Immuo-colloidal gold test paper for detecting pyloric helicobacter antigen and its prepn process

A Helicobacter pylori, colloidal gold test paper technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve other bacteria such as Campylobacter Bacteria are also found, false positives and other problems

Inactive Publication Date: 2006-10-18
LANZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Urease is a characteristic enzyme of Hp. Its main active part is located on the surface of the bacteria, which can decompose urea to produce a large amount of ammonia, which plays an important role in the colonization and survival of Hp. Campylobacter, Proteus, Morganella, and Providencia are also found, so only using urease monoclonal antibody to detect Hp infection will have false positives; cell vacuolation toxin (VacA) can cause cells to produce vacuoles Denatured protein, Mr87000, is one of the virulence factors of Hp causing ulcer; cytotoxin-associated protein (CagA), Mr120000-140000, has no effect on causing vacuolar degeneration, but has a good correlation with VacA

Method used

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  • Immuo-colloidal gold test paper for detecting pyloric helicobacter antigen and its prepn process
  • Immuo-colloidal gold test paper for detecting pyloric helicobacter antigen and its prepn process

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Embodiment 1 Helicobacter pylori antigen preparation

[0028] Hp international standard strain NCTC11637 was inoculated on Skirrow agar containing 10% defibrinated sheep blood after systematic identification, placed in the environment of 5% O2, 10% CO2, and 85% N2, cultured at 37°C for 3 days and harvested bacteria. Centrifuge at 4000 rpm for 10 minutes, discard the supernatant, add 1×PBS buffer to the precipitate, blow it away, crush it with ultrasonic waves, and concentrate it to obtain the antigen with a protein concentration of 5 mg / ml.

Embodiment 2

[0029] The preparation of embodiment two immune colloidal gold test paper 1

[0030] One end of the PVC backing is glued with water-absorbing paper and glass fiber membrane in turn, the middle is glued with nitrocellulose membrane, and the other end is glued with water-absorbing fiber. Among them, the monoclonal antibody mixture of gold-labeled urease and vacuolar toxin A is adsorbed on the glass fiber membrane, and the detection areas 1 and 2 on the nitrocellulose membrane are respectively coated with different epitopes of anti-urease and anti-vacuolar toxin A The monoclonal antibody of the goat anti-mouse polyclonal antibody was coated on the nitrocellulose membrane as the quality control area.

[0031] Prepare as follows:

[0032] (1) Preparation of Helicobacter pylori monoclonal antibodies: BALB / C mice were immunized with Helicobacter pylori antigens several times to obtain ten monoclonal antibodies. These ten monoclonal antibodies were analyzed by electrophoresis, titer...

Embodiment 3

[0040] The preparation of embodiment three immune colloidal gold test paper 2

[0041] One end of the PVC backing is glued with water-absorbing paper and glass fiber membrane in turn, the middle is glued with nitrocellulose membrane, and the other end is glued with water-absorbing fiber. Among them, the monoclonal antibody mixture of gold-labeled urease and cell-associated toxin A is adsorbed on the glass fiber membrane, and the detection areas 1 and 2 on the nitrocellulose membrane are respectively coated with different epitopes of anti-urease and anti-cell-associated toxin A The monoclonal antibody of the rabbit anti-mouse polyclonal antibody was coated on the nitrocellulose membrane as the quality control area, and other operations were the same as in Example 2.

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Abstract

The present invention relates to one kind of immuno-colloidal gold test paper for detecting pyloric helicobacter antigen and its preparation process. The test paper has coated urease monoclonal antibody, CagA or VacA monoclonal antibody and anti-mouse polyclonal antibody. It is used in detecting pyloric helicobacter related antigen existing in saliva, gastric juice, vomited matter, dental plaque and excrement of mammal so as to monitor the infection status of pyloric helicobacter, assist the diagnosis of gastritis, peptic ulcer and other diseases and predict gastric cancer probability. The present invention simple operation, high specificity and high sensitivity.

Description

technical field [0001] The invention relates to an immune colloidal gold reagent for detecting antigens and a detection method thereof, and also relates to a preparation method of the reagent. Background technique [0002] Helicobacter pylori (Helicobacter pylori, Hp) has a chronic infection rate of more than 50% in the population of the world, and the infection rate of the general population in my country is about 50%-80%, and it is still at an annual rate of 1%-2%. Increase. Hp is the main pathogenic factor of type B gastritis, peptic ulcer and gastric mucosa-associated lymphoid tissue lymphoma, and is closely related to the occurrence of gastric cancer. It has been listed as the first-class biological carcinogen by WTO. It can also cause other diseases such as: coronary heart disease angina pectoris, growth retardation or indigestion in children, urticaria, immune platelet purpura, hepatic coma, symptomatic hypoglycemia, high blood pressure, and gallstones. [0003] Epid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543
Inventor 李红玉肖曼李静贤
Owner LANZHOU UNIVERSITY
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