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Detection kit of helicobacter pylori emulsion method and preparation process thereof

A Helicobacter pylori detection kit technology, applied in the field of immunochromatography, can solve the problems of strict requirements for culture and pathological detection conditions, inability to identify past infection and current epidemic patients, and unsuitability for grassroots detection work, and achieve high sensitivity , high immunogenicity and reactogenicity, convenient storage and transportation

Active Publication Date: 2012-08-15
SHANGHAI CHEMTRON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (1) Enzyme-linked immunosorbent assay and western blot test: the presence of HP in the human body can be determined by detecting the HP antibody in the serum. This method has high specificity and can perform quantitative analysis, but it cannot distinguish between past infection and current epidemic patients
[0007] (2) Urea breath test and PCR detection test: the presence of HP is determined by measuring the metabolites or genes of urease produced by HP in the body. This method has high sensitivity, but the operation is complicated and the equipment is expensive, so it is not suitable for the grassroots Detection work, promotion and use are limited
[0008] (3) Bacterial culture and other pathological detection methods, etc. These methods also generally have the advantages of high sensitivity and strong specificity, but there are also limitations in the detection conditions of culture and pathological methods, which require strict requirements, complicated operations, and professional operations. sex

Method used

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  • Detection kit of helicobacter pylori emulsion method and preparation process thereof
  • Detection kit of helicobacter pylori emulsion method and preparation process thereof
  • Detection kit of helicobacter pylori emulsion method and preparation process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 Preparation of anti-gastric Helicobacter pylori urease antibody

[0068] 1. Design of antigen fusion protein

[0069] According to the immune mechanism of the body against gastric Helicobacter pylori and the immune properties of the gastric Helicobacter pylori urease epitope, select the specific fragment UreB of urease UreB 138 (UreB amino acid sequence GenBank: AAU21200.1) and a specific fragment of UreA UreA 65 (UreA amino acid sequence GenBank: AAK69750.1) is used for the construction of Hp antigen fusion protein. UreB 138 and UreA 65 There is a connecting peptide fragment RWYCR inserted between them, and urease UreB 138 The fragment is fused at the C-terminus of the protein, UreA 65 The fragment is fused to the N-terminus of the protein.

[0070] 2. Construction of recombinant expression vector

[0071] (1) Primer synthesis

[0072] Design UreB with Primer Premier V5.0 138 Gene and UreA 65 The primers of the gene are as follows, the underlined...

Embodiment 2

[0119] Embodiment 2 The preparation of gastric Helicobacter pylori latex method detection kit

[0120] 1. Reagent source:

[0121] Goat anti-rabbit IgG polyclonal antibody, anti-helicobacter pylori urease antibody II (purchased from MAXMED LABORATORIES INC.)

[0122] Nitrocellulose film, polyester film, polystyrene particles (purchased from Germany Sartorius company SARTORIUS) polystyrene particles (purchased from WHATMAN company)

[0123] 2. Preparation steps:

[0124] method 1:

[0125] 1. Preparation of sensitized latex particles:

[0126] a. Take 0.4ml of 10% carboxylated polystyrene latex into a centrifuge tube, add 1ml of pH 6.0 carbonic acid buffer solution containing 0.01% SDS 20mM to wash twice, centrifuge at 14000g / min for 10min, discard the supernatant to obtain carboxylation The polystyrene particles precipitated.

[0127] b. Suspend the carboxylated polystyrene pellet obtained in the previous step with 0.5ml of pH 6.0 20mM phosphate buffer solution, add 0.5ml...

Embodiment 3

[0166] Embodiment 3 The specificity experiment of gastric Helicobacter pylori latex method detection kit

[0167] Detection method:

[0168] 1. Take the gastric Helicobacter pylori latex method detection kit prepared in Example 2, and place the kit on a horizontal platform.

[0169] 2. Preparation of the sample to be tested: According to the four types of test objects in Table 1, use a special sampling stick to randomly sample from different parts of the tester's feces. The sampling amount is subject to the small ring at the front end of the special sampling stick; Put the processing tube vertically on the processing tube bracket, and add 0.6ml of distilled water into the sample tube; insert the sample taken from the special sampling stick into the distilled water in the sample tube and stir well as the sample to be tested.

[0170] 3. There are 100 subjects for each type of experiment, and the test results are shown in Table 1:

[0171] Table 1 Specificity test results of g...

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Abstract

The invention relates to the field of the emulsion method immune chromatography, and particularly relates to a detection kit of a helicobacter pylori emulsion method. The detection kit of the helicobacter pylori emulsion method comprises a reagent strip, wherein the reagent strip comprises a substrate, filter sample paper, a sensitized emulsion polyester film, an immune cellulose nitrate film and absorbent paper, wherein the filter sample paper, the sensitized emulsion polyester film, the immune cellulose nitrate film, and the absorbent paper are orderly connected end to end, and are fixed on the substrate, the immune cellulose nitrate film is covered with helicobacter pylori resisting urea enzyme antibodies I marked by colorful emulsion particles, and the immune cellulose nitrate film is provided with a detection line coated with helicobacter pylori resisting urea enzyme antibodies II and a quality control line coated by goat anti-rabbit serum IgG. The detection kit of the helicobacter pylori emulsion method, which is provided by the invention, has the advantages of convenience in detection, stability in detection results and cost conservation.

Description

technical field [0001] The invention relates to the technical field of immunochromatography, in particular to a latex method detection kit for gastric Helicobacter pylori and a preparation process thereof. Background technique [0002] Helicobacter Pylori (HP for short) is a spiral Gram-negative bacterium in the human gastric mucosa. This bacterium is the culprit that causes chronic gastritis, gastric ulcer, duodenal ulcer and even gastric cancer. It is generally believed that the clinical course of gastric Helicobacter pylori infection is as follows: gastric Helicobacter pylori colonizes and infects after reaching the gastric mucosa through the mouth, causing chronic and superficial gastritis after several weeks or months, and develops into gastritis after several years or decades. Duodenal ulcer, gastric ulcer, lymphoproliferative gastric lymphoma, chronic atrophic gastritis, etc., and the latter is the most dangerous factor leading to gastric cancer. Experts believe tha...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/573G01N33/558
Inventor 张国华
Owner SHANGHAI CHEMTRON BIOTECH
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