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Whole genome sequencing analysis method of pigling diarrhoea Escherichia coli

A whole-genome sequencing, E. coli technology, applied in the direction of microbial-based methods, microbial determination/inspection, biochemical equipment and methods, etc., to achieve the effect of reducing abuse

Inactive Publication Date: 2017-10-13
BEIJING VOCATIONAL COLLEGE OF AGRI
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Problems solved by technology

However, many strains currently isolated have multiple types of major virulence factors and are considered to be more pathogenic

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  • Whole genome sequencing analysis method of pigling diarrhoea Escherichia coli
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  • Whole genome sequencing analysis method of pigling diarrhoea Escherichia coli

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Embodiment Construction

[0032] The present invention will be further described in detail with reference to the accompanying drawings and embodiments.

[0033] The present invention provides a method for whole genome sequencing analysis of piglet diarrhea Escherichia coli, said method comprising the steps of:

[0034] The first step is sample collection.

[0035] 400 piglet diarrhea fecal samples were collected from 8 pig farms in Beijing area, each pig farm had more than 500 sows. Piglets are housed on concrete with perforated floors. Diarrhea can be diagnosed if piglets have watery stools or even bloody stools or obviously frequent defecation.

[0036] The second step is the isolation and identification of Escherichia coli from stool samples.

[0037] (1) Feces samples were cultured in tryptone medium containing 5% (mass percentage concentration) serum, and cultured at 37° C. for 20 h.

[0038] (2) Bacterial clones were picked from the tryptone medium and further cultured on MacConkey solid medi...

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Abstract

The invention discloses a whole genome sequencing analysis method of pigling diarrhoea Escherichia coli, and belongs to the technical field of biological gene detection. The whole genome sequencing analysis method of pigling diarrhoea Escherichia coli comprises following steps: sample collecting; separation and identification of fecal sample Escherichia coli; serotype identification; virulence factor result detection; whole genome sequencing analysis and whole genome sequence study of Escherichia coli containing a plurality of virulence factors. According to the whole genome sequencing analysis method, whole genome sequencing is adopted for more comprehensive knowing of the genome information of Escherichia coli ST4214, and analysis of the virulence factors contained by Escherichia coli ST4214; genome analysis of pigling diarrhoea Escherichia coli ST4214 possesses significant importance on study of pigling diarrhoea epidemic serotype; study on pigling diarrhoea Escherichia coli ST4214 drug resistance gene is capable of guiding applications of antibiotics based on the antibiotic sensitivity and drug resistance, and avoiding antibiotics abuse; and the whole genome sequencing analysis method is used for providing data base for development of pigling diarrhoea vaccines.

Description

technical field [0001] The invention belongs to the technical field of biological gene detection, and in particular relates to a whole-genome sequencing analysis method for piglet diarrhea Escherichia coli. Background technique [0002] Piglet diarrhea caused by Escherichia coli is a major loss to agriculture. Escherichia coli that causes porcine diarrhea can be divided into enteroaggregative Escherichia coli (EAEC) according to the colonization site of its host, mechanism of virulence factors, clinical symptoms and outcome, which can aggregate and adhere to cultured HEp-2 / HeLa cells in the intestinal tract. Mucous membranes form biofilms; pathogenic Escherichia coli (EPEC) attaches to enterocytes and causes intestinal cell damage; enterohaemorrhagic Escherichia coli (EHEC / STEC) can produce different forms of gag toxins; enterotoxigenic Escherichia coli ( ETEC), capable of producing thermostable or thermolabile toxins; enteroinvasive Escherichia coli (EIEC), producing toxin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12R1/19
CPCC12Q1/689
Inventor 王振玲郭艳杰王黎霞王金秋
Owner BEIJING VOCATIONAL COLLEGE OF AGRI
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