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Microorganism identification and typing method in combination with mass spectrums and FTIR spectrums based on matrix-assisted laser desorption ionization flight time

A technology of time-of-flight mass spectrometry and matrix-assisted laser, applied in the field of microbial detection and analysis, can solve problems such as water interference

Inactive Publication Date: 2020-01-14
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of collecting microbial infrared signals, they are often severely interfered by water

Method used

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  • Microorganism identification and typing method in combination with mass spectrums and FTIR spectrums based on matrix-assisted laser desorption ionization flight time
  • Microorganism identification and typing method in combination with mass spectrums and FTIR spectrums based on matrix-assisted laser desorption ionization flight time
  • Microorganism identification and typing method in combination with mass spectrums and FTIR spectrums based on matrix-assisted laser desorption ionization flight time

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Typing of Escherichia coli and Shigella (cultivation on tryptone soybean agar (TSA) solid medium)

[0036] 1. Collect microbial characteristic mass spectrum and infrared absorption spectrum respectively

[0037] 1) The strains frozen at -80°C were solid-cultured with TSA in a 37°C incubator for 14 hours for the subsequent collection of mass spectrometry and infrared spectrometry data. This bacterial culture method has no specificity, and only requires that it is suitable for bacterial growth and that the consistency of culture conditions should be ensured as much as possible throughout the identification process, so as to reduce the interference of unknown factors caused by different culture methods.

[0038] 2) For mass spectrometry, use an inoculation loop to pick an appropriate amount of bacteria on the solid medium and resuspend them in ethanol solution, and then use formic acid-acetonitrile extraction method (for general bacteria, direct coating method can ...

Embodiment 2

[0046] Types of Escherichia coli and Shigella (Cultivated on Columbia blood agar solid medium) (cluster analysis results see Figure 4 ) Figure 4 The green line in is the threshold line, and the red dots are the correctly clustered clusters. in Figure 4 A is the result of cluster analysis using MALDI-TOF data alone, a total of 19 strains, of which 9 strains are correctly clustered, and the clustering accuracy rate is 9 / 19; Figure 4 B is the result of cluster analysis using FTIR data alone, a total of 19 strains, of which 16 strains are correctly clustered, and the clustering accuracy rate is 17 / 19; Figure 4 C is the result of cluster analysis by fusion of MALDI-TOF and FTIR data. There are 19 strains in total, and 19 strains are correctly clustered, and the clustering accuracy rate is 100%. It shows that MALDI-TOF alone is used for typing Escherichia coli and Shigella, and its correct rate is low; FTIR is used alone for typing Escherichia coli and Shigella, and its typi...

Embodiment 3

[0048] Typing of Escherichia coli and Shigella (cultivated in LB broth liquid medium) (for cluster analysis results, see Figure 5 ) Figure 5 The green line in is the threshold line, and the red dots are the correctly clustered clusters. in Figure 5 A is the result of cluster analysis using MALDI-TOF data alone, a total of 19 strains, of which 9 strains are correctly clustered, and the clustering accuracy rate is 9 / 19; Figure 4 B is the result of cluster analysis using FTIR data alone, a total of 19 strains, of which 15 strains are correctly clustered, and the clustering accuracy rate is 1519; Figure 4 C is the result of cluster analysis by fusion of MALDI-TOF and FTIR data. There are 19 strains in total, and 19 strains are correctly clustered, and the clustering accuracy rate is 100%. It shows that MALDI-TOF alone is used for typing Escherichia coli and Shigella, and its correct rate is low; FTIR is used alone for typing Escherichia coli and Shigella, and its typing ac...

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Abstract

The invention discloses a microorganism identification and typing method in combination with mass spectrums and FTIR spectrums based on matrix-assisted laser desorption ionization flight time. The method comprises the following specific steps: (1) respectively collecting microorganism characteristic mass spectrums and infrared absorption spectrums; (2) preprocessing the collected characteristic mass spectrums and infrared absorption spectrums; (3) performing data fusion on the data of the preprocessed characteristic mass spectrums and infrared absorption spectrums; and (4) carrying out clusteranalysis on the fused data of the mass spectrums and the infrared absorption spectrums to realize identification and typing of microorganisms. A data set used for microbial typing contains more characteristic biological information, and effective distinguishing of extremely similar strains difficult to distinguish by simple means, such as Escherichia coli and Shigella, can be realized.

Description

technical field [0001] The invention belongs to the technical field of microbial detection and analysis, and in particular relates to a microbial identification and typing method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry combined with FTIR spectroscopy. Background technique [0002] In the past five years, the microbial identification system of matrix-assisted laser desorption time-of-flight mass spectrometry has been widely used in clinical practice. The reason is that compared with traditional culture and biochemical identification mass spectrometry, it has a huge time and cost advantage. Provide guidance for doctors' clinical treatment. At present, domestic tertiary hospitals are basically equipped with a microbial identification system using mass-assisted laser analysis time-of-flight mass spectrometry. The identification of microorganisms by mass spectrometry is mainly based on the differences in microbial ribosomal proteins,...

Claims

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Application Information

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IPC IPC(8): G01N27/64
CPCG01N27/64
Inventor 余绍宁施海梅吴芳玲
Owner NINGBO UNIV
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