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Kit and method for detecting helicobacter pylori

A technology of Helicobacter pylori and detection kit, applied in the field of medical detection, can solve the problems of high price, complicated preparation method, long time consumption, etc., and achieve the effects of simple operation, convenient and fast production, and cost reduction.

Inactive Publication Date: 2018-07-20
李梅秀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Existing detection kits for Helicobacter pylori have problems such as complicated preparation methods, long time-consuming, and expensive

Method used

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  • Kit and method for detecting helicobacter pylori
  • Kit and method for detecting helicobacter pylori
  • Kit and method for detecting helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Human peripheral blood (PBMC) samples were obtained from patients with Helicobacter pylori infectious gastritis and non-H.

[0059] 10mL of blood was collected from each sampled individual with a heparin sodium collection tube, and transported to the laboratory within 6 hours at room temperature.

[0060] The detection steps of each sample to be tested are as follows:

[0061] (1) Separate the PBMC with the separation medium, resuspend the separated cells in serum-free medium, and the cell concentration is 1.5×10 5 Cells / mL; Add the resuspended cells into a PVDF96-well plate, add 100 μL to each reaction well, add each sample to 10 wells, and make 10 repetitions;

[0062] (2) Add 50 μL of a mixture consisting of serum-free medium and 7 polypeptides with a final concentration of 5 μM to each reaction well, mix well and carry out co-incubation; set up a positive control and a negative control, and add high Specific mouse anti-human IgG antibody for antigen-antibody react...

Embodiment 2

[0071] Human peripheral blood (PBMC) samples were obtained from patients with Helicobacter pylori infectious gastritis and non-H.

[0072] 10mL of blood was collected from each sampled individual with a heparin sodium collection tube, and transported to the laboratory within 6 hours at room temperature.

[0073] The detection steps of each sample to be tested are as follows:

[0074] (1) Separate the PBMC with the separation medium, resuspend the separated cells in serum-free medium, and the cell concentration is 3.5×10 5 Cells / mL; Add the resuspended cells into a PVDF96-well plate, add 100 μL to each reaction well, add each sample to 10 wells, and make 10 repetitions;

[0075] (2) Add 50 μL of a mixture consisting of serum-free medium and 7 polypeptides with a final concentration of 5 μM to each reaction well, mix well and carry out co-incubation; set up a positive control and a negative control, and add high Specific mouse anti-human IgG antibody for antigen-antibody react...

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Abstract

The invention provides a kit and a method for detecting helicobacter pylori. The kit comprises seven polypeptides with amino acid sequences shown in SEQ ID No. 1 to SEQ ID No. 7. The detection methodcomprises the following steps: (1) separating a sample to be detected by using a separating medium, and resuspending the separated cells in a serum-free culture medium; (2) adding the seven polypeptides with the amino acid sequences shown in SEQ ID No. 1 to SEQ ID No. 7 into the resuspended cells for co-incubation, and then adding a highly specific murine anti-human IgG antibody into a co-incubation system for an antigen-antibody reaction; (3) washing the system after the antigen-antibody reaction, adding a color developing substrate for a color developing reaction, and counting spots generated in the reaction. The kit provided by the invention can be synthesized manually in complete sequence, is convenient and fast to produce, has high stability and specificity, is simple and convenient to operate, and greatly reduces the cost.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a detection kit for Helicobacter pylori and a detection method thereof. Background technique [0002] Helicobacter pylori (Hp) is a microaerophilic Helicobacter, which mainly enters the digestive tract through the oral cavity to infect the body. The infection caused by Helicobacter pylori mainly causes gastric and duodenal ulcers and gastric cancer. An important cause, in 1994, the World Health Organization listed Helicobacter pylori as the first type of high-risk malignant transformation factor. It is a high-incidence area of ​​Hp, and the average infection rate is as high as 40%-80%, so the detection of Helicobacter pylori has become a clinical need. [0003] Now there are various diagnostic methods for Hp infection, involving fields such as pathology, bacteriology, serology, and isotope tracer. However, the routine detection methods in the diagnosis of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/005
Inventor 李梅秀
Owner 李梅秀
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