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Immunochromatographic assay kit for determinating lipoprotein-associated phospholipase A2 and preparation method thereof

A technology of immunochromatography detection and phospholipase, which is applied in the field of immunochromatography in medical immunology, can solve the problems of qualitative result deviation, long time to obtain results, and inability to obtain the severity of the disease, and achieve rapid results, easy operation, and measurement accurate effect

Pending Publication Date: 2018-12-14
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Quantitative methods generally require expensive instruments, and it takes a long time to obtain results, and Lp-PLA2 detection-related diseases such as myocardial infarction and heart failure urgently need to obtain test results as soon as possible; while the existing qualitative tests can be used for a short time, but Qualitative results are prone to bias, and the severity of the disease cannot be obtained when positive results are obtained

Method used

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  • Immunochromatographic assay kit for determinating lipoprotein-associated phospholipase A2 and preparation method thereof
  • Immunochromatographic assay kit for determinating lipoprotein-associated phospholipase A2 and preparation method thereof
  • Immunochromatographic assay kit for determinating lipoprotein-associated phospholipase A2 and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The mouse anti-human Lp-PLA2 monoclonal antibody combined with the colored fluorescent microspheres on the reagent pad in the present invention is prepared by the following measures:

[0033] 1. Acquisition of monoclonal antibody cell lines: immunize mice with Lp-PLA2 protein, and prepare at least 100 specific and high-affinity monoclonal antibody cell lines using standard monoclonal antibody preparation methods. The corresponding anti-monoclonal antibodies were prepared from the cell lines, paired in pairs with 96-well plates and ELISA, and more than 100 monoclonal antibodies were initially screened, and 10 pairs of antibody pairs with the best response were selected, with a total of 13 cell lines. 5 strains are used for reagent pads and 8 strains are used for nitrocellulose membranes. Further use the above 13 strains of monoclonal antibodies, refer to the aforementioned kit preparation scheme according to the preliminary screening results, prepare 40 sets of detection...

Embodiment 2

[0036] Embodiment 2: color microsphere qualitative test and fluorescent immunoassay accuracy test

[0037] Select the above kit and fluorescence immunochromatography analyzer (model: NEO-007),

[0038] Setting of the parameters of the fluorescence immunoassay analyzer: After setting the process parameters of the kit on the fluorescence immunoassayer, take the above assembled kit and use 10, 25, 100, 200, 400, 600, 800, 1000ng / The Lp-PLA2 calibrator in ml was measured with a kit to obtain the fluorescence intensity value of each calibrator, and the results were input into the parameters of the analyzer to complete the final setting of the parameters of the analyzer.

[0039] Main testing materials: 200 clinical samples were obtained from relevant hospitals, and the lipoprotein-related phospholipase A2 assay kit of Desai Diagnostic System was used to determine the value, including 100 serum samples, 100 whole blood samples, and the distribution range of Lp-PLA2 content Between...

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Abstract

The invention relates to an immunochromatographic assay kit for determinating lipoprotein-associated phospholipase A2. The immunochromatographic assay kit comprises an immunochromatographic test stripand a plastic shell, wherein the immunochromatographic test strip is a test strip consisting of a bottom plate, a sample pad, a reagent pad, a nitrocellulose membrane and an absorbent pad; the samplepad, the reagent pad, the nitrocellulose membrane and the absorbent pad are sequentially fixed to the bottom plate in a staggered manner; the reagent pad is coated with colored fluorescent microspheres labeled by a mouse anti-human Lp-PLA2 monoclonal antibody; a test line on the nitrocellulose membrane is coated with the mouse anti-human Lp-PLA2 monoclonal antibody, and a quality control line thereon is coated with a goat anti-mouse polyclonal antibody; the kit has the qualitative determination boundary concentration of 200ng / ml, a fluorescent immunoassay analyzer has the detection sensitivity of 10ng / ml, and the linear range is 10-1000ng / ml; through use of the colored fluorescent microspheres, a qualitative result can be rapidly obtained and a quantitative result can be obtained later. The immunochromatographic assay kit has the characteristics of simple operation, rapid response, high sensitivity, qualitative and quantitative determination and the like, and can help to meet needs ofdifferent inspection scenes.

Description

technical field [0001] The invention relates to the technical field of immunochromatography in medical immunology, in particular to an immunochromatographic detection kit for detecting lipoprotein-associated phospholipase A2 which can quickly and accurately perform simultaneous qualitative and quantitative analysis of Lp-PLA2. Background technique [0002] 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 attention in recent years. More and more studies have shown that Lp-PLA2 is closely related to coronary atherosclerotic heart disease (Coronary heart disease) is closely related to the occurrence of stroke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 王鹏浩宋璐琳王文亮刘衍亮陈萍萍孙宁宁
Owner WEIHAI NEOPROBIO
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