Method for rapidly identifying strain-level pathogenic bacteria in food through double adsorption

A pathogenic bacteria and double-adsorption technology, which is applied in the field of detection and identification of pathogenic bacteria at the strain level in liquid food, can solve the problems of multiple detection steps and time-consuming and labor-intensive detection of pathogenic bacteria at the strain level, and overcome the time-consuming and labor-intensive problems. Effect

Inactive Publication Date: 2018-05-11
TIANJIN MODERN VOCATIONAL TECH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the problems of time-consuming and laborious and many detection steps in the identification of strain-level pathogens in the prior art, and provide a method for quickly identifying strain-level pathogens in food by using double adsorption

Method used

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  • Method for rapidly identifying strain-level pathogenic bacteria in food through double adsorption
  • Method for rapidly identifying strain-level pathogenic bacteria in food through double adsorption
  • Method for rapidly identifying strain-level pathogenic bacteria in food through double adsorption

Examples

Experimental program
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Effect test

Embodiment 1

[0037] At 37°C, Escherichia coli O157:H7 was inoculated in 25 mL of sterilized TSB medium, and the shaker was 120 r min -1 After culturing for 16 h, 1 mL of bacterial suspension was centrifuged (12000 r min -1 ) for 5 min, pour out the supernatant, and wash twice with PBS buffer solution, then, dilute Escherichia coli with different concentrations, and count them by counting viable bacteria. At the same time, MIL-101 magnetic particles (Fe 3 o 4 -COOH@MIL-101) can be synthesized independently or purchased from the market ( figure 2 is the transmission electron microscope image of MIL-101 magnetic particles).

[0038] Using carbonated beverages, add different concentrations (5 × 10 5 cfu mL -1 ) of Escherichia coli O157:H7 bacterial suspension, and then utilize Fe 3 o 4 - COOH@MIL-101 enrichment and microwave-assisted wall breaking, MALDI / TOF MS detection, and ribosomal protein database for strain level identification. image 3 Escherichia coli Fe in carbonated beverag...

Embodiment 2

[0040] Using carbonated beverages, add different concentrations (5 × 10 3 cfu mL -1 ) of Escherichia coli O157:H7 bacteria suspension, after Fe 3 o 4 - COOH@MIL-101 enrichment and microwave-assisted wall breaking, MALDI / TOF MS detection, and ribosomal protein database for strain level identification. There are 9 ribosomal proteins searched by Tagident (see Table 1), and Escherichia coli O157:H7 can be identified through the ribosomal protein database (Rapid Microorganism Identification Database; http: / / rmidb.org), Figure 5 Escherichia coli O157:H7 (bacterial concentration is 5×10 3 cfu mL -1 ) ribosomal protein database identification results.

[0041] Table 1 Escherichia coli O157:H7 protein ribosomal protein search results in Swiss-Prot / TrEMBL database a

[0042]

[0043] a The protein database search matching error is ±3Da.

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Abstract

The invention discloses a method for rapidly identifying strain-level pathogenic bacteria in food through double adsorption. The method comprises main steps as follows: a) strain-level pathogenic bacteria in liquid food are adsorbed by MIL-101 magnetic particles, and wall breaking is performed under microwave assistance; after wall breaking, to-be-identified pathogenic bacterium protein is rapidlyenriched through the MIL-101 magnetic particles; b) a mass spectrum graph of the pathogenic bacterium protein is collected through MALDI / TOF MS (matrix-assisted laser desorption ionization / time of flight mass spectrometry); c) a Tagident search tool is used for performing protein database searching on mass spectrum peak, and ribosomal protein is selected; d) a rapid microorganism identification database is sought by use of ribosomal protein obtained through searching, and the attributes of the strain-level pathogenic bacteria are determined. The method has the advantages of simplicity and rapidness. Enrichment of pathogenic bacteria, wall breaking and mycoprotein enrichment are integrated, finally, a ribosomal protein database is used for rapidly identifying the pathogenic bacteria, and the strain level of pathogenic bacteria in the liquid food is rapidly identified.

Description

technical field [0001] The invention belongs to the technical field of food detection, and relates to a method for rapidly enriching and identifying strain-level pathogenic bacteria in food by using MIL-101 magnetic particles, and is particularly suitable for the detection and identification of strain-level pathogenic bacteria in liquid food. Background technique [0002] Facing the current severe food safety problems, the rapid analysis and identification of pathogenic bacteria at the level of food-borne strains has become a multidisciplinary research hotspot. The ability to rapidly detect and identify pathogenic bacteria at the "strain" level is very important in food safety, biodefense, and clinical diagnostics. Rapid and accurate detection and identification can provide important information for pathogenic bacteria in terms of pathogenicity, prevalence, variability and drug resistance, and is of great significance for quickly proposing solutions, controlling the spread o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848
Inventor 魏纪平黄艳凤黄艳玲刘鑫龙张乐石琛孙波安娜刘宇豪陈珊汤卫华程秀玮
Owner TIANJIN MODERN VOCATIONAL TECH COLLEGE
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