Fluorescence immunochromatography kit for simultaneously detecting CRP, lipoprotein phospholipase A2 and D-dimers and preparation method thereof

A technique of fluorescence immunochromatography and immunochromatography test strips, applied in the field of medical testing, can solve the problems of multiple samples, missed diagnosis and treatment opportunities, high testing costs and labor costs, and achieves reduction of industrial production costs, simple structure, and improved detection and efficiency. The effect of diagnostic efficiency

Inactive Publication Date: 2018-11-23
江苏恒易生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This requires the hospital to invest in longer testing time, more samples, higher testing costs and labor costs, but often when clinicians get the test report, they have missed the best time for diagnosis and treatment

Method used

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  • Fluorescence immunochromatography kit for simultaneously detecting CRP, lipoprotein phospholipase A2 and D-dimers and preparation method thereof
  • Fluorescence immunochromatography kit for simultaneously detecting CRP, lipoprotein phospholipase A2 and D-dimers and preparation method thereof

Examples

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Embodiment 1

[0052] A preparation method of a fluorescent immunochromatography kit for simultaneously detecting ultrasensitive CRP, lipoprotein phospholipase A2 and D-dimer, the preparation method comprising the following steps:

[0053] (a) Treatment of fluorescent substances:

[0054] Dissolve carboxyfluorescein in morpholineethanesulfonic acid (MES) buffer solution with a pH of 5.0. After carboxyfluorescein is fully dissolved, add the activator 1-(3-dimethylaminopropyl)-3-ethane Carbodiimide (EDC), mix evenly, then add the activator N-hydroxysuccinimide (NHS) equal to EDC, mix evenly, incubate for 0.5h at room temperature in the dark, and then centrifuge Centrifuge for 5 minutes, take out the supernatant, set aside, wash and redissolve the precipitate with morpholineethanesulfonic acid (MES) buffer solution with pH 5.0, the concentration of EDC and NHS is 1mg / mL;

[0055] (b) Antibody labeling:

[0056] Add the hs-CRP monoclonal antibody to the carboxyfluorescein solution obtained in ...

Embodiment 2

[0068] A preparation method of a fluorescent immunochromatography kit for simultaneously detecting ultrasensitive CRP, lipoprotein phospholipase A2 and D-dimer, the preparation method comprising the following steps:

[0069] (a) Treatment of fluorescent substances:

[0070] Dissolve carboxyfluorescein in morpholineethanesulfonic acid (MES) buffer solution with a pH of 5.0. After carboxyfluorescein is fully dissolved, add the activator 1-(3-dimethylaminopropyl)-3-ethane Carbodiimide (EDC), mix evenly, then add the activator N-hydroxysuccinimide (NHS) equal to EDC, mix evenly, incubate for 0.5h at room temperature in the dark, and then centrifuge Centrifuge for 5 minutes, take out the supernatant, set aside, wash and redissolve the precipitate with morpholineethanesulfonic acid (MES) buffer solution with pH 5.0, the concentration of EDC and NHS is 1mg / mL;

[0071] (b) Antibody labeling:

[0072]Add the hs-CRP monoclonal antibody to the carboxyfluorescein solution obtained in s...

Embodiment 3

[0083] A preparation method of a fluorescent immunochromatography kit for simultaneously detecting ultrasensitive CRP, lipoprotein phospholipase A2 and D-dimer, the preparation method comprising the following steps:

[0084] (a) Treatment of fluorescent substances:

[0085] Dissolve fluorescein cy5 in morpholineethanesulfonic acid (MES) buffer solution with a pH of 6.0. After fluorescein cy5 is fully dissolved, add the activator 1-(3-dimethylaminopropyl)-3-ethane Carbodiimide (EDC), mix evenly, then add the activator N-hydroxysuccinimide (NHS) equal to EDC, mix evenly, incubate for 1.5h at room temperature in the dark, and then centrifuge Centrifuge for 15 minutes, take out the supernatant, set aside, wash and redissolve the precipitate with morpholineethanesulfonic acid (MES) buffer solution with pH 6.0, the concentration of EDC and NHS is 1mg / mL;

[0086] (b) Antibody labeling:

[0087] Add the hs-CRP monoclonal antibody to the fluorescein cy5 solution obtained in step (a)...

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Abstract

The invention discloses a fluorescence immunochromatography kit for simultaneously detecting CRP, lipoprotein phospholipase A2 and D-dimers and a preparation method thereof. The kit consists of an immunochromatography test paper strip and a plastic card case, wherein the immunochromatography test paper strip comprises a sample pad, a marker pad, a immunochromatography film, a water suction pad anda bottom plate; a fluorescent marker hs-CRP monoclonal antibody-1, a fluorescent marker Lp-PLA2 monoclonal antibody-1 and a fluorescent marker D-Dimer monoclonal antibody-1 are coated on the marker pad; hs-CRP monoclonal antibody-2, Lp-PLA2 monoclonal antibody-2 and D-Dimer monoclonal antibody-2 are respectively coated on a first detection line, a second detection line and a third detection lineon the immunochromatography film. According to the preparation method, the kit is obtained through fluorescent material processing, antibody marking, marker pad preparation, and immunochromatography film preparation and assembly. The kit realizes the combined detection of hs-CRP, Lp-PLA2 and D-Dimer; the detection and diagnosis efficiency is greatly improved; the structure of the kit is simple; the materials are popularized materials on the market; the industrial production cost can be greatly reduced.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a fluorescent immunochromatography kit and a preparation method for simultaneously detecting ultrasensitive CRP, lipoprotein phospholipase A2 and D-dimer. Background technique [0002] Acute coronary syndrome (acute coronary syndrome, ACS) is a common clinical cardiovascular disease, and atherosclerosis is the case basis of acute coronary syndrome. Studies have found that inflammatory mediators are involved in the formation of atherosclerosis, and the inflammatory response is closely related to the occurrence, development and prognosis of acute coronary syndrome. [0003] Hypersensitive C-reactive protein (hs-CRP) is currently the most commonly used clinical inflammatory response marker. It is a protein that rises sharply in the plasma when the body is infected or tissue damage, activates complement and strengthens the phagocytosis of phagocytic cells to regulate Functio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/533
CPCG01N33/533G01N33/558G01N33/577
Inventor 邱又彬欧阳文徐玉王宇
Owner 江苏恒易生物科技有限公司
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