Quantitative determination of Helicobacter pylori live bacteria, drug susceptibility determination kit and determination method

A technology for quantitative determination of Helicobacter pylori, which is applied in the field of rapid screening methods for anti-Helicobacter pylori sensitive drugs and the kits thereof, can solve the problems of high pain and suffering for patients, can only be qualitative and expensive, and achieves cheap reagents and methods. Simple, cost-saving effect

Inactive Publication Date: 2016-01-20
SICHUAN VACCINE TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Based on the current detection situation, the present invention provides a quantitative detection method using live Helicobacter pylori, which uses live Helicobacter pylori as a detection sample, utilizes the characteristic that live Helicobacter pylori can produce urease, and combines the standard The curve can quickly and accurately read the number of Helicobacter pylori without destroying the Helicobacter pylori. Injury and pain, time-consuming and labor-intensive issues

Method used

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  • Quantitative determination of Helicobacter pylori live bacteria, drug susceptibility determination kit and determination method
  • Quantitative determination of Helicobacter pylori live bacteria, drug susceptibility determination kit and determination method
  • Quantitative determination of Helicobacter pylori live bacteria, drug susceptibility determination kit and determination method

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Embodiment 1

[0027] Quantitative determination method of viable bacteria of Helicobacter pylori

[0028] Add 8-10 parts of wt2% urea, 0.1-0.5 parts of wt0.04% phenol red, wt0.04% NaH 2 PO 4 ﹒ H 2 O3-5 parts, wt 0.1%Na 2 HPO 4 2-3 parts are configured as sterile urease reagent.

[0029] Draw the standard curve of the CFU / ml value corresponding to the OD value, that is, the number of individual bacteria in each ml of bacterial solution. The standard curve of the CFU corresponding to the OD value is determined by the gold standard of bacteriological detection (colony plate count), and the specific operation of the colony plate count is briefly described as follows:

[0030] Dilute a certain concentration of bacterial solution by a 10-fold gradient, take 100 microliters of the diluted bacterial solution and spread it on the medium, parallel 3 plates, count the number of colonies after 72 hours of cultivation, and dilute the gradient until there are only 1-5 colonies on the plate

[00...

Embodiment 2

[0043] Helicobacter pylori drug susceptibility testing method

[0044] Add 8-10 parts of wt2% urea, 0.1-0.5 parts of wt0.04% phenol red, wt0.04% NaH 2 PO 4 ﹒ H 2 O3-5 parts, wt0.1%Na 2 HPO 4 2-3 parts are configured as sterile urease reagent.

[0045] Draw the standard curve of the CFU / ml value corresponding to the OD value, that is, the number of individual bacteria in each ml of bacterial solution.

[0046] The standard curve of the CFU corresponding to the OD value is determined by the gold standard of bacteriological detection (colony plate count), and the specific operation of the colony plate count is briefly described as follows:

[0047] Dilute a certain concentration of bacterial solution by a 10-fold gradient, and take 100 microliters of the diluted bacterial solution and smear it on the culture plate.

[0048] On the base, parallel 3 plates, count the number of colonies after 72 hours of culture, and dilute the gradient until there are only 1-5 colonies on ...

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Abstract

The invention discloses quantitative determination of live Helicobacter pylori bacteria, a drug susceptibility determination kit and a determination method. The method uses live Helicobacter pylori as a sample to be tested, and utilizes the characteristic that the live Helicobacter pylori can produce urease. The standard method of Helicobacter pylori colony counting (CFU / ml, that is, the number of individual bacteria in each ml of bacterial liquid, the standard curve of the number of colonies) can quickly and accurately read the number of Helicobacter pylori, and the detection results are accurate, sensitive and reliable. It solves the difficult counting problem of the existing Helicobacter pylori clinical microbiological identification due to difficult cultivation and complicated operation, and the expanded drug susceptibility assay kit and preparation method of the present invention can carry out multiple drug susceptibility tests at the same time, and clinicians can quickly and conveniently screen Appropriate anti-Helicobacter pylori drugs can save time, labor and cost, and provide a beneficial technical solution for clinical scientific treatment.

Description

technical field [0001] The present invention relates to a measurement method of Helicobacter pylori and a preparation method thereof, in particular to a simple, fast and sensitive quantitative measurement method of Helicobacter pylori, a kit thereof and an improved anti-Helicobacter pylori sensitive drug Rapid screening method and its kit. Background technique [0002] 1983 Helicobacter pylori ( Helicobacter pylori Hp for short) was discovered by Warren and Warshail in Australia and has been identified as one of the most common pathogens of chronic infectious diseases. At present, the clinical detection of Hp is mainly divided into two categories: invasive and non-invasive. The former includes gastroscopy and serum antibody detection; the latter includes "C-UBT" detection and Helicobacter pylori antigen detection, but both are inconvenient to detect , The detection has some shortcomings such as trauma and detection hysteresis. [0003] There are many inspection methods fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/06C12Q1/18
Inventor 祝洁周永君张勇
Owner SICHUAN VACCINE TECH
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