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A modified latex immunoturbidimetric assay kit for improving the detection rate of antibodies against Helicobacter pylori pathogenic strains

A technology of Helicobacter pylori and pathogenic bacteria, applied in the field of medical testing, can solve the problems of low specificity and accuracy, affecting sensitivity, stability of cross-linking agents and latex microspheres, specificity and coupling efficiency, etc. , to achieve the effect of increasing linearity and sensitivity, improving sensitivity and shortening activation time

Active Publication Date: 2019-08-09
北京万泰德瑞诊断技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, this method still has the following technical defects: 1) Compared with the general clinical diagnostic criteria, the sensitivity, specificity and accuracy are low; 2) The stability and specificity of the cross-linking agent and latex microspheres in the antigen cross-linking process and coupling efficiency still needs to be improved; 3) The selection, proportioning and synergy with microspheres of the protective agent, stabilizer and surfactant in the reagent directly affect the sensitivity, linearity, specificity and stability of the reagent detected; In order to facilitate clinical promotion and ensure the reliability of test results, it is necessary to further improve the sensitivity, linearity, specificity and stability of existing technologies

Method used

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  • A modified latex immunoturbidimetric assay kit for improving the detection rate of antibodies against Helicobacter pylori pathogenic strains
  • A modified latex immunoturbidimetric assay kit for improving the detection rate of antibodies against Helicobacter pylori pathogenic strains
  • A modified latex immunoturbidimetric assay kit for improving the detection rate of antibodies against Helicobacter pylori pathogenic strains

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0052] 1. Preparation of antibody assay kit for Helicobacter pylori pathogenic strains

[0053] Preparation of Helicobacter pylori Antigen in East Asia

[0054] 1) Isolation, cultivation and identification of gastric Helicobacter pylori: The gastric mucosal tissue of patients infected with Helicobacter pylori was bitten and examined through a gastroscope, and then the gastric mucosal tissue was ground into a slurry in a glass grinder containing 0.5 mL of Brooke's broth, and absorbed Spread 100 μL on the Columbia selection medium plate (containing 8% defibrinated sheep blood, 5mg / LTMP, 5mg / L polymyxin B, 5mg / L amphotericin B, 10mg / L vancomycin) , and then place the plate in a three-gas incubator (10% CO 2 , 85%N 2 , 5%O 2 ) at 37°C for 3 to 7 days, and carry out Gram staining microscopic examination, urease, catalase, and oxidase tests on the suspicious colonies cultured. The results are Gram-negative, S-type or arc-type. It was initially identified as Helicobacter pylori (...

Embodiment 2

[0129] 1. Preparation of antibody assay kit for Helicobacter pylori pathogenic strains

[0130] The preparation of East Asian Helicobacter pylori antigen is the same as that in Example 1 "Preparation of East Asian Helicobacter pylori antigen".

[0131] Preparation of reagent 1: 50mmol / L Tris-HCl buffer, add 0.8% sodium chloride, 0.5% BSA, 0.1% TRITON X-100, 5% sucrose, 0.05% dodecyl glucoside, 0.05% Proclin300 , each raw material should be stirred evenly after the addition of the raw materials, until the raw materials are added, continue to stir for 5 minutes, stand still for 2 minutes, adjust the pH to 8.0, and obtain reagent 1.

[0132] Preparation of Reagent 2:

[0133] 1) Modification of latex microspheres: dissolving hyperbranched polyglycidyl ether in pyridine, and modifying with succinic anhydride (Suc) to obtain hyperbranched polyglycidyl ether containing a large number of carboxyl groups.

[0134] 2) Latex particle labeling: Dilute the latex particle to 5mg / mL with ...

Embodiment 3

[0186] 1. Preparation of antibody assay kit for Helicobacter pylori pathogenic strains

[0187] The preparation of East Asian Helicobacter pylori antigen is the same as that in Example 1 "Preparation of East Asian Helicobacter pylori antigen".

[0188]Preparation of reagent 1: 50mmol / L Tris-HCl buffer, add 0.8% sodium chloride, 0.8% BSA, 0.5% TRITON X-100, 2% sucrose, 0.01% dodecyl glucoside, 0.05% Proclin300 , each raw material should be stirred evenly after the addition of the raw materials, until the raw materials are added, continue to stir for 5 minutes, stand still for 2 minutes, adjust the pH to 8.4, and obtain reagent 1.

[0189] Preparation of Reagent 2:

[0190] 1) Modification of latex microspheres: dissolving hyperbranched polyglycidyl ether in pyridine, and modifying with succinic anhydride (Suc) to obtain hyperbranched polyglycidyl ether containing a large number of carboxyl groups.

[0191] 2) Latex particle labeling: Dilute the latex particle to 5mg / mL with 1...

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Abstract

The invention relates to a modified latex immunonephelometry kit for improving a detection rate of pathogenic strain antibodies of helicobacter pylori. The kit is formed by a reagent 1, a reagent 2, a calibration product, a low-value quality control product and a high-value quality control product which are placed. Through the identification and classification of the helicobacter pylori strains in the earlier stage, the detection rate of the pathogenic strains is improved. In the antigen coating process, 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) is selected as a cross-linking agent for binding of latex microspheres decorated by hyperbranched polyglycerol and antigens, coupling antigen conformation is protected by using the hydrophilic property of the hyperbranched polyglycerol while the coupling efficiency is increased, and the specific binding of the antigens and the antibodies is improved. In the preparation process of the reagents, a novel surfactant of decyl polyglucoside is selected for effectively improving the scavenging activity of reagent lipid impurities and protein impurities under the combined action with other active agents, stabilizers, protective agents and preservatives.

Description

technical field [0001] The invention belongs to the technical field of medical inspection, and relates to a modified latex immunoturbidimetric assay kit for improving the detection rate of antibodies to pathogenic strains of Helicobacter pylori. Background technique [0002] Helicobacter pylori (H. pylori for short) is a unipolar, multi-flagellate, blunt-rounded end, spiral-curved bacterium, which is negative for Gram stain, and often presents a typical spiral or helical pattern on the surface of gastric mucosal epithelial cells arc. [0003] Helicobacter pylori is one of the risk factors for gastric cancer. In 1994, the World Health Organization / International Agency for Research on Cancer (WHO / IARC) classified it as a class I carcinogen. Epidemiology shows that almost half of the world's population is infected with this bacterium, even as high as 60% to 70% in developing countries. Therefore, Helicobacter pylori infection is a public health problem that countries all ove...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/544G01N33/543G01N33/531G01N21/82C12Q1/689C12Q1/04C12N1/20C12R1/01
CPCC12N1/20C12Q1/689G01N21/82G01N33/531G01N33/54313G01N33/54393G01N33/544G01N33/56922G01N2021/825
Inventor 李雪潘玥许泼实
Owner 北京万泰德瑞诊断技术有限公司
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