Gene microfluidic chip for detecting 26 animal-derived components at same time and application thereof

A technology of animal-derived components and microfluidic chips, applied in bioreactor/fermenter combinations, specific-purpose bioreactor/fermenter, biomass post-processing, etc., can solve the problem that probes are susceptible to environmental interference, reduce Sensitivity and specificity, unfavorable long-term storage and other issues, to achieve high species conservation, increase throughput, and high inter-species specificity

Active Publication Date: 2017-10-13
北京市食品安全监控和风险评估中心
View PDF2 Cites 26 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are reports of 16 kinds of animal-derived components using aldehyde-based glass slides as solid-phase carriers to make gene chips (Zong Hui, 2011 application number: 201110110348.2). This method requires PCR reaction first, and then the probes on the chip Perform hybridization, and finally perform fluorescence scanning and data processing on the hybridized chip, with many steps and cumbersome

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene microfluidic chip for detecting 26 animal-derived components at same time and application thereof
  • Gene microfluidic chip for detecting 26 animal-derived components at same time and application thereof
  • Gene microfluidic chip for detecting 26 animal-derived components at same time and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The design of embodiment 1 primer and the preparation of gene microfluidic chip

[0035] Primer-probe design is performed in three steps:

[0036] 1. For 26 kinds of animals: pig, cow, sheep, goat, chicken, duck, cat, mouse, dog, turkey, deer, pigeon, fox, mink, camel, ostrich, rabbit, goose, roe deer, horse, quail , grass carp, salmon, perch, rainbow trout and crucian carp, select COX1, ND1, ND2, 16S and other genes on the mitochondria as target genes, design primers through primer and probe analysis software, and effectively screen out the genes that can be obtained under the same reaction conditions on the chip. Primer probes with good amplification efficiency. The obtained candidate probes need to be analyzed by BLAST on NCBI, and the probes with the best specificity between species are selected for chip design. The present invention has designed 32 pairs of primers and corresponding Taqman probes, the nucleotide sequences of which are shown in SEQ ID NO.1-96 resp...

Embodiment 2

[0042] Example 2 Establishment of a method for detecting 26 kinds of animal-derived components using the gene microfluidic chip prepared in Example 1

[0043] 1. Extract sample DNA

[0044] (1) Take 26 kinds of animals (pig, cow, sheep, goat, chicken, duck, cat, mouse, dog, turkey, deer, pigeon, fox, mink, camel, ostrich, rabbit, goose, roe deer, horse , quail, grass carp, salmon, perch, rainbow trout and crucian carp) muscle sample 0.2g, cut as much as possible. Place in a 1.5ml centrifuge tube, add 1ml of cell lysis buffer, 20μl proteinase K (500μg / ml), and mix well. Water-bath in a constant temperature water bath at 65°C for 30 minutes, and shake the centrifuge tube several times intermittently. Centrifuge at 12,000 rpm for 5 minutes in a tabletop centrifuge, and transfer the supernatant to another centrifuge tube.

[0045] (2) Add an equal volume of phenol:chloroform mixture (1:1), shake and mix, and centrifuge at 12,000rpm for 10min.

[0046] (3) Take the supernatant ...

Embodiment 3

[0068] Embodiment 3 specificity test

[0069] Adopt the method of embodiment 2 to detect above-mentioned 26 species and rat, Chinese grass shrimp, grouper genomic DNA, its experimental result is: the Ct value of positive quality control is 24.54, the Ct value of positive internal reference is 25.49, 26 The CT values ​​of each species are all less than 30, and the Ct values ​​of rat, grass shrimp and grouper are all greater than 30. This experiment proves that the gene microfluidic chip of the present invention has good species specificity.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a gene microfluidic chip for detecting 26 animal-derived components at the same time and application thereof, and belongs to the technical field of animal food detection. 32 combinations each of which consists of a pairs of primers and a probe are fixed to the gene microfluidic chip, and the nucleotide sequences are shown as SEQ ID NO.1-96; fluorescence quantitative PCR detection is performed on a sample to be detected, and according to a Ct value, whether the sample to be detected contains any of the 26 animal-derived components is judged. By a detection method provided by the invention, rapid identification detection of the 26 animal-derived components can be completed within 3 hours; the detection method has the advantages of accurate detection, high sensitivity, strong specificity, simpleness, convenience and rapidness; the lowest detection limit is 1pg; requirements on large-batch and rapid identification detection of the 26 animal-derived components can be met; adulteration of meat products can be effectively resisted so as to enhance protection of the interests of consumers, so that the social benefits are relatively high.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a gene microfluidic chip for detecting 26 kinds of animal-derived components in food and its application. Background technique [0002] The identification of animal-derived ingredients in food is an important part of today's food safety testing. In 2013, the incident of beef mixed with horse meat caused a sensation in Europe. Maternity fakes are also common in the Chinese market. Duck meat is often used. Cheap meat such as chicken is sold as beef and mutton, and there have even been incidents of poisoning consumers by mixing rat meat poisoned by rat poison in meat products. These incidents have seriously disrupted the order of the food market and violated the interests of consumers. In severe cases, they may cause ethnic problems and disrupt social harmony. [0003] With the continuous development of the market, the technical means of meat adulteration are also constantly...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12N15/11C12M1/34B01L3/00
CPCB01L3/5027B01L2200/10C12Q1/6851C12Q1/6888C12Q2531/113C12Q2561/101C12Q2565/629
Inventor 赵琳娜杨昕霆薛晨玉王丹黄华郭淼胡智恺杨红莲韩月贝刘剑李赫婧
Owner 北京市食品安全监控和风险评估中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap