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Kit for determining lipoprotein-associated phospholipase A2 and manufacturing method

A phospholipase and lipoprotein technology, applied in the field of fluorescence immunochromatography, can solve the problems of high intensity and strong interference, and achieve the effects of rapid reaction, high sensitivity and strong specificity

Inactive Publication Date: 2017-05-10
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the usual fluorescence measurement, because the test sample contains a variety of fluorescent components, the background fluorescence (scattered light caused by colloidal particles and solvent molecules in the sample and non-specific fluorescence emitted by proteins and other compounds in the serum) has a large intensity and interference. Strong, become the bottleneck of large-scale promotion of fluorescence analysis

Method used

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  • Kit for determining lipoprotein-associated phospholipase A2 and manufacturing method
  • Kit for determining lipoprotein-associated phospholipase A2 and manufacturing method
  • Kit for determining lipoprotein-associated phospholipase A2 and manufacturing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The various components of the test paper card in the lipoprotein-associated phospholipase A2 assay kit can be prepared by the following measures:

[0044] 1. Preparation of sample pad 2:

[0045] Soak the glass fiber membrane in the treatment solution containing 1.0% Triton X-100, 2.5% BSA, 0.15M Tris buffer, pH7.5, soak at 4°C for 4 hours, then place it in an oven and dry it at 37°C 2 hours. 2. Preparation of binding pad 3 for absorbing fluorescent microsphere-labeled antibody:

[0046] Soak the glass fiber membrane in 150mM Tris-HCL treatment solution (containing 1.0% Triton X-100, 2.5% BSA, pH7.4), soak for 2 hours at 4°C, then take it out of the oven at 37°C and dry it for 4 hours, and set aside. The glass fiber membrane was placed on the Bio-DotXYZ3050 three-dimensional spraying platform, and the rare earth Eu 3+ The conjugated complex of anti-lipoprotein-associated phospholipase A2 monoclonal antibody labeled with fluorescent microspheres is sprayed onto the gl...

Embodiment 2

[0055] Embodiment 2: accuracy test

[0056] Select the above-mentioned test paper card and fluorescent immunochromatography analyzer (model: NEO-007), the setting of the parameters of the fluorescent immune analyzer: after setting the process parameters of the test paper card on the fluorescent immune analyzer, take the above-mentioned assembled test paper card , use 10, 100, 200, 400, 600, 800, 1000ng / mL lipoprotein-related phospholipase A2 calibrator respectively, measure with test paper card, get the fluorescence intensity value of each calibrator, and input the result into the analyzer’s In Parameters, complete the setting of the parameters of the analyzer.

[0057] Main testing materials: clinical samples were obtained from relevant hospitals, a total of 300 latex-enhanced immunoturbidimetric value samples, including 100 serum samples, 100 plasma samples, and 100 whole blood samples, the content distribution range of lipoprotein-related phospholipase A2 was Between 10.0-...

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Abstract

The invention relates to the technical field of fluorescence immunochromatography in medical immunology and in particular relates to a kit for determining lipoprotein-associated phospholipase A2 and a manufacturing method. The kit is provided with a test paper card and is characterized in that the test paper card is provided with a PVC (Polyvinyl Chloride) plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad in sequence from bottom to top, wherein a lipoprotein-associated phospholipase A2 monoclonal antibody marked by a rare earth Eu<3+> fluorescent microsphere is adsorbed on the combination pad; the diameter of the fluorescent microsphere is 150nm; the rare earth fluorescent microsphere contains a rare earth lanthanide element Eu<3+> and is stable under a base state; the rare earth fluorescent microsphere sends out fluorescent light with the wavelength of 615nm under the action of a 337nm excitation light source; the monoclonal antibody is a purified and mixed monoclonal antibody and is selected from a monoclonal antibody cell strain aiming at 2 to 6 different antigenic epitopes of the lipoprotein-associated phospholipase A2; the kit has the advantages of simplicity and convenience for operation, rapid reaction, high sensitivity, high specificity and the like.

Description

[0001] Technical field: [0002] The invention relates to the technical field of fluorescence immunochromatography in medical immunology, including protein cross-linking technology, membrane chromatography technology, marker immunoassay technology and the like. Specifically, it is a lipoprotein-associated phospholipase A2 assay kit and a production method capable of quickly and accurately performing quantitative analysis of the lipoprotein-associated phospholipase A2 in samples such as serum, plasma, and whole blood. [0003] Background technique: [0004] With the in-depth research on the pathogenesis of atherosclerosis, people have found that in the process of the occurrence and development of atherosclerosis, there are always various inflammatory cells and a large number of inflammatory mediators participating in it. core factor. Lp-PLA2 is a phospholipase A2 superfamily closely related to atherosclerotic heart and cerebrovascular diseases that has attracted widespread atte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/573G01N33/558G01N33/543G01N33/533
CPCG01N33/577G01N33/533G01N33/54313G01N33/558G01N33/573
Inventor 王鹏浩李红江宋璐琳王文亮刘衍亮于鸿翔
Owner WEIHAI NEOPROBIO
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