Method and reagent kit for detecting phosphatidylinositol proteoglycan 3 with fluorescence immunochromatographic method

A technology of fluorescence immunochromatography and phosphatidylinositol, which is applied in the field of medical immunology, can solve the problems of low degree of automation, inaccurate quantitative detection, long operation time of ELISA method, etc., and achieve simple operation, fast response and high sensitivity Effect

Inactive Publication Date: 2016-06-29
SHANDONG KANGLI MEDICAL EQUIP TECH
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Problems solved by technology

[0004] Glypican 3 detection methods mainly include double-antibody sandwich method (ELISA) and immunohistochemical method. ELISA method takes a long time to operate, has low degree of automation, and quantitative detection is not accurate enough; immunohistochemical method requires tissue sections, and Impossibility of tissue sampling in the early stages of PHC and inability to obtain results at an early stage for early diagnosis

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  • Method and reagent kit for detecting phosphatidylinositol proteoglycan 3 with fluorescence immunochromatographic method

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1: Preparation of Glypican 3 Fluorescence Immunochromatography Detection Kit

[0036] 1 main material:

[0037]1.1 Glypican 3-specific paired antibody: Abcam Shanghai Trading Co., Ltd.; Glypican 3 quality control product: Beijing Yiqiao Shenzhou Biotechnology Co., Ltd.; rare earth fluorescent microspheres : Shanghai Zhenzhun Biotechnology Co., Ltd.; nitrocellulose (NC) membrane: product of Millipore; bovine serum albumin (BSA), polyethylene glycol PEG20000, hydrolyzed casein: product of Sigma. Other commonly used reagents are analytical reagents.

[0038] 1.2. Fluorescence Immunochromatography Analyzer: Model: HF201, Shenzhen Huakerui Technology Co., Ltd.

[0039] 2 methods

[0040] 1) Aldehydization of rare earth fluorescent nanospheres:

[0041] Take 3 mg of rare earth fluorescent microspheres, wash them three times with 50 mM, pH9.5 carbonate buffer, centrifuge at 12,000 rpm for 10 minutes, and finally resuspend in 100 μl of the above carbonate buffer . ...

Embodiment 2

[0055] Embodiment 2: sample accuracy test

[0056] 1. Main materials: clinical samples obtained by our company in relevant hospitals, a total of 200 ELISA-valued samples, including 100 serum samples, 100 whole blood samples, and the content distribution range of Glypican 3 is 0-800ng / mL.

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Abstract

The invention provides a method and a reagent kit for detecting phosphatidylinositol proteoglycan 3 with a fluorescence immunochromatographic method. The method for detecting phosphatidylinositol proteoglycan 3 with the fluorescence immunochromatographic method comprises the steps that the fluorescence characteristic of rare earth fluorescent microspheres is utilized and combined with a mature immunochromatography technology, and fluorescent quantitative determination of phosphatidylinositol proteoglycan 3 is achieved by optimizing all steps. The reagent kit is characterized by being composed of a test paper card and an immunofluorescence chromatography analysis meter. A preparing method of the reagent kit comprises the steps of preparation of a specificity high-appetency monoclonal antibody, antibody labeling with rare earth fluorescent microspheres, preparation of a fluorescent microsphere combined pad and a nitrocellulose membrane sprayed with the antibody, assembling and wrapping of test paper strips, and the like. In the detection process, a sample is operated as a specification, a result is subjected to quantitative determination with the immunofluorescence chromatography analysis meter, and the advantages of being easy, convenient and fast to implement, suitable for field detection and the like are achieved.

Description

Technical field: [0001] The present invention belongs to the field of medical immunology, and relates to a fluorescent immunochromatography technique. Further, the present invention relates to a method for detecting glypican 3 by rare earth fluorescent immunochromatography and a kit thereof. Achieve fast and accurate quantitative analysis of Glypican 3. Background technique: [0002] Primary liver cancer (primary hepatic carcinoma, PHC) is one of the malignant tumors with the highest mortality rate in the world. The five-year survival rate after diagnosis is less than 12%. In recent years, its incidence has been increasing worldwide. Surgical resection is still the most commonly used treatment for PHC at present, but most patients are already in the middle and advanced stages when they seek treatment, and the best time for surgical treatment has been lost. Therefore, improving the early diagnosis of PHC is the key to improving the treatment and prognosis of liver cancer. Al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/558
CPCG01N33/68G01N33/558G01N33/57438G01N2333/4722
Inventor 钱承玉钱琛吕作文刘壮
Owner SHANDONG KANGLI MEDICAL EQUIP TECH
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