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Method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology

A nucleic acid and peptide nucleic acid technology, applied in the field of detection of food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology, to achieve the effects of improving detection sensitivity and stability, efficient capture, and expanding the scope of application

Inactive Publication Date: 2017-10-20
CHINA AGRI UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RPA technology is a recombinant polymerase amplification technology that can accumulate products in a short period of time, but the product is double-stranded nucleic acid, which cannot be visually detected with single-stranded nucleic acid test strips

Method used

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  • Method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology
  • Method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology
  • Method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology

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Embodiment 1

[0068] Embodiment 1 Method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology

[0069] In this example, a total of 16 microorganisms were used for the construction of the biosensor, and Salmonella typhimurium was used as the model strain for the construction of the biosensor (Table 2).

[0070] Table 2

[0071]

[0072] All strains were activated with LB medium and cultured at 37°C for 16h, in which Salmonella typhimurium was serially diluted with 0.8% normal saline to ensure the final concentration was 1-10 6 CFU mL -1 .

[0073] 1. Primer and probe design

[0074] The invA gene of Salmonella was selected as the target sequence (GenBank: DQ644633.1), according to TwistAmp TM Guided by the reaction kit, self-designed RPA amplification primers (RPA-F, RPA-R). A universal adaptor and a C3 blocker were ligated to the 5' end of the RPA primers for the design of primers (RPA-UF, RPA-UR) for universal adaptor blocking recomb...

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Abstract

The invention relates to a method for detecting food-borne pathogenic bacteria based on a nucleic acid chromatography biosensing technology. The method comprises the following steps: screening out a high-specific virulence gene according to pathogenic bacteria, establishing a novel universal adaptor recombinant polymerase amplification technology, assembling a single strand nucleic acid chromatography test strip based on peptide nucleic acid, and establishing a universal adaptor blocking RPA biosensor detection technology based on functional nucleic acid. The method is applied to the on-site rapid detection of food-borne pathogenic bacteria, can efficiently capture single strand nucleic acid, improve the detection sensitivity and stability and realize visual detection.

Description

technical field [0001] The present invention relates to the technical field of biosensing detection, in particular to a method for detecting food-borne pathogenic bacteria based on nucleic acid chromatography biosensing technology. Background technique [0002] In recent years, food safety incidents caused by food-borne pathogens have occurred frequently, and the corresponding testing technical requirements have become increasingly strict. Traditional detection techniques mainly rely on plate culture and physiological and biochemical identification. For some strains that are difficult to cultivate, false negatives are likely to occur. Rapid detection technologies emerging in recent years include enzyme-linked immunosorbent technology, PCR and quantitative PCR technology, isothermal amplification technology, and sensor technology. Advantage. [0003] With the development of technology, a variety of sensors have been applied to the detection of pathogenic bacteria, such as o...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6844C12Q2525/107C12Q2525/191C12Q2521/101C12Q2521/507C12Q2563/137C12Q2565/625Y02A50/30
Inventor 罗云波许文涛徐瑗聪黄昆仑程楠王蔚然卫昱君
Owner CHINA AGRI UNIV
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