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Immunological microball method for detecting pyloric helicobacterium in stool sample

A Helicobacter pylori and anti-Helicobacter pylori technology, which is applied in the directions of measuring devices, biological tests, material inspection products, etc., can solve the problems of high cost of reagents and instruments, time-consuming operation, etc., and achieves reliable results, low cost, and convenient and fast operation. Effect

Inactive Publication Date: 2002-11-06
王滔
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The purpose of the present invention is to solve the problems of high cost and time-consuming operation of reagents and instruments used in the prior art

Method used

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  • Immunological microball method for detecting pyloric helicobacterium in stool sample

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Experimental program
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Effect test

Embodiment 1

[0044] Example 1: Physical method sensitization of microspheres

[0045] Material preparation: polystyrene microspheres with a diameter of 0.6-1.2 microns (available from SIGMA, USA), goat polyclonal antibody against Helicobacter pylori, Helicobacter pylori antigen (antigen standard or soluble antigen) (available from KPL, USA) Company, Shenzhen Jingmei Biological Engineering Co., Ltd., China).

[0046] Buffer preparation:

[0047] 0.27mol / L glycine buffer solution: 7.51g glycine and 9.95g sodium chloride were dissolved in distilled water, adjusted to pH 8.2 with 1mol / L sodium hydroxide, and made up to 1 liter with distilled water.

[0048] 0.27mol / L glycine buffer + 1% human serum albumin: 1 g of human serum albumin is dissolved in 100 ml of 0.27mol / L glycine buffer.

[0049] 0.054mol / L glycine buffer solution: Dilute 0.27mol / L glycine buffer solution 1:5 with distilled water.

[0050] First dilute 1 milliliter of 10% latex suspension to 1-2% with distilled water, wash and...

Embodiment 2

[0051] Embodiment 2: chemical cross-linking method sensitization microspheres

[0052] Polyaldehyde-based polystyrene microspheres with aldehyde groups Add 0.1 mg of antibody or antigen to 1 ml of 1% latex, add 10 ml of 0.1 mol / L pH5.3 acetate buffer. The vessel was allowed to stir for 10 hours at room temperature, then centrifuged at 10,000g for 20 minutes and the supernatant removed. Add 10 ml of pH9.5 1mol / L ethanolamine / 0.1% Tween-20 to the precipitate. Stir for 3 hours, then centrifuge at 10,000 g for 20 minutes, and remove the supernatant. Add 10 ml of Tris buffer to the precipitate, stir for 24 hours, centrifuge at 10,000 g for 20 minutes, and remove the supernatant. Add 10 ml of phosphate buffer solution containing 1% human serum albumin, resuspend; add 0.04% sodium azide, store at 4°C, and set aside.

[0053] 0.1mol / L pH5.3 acetate buffer solution: 10ml of 0.1mol / L acetic acid, 40ml of 0.1mol / L sodium acetate.

[0054] Tris buffer: 0.05mol / LTris 0.1mol / sodium chlo...

Embodiment 3

[0055] Example 3: Fecal sample preparation

[0056] Dilute the stool sample 1:4 or 1:6, add the appropriate stool sample into the sample diluent, and mix thoroughly. Centrifuge at 200 rpm for 3 minutes. Take the supernatant for testing. When using agglutination reaction and agglutination inhibition reaction to detect Helicobacter pylori, in order to improve the detection sensitivity, the sample can be concentrated, the supernatant is centrifuged at 12000g for 15 minutes, the supernatant is removed, and the precipitate is dispersed and mixed. to be tested.

[0057] Sample diluent: 100 ml of 0.27mol / L glycine buffer, add 1 g of human serum albumin, add 3 ml of Tween-20. At the same time, add 0.04% sodium azide.

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Abstract

The immunological microsphere method of detecting pyloric helicobacterium in stool sample includes labeling microsphere to form immunological microsphere; suspending the tested stool sample containing pyloric helicobacterium in buffering liquid; mixing the immunological microsphere and the sample buffering liquid or mixing pyloric helicobacterium resisting antibody with the buffering liquid before adding pyloric helicobacterium resisting antigen labeled microsphere; and detecting pyloric helicobacterium in the stool sample. The method has the advantages of low cost, fast and convenient operation and reliable result.

Description

(1) Technical field: [0001] The invention relates to an immune microsphere method for detecting Helicobacter pylori in stool samples. (two) background technology: [0002] Helicobacter pylori belongs to the genus Helicobacter. It is a bacterium found in the human gastrointestinal tract, and it has been found to be closely related to human gastroduodenal diseases such as gastritis, peptic ulcer, gastric cancer and other diseases. [0003] A number of different techniques have been used for the detection of H. pylori. According to whether or not the samples are invasive, the current clinical diagnostic methods for Helicobacter pylori infection can be divided into two categories: ① Non-invasive examination: including serological methods, breath test of isotope-labeled urea, and gastric juice polymerase chain reaction. ②Invasive examination: all are gastroscopy-dependent methods, including a variety of morphological, microbiological and molecular biological techniques. [000...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/569G01N33/574G01N33/577
Inventor 王滔
Owner 王滔
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