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A method for rapid detection of bacterial activity based on laser desorption ionization mass spectrometry

A technology of ionization mass spectrometry and bacterial activity, which is applied in the field of rapid detection of bacterial activity based on laser desorption ionization mass spectrometry technology, which can solve the problems of changes in bacterial metabolic activity, insufficient sensitivity, and susceptibility to interference, and achieves time-saving and high-throughput bacteria. The effect of activity detection

Active Publication Date: 2021-09-24
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still some defects in the traditional colorimetric method. First, relying on ultraviolet absorption or fluorescence detection, the sensitivity is insufficient and susceptible to interference. Second, the metabolic activity of bacteria may change during the long-term incubation with the indicator.

Method used

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  • A method for rapid detection of bacterial activity based on laser desorption ionization mass spectrometry
  • A method for rapid detection of bacterial activity based on laser desorption ionization mass spectrometry
  • A method for rapid detection of bacterial activity based on laser desorption ionization mass spectrometry

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Embodiment 1～4

[0032] The following examples provide a method for rapidly detecting bacterial activity based on a laser desorption ionization mass spectrometry platform, comprising the following steps:

[0033] Prepare resazurin with sterile water to a concentration of 0.05 mM solution, add 1 microliter of the solution to each sample point on the target plate with a pipette gun, and let it dry naturally.

[0034] Ceftriaxone sodium was prepared into a 10 mg / mL solution with sterile water, aliquoted, and stored at -20°C until use.

[0035] Take 3mL of bacterial liquid, centrifuge at 7000g for 3 minutes, and discard the supernatant. Resuspend in 1 mL sterile water, centrifuge at 7000 g for 3 minutes, and wash twice. Resuspended in culture medium.

[0036] Prepare 6 centrifuge tubes, add 50 μL of ceftriaxone sodium to each centrifuge tube, add 450 μL of the above bacterial solution, incubate in a shaker at 37°C and 170 rpm, and start timing.

[0037] At 0, 1, 2, 4, 6, and 8 hours, take a cen...

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Abstract

The invention relates to a method for rapidly detecting bacterial activity based on laser desorption ionization mass spectrometry technology, comprising: S1: dropping a bacterial activity indicator solution on a MALDI target plate; S2: configuring a bacterial suspension to be tested; Bacterial suspension is added to the target containing the indicator; S4: The sample point is analyzed by MALDI-TOF MS, and the mass spectrum of the peak intensity change of the bacterial indicator and its reduction product is obtained; S5: The indicator peak in the mass spectrum is compared with its peak intensity. The peak intensity ratio of the reduction product peak is calculated, and the change degree of bacterial activity is reflected by the change in the degree of reduction of the indicator. Compared with the prior art, the present invention utilizes laser desorption ionization time-of-flight mass spectrometry technology as a detection means, and realizes the analysis and detection of bacterial activity by detecting the change of molecular weight before and after the indicator is reduced by live bacteria, and calculating the ratio of its peak intensities. The detection method can rapidly, accurately and efficiently detect the bacterial activity in complex samples.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a method for rapidly detecting bacterial activity based on laser analysis ionization mass spectrometry technology. Background technique [0002] Bacterial viability detection is crucial in applications in various fields such as environmental monitoring, disease diagnosis, food safety, and biological research. Among the bacterial viability detection methods, the most widely used method is the plate count method. However, the method based on plate culture often takes 24-48 hours or even longer, which is very time-consuming and laborious, and is not suitable for large-scale sample testing. [0003] At present, some non-culture detection methods are more and more widely used in the detection of bacteria in various fields. Various colorimetric and fluorescence-based microbial activity assays have been reported, among which resazurin salts and tetrazolium salts are widely used as ind...

Claims

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Application Information

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IPC IPC(8): G01N27/64
CPCG01N27/64
Inventor 乔亮吴恩慧
Owner FUDAN UNIV
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