92 results about "Hook effect" patented technology

A lateral flow assay device for detecting the presence or quantity of an analyte within a test sample is provided. The device utilizes multiple zones, one of which serves as an indicator of whether or not the analyte in the test sample is within the “hook effect” region. Based on this indication, a technique may be selected for correlating a measured signal intensity to an analyte concentration or range of concentrations. For example, when it is determined that the test sample falls outside the “hook effect” region, the analyte concentration may be determined using one portion of a dose response curve. On the other hand, when it is determined that the test sample falls within the “hook effect” concentration, the analyte concentration may be determined using another portion of the dose response curve. Alternatively, the sample may simply be diluted for re-performing the assay. The present inventor has discovered that such a detection technique may simply, quickly and accurately detect an analyte present at any concentration.

The invention provides a diagnostic kit for determination of serum total IgE, a preparation method and an application method. The kit comprises an IgE standard substance, a biotin labeled rate anti-human IgE monoclonal antibody solution, a rabbit anti-human IgE polyclonal antibody coated acceptor microspheres solution and a streptavidin donor microspheres solution. The kit provided by the present invention solves a tedious washing step of current heterogeneous immunoassay kits, overcomes the defects of environmental pollution caused by radioimmunoassay and short shelf life, and overcomes the defects of poor repeatability of enzyme immunoassay analysis and easy hook effect generation, and the detection precision is higher than that of immuno-turbidimetric analysis.

The invention discloses a method for qualitatively and quantitatively detecting a target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay, and the method comprises the following steps of placing luminous microspheres coating a substance antibody, biotin-coated target substance antibody and optical sensitization microspheres wrapped by streptavidin (SA) into a specimen to be detected to perform a primary immunity reaction and detection, and the method also comprises the step of placing anti-He agent into the reaction system to facilitate the target substance immune superimposition reaction, and carrying out the secondary optical excitation chemicalluminescence immune detection. A purpose for comprehensively correcting a hooks effect can be realized by comprehensively analyzing detection results of twice light initiated chemiluminescence assay (LiCA), and the analysis process comprises the following steps of classifying the specimen to be detected into five concentration intervals such as cathode, low anode, middle anode, high anode and ultrahigh anode according to the signal characteristics of the primary LiCA detection and the secondary LiCA detection; and quantitatively analyzing the specimen in the low anode concentration interval and the ultrahigh anode concentration interval according to the primary LiCA detection. The method has the characteristics of accuracy in result, simplicity and convenience in operation, wide application range and the like.

The present invention is directed towards the synthesis and / or use of a mixture of analyte capture reagents comprising antibodies and a non-immunological reagent provided on a solid phase assay, selected to screen for very high and low levels of an analyte. The non-immunological reagent comprising a PC-conjugate that contains multiple copies of covalently coupled phosphorylcholine (PC) moieties, particularly towards a phosphorylcholine-thyroglobulin conjugate assaying for C-reactive protein (CRP), a known inflammatory marker. The mixture of capture reagents function to eliminate hook effect (i.e. false negatives) when CRP is present in sample at high concentrations.

A membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample is provided. The device utilizes a chromatographic zone on which is disposed a plurality of microporous particles. The chromatographic zone can effectively reduce the “hook effect” in a simple, efficient, and relatively inexpensive manner. In particular, the plurality of microporous particles allows larger-sized analyte / probe complexes to reach the detection zone before the uncomplexed analyte. Because the uncomplexed analyte is substantially inhibited from competing with the complexes for the binding sites at the detection zone, the incidence of “false negatives” may be limited, even at relatively high analyte concentrations.

The invention relates to a test strip and a method for fast quantitative detection of human chorionic gonadotropin (HCG). The test strip comprises a sample loading pad, a marker pad, an NC membrane, a sample suction pad and a plastic plate. The sample loading pad, the marker pad, the NC membrane and the sample suction pad are orderly adhered to the plastic plate. The marker pad is coated with colloidal gold-marked mouse anti-human HCG-beta monoclonal antibodies. The NC membrane is coated with a detection line T composed of mouse anti-human HCG-alpha antibodies and a quality control line C composed of goat anti-mouse IgG antibodies. On the NC membrane, a detection line T side close to the marker pad is provided with a sample loading indication line for indicating a sample loading position. Through loading of a sample on the NC membrane, the test strip can solve the problem that the existing detection reagent produces a hook effect because of too high HCG content, and can realize quantitative detection of HCG in blood or urine by a matched immunochromatographic assay interpretoscope.

A lateral flow assay device for detecting the presence or quantity of an analyte within a test sample is provided. The device utilizes multiple zones, one of which serves as an indicator of whether or not the analyte in the test sample is within the “hook effect” region. Based on this indication, a technique may be selected for correlating a measured signal intensity to an analyte concentration or range of concentrations. For example, when it is determined that the test sample falls outside the “hook effect” region, the analyte concentration may be determined using one portion of a dose response curve. On the other hand, when it is determined that the test sample falls within the “hook effect” concentration, the analyte concentration may be determined using another portion of the dose response curve. Alternatively, the sample may simply be diluted for re-performing the assay. The present inventor has discovered that such a detection technique may simply, quickly and accurately detect an analyte present at any concentration.

The invention discloses a kit for detecting a hepatitis B surface antigen and a detection method and application of the kit, belonging to the technical field of in vitro diagnosis and detection. The kit comprises the following components: (1) a magnetic micro-spherical system which comprises magnetic microspheres directly connected or indirectly connected with an antiBsAg antibody 1, (2) a first marker system which comprises an anti-HBsAg antibody 2 directly or indirectly connected with a first tracer or a second tracer, and (3) a second marker system which comprises an anti-HBsAg antibody 2 directly or indirectly connected with the first tracer or the second tracer or an anti-HBsAg antibody 3 directly or indirectly connected with the first tracer or the second tracer. The kit and the detection method can be used for detecting the hepatitis B surface antigen and have the advantages of high sensitivity and no HOOK effect.

The present invention relates to systems and methods for the rapid in situ determination of the existence of a hook effect and expansion of the dynamic range of a point of care immunoassay. For example, a system for identifying a hook effect and expanding the dynamic range of an immunoassay is provided that may include a primary sensor having first immobilized antibodies that may be configured to generate a first signal based on a presence or absence of a target analyte in a sample. The system may further include an attenuated sensor having second immobilized antibodies at a reduced concentration relative to a concentration of the first immobilized antibodies on the primary sensor and that may be configured to generate a second signal based on the presence or absence of the target analyte in the sample. The system may further include a processor configured to determine a presence of a hook effect in the immunoassay based on relative values of the first and second signals and optionally determine the target analyte concentration of the sample.

A method is disclosed for accurately determining the concentration of a target analyte utilizes reagent pairs having different affinity for the target. The different affinity provides distinct binding profiles that can be analyzed to absolutely determine the analyte concentration. The method provides an assay system having expanded dynamic range to cover a wider range of analyte concentration and can overcome the hook-effect that commonly exists in homogenous assay systems. The method utilizes distinguishable signals that allows for the analysis of multiple binding profiles and multiplex analysis.

The invention provides a kit for measuring microalbuminuria by adopting immune competition turbidimetry. The kit comprises an anti-human albumin antibody, human albumin marked leatexbeads and a buffersolution. The anti-human albumin antibody and the human albumin marked leatexbeads are separately placed. The particle size of the leatexbeads is 80-267 nm. When the kit is used for measuring microalbuminuria in human urine, the maximum linear range is 5 mg / L-1500 mg / L. The kit can meet screening of clinical normal people, and can also avoid result underestimation or even false negative result arising from a hook effect caused by antigen excess.

The invention discloses a reagent card for accurately detecting a test object, a kit and application. A sample pad (1), an antibody carrier film (2) and water absorption paper (3) are successively adhered to a PVC bottom plate (6) from one end to the other end, wherein the sample pad is a sample loading area, and the sample pad is sprayed with a test object antibody-1 coupling marker and a biotin coupling marker; the antibody carrier film is provided with three lines which are successively a line T1 (4), a line T2 and a line C (5) from the sample pad to the water absorption paper, or are the line T1 (4), the line C (5) and the line T2; the line T1 is a detection line, and a test object antibody 2 is smeared on the position of the line T1; an antigen of the test object is smeared on the position of the line T2; streptavidin is smeared on the position of the line C. By adopting the reagent card, the false negative rate caused by the high-dose hook effect can be greatly reduced, the clinical detection accuracy is improved, and the adverse impact of the hook can be effectively solved.

The invention discloses a method for monitoring HOOK effect in an immune gold experiment. The method comprises the steps of determining T line gray values A50 and A at t50 and t by using a vaccination effect identification recorder; when A / A50 is smaller than 1.05, determining that the HOOK effect exists, and indicating that a sample needs to be detected after dilution and the result multiplies by the dilution ratio; and when A / A50 is greater than 1.05, determining that the HOOK effect does not exist. By adopting the method disclosed by the invention, whether all the immune gold experiments have the HOOK effect or not is rapidly judged, and the detection result is corrected in time, the sample with the HOOK effect is diluted to be redetermined, the result multiplies by the dilution ratio to obtain a final detection result; if the HOOK effect does not exist, the detection result can be directly reported and the sample dilution redetermination is not needed, so that the immune gold detection efficiency and accuracy rate can be greatly improved.

The invention belongs to the field of clinical medical diagnosis and particularly relates to an HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and a detection method thereof. The HCG colloidal gold immunoassay lateral chromatographic test strip consists of a nitrocellulose film, a Fusion 5 film and water-sucking paper, wherein a detection line T line and a quality control line C line are arranged on the nitrocellulose film; a T line solution is sprayed onto the detection line T line; a C line solution is sprayed onto the detection line C line. A standard curve line is made through different concentration values and corresponding T / C average value of a standard product, the standard curve line is subjected to interpolation fit by sample strips for three times, the curved line which is relatively good in linearity and completely raises can be obtained, HOOK effect can be overcome, and detection on high-concentration HCG is realized. The detection range is up to 10 to 200,000mIU / ml, a to-be-detected sample is not required to be diluted, the detection process is simple and convenient, and the detection cost is low.

The invention discloses a biochip utilizing the hook effect to enlarge the detection range and a detection method thereof. The biochip comprises at least one target area, a capturing antibody is fixed in the target area, the capturing antibody is combined with an analyte, through the labeling antibody combined with the analyte, detectable signals can be generated and can be used to quantitatively analyze the analyte in a sample; the biochip also comprises at least one indicating area, an antigen is fixed in the indicating area; and the fixed antigen is not combined with the analyte and is combined with labeling antibody that is not combined with the analyte to generate a detectable signal, which can be used to judge whether a hook effect is generated or not. The biochip has the advantages of flexible and convenient operation, time saving, and high sensitivity, largely enlarges the detection range of biological molecules, and avoids the false negative results.

The invention discloses a one-step semi-double-antigen sandwich immunological detection method, which comprises the following steps of: adding a sample to be detected into a solid-phase support enveloped with an envelope antigen; keeping the temperature at the preset temperature, fully reacting for the first preset time, not removing the sample to be detected, and directly adding a labeled antigen; reacting for the second preset time, and completely washing an unbonded sample and the labeled antigen; and adding a substrate to react for the third preset time, detecting a display result and finishing the detection. Compared with the traditional one-step method, the detection method has the advantages of obviously improving the probability of forming an 'envelope antigen-antibody-labeled antigen' sandwich complex by bonding the labeled antigen and an 'envelope antigen-antibody' complex and effectively reducing adverse effect of the hook effect. Compared with the traditional two-step method, the method obviously improves the specificity. In the reaction of an antibody to be detected and a solid-phase antigen, multiple kinds of buffer solution and a certain inert protein are taken as diluted samples in the traditional two-step method, while the sample to be detected is used as a natural diluted sample in the invention, and the natural diluted sample can effectively eliminate non-specific binding.

The invention relates to a double-antigen sandwich antibody detection method. According to the method, detection is finished in a manner of forming a solid-phase support-a first antigen Ag1-a to-be-detected antibody-a second antigen Ag2, wherein the Ag2 is coupled with a marker for displaying signal strength; the method comprises the following steps: 1), enabling the Ag1 and the Ag2 to be in contact with a to-be-detected object under the condition that antigen / antibody reaction can occur with enough Ag1, Ag2 and the to-be-detected object, so as to form an immune compound, wherein the content of the Ag1 is greater than that of the Ag2 based on mol number; 2) washing the unbound to-be-detected antibody; 3) adding the Ag2 to bind the Ag2 with residual antigen binding sites in the immune compound; 4) detecting the markers to indicate the existence and / or content of the to-be-detected antibody. Compared with the prior art, the method provided by the invention ensures the low-value sensitivity of detection, also can solve a hook effect and reduce the miss rate.

The invention discloses a preparation method of a colloidal gold immunity lateral chromatography test paper strip for detecting HCG concentration in a urine sample. A HCG antigen serves as a material of a quality control line C, the HCG antigen on the quality control line C and a detecting line T react, and a gold-labelled antibody is left. Along with increase of HCG concentration in the sample, a typical HOOK effect happens to the detecting line T, and within the concentration range where the HOOK effect happens, the color of a quality control line C strip becomes lighter. By means of color changes of a T line strip and the C line strip, quantitative detection can be performed on HCG urine samples within the concentration range of 0-200000 mIU / mL, the false negative phenomenon caused by the HOOK effect is avoided, and clinical detection accuracy is improved.

A patient or animal side method and assay for eliminating the hook effect in the detection of a target analyte such as an acute phase protein in a bodily fluid in which the target analyte comprises a member of a specific binding pair comprising applying the sample to a solid phase carrier material, generating a signal in accordance with downstream movement of the labelled first or second members and the target analyte to bind with the complimentary immobilised first or second members, and detecting the presence of the target analyte in accordance with the signal generated at the complimentary immobilised first or second members.

The invention provides a joint test strip and a preparation method thereof. The test strip comprises a bottom plate, a sample pad, a coating film and absorbent paper, wherein the sample pad, the coating film and the absorbent paper are sequentially lapped and adhered on the bottom plate; the sample pad is sprayed with a marked first-type analyte antibody, a second-type analyte antibody and a marked quality control protein; the coating film comprises a first-type analyte detection area, a second analyte detection area and a quality control area; the first-type analyte detection area is sprayedwith a protein capable of specifically binding the marked first-type analyte antibody; the second-type analyte detection area is sprayed with another second-type analyte antibody which is located at an epitope different from the epitope of the marked second-type analyte antibody; and the quality control area is sprayed with a protein capable of binding the quality control protein. The joint test strip is capable of realizing the simultaneous detection of two types of analytes under a same high dilution ratio and a long reaction time in the same detection system, and effectively avoiding the hook effect, and is accurate and reliable in detection.

The invention provides a method for widening a detection range of immune colloidal gold. According to the method, signal values GOD0.5 and GOD value of a reaction line (line T) of a colloidal gold test strip are determined by an immunochromatographic interpretation recorder at time t0.5 and time t, t represents time for complete reaction, t0.5 represents time of half of the signal value of complete reaction, when the quotient obtained by dividing GOD by GOD0.5 is larger than 1.5, the condition that no hook effect is produced is determined, a signal value of a detection result is subjected to concentration value calculation according to a linear regression equation, when the quotient obtained by dividing GOD by GOD0.5 is smaller than 1.5, the condition that the hook effect is produced is determined, and the signal value of the detection result is subjected to concentration value calculation according to a logistic curve-fitting four-parameter equation. With the adoption of the method, whether the hook effect exists in detection samples can be judged, and the sample with the hook effect can be directly intercepted and detected, reagents are not wasted, the detection result can be determined only by one-time detection, the detection is fast and easy, and the accuracy is high.

The invention provides a kit for expanding a quantitative detection range and a detection method, the kit comprises a carrier, the carrier is provided with a labeled site, a T line and a C line, the labeled site contains a labeled antibody which is subjected to a specific reaction with a sample antigen and is labeled by signal particles, and the T line is connected with the C line. A capture antibody which is subjected to double-antibody sandwich reaction with a sample antigen and a labeled antibody is fixed on the T line, the capture antibody is formed by mixing two antibodies with different combining capacities with the sample antigen, and an antigen-antibody conjugate which is subjected to double-antibody sandwich reaction with the labeled antibody is fixed on the C line. A capture antibody on the T line is formed by mixing two antibodies with different binding capacities, generation of the HOOK effect is delayed, a directional coating method is adopted on the C line, for a to-be-detected sample with higher concentration, an antigen-antibody conjugate can show a more remarkable competition gradient when the antigen and a sample antigen compete for a labeled antibody, the descending range of the C line is widened, and the detection accuracy is improved. And the quantitative detection range of macromolecular proteins is expanded.

Related to is the technical field of light initiated chemiluminescence technology, and in particular, to a method for determining HD-Hook-effect sample, a system, a reagent kit, and a device for determining HD-Hook effect in immunoassay, an immunoassay method, a system, a reagent kit, and a device for determining immunoassay.

The invention discloses a latex-enhanced two-antibody competitive immunoturbidimetric assay kit as well as preparation and application methods thereof. According to the precipitation or agglutinationphenomenon of an immune complex formed by an antigen-antibody reaction and the principle of latex-enhanced immunological turbidimetry, on the basis of precipitation or agglutination formed by secondary antibody and primary antibody reaction, a secondary antibody-latex connector is prepared, antibody titer of primary antibodies is determined by the secondary antibody-latex connector, the quantity of the primary antibodies is limited, and during measuring of a reaction system, when antigens exist, the antigens compete with secondary antibodies for binding sites of the primary antibodies, so thatthe primary antibodies and the secondary antibodies are inhibited from forming the immune complex and agglutination or precipitation effect is reduced. A latex particle preparation process can be standardized by universal secondary antibody-latex, the product quality difference of the kits prepared from different kinds of latex and different antibodies with different methods is reduced, and the method is particularly suitable for clinical kit preparation; when the antigens are excessive, a false-negative result caused by hook effect can be eliminated.

The invention provides a reagent or a kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of the reagent or the kit, and belongs to the technical field of in-vitro diagnostic reagents. The reagent comprises a magnetic particle reagent and an acridinium ester reagent, wherein the magnetic particle reagent is composed of 2-8 [mu] g / ml of a magnetic particle labeled KL-6 antibody 1, 20%-30% of newborn bovine serum, 0.01%-0.1% of a preservative, 20%-30% of glycerin, 0.01%-0.1% of Tween 20, 0.1%-1% of BSA and 50%-70% of a PBS buffer solution, and the acridinium ester reagent is composed of 6-10 [mu] g / ml of an acridinium ester labeled KL-6 antibody 2, 10%-40% of glycerin, 0.5%-5% of BSA and 60%-90% of a PBS buffer solution. The KL-6 detection kit has the characteristics of high sensitivity, high precision and high accuracy, has excellent anti-interference capability, and does not have an obvious HOOK effect under a high-concentration sample of 40000IU / mL.

The invention provides an immunoassay method which is characterized by comprising the following steps: (1) performing chemiluminiscence immunoreactions on a sample to be tested with antigen (or an antibody) to be tested, executing and recording a first reading and a second reading of chemiluminiscence, and recording a difference increase amplitude between the first reading and the second reading as A; (2) making a standard curve according to the increase amplitude A of two readings of a series of known standard substances of the target antibody (or the antibody) to be tested, wherein the concentrations of the standard substances are lower than that for an HOOK effect; (3) if the increase amplitude A of the two readings of the sample to be tested with the target antibody (or the antibody) to be tested is greater than the maximum value of the standard curve, testing the sample for another time after the sample is diluted. The invention further relates to a system and a kit for identifying immunoassay. The invention further relates to another immunoassay method, another system and another kit for identifying immunoassay.

The invention relates to a method for judging the hook effect of homogeneous phase time-resolved fluoroimmunoassay, which comprises the following steps: (1) measuring the fluorescence intensity of a fluorescence donor and a fluorescence receptor at two detection wavelengths under the same excitation wavelength, and taking the fluorescence intensity ratio of the two wavelengths as a signal value; (2) respectively establishing standard curves of the absolute value of the signal intensity, the rate value of the signal change and the sample concentration by measuring the signal values of at leasttwo moments in the reaction initial stage through linear fitting and four-parameter fitting; and (3) detecting the signal value of the unknown sample, and obtaining the concentration of the unknown sample according to the fluorescence intensity of the unknown sample through the standard curve, thereby avoiding the hook effect; and calculating the relative value of the relative signal intensity andthe signal change rate by utilizing the absolute value of the signal intensity and the ratio of the signal change rate value to the background signal, and realizing consistent results on different instruments. And concentration interpretation of more samples can be completed within set time, so that the system has higher detection flux.

The invention provides a GP73 antigen silicon-based magnetic bead conjugate as well as a competition method GP73 quantitative determination kit formed by taking the conjugate as one of the components and matching with other components. The kit disclosed by the invention consists of a Golgi protein 73 (GP73) antigen silicon-based magnetic bead conjugate working solution, a Golgi protein 73 (GP73) calibration material, an alkaline phosphatase labeled Golgi protein 73 (GP73) monoclonal antibody working solution, a cleaning solution and a chemiluminescence substrate. Meanwhile, the invention provides a method for preparing the GP73 antigen silicon-based magnetic bead conjugate and a method for preparing each component in the kit. The method comprises the following steps: preparing the GP73 antigen silicon-based magnetic bead conjugate working solution, preparing an enzyme-labeled antibody, preparing the calibration material, preparing the cleaning solution and preparing the chemiluminescence substrate. The kit provided by the invention can be mainly applied to diagnosis of primary hepatic carcinoma and has the advantages of high sensitivity, wide linear range, hook effect avoidance, low cost, short detection time consumption and the like.

The invention discloses a kit for detecting a hepatitis B surface antigen and a detection method and application of the kit, belonging to the technical field of in vitro diagnosis and detection. The kit comprises the following components: (1) a magnetic micro-spherical system which comprises magnetic microspheres directly connected or indirectly connected with an antiBsAg antibody 1, (2) a first marker system which comprises an anti-HBsAg antibody 2 directly or indirectly connected with a first tracer or a second tracer, and (3) a second marker system which comprises an anti-HBsAg antibody 2 directly or indirectly connected with the first tracer or the second tracer or an anti-HBsAg antibody 3 directly or indirectly connected with the first tracer or the second tracer. The kit and the detection method can be used for detecting the hepatitis B surface antigen and have the advantages of high sensitivity and no HOOK effect.