MALDI-TOF MS (Matrix-assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry)-assisted identification method for listeria monocytogenes

A MALDI-TOF-MS, Listeria technology, applied in the field of detection of Listeria monocytogenes, to achieve the effect of simple sample preparation and accurate identification

Inactive Publication Date: 2011-11-23
曹际娟 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the lack of resources of the standard database for bacterial comparison is one of its limitations (Rapid typing of bacteria using matr

Method used

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  • MALDI-TOF MS (Matrix-assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry)-assisted identification method for listeria monocytogenes
  • MALDI-TOF MS (Matrix-assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry)-assisted identification method for listeria monocytogenes

Examples

Experimental program
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Effect test

Embodiment 1

[0053] (1) Select standard Listeria monocytogenes strains

[0054] 37 strains of Listeria monocytogenes collected and preserved by Liaoning Entry-Exit Inspection and Quarantine Bureau were selected, and the specific information is shown in Table 1. The Listeria monocytogenes strains involved in Table 1 were further confirmed to be Listeria monocytogenes by API-20E biochemical identification and automatic bacterial biochemical analyzer, and could be used to establish a MALDI-TOF MS identification standard map.

[0055] Table 1

[0056] serial number

Strain number

serial number

Strain number

serial number

Strain number

serial number

Strain number

1

DD-L20

11

DD-L37

21

SD-L-A5-6

31

GD-L16

2

DD-L21

12

DD-L44

22

SD-L-DAH1-11

32

GD-L17

3

DD-L25

13

DD-L51

23

SD-L-F1-1

33

GD-L18

4

DD-L26

14

DD-L52 ...

Embodiment 2

[0077] MALDI-TOF-MS Clustering Analysis of 37 Strains of Listeria monocytogenes

[0078] On the basis of Example 1, the similarity of the main spectra of 37 strains of Listeria monocytogenes in the obtained Listeria monocytogenes MALDI-TOF-MS database was calculated by pattern matching, and the similarity score was used to construct a phylogenetic tree. 37 strains of bacteria were analyzed by clustering and typing, and the obtained clustering and typing dendrogram is shown in the attached figure 2 shown. The level of difference represents the relatedness to each other with a value between 0 and 1.6. A difference of 0 means that the two MALDI-TOF-MS mass spectra are identical and belong to the same class; a difference of 1.6 means that the relationship between the two strains is the farthest. Depending on the level of difference selected, the number of Listeria monocytogenes that was typed varied. When the difference level was between 0.7 and 0.8, the MALDI-TOF-MS typing res...

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Abstract

This invention relates to an MALDI-TOF MS (Matrix-assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry)-assisted identification method for listeria monocytogenes, comprising the following steps of: (1) selecting 35 to 40 strains of fresh cultures of listeria monocytogenes, pre-treating the samples; (2) collecting maps of all bacterial strain samples; (3) analyzing and unifying the obtained maps by using BioTyper software to obtain a standard map of listeria monocytogenes; (4) preparing a sample of a microorganism to be detected by adopting the method in the step (1); (5) collecting the MALDI-TOF-MS map of the sample to be detected according to the method in the step (2); comparing the obtained MALDI-TOF-MS map of the sample to be detected in step (5) with the obtained standard map of listeria monocytogenes in the step (3), judging the detection results according to matching fractions. According to the invention, a mass spectrogram database and the standard map in listeria monocytogenes identification are created successfully, and the accurate identification with convenience, microscale and high automation for listeria monocytogenes can be realized.

Description

technical field [0001] The invention relates to a new detection method for Listeria monocytogenes, in particular to a method for assisting identification by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF MS). Background technique [0002] Listeria monocytogenes is a zoonotic pathogen. The bacteria can cause meningitis, encephalitis, sepsis, miscarriage, stillbirth and other diseases in pregnant women and animals. Pregnant women, newborns, the elderly and immunocompromised persons are susceptible groups. Although the incidence of foodborne Listeria is not high, the mortality rate can sometimes reach 20% to 50%. WHO listed it as one of the four major food pathogens in the 1990s. The fungus is widely distributed in nature, with livestock and poultry as the main host, and it is easy to contaminate food and cause food poisoning and listeria outbreaks [4] . Meat, eggs, poultry, seafood, dairy products, vegetables, etc. have all...

Claims

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Application Information

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IPC IPC(8): G01N27/64
Inventor 曹际娟曹冬梅
Owner 曹际娟
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