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A method for culturing Helicobacter pylori without a microaerobic device

A Helicobacter pylori, aerobic technology, applied in the field of cell culture, can solve problems such as difficulty in Hp culture, achieve the effect of simple operation and overcome the defects of culture

Active Publication Date: 2021-02-19
北京健为医学检验实验室有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the current difficult situation of Hp cultivation, the present invention provides an oily liquid mixture specially used for Hp cultivation

Method used

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  • A method for culturing Helicobacter pylori without a microaerobic device
  • A method for culturing Helicobacter pylori without a microaerobic device
  • A method for culturing Helicobacter pylori without a microaerobic device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Hp solid culture method

[0027] (1) Preparation of oily liquid mixture

[0028] Mix 42ml oleic acid, 38ml linoleic acid and 20ml arachidic acid to prepare 100ml oily liquid mixture;

[0029] (2) Preparation of Hp liquid medium and Hp solid medium

[0030] Liquid culture medium: Weigh 11.1g brain heart infusion culture medium and dissolve it in 270ml deionized water, after autoclaving and cooling down to room temperature, add 30ml fetal bovine serum, amphotericin B (final concentration 10 mg / L), trimethoprim (final concentration 5mg / L), vancomycin (final concentration 10 mg / L), cefsulodin (final concentration 10 mg / L);

[0031] Solid medium: Weigh 13.2 grams of Columbia blood agar basal medium and dissolve it in 240ml deionized water, after autoclaving, cool to about 55°C, add 30ml sterile defibrillated horse blood, 30ml fetal bovine serum and amphotericin B (final concentration 10 mg / L), trimethoprim (final concentration 5 mg / L), vancomycin (final con...

Embodiment 2

[0042] Embodiment 2: Hp liquid culture method

[0043] Draw 4ml of the Hp liquid medium prepared in step (2) of Example 1 and add it to a 15ml glass test tube, pick the Hp monoclonal grown on the solid medium of Example 1 and inoculate it in the liquid medium, and in the liquid medium Add 500 μl of the oily liquid mixture to the surface; incubate on a shaker (150 rpm) at 37°C for 7 days and observe the results; the results are identified using the method in step (5) of Example 1.

[0044] The results show that after inoculating Hp with liquid culture medium sealed with oily substances, shake culture for 7 days and use Hp colony PCR to identify, confirming that Hp liquid culture can reach the same culture effect as that in microaerophilic devices after being sealed with oily substances (see image 3 ).

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Abstract

The invention relates to an oily liquid mixture and a method for cultivating Helicobacter pylori using the mixture. The oily liquid mixture of the invention can not only isolate the air, provide a micro-anaerobic environment for the cultivation of the Helicobacter pylori, but also simulate the growth environment of the Helicobacter pylori in the body to promote its growth. The invention of the oily liquid mixture of the present invention relieves the special requirements for culturing Helicobacter pylori on the microaerophilic device, and the method is easy to operate, does not affect the growth of Helicobacter pylori, and is easy to observe the growth state of Helicobacter pylori at any time during the cultivation process .

Description

technical field [0001] The invention relates to an oily liquid mixture for cell culture and a cell culture method thereof, in particular to a method for cultivating Helicobacter pylori using the oily liquid mixture, and belongs to the technical field of cell culture. Background technique [0002] Helicobacter pylori (Hp) is a Gram-negative, spiral-shaped, micro-anaerobic bacteria that requires very harsh growth conditions. Since Australian scholars discovered Hp in 1982 and separated it from the gastric mucosa, the research on Hp has become the focus of gastroenterology research. The research center has gradually discovered that Hp infection can cause gastritis, peptic ulcer, gastric lymphoma and gastric cancer. The infection rate of Hp in the global population exceeds 50%, and the infection rate of Hp in my country ranges from 42% to 64%. Hp culture is a method for detecting Hp infection by culturing gastric or duodenal mucosal bacteria, with a sensitivity of 98% and a sp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 刘勇王春香
Owner 北京健为医学检验实验室有限公司