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PG (pepsinogen) and H.pylori antibody detection method and kit

A technology of pepsinogen and Helicobacter pylori, applied in the field of biomedicine, can solve the problems of inaccurate quantification, high detection sensitivity, and complicated operation process, and achieve rapid and highly sensitive determination, high sensitivity, and strong specificity

Inactive Publication Date: 2017-11-07
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • PG (pepsinogen) and H.pylori antibody detection method and kit
  • PG (pepsinogen) and H.pylori antibody detection method and kit
  • PG (pepsinogen) and H.pylori antibody detection method and kit

Examples

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preparation example Construction

[0044] The general preparation method for detecting pepsinogen Ⅰ, pepsinogen Ⅱ and anti-Helicobacter pylori antibody lateral flow chromatography detection reagent comprises the following steps:

[0045] (1) Preparation of carboxyl-modified fluorescent microsphere-labeled probes

[0046] 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.

[0047] After activating the carboxyl groups on the sur...

Embodiment 1

[0060] Example 1 Preparation of Lateral Flow Detection Reagent for Gastric Cancer Screening

[0061] (1) Preparation of fluorescent microsphere-labeled pepsinogen Ⅰ and pepsinogen Ⅱ-labeled antibodies, Helicobacter pylori-labeled antigen and mouse IgG

[0062] 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.

[0063] Take 20 mg of the above-mentioned carboxyl-modified fluorescent microspher...

Embodiment 2

[0069] Example 2 Evaluation of Lateral Flow Detection Reagents for Gastric Cancer Screening

[0070] (1) Detection sensitivity

[0071] Use pepsinogen I, pepsinogen II antigen, and anti-Helicobacter pylori antibody as the samples to be tested to determine the detection of pepsinogen I, pepsinogen II, and anti-Helicobacter pylori antibody lateral flow detection reagent in Example 1. sensitivity.

[0072] Pepsinogen Ⅰ, pepsinogen Ⅱ antigen, and anti-Helicobacter pylori antibody were prepared with 0.02M PBS buffer solution at pH 7.4 containing 5% calf serum to a series of concentrations (0, 0.5, 1, 5, 10, 50, 100, 500 ng / mL), respectively added to the loading end of the detection pepsinogen I, pepsinogen II antigen, and anti-Helicobacter pylori antibody lateral flow detection reagent obtained in Example 1, and detected by fluorescence The instrument detects the fluorescence intensity. Detection steps: return the sample to be tested to room temperature (25°C) before the test, a...

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Abstract

The present invention proposes a kit, the use of the kit in detecting pepsinogen I, pepsinogen II and anti-Helicobacter pylori antibodies, and the method of using the kit to detect pepsinogen I, pepsinogen II and anti-Helicobacter pylori antibodies method. The kit comprises: a first coating membrane; and a second coating membrane, one end of the first coating membrane is connected to one end of the second coating membrane; at least one area of ​​the first coating membrane is coated with pepsinogen labeled with fluorescent microspheres Ⅰ antibody pepsinogen Ⅰ antibody, fluorescent microsphere-labeled pepsinogen Ⅱ antibody and fluorescent microsphere-labeled first Helicobacter pylori antigen, the second coating membrane contains the separated first area, second area, third area and In the fourth area, the materials for forming fluorescent microspheres include: polystyrene-methyl methacrylate copolymer. The kit and method of the invention have the advantages of high sensitivity, strong specificity, rapidity, simplicity, objective determination 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 and anti-helicobacter pylori antibody and a kit thereof. More specifically, the present invention relates to a kit, the use of the kit in detecting pepsinogen I, pepsinogen II and anti-H. Bacterial antibody 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 upper duodenum also produce PGII, and a small amount of PGII is also produced by the prostate and pancreas. Under normal circumstances, about ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/573G01N33/569G01N33/558
CPCG01N33/577G01N33/558G01N33/56911G01N33/573
Inventor 马岚
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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