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A Rapid Antibiotic Susceptibility Testing Method Combining Deuterium Water Culture and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

A technology of matrix-assisted laser and time-of-flight mass spectrometry, which is applied in the field of medical testing, can solve problems such as time-consuming, and achieve strong operability and practicability

Active Publication Date: 2022-02-15
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is exactly to provide a kind of combination deuterium water culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis (D 2 O-MALDI) rapid antibiotic susceptibility test (AST) method, overcome the time-consuming deficiency of the existing clinical AST method and further expand the clinical application of MALDI-TOF MS, the present invention realizes the minimum inhibitory concentration (MIC) under the presence of different antibiotics ) and the detection of bacterial metabolic activity, the operation is simple, efficient, low cost, and the required consumables are easy to obtain

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  • A Rapid Antibiotic Susceptibility Testing Method Combining Deuterium Water Culture and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry
  • A Rapid Antibiotic Susceptibility Testing Method Combining Deuterium Water Culture and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry
  • A Rapid Antibiotic Susceptibility Testing Method Combining Deuterium Water Culture and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

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Embodiment 1

[0038] This embodiment provides a method for rapidly determining the sensitivity of Escherichia coli to chloramphenicol in combination with deuterium water culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, comprising the following steps:

[0039] Preparation of deuterium-containing water medium: According to the instructions of the purchased broth medium, weigh a certain amount of medium dry powder, add it to a certain volume of deuterium water, and perform high-pressure sterilization.

[0040] Preparation of chloramphenicol stock solution: follow the EUCAST standard test method.

[0041] Preparation of working solution: follow the EUCAST standard test method, and the diluent is broth medium prepared with deuterium water.

[0042] Aspirate the culture solution of Escherichia coli (ATCC 25922), determine its concentration by ultraviolet spectrophotometer, add it into the working solution after dilution. The operation is as follows:

[0...

Embodiment 2

[0056] This embodiment provides a method for rapidly determining the sensitivity of Escherichia coli to gentamicin in combination with deuterium water culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, comprising the following steps:

[0057] Preparation of deuterium-containing water medium: According to the instructions of the purchased broth medium, weigh a certain amount of medium dry powder, add it to a certain volume of deuterium water, and perform high-pressure sterilization.

[0058] Preparation of gentamicin stock solution: follow the EUCAST standard test method.

[0059] Preparation of working solution: follow the EUCAST standard test method, and the diluent is broth medium prepared with deuterium water.

[0060] The culture solution of Escherichia coli (CICC 10663) was absorbed, its concentration was determined by an ultraviolet spectrophotometer, and after dilution, it was added to the working solution. The operation is as fol...

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Abstract

The invention relates to a rapid antibiotic susceptibility testing method combining deuterium water culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis. Bacteria are incubated in a liquid medium containing deuterium water, and the bacteria incorporate deuterium into newly synthesized antibiotics through their own metabolism. Among the proteins, the bacterial fingerprints after incubation were characterized using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and the peak intensity (I D ) and the peak intensity (I ND ), I D / I ND Reflects the activity of bacterial metabolism; in the presence of antibiotics, deuterium incorporation will be inhibited to a certain extent, so use I D / I ND value to characterize the antibiotic susceptibility of bacteria. Compared with the prior art, the method of the present invention can perform AST on bacteria within 2 hours, and has strong operability and practicability; and the method is applicable to different bacteria, and is a general fast AST method.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a combination of deuterium water culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (D 2 O-MALDI) rapid antibiotic susceptibility testing (AST) method. Background technique [0002] Bacterial infection is one of the diseases with high morbidity and mortality in the world today, which poses a huge threat to global public health. The latest research shows that the absolute number of infections caused by drug-resistant microorganisms is increasing globally. It is reported that 700,000 people die from drug-resistant bacterial infections every year in the world. If effective measures are not implemented to reduce the spread of drug resistance and to invent new antibiotics, this number will reach 10 million in 2050. [0003] The development of rapid and accurate antibiotic susceptibility testing (AST) methods is the key to address the increasing ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/64C12Q1/18C12R1/19C12R1/01
CPCG01N27/64C12Q1/18G01N2333/25G01N2333/245
Inventor 乔亮易佳
Owner FUDAN UNIV