Multiplex PCR (Polymerase Chain Reaction) detection primer group for avian pathogenic escherichia coli and the like, method and kit

A technology for the pathogenicity of Escherichia coli and poultry, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, recombinant DNA technology, etc., to achieve the effects of high sensitivity, simple operation and strong specificity

Active Publication Date: 2018-07-06
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

PCR technology is widely used because of its strong specificity, high sensitivity, simple and fast operation, but these methods usually only detect one or a few pathogenic bacteria in each experiment

Method used

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  • Multiplex PCR (Polymerase Chain Reaction) detection primer group for avian pathogenic escherichia coli and the like, method and kit
  • Multiplex PCR (Polymerase Chain Reaction) detection primer group for avian pathogenic escherichia coli and the like, method and kit
  • Multiplex PCR (Polymerase Chain Reaction) detection primer group for avian pathogenic escherichia coli and the like, method and kit

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Embodiment 1

[0044] Embodiment 1 multiplex PCR detection method

[0045] In this embodiment, 6 7-day-old three-yellow chickens were respectively used with avian pathogenic Escherichia coli (Avia n pathogenic Escherichia coli, APEC), Pasteurella multocida (Pasteurella multocida), Proteus mirabilis (Proteus mirabilis), Pseudomonas aeruginosa (Pseudom onas aeruginosa), Salmonella (Salmonella) and Staphylococcus aureus (Staphylococcus aureus) were challenged. After 24 hours, the blood, spleen, liver and kidney of the challenged chickens were aseptically collected, and multiplex PCR was used to Detection of avian pathogenic Escherichia coli, Pasteurella multocida, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella and Staphylococcus aureus, including the following steps:

[0046] Step 1, prepare a PCR template based on the bacteria to be detected

[0047] Aseptically take the liver of the challenged three-yellow chicken and grind it, filter it with a 200-mesh copper mesh, take the filtrate ...

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Abstract

The invention provides a multiplex PCR (Polymerase Chain Reaction) primer group for avian pathogenic escherichia coli, pasteurella multocida, proteus mirabilis, pseudomonas aeruginosa, salmonella andstaphylococcus aureus, a multiplex PCR detection kit comprising the primer group, and a multiplex PCR detection method using the primer group. The primer group comprises primer pairs PhoA-F and PhoA-Rfor specifically amplifying a gene phoA of the avian escherichia coli, primer pairs KMT-F and KMT-R for specifically amplifying a gene KMT1 of the pasteurella multocida, primer pairs AtpD-F and AtpD-R for specifically amplifying a gene AtpD of the proteus mirabilis, primer pairs PETA-F and PETA-R for specifically amplifying a gene PETA of the pseudomonas aeruginosa, primer pairs InvA-F and InvA-Rfor specifically amplifying a gene InvA of the salmonella, and prier pairs NuC-F and NuC-R for specifically amplifying a gene nuc of the staphylococcus aureus.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a multiple PCR primer set for detecting avian pathogenic Escherichia coli, Pasteurella multocida, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella and Staphylococcus aureus, including The multiple PCR detection kit of the primer set and the multiple PCR detection method using the primer set. Background technique [0002] Avian pathogenic Escherichia coli (APEC), Pasteurella multocida, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella and golden yellow Staphylococcus (Staphylococcus aureus) is the main pathogenic microorganism that endangers the poultry industry. [0003] After infecting poultry with avian pathogenic Escherichia coli, symptoms such as hepatomegaly, intestinal mucosal bleeding and ulcers, and pericardial inflammation appear. Extensive use will easily cause drug residues in poultry products, seriously affecting human health and causing public health problems....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/14C12Q1/10C12N15/11C12Q1/686C12R1/19C12R1/42C12R1/445C12R1/37C12R1/01
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2537/143
Inventor 韩先干王志豪祁克宗左佳坤龚建森苗晋锋陈兆国蒋蔚王少辉米荣升黄燕
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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