Microfluidics detecting chip based on nucleic acid amplification technology and preparation method thereof

A detection chip and microfluidic technology, applied in biochemical equipment and methods, microbial determination/inspection, enzymology/microbiology devices, etc., can solve the problem that the supporting products are not perfect, the method system is not perfect, and the problem is not comprehensive enough And other problems, to achieve the effect of wide application, easy to carry, small size

Active Publication Date: 2016-02-17
TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Molecular detection technology promotes the quality and safety of agricultural products and food, but there are still problems such as insufficient method system, insufficient solution to problems, and inadequate supporting products

Method used

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  • Microfluidics detecting chip based on nucleic acid amplification technology and preparation method thereof
  • Microfluidics detecting chip based on nucleic acid amplification technology and preparation method thereof
  • Microfluidics detecting chip based on nucleic acid amplification technology and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Application: It is used for the detection of genetically modified ingredients in food. The target genes are endogenous gene ZSSIIb in corn, Lectin endogenous gene in soybean, SPS endogenous gene in rice, CAMV35S promoter, NOS terminator and BAR gene.

[0053] (2) Preparation process: Use computer software to draw the structure and channel graphics of each layer of the microfluidic detection chip for later inkjet printing, printing and etching; inkjet 20 serial number marking points and 20 on the bottom plate layer Reaction area: Apply printing technology on the primer layer to inkjet print oligonucleotide primer sequences in 20 reaction areas, 12 reaction chambers in each reaction area, a total of 240 areas, respectively endogenous gene ZSSIIb in corn, endogenous soybean Gene Lectin, rice endogenous gene SPS, CAMV35S promoter, NOS terminator and BAR gene, each primer was repeated twice, and stored in freeze-dry; 20 sample chambers, channels and 240 reaction chambers ...

Embodiment 2

[0058] (1) Application: It is used for the detection of animal-derived components in food. The targets are eukaryotic 18S, pigs, goats, sheep, cattle, buffaloes, chickens, ducks, geese, mice, foxes, and cats, a total of 12 targets.

[0059] (2) Preparation process: Use computer software to draw the structure and channel graphics of each layer of the microfluidic detection chip for later inkjet printing, printing and etching; inkjet 20 serial number marking points and 20 on the bottom plate layer Reaction area: Apply printing technology on the primer layer to inkjet print oligonucleotide primer sequences in 20 reaction areas, 12 reaction chambers in each reaction area, a total of 240 areas, respectively eukaryotic 18S, pig, goat, Sheep, cow, buffalo, chicken, duck, goose, mouse, fox, cat, 12 in total, freeze-dried and preserved; 20 sample chambers, channels and 240 reaction chambers were etched on the reaction layer, each reaction chamber volume was about 1 μL; print out the in...

Embodiment 3

[0064] (1) Application: used for the detection of foodborne pathogenic microorganisms in food, the targets are prokaryotic 16S, Escherichia coli, Salmonella, Staphylococcus aureus, Shigella, Listeria monocytogenes, a total of 6 .

[0065] (2) Preparation process: Use computer software to draw the structure and channel graphics of each layer of the microfluidic detection chip for later inkjet printing, printing and etching; inkjet 20 serial number marking points and 20 on the bottom plate layer Reaction area: Apply printing technology on the primer layer to inkjet print oligonucleotide primer sequences in 20 reaction areas, 12 reaction chambers in each reaction area, a total of 240 areas, respectively prokaryotic 16S, Escherichia coli, Salmonella, 6 sets of Staphylococcus aureus, Shigella, and Listeria monocytogenes, 2 replicates of each primer, freeze-dried and stored; 20 sample chambers, channels, and 240 reactions were etched on the reaction layer The volume of each reactio...

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Abstract

The invention discloses a microfluidics detecting chip based on a nucleic acid amplification technology and a preparation method thereof. The chip has four layers, and is in a disc centrifugal design, one chip can be used for detecting 20 samples, and each sample has 12 parameters. Through an optical driving type centrifugal machine, temperature control equipment and nucleic acid amplification reagent consumable items which are matched with the microfluidics detecting chip, requirements on detection of transgenic ingredients in agricultural products and food, pathogenic bacteria, animal and plant species identification and the like are met, and the microfluidics detecting chip has the features of quickness, convenience, high flux and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular detection and application of agricultural products and food quality and safety, and in particular relates to a microfluidic detection chip based on nucleic acid amplification technology and a preparation method thereof. The chip is a 4-layer disc centrifugal design. Each chip is divided into 20 sample areas, and each sample area contains 12 detection chambers. It is equipped with CD-ROM centrifuge, temperature control equipment and nucleic acid amplification reagent consumables, which can meet the molecular detection needs of agricultural products, genetically modified ingredients in food, pathogenic bacteria, animal and plant species, etc., and has the characteristics of fast, convenient and high throughput. Background technique [0002] Molecular detection technology is a new type of detection technology that targets nucleic acid differences in agricultural products and foods. It overcomes the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/68
Inventor 兰青阔徐石勇赵新李欧静刘娜王永朱珠陈锐
Owner TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS
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