is prepared by mixing a model number of UDA 9107-1, a model number of UDA 9107-2, a model number of UDA 9107-3 and a model number of UDA 9107-4 according to a mass ratio of (0.8 to 1.2): (0.8 to 1.2): (0.8 to 1.2): (0.8 to 1.2). A mixed antibody of four mALB monoclonal antibodies is specifically combined with a urine microalbumin antigen in a sample to form an antigen-antibody compound, and the antigen-antibody compound has high specificity and high sensitivity; the method has a relatively high detection range, can effectively avoid a hook effect of an ultrahigh-value sample, and can realize rapid and convenient automatic detection.

The invention belongs to the technical field of medicine detection, and particularly relates to a method for reducing a HOOK effect in colloidal gold immunochromatography detection and application. The upstream of the T line is provided with the B line, the B line is coated with the first antibody which is not marked, the combination pad is coated with the first antibody which is marked by colloidal gold, and the first antibody which is marked by colloidal gold and the first antibody which is not marked can be specifically combined with an analyte antigen in a sample to be detected; in addition, the T line is coated with a second antibody, the second antibody and a first antibody marked by colloidal gold, and the unmarked first antibody is a pair of antibodies capable of performing specific immunoreaction with different epitopes of the same antigen. According to the invention, due to the arrangement of the B line, the competitive strength of a large number of free analyte antigens and gold-labeled antibody-analyte antigen compounds on the binding site of the T line is favorably reduced, so that the HOOK effect is reduced, and the detection accuracy is remarkably improved.

The present invention provides a quantitative measurement method using competitive immunochromatography, which has not been previously reported. The quantitative measurement method comprises: (1) creating a calibration curve based on the signal intensity of a test line according to the concentration of a standard material by using multiple strips, and calculating the concentration of a target analyte by using the calibration curve; (2) when the concentration of the target analyte increases to a significant level on the calibration curve of the standard material, a labeled signal of a detection ligand on a control line remains unchanged in a case where a detection ligand of a target analyte-detection ligand conjugate binds to and saturates a capture ligand on the control line, making it impossible to estimate the concentration beyond that level; however, by designing a binding site for the capture ligand on the control line to be the target analyte of the target analyte-detection ligand conjugate, a high concentration of the target analyte breaks down the triple conjugate of detection ligand-target analyte-capture ligand, leading to dissociation of the labeled detection ligand and thereby weakening the signal (hook effect); and (3) comparing the signal intensity patterns of the test line and the control line according to the calibration curve of the standard material with the signal intensity pattern of a sample target analyte, and determining how far the target analyte is from the saturation point of the detection ligand. This method is characterized in that, in cases where the target analyte is present in an excessively large amount and exceeds the maximum value of a measurement range (dynamic range) of the calibration curve, the calibration curve using multiple strips and the configurations (test line and control line) of an immunochromatographic device are designed differently to distinguish such cases.

A method, device and computer readable medium for identifying hook effect in immunoturbidimetry. The method generates a reaction curve by measuring optical signals of an immune reaction of a to-be-measured substance in a sample within a predetermined time period, and determines whether the sample has hook effect by using distribution information of the reaction curve within the predetermined time period. The method only uses measurement information of a small predetermined time period in the entire reaction time to identify samples with hook effect, and terminates the detection in time, thereby accelerating the speed of immunoturbidimetry detection of samples with hook effect.

Devices and methods for the hook effect detection associated with analytes of interest in the conductance of one or more diagnostic assays, including, without limitation, immunoassays.

The utility model provides an immunodetection test strip capable of avoiding a HOOK effect. The immunodetection test strip comprises a bottom plate, a sample pad, a combination pad, an NC (nitrocellulose) membrane and a water absorption pad are sequentially arranged on the bottom plate, at least two detection lines and at least one quality control line are arranged on the NC membrane, and a detection line and a quality control line are sequentially arranged on the NC membrane along the chromatography direction of the test paper. The design of the test strip allows rapid detection, the result can be obtained in a short time, the test strip is suitable for on-site rapid screening, the whole detection process is simple, a user only needs to add a sample, and complex operation is not needed. The C line serves as a built-in contrast, the reliability of each test is ensured, and a user can conveniently judge whether the test is effective or not.

This invention provides an immunoassay method comprising the following steps: S1: performing two parallel immunoassays on a sample containing the target molecule, recording the results of the two parallel immunoassays as the first and second values, respectively; S2: calculating the ratio of the first value to the second value; S3: determining the concentration of the target molecule in the sample. The assay method of this invention can solve the problem of HOOK effect samples, and the method is not limited by the detection range, capable of directly measuring up to 10... 6 High sample concentrations at the ng / ml level; good repeatability and fast detection speed.

This invention also provides a creatine kinase isoenzyme MM detection kit, comprising reagent 1 and reagent 2. Reagent 1 comprises the following components: buffer solution 20-50 mmol / L; creatine kinase isoenzyme B subunit antibody 10-100 mmol / L; creatine kinase MM isoenzyme coated modified magnetic microspheres 0.2-2 g / L; blocking agent 0.5-2 g / L; protective agent 0.5-10 g / L; complex 1-50 mmol / L; stabilizer 1-10 g / L; anti-crystallization component 2-5‰; and preservative 0.05-0.1 g / L. The beneficial effects of this invention are: the kit lowers the limit of detection for creatine kinase isoenzyme MM in serum, enhances specificity, improves repeatability, broadens the linear range, and enhances the hook effect.

The present application relates to the technical field of in vitro diagnosis, in particular to a chemiluminescence-based adiponectin detection method, which uses MQ silicon resin-coated Fe3O4 magnetic microspheres as a solid-phase carrier, captures adiponectin in a sample through a double-antibody sandwich immunoreaction, and uses HRP enzyme-labeled detection antibodies to combine with a luminol-phenolic enhanced chemiluminescence system for signal detection. The key lies in significantly reducing non-specific background by optimizing washing conditions and substrate formula, and regulating luminescence kinetics to adapt to the reading window of mainstream chemiluminescence instruments. The present application uses HRP enzyme-catalyzed cascade amplification and an optimized luminol enhancement system, and the detection lower limit is better than that of conventional methods, meeting the needs of early metabolic risk screening for ultra-low concentration detection, the linear detection range covers 0.01 mg / L to 100 mg / L, effectively avoiding the common hook effect in high-concentration samples, and the signal-to-noise ratio is greatly improved, ensuring the reliability of low-value sample detection.

The invention relates to the technical field of porcine ileitis detection, and particularly discloses a porcine ileitis antibody detection kit, the kit comprises a colloidal gold immunochromatography test strip, a sample treatment liquid and a negative / positive control, a detection line T line of the test strip is coated with a recombinant Lawsonia intracellularis LsaA protein antigen, a quality control line C line is coated with a goat anti-Protein A antibody, and the detection line C line is coated with a goat anti-Protein A antibody. And the colloidal gold labeling pad is coupled with a Protein A-colloidal gold compound. According to the present invention, the prokaryotically expressed recombinant LsaA antigen is adopted to improve the specificity, the ProteinA is adopted to replace the traditional secondary antibody to simplify the labeling process, the sample treatment liquid formula is optimized to reduce the non-specific reaction, and the HOOK effect is inhibited by adjusting the antigen coating concentration and the marker consumption. Rapid detection of the porcine ileitis antibody is realized, sensitivity and specificity are greatly improved, operation is simple and convenient, professional equipment is not needed, and the problems that an existing detection method is time-consuming, low in sensitivity and high in cost are solved.

Embodiments of the present invention disclose a hook effect identification method and related devices. In the embodiments of the present invention, by obtaining the degree of curve bending of the antigen-antibody reaction curve of a sample, or by obtaining the change time of the antigen-antibody reaction curve of the sample; the reaction curve is the original sampling curve or the fitted curve of the optical detection value of the sample changing with time, and the change time is the time length corresponding to the change from the first preset optical detection value to the second preset optical detection value in the reaction curve; when the degree of curve bending meets the preset conditions, or when the change time is less than the change time threshold, it is determined that the sample has a hook effect. The method of the embodiments of the present invention does not need to consider the selection of time periods like the rate judgment method, but uses the degree of curve bending or the change time to judge the hook effect. Therefore, the identification accuracy is high and the specificity is strong.

The present application relates to an immunoassay kit, method and system. The kit comprises reagent 1 and reagent 2 with the same components, and the total amount of specific capture molecules in reagent 1 is different from that in reagent 2, which can specifically bind to the target molecules to be detected. The method for performing immunoassay using the kit of the present application can solve the problem of the hook effect sample, the method is not limited by the detection range, and can directly measure the high value sample concentration up to 10 6 ng / ml level with good repeatability.

According to the immunodetection kit and the application thereof, the method is used for detecting the concentration of a sample to be detected, and the sample to be detected does not need to be diluted, so that more time and labor are saved; compared with direct detection of a traditional method, the detection range is widened, and the aim of improving the HOOK effect can be achieved.

The invention belongs to the technical field of immunodetection, and particularly relates to an immunochromatography test strip capable of eliminating HOOK effect and realizing naked eye semi-quantification and a preparation method of the immunochromatography test strip. The test strip is provided with a T1 detection line and a T2 detection line which are respectively coated with a to-be-detected substance and a specific recognition substance of the to-be-detected substance. Interpretation and semi-quantification are performed according to a developing result of T2 / T1: in a blank sample, T2 (-) / T1 (+) is negative; in a low-concentration sample, T2 (+) / T1 (+) is positive; for a high-concentration sample, T2 (strong +) / T1 (-) is strongly positive; over-high sample concentration, T2 (weak or no) / T1 (-) indicates the false negative risk caused by the HOOK effect. The concentration of the object to be detected is T2 (-) / T1 (+) when being smaller than the target concentration 1, is T2 (+) / T1 (+) when being between the target concentration 1 and the target concentration 2, is T2 (+) / T1 (-) when being between the target concentration 2 and the target concentration 3, and is T2 (-) / T1 (-) when being larger than the target concentration 3.

The utility model provides a colloidal gold immunochromatography detection test strip, a detection card and a kit, the test strip comprises a strip-shaped bottom plate and a membrane layer covering the strip-shaped bottom plate, the membrane layer comprises a sample pad, a gold label pad, an NC membrane and absorbent paper which are sequentially adjacent in the axial direction of the test strip; the NC membrane is provided with a detection line and a quality control line which are parallel to the bottom edge of the test strip, are parallel to each other and are separated from each other, the detection line is positioned on one side, close to the gold label pad, of the NC membrane, and the quality control line is positioned on one side, far away from the gold label pad, of the NC membrane; non-circulation area blocks are respectively arranged at the junctions of the two side edges of the NC membrane and the two ends of the detection line, and the two non-circulation area blocks are symmetrical along the central axis of the test strip. Symmetrical non-circulation area blocks with certain areas are arranged at the two ends of the side of the detection line, so that the sensitivity of the detection line is improved, and meanwhile, the HOOK effect of the quality control line is delayed.

The preparation method comprises the following steps: dissolving a buffer salt, a surfactant, a protein stabilizer and a heterotropism antibody blocking agent in a solvent, and uniformly mixing to obtain a reagent R1 with the pH value of 6.0-8.0; the method comprises the following steps: respectively coupling at least two monoclonal antibodies for recognizing different epitopes of myoglobin to the surfaces of a first latex microsphere with the average particle size of 50-100nm and a second latex microsphere with the average particle size of 100-300nm; and mixing the two kinds of coupled latex microspheres according to the quantity or mass ratio of the second latex microspheres to the first latex microspheres of 1: (0.05-0.1), and dispersing the obtained mixture in a buffer system containing a stabilizer to obtain a reagent R2. The method has the effects of high determination accuracy of human serum myoglobin, low false positive rate and false negative rate, good low-value repeatability, excellent HOOK resistance effect and good stability.

The invention belongs to the field of tumor screening reagents, and particularly discloses a broad-spectrum tumor screening kit for a carcino-embryonic antigen (CEA) and a preparation method of the broad-spectrum tumor screening kit. The kit comprises a solid phase carrier coated with a high-affinity mouse anti-human CEA monoclonal antibody, a horse radish peroxidase labeled goat anti-human CEA polyclonal antibody, a chemiluminescent substrate solution, a calibration product and a quality control product. The preparation method is characterized in that antibody pairs which are high in affinity and capable of recognizing different epitopes are adopted, and the antibody pairs are prepared through the processes of optimized coating, enzyme labeling and freeze drying. The core of the invention lies in that the affinity constant KD value is less than 1 * 10 < M > when the antibody pair is used in compatibility, and a chemiluminescence detection system is adopted. The kit has ultrahigh sensitivity, wide linear range and excellent stability, can accurately quantify low-concentration and high-concentration CEA in human serum, effectively avoids the hook effect, is suitable for early screening, curative effect monitoring and prognosis evaluation of various malignant tumors such as colorectal cancer, lung cancer and the like, and has the detection performance obviously superior to that of a traditional ELISA kit.

The invention discloses a preparation method of photosensitive microspheres, a homogeneous chemiluminiscence detection kit and a detection method thereof, and belongs to the technical field of homogeneous chemiluminiscence detection.The preparation method of the photosensitive microspheres comprises the steps that copper phthalocyanine is pretreated, then an antibody-acrylate conjugate solution is prepared, and the antibody-acrylate conjugate solution is prepared; finally, styrene, alkyl substituted methyl acrylate, a copper phthalocyanine photosensitive monomer solution and the antibody-acrylate conjugate are subjected to photopolymerization under the protection of a nitrogen atmosphere. The kit comprises a photosensitive microsphere reagent, a luminous microsphere reagent and a buffer solution. During detection, a homogeneous chemiluminescence detection method is adopted, an antigen-antibody-microsphere bridge type compound can be forcibly formed through molecular design, the HOOK effect is fundamentally avoided, and meanwhile the high sensitivity characteristic of low-concentration detection is kept.

Methods of minimizing hook effect interference in an immunoassay are disclosed. Also disclosed are reagents, kits, and immunoassay devices that may be utilized in accordance with the method.

The application provides an immunoassay method, which comprises the following steps: S1: detecting two parallel immuno-reactions of a sample to be tested containing target molecules to be tested, recording the detection results of the two parallel immuno-reactions, and taking the first detection result and the second detection result as the first measured value and the second measured value respectively; S2: calculating the ratio of the first measured value / second measured value; and S3: determining the concentration of the target molecules to be tested in the sample to be tested. The determination method of the application can solve the problem of the sample with the hook effect, the method is not limited by the detection range, and the concentration of the sample with a high value of up to 10 6 ng / ml level can be directly measured; the repeatability is good, and the detection speed is fast.

This invention belongs to the field of biodetection technology, specifically relating to a monoclonal antibody combination against feline serum amyloid A (fSAA) and its application. The combination consists of monoclonal antibodies 10G3 and 1E3, with the amino acid sequences of the complementarity-determining regions (CDRs) of the heavy and light chain variable regions as shown in SEQ ID NO. 1-12. This antibody combination possesses clear sequence information, high specificity, and high affinity, effectively recognizing fSAA protein and avoiding detectable cross-reactivity with equine SAA and other analogues. This invention also provides a colloidal gold test strip containing this antibody combination, wherein 10G3 is used as a capture antibody coated on the detection line, and 1E3 is used as a labeled antibody coated on the gold-labeled binding pad. The test strip has a detection limit of 50 μg / mL, reducing the risk of hook effect. This invention provides a stable and reliable tool for rapid, accurate, and sensitive on-site detection of feline inflammatory responses, and has significant clinical application value.

The application discloses a SP10 / ACP / PSA triple detection test strip and a preparation process thereof, wherein a nitrocellulose membrane in the test strip is provided with a first detection line T1, a second detection line T2, a third detection line T3 and a quality control line C; the first detection line T1 is marked with SP10 monoclonal antibody 1; the second detection line T2 is marked with ACP monoclonal antibody 1; the third detection line T3 is marked with PSA monoclonal antibody 1; the quality control line C is marked with avidin SA; and the combination pad is provided with quantum dot microspheres marked with SP10 monoclonal antibody 2, ACP monoclonal antibody 2 and PSA monoclonal antibody 2; the test strip can simultaneously detect three indexes, and has simple product structure and convenient operation; the marking process in the preparation process adopts a competitive inhibitor and a quencher, solves the hook effect, and simultaneously obviously improves the specificity of each index.

The invention relates to an immunoassay kit, an immunoassay method and an immunoassay system. The kit comprises a reagent 1 and a reagent 2 which are the same in component, the total content of specific capture molecules in the reagent 1 is different from the total content of specific capture molecules in the reagent 2, and the specific capture molecules can be specifically combined with target molecules to be detected. According to the present invention, with the immunoassay method using the kit, the HOOK effect sample problem can be solved, the method is not limited by the detection range, the high-value sample concentration reaching the level of 106 ng / ml can be directly measured, and the repeatability is good.

The invention discloses a kappa light chain detection kit and a preparation method, and relates to the technical field of clinical in-vitro detection reagents, the kit is in a double-reagent form, the double reagents comprise a reagent R1 and a reagent R2, the reagent R2 comprises a capture antibody and a detection antibody which can be specifically combined with different epitopes of a kappa light chain, and the detection antibody comprises a reagent R1 and a reagent R2 which can be specifically combined with different epitopes of the kappa light chain. A capture antibody-antigen-detection antibody sandwich structure is formed to improve the hook effect in detection. The kappa light chain detection reagent provided by the invention solves the problems that the existing kappa light chain detection reagent mostly uses a single antibody and a hook effect is easy to occur due to reduction of a HOOK point, a capture antibody-antigen-detection antibody sandwich structure is formed by two antibodies (capture antibody and detection antibody) aiming at different epitopes of an antigen, and the core is a specific binding mode, so that the recognition precision of the antigen can be improved, and the detection accuracy of the kappa light chain detection reagent is improved. And non-specific binding is reduced, the detection sensitivity and HOOK point are greatly improved, and further popularization and application in the market are facilitated.

The invention discloses a novel immunodetection method and application. The immunodetection method comprises the following steps: coating a captured antibody and a single-site recognition antigen on a solid-phase carrier; adding a to-be-detected sample and a biotin-labeled mixed detection antibody for incubation, so that the target antigen forms a double-antibody sandwich compound, and the single-site recognition antigen and the detection antibody form a competitive compound; after streptavidin marked by chemiluminescence is added for incubation, signal values of the two compounds are detected; whether interference exists or not is judged and detected by comparing the double signals, and the content of the target antigen is determined. The immunodetection method can effectively identify cross interference and hook effect, and improves the accuracy and reliability of multi-index detection.

The invention relates to the technical field of biological medicine, in particular to a preparation method and application of a p65 protein degradation agent, the structural formula of the compound is as shown in formula I: # imgabs0 # experiment results show that the compound (formula I) provided by the invention can significantly down-regulate the p65 protein level and presents a dose-dependent effect. The HOOK effect observed under the condition of high concentration prompts that the formula I possibly forms a ternary complex with p65 and E3 ubiquitin ligase CRBN to mediate degradation of p65, and important evidence is provided for the action mechanism of the compound. In addition, the compound (formula I) can effectively reduce gene expression of key inflammatory factors (such as IL-1beta, IL-6, TNF-alpha, iNOS and MCP-1) in LPS-induced RAW 264.7 macrophages, and shows potential application value in the aspects of regulating inflammatory response and improving lipid metabolism.

The present invention relates to a photochemiluminescence detection device, and relates to the technical field of chemiluminescence immunoassays. The device is used to address the technical problem in the prior art of false positive detection results due to the hook effect. The photochemiluminescence detection device of the present invention comprises an incubation device and a detection mechanism. The incubation device rotates to move the same test substance multiple times to a detection position, allowing the detection mechanism to perform multiple tests on the test substance, thereby determining whether the immunoassay has a hook risk. This avoids the possibility that the test sample cannot be correctly distinguished due to the hook effect, whether its concentration exceeds the linear range of the detection kit or is within the linear range, thereby avoiding experimental misdiagnosis.

The invention relates to an immunoassay device for detecting the concentration of a target molecule to be detected. The device comprises an immunoreaction assembly, the immunoreaction assembly comprises at least two reaction containers, and the two reaction containers are used for carrying out two parallel immunoreaction detections on the same sample to be detected; wherein the ratio of the content of the specific capture molecules in the reaction container 1 to the content of the to-be-detected target molecules in the to-be-detected sample for detecting the same to-be-detected sample is different from the ratio of the content of the specific capture molecules in the reaction container 2 to the content of the to-be-detected target molecules in the to-be-detected sample. The device disclosed by the invention can solve the problem of HOOK effect samples, is not limited by a detection range, and is good in repeatability.

The application discloses a sample analysis method, device and computer readable storage medium, wherein the sample analysis method comprises: obtaining a to-be-tested reaction curve of a mixed solution; wherein a to-be-tested sample solution and a reagent solution react to form the mixed solution, and the to-be-tested reaction curve represents a corresponding relationship between an absorbance value of the mixture and time; based on a characteristic parameter of the to-be-tested reaction curve, determining an estimated concentration value of the to-be-tested sample solution in a pre-established calibration curve; based on the estimated concentration value, determining a corresponding true concentration value in a pre-established hook effect curve; and determining a serum concentration of the to-be-tested sample solution according to the true concentration value. In this way, the detection cost can be reduced, and the detection efficiency can be improved.

Methods of minimizing hook effect interference in an immunoassay are disclosed. Also disclosed are reagents, kits, and immunoassay devices that may be utilized in accordance with the method.