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DNA chip for detecting virulence gene expression abilities of APEC and UPEC and construction method thereof

A DNA chip and virulence gene technology, applied in the field of zoonosis, can solve the problem that the pathogenesis is not completely clear and so on

Inactive Publication Date: 2008-01-02
YANGZHOU UNIV
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Problems solved by technology

The virulence factors associated with APEC infection have been reported successively, but their pathogenesis has not been fully understood

Method used

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  • DNA chip for detecting virulence gene expression abilities of APEC and UPEC and construction method thereof
  • DNA chip for detecting virulence gene expression abilities of APEC and UPEC and construction method thereof

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0134] 1. Construction of virulence gene expression chip of avian pathogenic Escherichia coli and human urinary tract pathogenic Escherichia coli:

[0135] 1. Target gene and primer design:

[0136] The sequences of virulence genes and drug resistance genes of APEC, UPEC and some other Escherichia coli included in GenBank were analyzed by multiple alignment, the conserved regions of each virulence gene were selected, and the primer design software of Primer Premier 5.0 was used to design each gene. For the primers of the target gene, the Tm value of the primers is controlled at about 55±5°C, and the size of the target gene fragment is between 200-900bp. The design results of the primers are shown in Table 1. The primers were synthesized by Shanghai Sangon Bioengineering Company. Some gene clones are from APEC-specific clones obtained by SSH and SCOTS methods. Among them, isolates of avian pathogenic Escherichia coli, porcine Escherichia coli TB1, C83907, 107 / 86 porcine Escher...

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Abstract

The invention discloses a DNA chip and constructing method to detect the expression level of APEC and UPEC virulent gene in the sick human-animal technical domain, which comprises the following steps: augumenting the 97 gene segments of the known virulent gene and partial drug tolerant gene of escherichia coli; using SSH method to acquire 44 gene segments of APEC specific gene; adopting SCOTS method to obtain 13 gene segments of APEC specific gene; making the recombinant plasmid as mould; recycling and purifying each gene segment; using SmartArrayTM microarrayer to spot on chemically decorative the slide; setting positive and negative contrast; constructing the DNA chip with 154 target gene probes; fitting for detecting the expression level of APEC and UPEC virulent gene.

Description

technical field [0001] The invention relates to the research on the virulence gene expression chip of avian pathogenic Escherichia coli and human urinary tract pathogenic Escherichia coli, and belongs to the technical field of zoonoses. Background technique [0002] Avian colibacillosis (Avian Colibacillosis) refers to the disease of local or systemic infection caused partially or entirely by avian pathogenic Escherichia coli (APEC), including Escherichia coli sepsis, Escherichia coli granuloma (Hjarre disease), air sac disease (chronic respiratory disease, CRD), avian cellulitis, swollen head syndrome, peritonitis, salpingitis, synovitis, panphthalmitis and omphalitis / yolk sac infection. Avian colibacillosis is a common infectious disease in chickens and turkeys aged 2 to 12 weeks, causing serious economic losses to the poultry industry. The virulence factors associated with APEC infection have been reported successively, but their pathogenesis has not been fully understoo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 高崧刘秀梵宦海霞陈祥赵李祥
Owner YANGZHOU UNIV
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