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Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof

A technology for enhancing Helicobacter pylori and latex, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of easy pollution, slow detection speed, and less cultured bacteria, and achieves good detection accuracy and precision and improved detection speed. , the effect of a wide linear range

Active Publication Date: 2012-09-12
BEIJING MOKOBIO LIFE SCI CO LTD
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

This method is simple, rapid and sensitive, but it may also produce false positives due to the presence of other bacteria containing urease
In addition, samples with recent use of drugs that reduce gastric bacterial load or directly inhibit urease activity can produce false negatives
[0008] 2. Pathological detection method: by carrying out section staining inspection on gastric mucosa tissue, it can be confirmed by directly displaying HP thalline, wherein the detection rate of silver staining method is high, but the differences observed by different pathologists have certain significance for the diagnosis of gastric atrophy samples. certain difficulty
[0009] 3. The bacterial culture method takes gastric mucosal biopsy tissue for HP culture, but it is not suitable for general application due to less cultured bacteria, more complicated operation process, longer time period, and high cost
This method has high sensitivity and can be quantified, but there is a certain risk of radioactivity, and it is difficult to be widely used due to equipment limitations.
[0013] 2. PCR detection method: mainly detect HP urease (UReA) gene and toxin-related protein A (CagA) gene in gastric juice and mucous membrane, but the operation is complicated and easy to be polluted, and the professional knowledge and operation technology requirements of the inspectors are relatively high. And greatly increase the economic expenditure of patients, so the clinical application is relatively limited
[0014] 3. Immuno-serological method: At present, existing methods include enzyme-linked immunosorbent assay, western blotting and colloidal gold method, which provide a favorable and convenient means for the epidemiological investigation of HP, but the detection speed is slow, except enzyme-linked immunosorbent assay can be used Except for the automatic enzyme immunoassay analyzer (which takes 2-3 hours and has many steps), the rest need to be operated manually. The test results obtained by patients are qualitative analysis, which is not suitable for treatment monitoring and efficacy evaluation
[0015] At present, although there are many methods for detecting Helicobacter pylori infection, there is no suitable biochemical analyzer that can be easily, quickly, mass-produced and quantitatively detected

Method used

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  • Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof
  • Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof
  • Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Preparation of embodiment 1 Helicobacter pylori antigen (taking the whole bacterium protein antigen as an example)

[0073] (1) Bacterial culture

[0074] (a) Solid-state culture: inoculate HP strains in chocolate medium and culture at 37°C for 3-4 days to produce flat, slightly transparent and jelly-like colonies, which are Helicobacter pylori. After the bacteria were drawn on the chocolate medium with the three-section line method, they were cultured at 37°C for 3-4 days, and they were subcultured every four days to maintain the activity of the bacteria.

[0075] (b) Small amount of liquid culture: hang several colonies of Helicobacter pylori cultured in the back state with an inoculation loop, and shake them off in a 125ml Erlenmeyer flask, which contains 50ml of BHI culture solution, which contains 1ml Newborn calf serum and 0.1ml Helicobacter pylori screening solution, the mouth of the Erlenmeyer flask was tightly covered with a cotton plug, and cultured at 37°C f...

Embodiment 2

[0082] Embodiment 2 Assembly of the kit of the present invention

[0083] 1 buffer configuration

[0084] 1.1 Preparation of diluent (reagent R1)

[0085] Weigh the reagents in Table 1 below and put them into a clean container, add purified water and mix well, measure the pH value to 7.4, and set the volume to 1000ml.

[0086] Preparation of Table 1 Diluent (Reagent R1)

[0087] Reagent

Amount of liquid per liter

Disodium phosphate

2.88g

Sodium dihydrogen phosphate

2.4g

Sodium chloride

116.88g

PEG-6000

2g

PVP-k30

20g

BSA

10.0g

Tween-20

0.5ml

Proclin-300

0.5ml

Add purified water to

1000ml

[0088] 1.2 Preparation of standard diluent

[0089]Weigh the reagents in Table 2 below and put them into a clean container, add purified water to dissolve and mix well, measure the pH value to 7.4, and set the volume to 1000ml.

[0090] Table 2 Prepa...

Embodiment 3

[0129] Embodiment 3 utilizes kit of the present invention to the mensuration of Helicobacter pylori antibody

[0130] 1. According to the method of Example 5, the kit of the present invention was used to analyze 280 people without gastropathy (after the physical examination, it was confirmed that there were no digestive tract diseases, liver and kidney diseases, and no history of stomach pain) and 329 cases of gastropathy group (all were examined by gastroscopy). , Pathologically confirmed. Divided into 5 groups: duodenal ulcer group 75 cases; gastric ulcer group 55 cases; atrophic gastritis group 61 cases; gastric cancer group 113 cases; cardia cancer group 5 cases. The results are shown in Table 4:

[0131] Table 6 utilizes kit of the present invention to the mensuration of Helicobacter pylori antibody

[0132] group

Number of cases

HP (AU / ML)

control group

280

5.2±4.4

duodenal ulcer group

75

10.5±3.6

gastric ulce...

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Abstract

The invention discloses a latex-enhanced immunoturbidimetry kit for measuring a Helicobacter pylori antibody as well as a preparation method and application thereof. The kit comprises a diluent, latex reagent and blank liquid of Helicobacter pylori antigen, a standard product and a quality control product, wherein the Helicobacter pylori antigen can be one or more of full-tropina antigen of Helicobacter pylori, urease antigen of Helicobacter pylori, cytotoxin-related protein A antigen of Helicobacter pylori or cell vacuole toxin A antigen of Helicobacter pylori; the latex reagent is a mixture of two latex granules which have different particle sizes and are adsorbed by Helicobacter pylori; and the Helicobacter pylori antibody for preparing the standard product and the quality control product is derived from IgM (immunoglobulin M) and / or IgG (immunoglobulin G) in serum of a patient infected by Helicobacter pylori. The kit is suitable for various full-automatic biochemical analyzers and semiautomatic biochemical analyzers, and has the advantages of rapid detection, high sensitivity, strong specifically, good accuracy and the like.

Description

technical field [0001] The invention relates to a detection kit, in particular to a latex immune kit for detecting Helicobacter pylori antibody in human serum and a preparation method thereof. The invention belongs to the technical field of biological detection Background technique [0002] Helicobacter pylori (HP for short) is a unipolar, multi-flagellated, blunt-rounded end, spiral-curved bacterium, Gram-negative, and often presents a typical spiral or arc on the surface of gastric mucosal epithelial cells . [0003] Helicobacter pylori infection is the main pathogenic factor of chronic active gastritis, peptic ulcer and gastric moving membrane-associated lymphoid tissue (MALT) lymphoma, and is closely related to the occurrence of gastric cancer. 1994 World Health Organization / International Cancer Research The agency (WHO / IARC) defines Helicobacter pylori as a class I carcinogen. The detection rate of Helicobacter pylori in gastric mucosal biopsy specimens of patients w...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/574
Inventor 金鑫
Owner BEIJING MOKOBIO LIFE SCI CO LTD
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