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Immunoassay method for helicobacter pylori in gastric mucosa sample

A technology for Helicobacter pylori and anti-Helicobacter pylori is applied in the field of immunoassay of Helicobacter pylori, can solve problems such as affecting the urease test, and achieve the effects of intuitive detection results, low cost and fast reaction speed

Inactive Publication Date: 2012-07-11
王滔
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has been found that acid-suppressing drugs, some antibiotics, etc. will affect the urease test, and acid-suppressing drugs may be used for patients
(3) pH method, because there are false positives due to other mixed urease, visual errors in color judgment, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment one: ELISA test

[0067] (1), ELISA test alkaline solution preparation

[0068] The preparation method of the alkaline biological buffer that the present invention relates to is as follows:

[0069] 1. Hydroxyethylpiperazineethanesulfonic acid (HEPES) buffer

[0070] HEPES molecular formula: C 8 h 18 N 2 o 4 S molecular weight: 238.3

[0071] Add 2.38 grams of HEPES to every 1000ml of deionized water, adjust the pH to 8.2 with 1N NaOH after dissolution, filter and sterilize before use.

[0072] 2. Preparation of sample diluent

[0073] The preparation method of the sample diluent is as follows: add 1g bovine serum albumin (BSA) to 100ml of hydroxyethylpiperazine ethylsulfuric acid (HEPES) buffer solution, add an appropriate amount (final concentration is 0.02%) thimerosal, add 0.1% Tween-20.

[0074] 3. Coating buffer is used to coat anti-Helicobacter pylori antibody to microwell plate in ELISA test

[0075] 0.05mol / L carbonate buffer solution (pH9....

Embodiment 2

[0100] Embodiment 2 dot-ELISA test

[0101] (1), the solution solution preparation required for this test

[0102] 1. The preparation of the alkaline buffer solution is the same as before, and the borax-calcium chloride buffer solution (pH8.0) is used as an illustration.

[0103] Borax-Calcium Chloride Buffer (pH8.0)

[0104] Take 0.572 g of borax and 2.94 g of calcium chloride, add about 800 ml of water to dissolve, adjust the pH value to 8.0 with about 2.5 ml of 1mol / L hydrochloric acid solution, add water to dilute to 1000 ml, and obtain the product.

[0105] 2. Preparation of sample diluent

[0106] The preparation method of sample diluent is as follows: add 1g bovine serum albumin (BSA) in 100ml of borax-calcium chloride buffer (pH8.0), add appropriate (final concentration is 0.01%) thimerosal, add 0.1% Tween-20.

[0107] 3. Coating buffer

[0108] 0.05mol / L carbonate buffer solution (pH9.6): 0.75g sodium carbonate, 1.46g sodium bicarbonate, add deionized water to 50...

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Abstract

An immunoassay method for Helicobacter pylori (H. pylori) in a gastric mucosa sample comprises (a) suspending the gastric mucosa sample doubted about containing the H. pylori into sample diluent; (b) enabling the gastric mucosa sample in the diluent to be contacted with a first polyclonal antibody of an antigen resisting to the H. pylori to form a compound of the antibody and the antigen; (c) separating the sample and the compound; (d) enabling the compound to be exposed on a second polyclonal antibody of the antigen, enabling one part of the antibody to react with the compound, combining one of the first polyclonal antibody and the second polyclonal antibody onto a solid carrier, and marking the other one of the first polyclonal antibody and the second polyclonal antibody through a detection agent; and (e) assaying volume of the marked antibody and sequentially assaying existence of H. pylori antibodies in the gastric mucosa sample. The immunoassay method for the H. pylori in the gastric mucosa sample has the advantages of being low in cost, visual and accurate in detection result and fast in reaction speed.

Description

technical field [0001] The invention relates to an immunoassay method for Helicobacter pylori in a gastric mucosa sample. Background technique [0002] Helicobacter pylori (H. pylori), a bacterium found in the upper gastrointestinal tract of humans, has been found to be associated with gastroduodenal diseases such as peptic ulcer, gastritis and other diseases. The bacterium was classified by origin into the genus Campylobacter and then on the basis of more detailed information on its ultrastructure and fatty acid composition in the genus Helicobacter. [0003] There are many detection methods for Helicobacter pylori, commonly used methods are: (1) Bacterial culture method. This method is the "gold standard" for the diagnosis of H. pylori. The method can directly prove the existence of H. pylori without false positive, and can also be used for drug sensitivity test. Genetic factor studies on different strains. However, the technical conditions for isolation and culture ar...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 王滔
Owner 王滔
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