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Pepsinogen, helicobacter pylori antibody and gastrin 17 detection method and kit thereof

A technology of pepsinogen and Helicobacter pylori, which is applied in the field of biomedicine, can solve the problems of low sensitivity, high detection sensitivity, long detection time, etc.

Inactive Publication Date: 2017-10-20
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Latex-enhanced immunoturbidimetric method, enzyme-linked immunoassay, and time-resolved fluorescent immunoassay, the operation process is complicated, requiring a large number of instruments and equipment and professional personnel to operate, the detection time is long, and the detection sensitivity is high
Colloidal gold immunochromatography is simple to operate, but the sensitivity is not high, and it cannot be accurately quantified

Method used

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  • Pepsinogen, helicobacter pylori antibody and gastrin 17 detection method and kit thereof
  • Pepsinogen, helicobacter pylori antibody and gastrin 17 detection method and kit thereof
  • Pepsinogen, helicobacter pylori antibody and gastrin 17 detection method and kit thereof

Examples

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preparation example Construction

[0040] The general preparation method for detecting pepsinogen Ⅰ, pepsinogen Ⅱ, gastrin 17 and anti-Helicobacter pylori antibody lateral flow chromatography detection reagent comprises the following steps:

[0041] (1) Preparation of carboxyl-modified fluorescent microsphere-labeled probes

[0042] Mix styrene and methyl methacrylate at a ratio of 1:1, add 1% rare earth complex Eu(TTA)3Phen or 0.5% CdSe / ZnS quantum dots, and ultrasonically mix to obtain liquid a. 0.05% carboxylated polyvinyl alcohol and 0.05% sodium bicarbonate were dissolved in water to obtain liquid b. Add liquid a to liquid b and ultrasonicate for 15 minutes, blow nitrogen for 30 minutes, stir to remove oxygen, and then heat to 80 degrees. Add 0.01-0.1% potassium persulfate and react for 12 hours to obtain polymer fluorescent microspheres, which are filtered, centrifuged and washed with deionized water to obtain purified functionalized fluorescent microspheres.

[0043] After activating the carboxyl group...

Embodiment 1

[0056] Example 1 Preparation of Lateral Flow Chromatography Detection Reagent for Gastric Function Detection

[0057] (1) Preparation of fluorescent microsphere-labeled pepsinogen Ⅰ and pepsinogen Ⅱ-labeled antibodies, Helicobacter pylori-labeled antigen and gastrin-17-labeled antibody

[0058] Mix styrene and methyl methacrylate at a ratio of 1:1, add 1% rare earth complex Eu(TTA)3Phen or 0.5% CdSe / ZnS quantum dots, and ultrasonically mix to obtain liquid a. 0.05% carboxylated polyvinyl alcohol and 0.05% sodium bicarbonate were dissolved in water to obtain liquid b. Add liquid a to liquid b and ultrasonicate for 15 minutes, blow nitrogen for 30 minutes, stir to remove oxygen, and then heat to 80 degrees. Add 0.01-0.1% potassium persulfate and react for 12 hours to obtain polymer fluorescent microspheres, which are filtered, centrifuged and washed with deionized water to obtain purified functionalized fluorescent microspheres.

[0059] Take 20 mg of the above-mentioned carbo...

Embodiment 2

[0065] Example 2 Evaluation of Lateral Flow Detection Reagents for Gastric Function Testing

[0066] (1) Detection sensitivity

[0067] Pepsinogen Ⅰ, pepsinogen Ⅱ, gastrin 17 antigen and anti-helicobacter pylori antibody were used as test samples to determine the detection of pepsinogen Ⅰ, pepsinogen Ⅱ, gastrin 17 and anti-pyloric antibody in Example 1. Sensitivity of Helicobacter antibody lateral flow detection reagents.

[0068] The pepsinogen Ⅰ, pepsinogen Ⅱ antigen and anti-Helicobacter pylori antibody were prepared with 0.02M PBS buffer at pH 7.4 containing 5% calf serum to a series of concentrations (0, 0.5, 1, 5, 10, 50, 100, 500ng / mL), gastrin 17 was prepared into a series of concentrations (0, 0.5, 1, 5, 10, 50, 100 , 500ng / L), respectively added in the loading end of detection pepsinogen Ⅰ, pepsinogen Ⅱ antigen, gastrin 17 and anti-helicobacter pylori antibody lateral flow detection reagent obtained by embodiment 1, and adopt A fluorescence detector detects the fl...

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Abstract

The invention provides a kit, application of the kit in detection of pepsinogen I, pepsinogen II, a helicobacter pylori antibody and gastrin 17 and a method for detecting the pepsinogen I, the pepsinogen II, the helicobacter pylori antibody and the gastrin 17 by using the kit. The kit comprises a first coating film and a second coating film, wherein at least one region in the first coating film is coated with a fluorescent microsphere-labeled pepsinogen I antibody, a fluorescent microsphere-labeled pepsinogen II antibody, a first fluorescent microsphere-labeled helicobacter pylori antigen and a fluorescent microsphere-labeled gastrin 17 antibody; the second coating film comprises a first region, a second region, a third region, a fourth region and a fifth region which are separated; a formed fluorescent microsphere material comprises a polystyrene-methyl methacrylate copolymer. The kit and the method which are provided by the invention have the advantages of high sensitivity, high specificity, quickness, simplicity, capability of realizing objective measurement and the like.

Description

technical field [0001] The invention relates to the field of biomedicine. Specifically, the present invention relates to a detection method for pepsinogen, anti-Helicobacter pylori antibody and gastrin 17 and a kit thereof. More specifically, the present invention relates to a kit, the use of the kit in the detection of pepsinogen I, pepsinogen II, anti-Helicobacter pylori antibodies and gastrin 17, and the detection of pepsinogen I, pepsinogen Ⅱ. Anti-Helicobacter pylori antibody and gastrin-17 method. Background technique [0002] Pepsinogen (PG), the precursor of pepsin, is divided into two subgroups based on their biochemical properties and immunogenicity. Components 1-5 have the same immunogenicity and are called PGⅠ, which are mainly secreted by the principal cells and mucus neck cells of the gastric glands. The mucous neck cells of the acid glands, the mucous cells of the cardia glands and the pyloric glands of the gastric antrum, and the Brunner glands of the uppe...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/569G01N33/74G01N33/574G01N33/577G01N33/533G01N33/68G01N21/64
CPCG01N33/573G01N21/6402G01N21/6428G01N33/533G01N33/56911G01N33/574G01N33/57446G01N33/577G01N33/6893G01N33/74G01N2021/6439G01N2333/20G01N2333/595G01N2333/96477G01N2800/06
Inventor 马岚
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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