Salmonella source tracing typing method based on gMLST (genome multilocus sequence typing) technology and application

A Salmonella, typing technology, applied in biochemical equipment and methods, microbial measurement/testing, and resistance to vector-borne diseases, etc., can solve the problem that no patent publications have been found, and the ability of MLST to distinguish cannot meet the needs of identifying non-outbreak pneumonia Cree To solve problems such as Burger bacteria strains, to achieve the effect of strong innovation and high resolution ability

Active Publication Date: 2020-01-21
中国海关科学技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In 2017, Chinese Center for Disease Control and Prevention Haijian Zhou and other scholars found that the discrimination ability of MLST could not satisfy the identification of non-outbreak Klebsiella pneumoniae strains
[0012] Through the search, no patent publications related to the patent application of the present invention have been found

Method used

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  • Salmonella source tracing typing method based on gMLST (genome multilocus sequence typing) technology and application
  • Salmonella source tracing typing method based on gMLST (genome multilocus sequence typing) technology and application
  • Salmonella source tracing typing method based on gMLST (genome multilocus sequence typing) technology and application

Examples

Experimental program
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Effect test

Embodiment 1

[0056] A method for traceability typing of Salmonella based on gMLST technology, the steps are as follows:

[0057] (1) Isolation and identification of Salmonella:

[0058] According to the national standard 4789.4-2016, more than 40 meat samples from the same area were collected for the isolation and identification of Salmonella. The 2 reference strains were the standard strains, which were Salmonella Enteritidis ATCC9184 and Salmonella Typhi CMCC 50071;

[0059] ⑵ DNA extraction, content detection:

[0060] Extract the DNA of the Salmonella sample isolated from the meat sample, melt the extracted DNA on ice, pipette, mix and centrifuge for detection; use agarose gel electrophoresis to detect the integrity of the sample and use it to construct a DNA library;

[0061] (3) Whole genome sequencing and g MLST typing

[0062] ① Whole-genome sequencing: de novo sequencing of the bacterial genome, constructing a library with qualified samples, using ultrasonic method to randomly i...

Embodiment 2

[0086] A method for traceability typing of Salmonella based on gMLST technology, the steps are as follows: (1) Isolation and identification of Salmonella.

[0087] According to the national standard 4789.4-2016, 285 meat samples collected in Beijing area were isolated and identified and the strains were preserved. 2 reference strains Standard strains are Salmonella enteritidis ATCC9184, Salmonella typhi CMCC 50071 (ATCC Culture Collection, USA)

[0088] (2) DNA extraction and content detection.

[0089] According to the extraction steps of the DNA extraction kit, the extracted DNA from 40 samples was melted on ice, then pipetted, mixed and centrifuged for detection. The integrity of the sample was detected by agarose gel electrophoresis (gel concentration 1%; voltage 150V, electrophoresis time 40min), and was used to construct a DNA library.

[0090] (3) Whole genome sequencing and g MLST typing

[0091] (1) Whole genome sequencing: bacterial genome de novo sequencing. The...

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Abstract

The invention discloses a salmonella source tracing typing method based on a gMLST (genome multilocus sequence typing) technology. The method comprises the steps as follows: (1) isolation and dentification of salmonella; (2) DNA extraction and content detection; (3) whole-genome sequencing and gMLST typing; (4) MLST typing; (5) drug resistance gene analysis and virulence gene analysis; and (6) cladogram analysis. The method establishes a salmonella gMLST technology for important food-borne pathogens such as salmonella, staphylococcus aureus or vibrio parahaemolyticus to be applied to epidemiological source tracing, so that the method is promoted to source tracing of food-borne pathogens.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method and application of Salmonella traceability typing based on gMLST technology. Background technique [0002] With the international development of food trade and the widespread use of broad-spectrum and high-efficiency antibacterial drugs, mutant strains continue to appear. In addition, the typing methods and types of foodborne pathogenic bacteria are complicated. Conventional phenotyping methods such as biochemical reactions, Serological typing and phage typing can no longer meet the requirements of bacterial typing, identification of mutant strains, and traceability of pathogenic bacteria, and cannot be used for correlation analysis between strains. At present, molecular typing based on DNA sequence analysis has become the main technical means. Common pathogenic microorganism typing methods include: Random Amplified Primer Polymorphism (RAPD), Amplified Fragment Length Poly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/10C12Q1/6869
CPCC12Q1/689C12Q1/6869C12Q2600/106C12Q2600/156C12Q2531/113C12Q2525/191C12Q2535/122Y02A50/30
Inventor 畅晓晖张捷赵琢高静杨磊石嵩杨向莹李小林高欣乔彬周熙成陈广全彭彦昆
Owner 中国海关科学技术研究中心
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