Method for detecting inhibition for bacteria by antibiotics through single-cell counting

A technology of bacteria inhibition and antibiotics, applied in the field of biomedicine, can solve problems such as complicated operation, expensive equipment, and difficult judgment of practical prospects

Active Publication Date: 2021-01-05
深圳艾尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these technologies are only in the research stage, only small sample analysis is required, and all of them require professional technicians to operate, expensive equipment, and non-traditional special equipment, complicated operation
Unstable performance, high cost, inconvenient to use, difficult to judge practical prospects

Method used

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  • Method for detecting inhibition for bacteria by antibiotics through single-cell counting
  • Method for detecting inhibition for bacteria by antibiotics through single-cell counting
  • Method for detecting inhibition for bacteria by antibiotics through single-cell counting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Objective: To search for the sensitive indicators of bacterial changes during bacterial culture in broth, to determine the theoretical basis for rapid drug susceptibility testing, and to establish a detection method for bacterial drug susceptibility.

[0041] Materials and methods:

[0042] 1. Strain preparation:

[0043]Three standard bacterial strains (ATOC 25922 Escherichia coli, ATOC 25923 Staphylococcus aureus, ATOC 27853 Pseudomonas aeruginosa) were transferred for use, and incubated at 37° C. for 18 hours.

[0044] 2. Broth preparation:

[0045] Prepare bacterial strain and grind on the bottle wall of AST (antibiotic susceptibility test) broth, mix well, cover the bottle cap and measure turbidity with a turbidimeter (BD company Phoeni xSpec Nephel omter), the turbidity is 0.5 McFarland units, spare. Inoculate the bacteria to be tested according to the inoculum concentration of the CLSI (American Clinical Laboratory Standards Organization) broth dilution method...

Embodiment 2

[0060] Ampicillin inhibition detection method for ATOC 25922 Escherichia coli (using resistance counting method):

[0061] Materials and methods:

[0062] 1. Strain preparation:

[0063] The standard strain ATOC 25922 Escherichia coli was transferred for use and incubated at 37°C for 18 hours.

[0064] 2. Broth preparation:

[0065] 10 of the drug susceptibility test tubes (or cups) contain the required antibiotics at the double dilution concentration (different drug concentrations refer to the US CLSI standard); the eleventh tube does not contain antibiotics, as a positive control (PC); there is no bacterial suspension The twelfth tube was used as a negative control (NC).

[0066] The prepared bacterial strains are ground on the bottle wall of MH (antibiotic susceptibility) broth, mixed evenly, and the bottle cap is closed and the turbidity is measured with a turbidimeter (BD company Phoeni xSpec Nephel omter), the turbidity is 0.5 McFarland units, and the .

[0067] Ino...

Embodiment 3

[0084] 1. Growth experiment

[0085] 1.1 Purpose of the experiment

[0086] According to the requirements of CLSI standard, inoculate the six common clinical strains (ATOC29212, ATOC29213, ATOC27853, ATOC25922, Klebsiella pneumoniae ATOC700603 and Acinetobacter baumannii) and record their growth trend to determine whether there is a possibility of their growth within 2 hours sex.

[0087] 1.2 Experimental method

[0088] 1.2.1 Negative control: test the uninoculated medium, use a resistance bacteria counter to test, and record the number of particles;

[0089] 1.2.2 Strain preparation: According to the requirements of CLSI standards, pick 0.5 McFarland fresh strains within 24 hours, take 100ul and add it to 10ml of culture medium;

[0090] 1.2.3 Record the results at 0h / 0.5h / 1h / 1.5h respectively, remove the negative background and analyze the results.

[0091] 2.3 Experimental results see Figure 4 , It can be seen from the results of the growth experiment that the bacter...

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Abstract

The invention provides a method for detecting inhibition for bacteria by antibiotics through single-cell counting. The method comprises the following steps of: after antibiotics of preset concentration are added into bacteria to be detected, setting into a bacteria antibiotic mixture, and meanwhile, setting bacteria to be detected, which are not added with the above antibiotics, as a positive control; after first preset duration is formed with a moment when the above antibiotics are added, obtaining a current amount of the above bacteria of the above bacteria antibiotic mixture and a current amount of the above bacteria of the above positive control; and determining the minimum inhibition concentration for the above bacteria by the above antibiotics, and determining an inhibition degree ofthe above antibiotics for the above bacteria. On the basis of implementation of drug sensitivity through the single-cell counting, the technical problem that a link between a single-cell counting value and a drug sensitivity experiment result can not be accurately determined is solved, a result is reliable, and therefore, the method disclosed by the invention has the beneficial effects that clinical practicality is high, cost is low and automation is easily realized.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a single cell counting method for detecting the inhibition of antibiotics on bacteria. Background technique [0002] The problem of bacterial drug resistance is becoming more and more serious, and it is rapidly spreading worldwide. Governments of all countries attach great importance to it. my country has also issued a large number of management regulations. Rational use of antibiotics is the core work to deal with bacterial resistance. Rapid antibiotic sensitivity test is the top priority. [0003] In view of the development of mass spectrometry technology and nucleic acid technology, the rapid identification of bacteria has basically been realized (the results can be obtained within 1-2 hours on the same day), so it is more urgent and practical to develop new rapid antibiotic sensitivity tests. [0004] Switching from empiric broad-spectrum antibiotic therapy to targeted therapy as s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18C12Q1/14C12Q1/10C12Q1/06C12R1/19C12R1/445C12R1/385C12R1/22C12R1/01
CPCC12Q1/18C12Q1/14C12Q1/10C12Q1/06G01N2333/21G01N2333/245G01N2333/31G01N2333/26G01N2333/212G01N2333/315Y02A50/30
Inventor 崔璟唐明忠张会翠
Owner 深圳艾尔生物科技有限公司
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