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Human extracellular matrix metalloproteinase inducible factor magnetic particle chemiluminescence immunoassay quantitative detection kit and detection method thereof

A technology of chemiluminescence immunity and metalloprotease, which is applied in the direction of chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, and measurement devices, can solve problems such as side effects, achieve easy dispersion, solve low sensitivity, and wide detection range Effect

Pending Publication Date: 2022-07-12
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if ACE2 is used as a therapeutic target, it may cause serious side effects

Method used

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  • Human extracellular matrix metalloproteinase inducible factor magnetic particle chemiluminescence immunoassay quantitative detection kit and detection method thereof
  • Human extracellular matrix metalloproteinase inducible factor magnetic particle chemiluminescence immunoassay quantitative detection kit and detection method thereof
  • Human extracellular matrix metalloproteinase inducible factor magnetic particle chemiluminescence immunoassay quantitative detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation of the kit

[0033]The magnetic particles of the present invention are used as the solid phase of the immune reaction, and the chemiluminescence immunoassay method is used in conjunction with a chemiluminescence analyzer to measure the content of human extracellular matrix metalloproteinase-inducing factors in human body samples. The kit includes streptavidin-coated magnetic particle suspension, alkaline phosphatase-labeled human extracellular matrix metalloproteinase-inducing factor (CD147) monoclonal antibody (referred to as enzyme-labeled monoclonal antibody), and biotin-labeled human cells Extracellular matrix metalloproteinase inducible factor (CD147) monoclonal antibody (referred to as biotin-labeled monoclonal antibody), human extracellular matrix metalloproteinase inducible factor (CD147) calibrator, etc. The kit according to the invention uses magnetic particles as the solid phase of the immune reaction, and uses the chemiluminescence imm...

Embodiment 2

[0059] Embodiment 2, the test method of test kit

[0060] (1) Sample addition and incubation process: Pipette 50uL of human extracellular matrix metalloproteinase-inducing factor (CD147) calibrator or fresh patient sample into the reaction tube, and then add alkaline phosphatase-labeled human extracellular matrix metalloproteinase-inducing factor ( CD147) monoclonal antibody 50uL and biotin-labeled human extracellular matrix metalloproteinase-inducing factor (CD147) monoclonal antibody 50uL, incubate at 37°C for 10 minutes; then add streptavidin-coated magnetic particle suspension 50uL, Incubate the reaction at 37°C for 10 minutes;

[0061] (2) Magnetic separation and cleaning process: put the reaction tube after the incubation reaction is completed on the magnetic separation rack for 1 minute, and remove the supernatant; add 300uL of magnetic bead coating buffer for the first time, and place it on the magnetic separation rack Let stand for 1 minute, remove the supernatant; a...

Embodiment 3

[0063] Example 3. Performance test results of the kit.

[0064] Kit performance evaluation indicators: The accuracy, linearity, precision and stability of the kit prepared by this method were determined. The standard curve is drawn after detection with protein calibrator. The standard curve is as follows: image 3 As shown, the standard curve formula y=1.0606x-0.1840, and the linearity of the test results is evaluated at the same time. The test results are shown in the following table:

[0065] 1. Accuracy test results

[0066]

[0067] 2. Linearity test results

[0068]

[0069] 3. Precision test results

[0070] repeat times low value samples 1 0.49 2 0.50 3 0.50 4 0.51 5 0.52 6 0.51 7 0.52 8 0.52 9 0.49 10 0.51 average value 0.51 Coefficient of Variation CV 2.38％

[0071] 4. Stability test results

[0072]

[0073] 5. Sensitivity test results

[0074]

[0075] It can be seen from...

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Abstract

The invention discloses a magnetic particle chemiluminescence immune quantitative detection kit for a human extracellular matrix metalloproteinase inducible factor and a detection method of the magnetic particle chemiluminescence immune quantitative detection kit for the human extracellular matrix metalloproteinase inducible factor. The kit comprises a magnetic particle suspension coated with streptavidin, a human extracellular matrix metalloproteinase inducible factor CD147 monoclonal antibody labeled by alkaline phosphatase, a human extracellular matrix metalloproteinase inducible factor CD147 monoclonal antibody labeled by biotin, and a magnetic particle suspension coated by the magnetic particle suspension. According to the kit, an antigen of a to-be-detected sample is coupled with a biotin-labeled CD147 monoclonal antibody and an alkaline phosphatase-labeled CD147 monoclonal antibody to form a compound, and after an enzymatic chemiluminescence substrate is added into the compound, a luminescence reaction is formed to detect the luminescence intensity so as to detect the concentration of the human extracellular matrix metalloproteinase-inducing factor in the to-be-detected sample. According to the kit, the defects of narrow linear range, complicated operation and the like of an enzyme linked immunosorbent assay kit are overcome, and the defects of low sensitivity, poor stability, high cost and the like are overcome.

Description

technical field [0001] The invention relates to a human extracellular matrix metalloproteinase-inducing factor magnetic particle chemiluminescence immunoquantitative detection kit and a detection method thereof, belonging to the field of immunodetection analysis, in particular to the field of chemiluminescence detection. Background technique [0002] The infectivity of coronaviruses is mainly determined by the S protein, which can bind to membrane receptors on the host cell membrane. This mediates the fusion of virus and cell membranes. The first key protein of SARS-CoV-2 infection - ACE2. In addition to ACE2, the research team of Academician Chen Zhinan recently found that CD147 can be used as another new potential way for SARS-CoV-2 to invade host cells in addition to ACE2 through surface plasmon resonance analysis, supplemented by competitive inhibition experiments, and further It was confirmed that the ability of the virus to invade host cells was enhanced after the S ...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/577
CPCG01N21/76G01N33/577Y02A50/30
Inventor 印明柱陈翔
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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