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Helicobacter pylori drug sensitivity detection method

A Helicobacter pylori and detection method technology, applied in the field of Helicobacter pylori drug sensitivity detection, can solve the problems of antibiotics that cannot include Helicobacter pylori, low reliability of quantification and drug sensitivity, long time of drug sensitivity detection, etc., and achieve consensus The effect of high sensitivity, improving the efficiency of drug sensitivity detection, and shortening the incubation time

Pending Publication Date: 2022-04-08
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular resistance testing currently only detects two antimicrobials, clarithromycin and levofloxacin, and does not include other antibiotics used to treat Helicobacter pylori
[0004] CN103757088A discloses quantitative determination of viable Helicobacter pylori bacteria, a drug susceptibility assay kit and a determination method. In this invention, feces or blood or secretions suspected of containing live Helicobacter pylori are used, and the cultured bacteria cannot be identified as Helicobacter pylori. The urease reaction is not a unique reaction of Helicobacter pylori, it may be other bacteria, the false positive rate is high, the reliability of quantification and drug sensitivity based on this is low, and the drug sensitivity test takes a long time
In addition, the traditional Helicobacter pylori drug susceptibility test needs to prepare a large number of culture plates, and the operators need to be professionally trained, and need to be cultured for 72-96 hours to obtain the drug susceptibility results

Method used

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  • Helicobacter pylori drug sensitivity detection method
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  • Helicobacter pylori drug sensitivity detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method for detecting drug susceptibility of Helicobacter pylori, comprising the steps of:

[0037] (1) Preparation of antibiotic-containing liquid medium: Dilute the antibiotic stock solution to 5120ug / mL, 2560ug / mL, 1280ug / mL, 640ug / mL, 320ug / mL, 160ug / mL, 80ug / mL, 40ug / mL , 20ug / mL, 10ug / mL, 5ug / mL, 2.5ug / mL, 1.25ug / mL, take 1mL of antibiotics of each diluted concentration, add them to 15mL BD tubes, and add 9mL Brucella to each BD tube Culture medium, containing 10% fetal bovine serum in the Brucella culture medium, mixes homogeneously, obtains containing antibiotic liquid medium:

[0038] (2) Helicobacter pylori susceptibility plate preparation: take 250 μ L of the antibiotic-containing liquid medium and add it to 48 wells of the plate, and set a negative control group and a positive control group at the same time, wherein, the control group is without Helicobacter pylori and without 500 μL of medium containing antibiotic liquid, and the positive control group is...

Embodiment 2

[0051] A method for detecting drug susceptibility of Helicobacter pylori, comprising the steps of:

[0052] (1) Preparation of antibiotic-containing liquid medium: Dilute the antibiotic stock solution to 5120ug / mL, 2560ug / mL, 1280ug / mL, 640ug / mL, 320ug / mL, 160ug / mL, 80ug / mL, 40ug / mL , 20ug / mL, 10ug / mL, 5ug / mL, 2.5ug / mL, 1.25ug / mL, take 1mL of antibiotics of each diluted concentration, add them to 15mL BD tubes, and add 9mL Brucella to each BD tube Culture medium, containing 8% fetal calf serum in the Brucelle culture medium, mix homogeneously, obtain containing antibiotic liquid medium:

[0053] (2) Helicobacter pylori susceptibility plate preparation: take 250 μ L of the antibiotic-containing liquid medium and add it to 48 wells of the plate, and set a negative control group and a positive control group at the same time, wherein, the control group is without Helicobacter pylori and without A culture medium containing antibiotic liquid, the positive control group is a culture...

Embodiment 3

[0058] A method for detecting drug susceptibility of Helicobacter pylori, comprising the steps of:

[0059] (1) Preparation of antibiotic-containing liquid medium: Dilute the antibiotic stock solution to 5120ug / mL, 2560ug / mL, 1280ug / mL, 640ug / mL, 320ug / mL, 160ug / mL, 80ug / mL, 40ug / mL , 20ug / mL, 10ug / mL, 5ug / mL, 2.5ug / mL, 1.25ug / mL, take 1mL of antibiotics of each diluted concentration, add them to 15mL BD tubes, and add 9mL Brucella to each BD tube Culture medium, containing 12% fetal calf serum in the Brucelle culture medium, mixes homogeneously, obtains containing antibiotic liquid medium:

[0060] (2) Helicobacter pylori susceptibility plate preparation: take 250 μ L of the antibiotic-containing liquid medium and add it to 48 wells of the plate, and set a negative control group and a positive control group at the same time, wherein, the control group is without Helicobacter pylori and without A culture medium containing antibiotic liquid, the positive control group is a cul...

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Abstract

The invention relates to a helicobacter pylori drug sensitivity detection method, which adopts a trace bouillon dilution method, can detect the MIC value of a clinical isolated strain to a plurality of antibacterial drugs at one time by only needing two 48 pore plates, greatly improves the drug sensitivity detection efficiency, has high consistency between the drug sensitivity result and a gold standard, namely an agar dilution method, is simple and convenient to operate, and can be used for detecting the drug sensitivity of helicobacter pylori. Semi-automation of drug sensitivity result detection is realized; the method is simple to operate and low in cost, compared with an agar dilution method, a large number of plates do not need to be configured, antibiotic liquid diluted in a gradient mode and a culture medium can be added through a workstation, the cost is lower than that of an E-test strip, the drug sensitivity result can be detected through a microplate reader, staining is conducted through a staining solution, and the result is visual and visible.

Description

technical field [0001] The invention belongs to the technical field of drug sensitivity detection, and in particular relates to a drug sensitivity detection method of Helicobacter pylori. Background technique [0002] Helicobacter pylori is a Gram-negative bacterium. Helicobacter pylori infection can cause gastritis, duodenitis and peptic ulcer, and is associated with gastric cancer and gastric mucosa-associated lymphoid tissue (MALT) lymphoma is closely related to the occurrence of HIV, infecting more than half of the world's population. However, due to the non-standard use of antibacterial drugs, the resistance rate of Helicobacter pylori to commonly used antibacterial drugs has increased year by year, resulting in a significant increase in the drug resistance rate of Helicobacter pylori and a significant decline in the eradication rate. The drug susceptibility test can determine the susceptibility of bacteria to antibiotics, and is currently recognized as a method for ba...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/18C12R1/01
CPCY02A50/30
Inventor 李红唐红申亚琳宋小娜唐小琼
Owner WEST CHINA HOSPITAL SICHUAN UNIV