Method for detecting Golgi protein 73, detection reagent and detection kit

An antibody and volumetric technology, applied in the field of medical testing, can solve problems such as difficulty in automation, and achieve the effects of improving detection efficiency, high sensitivity, and reducing detection costs

Inactive Publication Date: 2016-12-14
FUJIAN COSUNTER PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] not easy to automate

Method used

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  • Method for detecting Golgi protein 73, detection reagent and detection kit
  • Method for detecting Golgi protein 73, detection reagent and detection kit
  • Method for detecting Golgi protein 73, detection reagent and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1. Preparation of GP73 Magnetic Particle Separation Immunofluorescence Analysis Detection Reagent

[0089] 1. Preparation of GP73 antigen

[0090] (1) Construction of pColdⅡ-GP73 expression vector

[0091] The primers were designed according to the gene sequence of human GP73 in GenBank and the GP73 gene was synthesized. After PCR amplification, the GP73 amplified product was detected by 1% agarose gel electrophoresis, the target band was excised, and the target fragment was recovered using a gel recovery kit. It was ligated with the prokaryotic expression vector pColdII double digested with restriction enzymes NdeI and HindIII, transformed into E.coli.DH5α for amplification, and then transformed into E.coli.BL21(DE3) competent cells, inoculated In LB solid medium containing 100 mg / L ampicillin (Amp), the plasmids were extracted, identified and sequenced by NdeI and HindIII digestion. The recombinant plasmid pColdⅡ-GP73 was double digested by 5'NdeⅠand 3'HindⅢ...

Embodiment 2

[0152] Example 2. Composition of GP73 Magnetic Particle Immunofluorescence Detection Kit

[0153] The present invention designs a kit according to the method of the present invention, and the kit can be used for the detection of GP73. By using the kit, the operation is simple, time-saving and labor-saving, avoiding the tediousness of current use and preparation, and at the same time making Standardize operations.

[0154] Therefore, the present invention provides a kit.

[0155] The kit of the present invention includes magnetic microparticles, and optionally the following components: magnetic microparticles A coupled with GP73 antibody, magnetic microparticles B coupled with GP73 antigen, GP73 antigen, anti-GP73 antibody, fluorescein, and coupling buffer , washing buffer, eluent, blocking solution, preservation solution, magnetic stand.

[0156] Another kit of the present invention comprises: magnetic particles A coupled with GP73 antibody, magnetic particles B coupled with...

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Abstract

The invention discloses a method for detecting Golgi protein 73, a detection reagent and a detection kit used in the method. The detection method comprises the following steps: 1, preparation of a magnetic particle A with an anti GP73 antibody coupled on the surface and preparation of a magnetic particle B with a GP73 antigen coupled on the surface; 2, preparation of a fluorescent-labeled anti GP73 antibody; and 3, detection of GP73 content , to be more specific, a plasma or serum sample is reacted with the magnetic particle A, after the reaction, GP73 is eluted, the fluorescent-labeled anti GP73 antibody is added for incubation, then the magnetic particle B is added to continue incubation, magnetic separation is performed, sample fluorescence value is detected by a fluorescence spectrophotometer, and the GP73 content in the sample is calculated.

Description

technical field [0001] The invention relates to a medical detection method, in particular to a GP73 magnetic particle immunofluorescence detection method, as well as a GP73 magnetic particle immunofluorescence detection reagent and a detection kit. Background technique [0002] Liver cancer is one of the 10 common malignant tumors in the world. About 250,000 people die from liver cancer every year in the world, and the number of deaths due to liver cancer in my country accounts for about 45% to 50% of the global deaths. The incidence of liver cancer is relatively insidious, the treatment is difficult, the surgical complications are many, the curative effect is usually poor, and the patient's survival period is short. At present, the main reason for the poor efficacy of clinical treatment of liver cancer is the difficulty of diagnosis. Most of the clinical cases are in the advanced stage and the treatment effect is poor. Therefore, early detection and timely treatment are ver...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543G01N33/68
CPCG01N33/57438G01N33/54326G01N33/57484G01N33/68G01N2800/7028
Inventor 刘婕吴文强李国栋
Owner FUJIAN COSUNTER PHARMA CO LTD
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