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Difference reaction immune detection method

A technology of immunodetection and detection method, which is applied in the field of medical immunoassay to achieve the effect of reducing adsorption

Inactive Publication Date: 2009-04-15
陈纯美
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two methods have a hook effect when detecting high-concentration samples (too high concentration of the analyte is false negative), and both need repeated washing to remove unbound markers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment ( 1

[0015] Example (1): Detection of alpha-fetoprotein (AFP) by differential sandwich method (two wells)

[0016] Take 3-50 μl of diluent into the reaction well, take a dry and clean suction nozzle with a micro-sampler, put the suction nozzle into the specimen, make the outside of the suction nozzle tip wet with the specimen, remove the suction nozzle, and touch it on the tube wall After a while, insert it into the diluent in the reaction well, and then remove the nozzle; add reagent I (5-50 μl enzyme AKP-labeled monoclonal antibody, 5-50 μl fluorescein FITC-labeled monoclonal antibody), mix well, and undiluted after reacting at 37°C Add the sample into the well, mix well, add 10-100 μl of reagent II (magnetic particles coated with anti-FITC antibody) after reacting at 37°C, mix well, and apply a magnetic field at the bottom and side of the well after reacting at 37°C to make the sediment pile up. on the bottom side of the hole. Fix the precipitate in the magnetic field, remove a...

Embodiment ( 2

[0017] Embodiment (2): Differential Sandwich Method (Single Hole) Detection of C-peptide

[0018] Take reagent I (10-50 μl enzyme HRP-labeled monoclonal antibody, 10-50 μl fluorescein FITC-labeled monoclonal antibody) and add it to the reaction well (pre-sealed); use a micro-sampler to take a dry and clean nozzle, and put the nozzle into In the specimen, wet the tip of the suction nozzle with the specimen, remove the suction nozzle, touch the tube wall for a while, insert it into the reagent I in the reaction well, remove the suction nozzle, mix well, add reagent II (coated Magnetic particles of anti-FITC antibody) 20-100 μl, mix well, and after reacting at 37°C, apply an external magnetic field to the side of the bottom of the well to make the sediment precipitate on the side of the bottom of the well. Fix the precipitate in a magnetic field, remove the supernatant in the well, add the substrate solution, and measure the absorbance A value after reacting at 37°C; then remove ...

Embodiment ( 3

[0019] Example (3): Detection of thyrotropin (TSH) by differential competition method

[0020] Take 5-50 μl of specimen and add it to the reaction well, add reagent I (5-50 μl anti-TSH antibody (rabbit anti-human) and 5-50 μl enzyme-labeled TSH antigen), mix well, react at 37°C and add reagent II (coated sheep Magnetic particles of anti-rabbit antibody) 10-100 μl, mix well, and after reacting at 37°C, apply an external magnetic field to the side of the bottom of the well to make the sediment pile up on the side of the bottom of the well. Fix the precipitate in a magnetic field, remove all the supernatant in the original well (well 1), add the washing solution, and then transfer the washing solution in the well 1 to the well 2; add appropriate amount of the display substrate in the well 1 and the well 2 respectively After mixing the reaction, add an appropriate amount of stop solution, measure the absorbance A value, compare the A value of the two wells and the blank well A val...

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PUM

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Abstract

The invention relates to a particle immunodetection method which is characterized in that a sample is diluted into different concentration (the method can be used to dilute the sample in a reaction hole), and then respectively added into the reaction hole, so that the reaction result of each concentration can be superimposed; the supernatant liquid or the diluted supernatant liquid after reaction is inhaled into the other hole for comparable reaction and display, or in the same hole for step-by-step reaction and display, and then the displayed value of the sediment is calculated, and the concentration of the object to be tested can be identified. The method can reduce the hook effect and avoid washing.

Description

technical field [0001] The invention relates to a medical immunoassay method, in particular to a microparticle immunoassay method which uses a sample to react with different concentrations and controls to display reaction products. Background technique [0002] The principle of the existing microparticle solid-phase detection method is to use microparticles as the carrier to capture the analyte in the liquid phase to form a microparticle immune complex, then settle the microparticle immune complex, remove the supernatant, and detect the concentration of the precipitate after washing. Display the value and calculate the concentration of the analyte in the sample. The existing magnetic ELISA sandwich method uses a monoclonal antibody for the antigen to be tested to label the enzyme AKP, and another monoclonal antibody to label the fluorescein FITC; Anti-mixing combined to form a "sandwich sandwich". Add magnetic particles coated with goat anti-FITC antibody. After the reactio...

Claims

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Application Information

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IPC IPC(8): G01N33/539G01N33/543
Inventor 陈纯美
Owner 陈纯美
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