Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Real-time fluorescent PCR assay for detection of bovine-derived components in food and feed using single-copy nuclear genes

A bovine-derived, nuclear gene technology, applied in the field of real-time fluorescent PCR detection, can solve the problems of difficult quantitative detection, prone to false positive results, etc., and achieve the effect of small sample volume, protection of the right to know and choice, and a wide range of applications.

Active Publication Date: 2019-07-30
PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The primary purpose of the present invention is to provide a real-time fluorescent PCR detection method for bovine-derived components in food and feed, so as to overcome the deficiencies in the prior art that are difficult to quantitatively detect and prone to false positive results.

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
  • Real-time fluorescent PCR assay for detection of bovine-derived components in food and feed using single-copy nuclear genes
  • Real-time fluorescent PCR assay for detection of bovine-derived components in food and feed using single-copy nuclear genes
  • Real-time fluorescent PCR assay for detection of bovine-derived components in food and feed using single-copy nuclear genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The design of embodiment 1 primer pair and probe

[0043] (1) According to the bovine (Bos taurus) β-actin sequence published in NCBI (accession number: EH170825.1), select appropriate sequence fragments to design primers and probes.

[0044] The length of the target fragment for bovine-derived components is 62bp. The sequences of fluorescent PCR primer pairs and probes are as follows:

[0045] Bovine-62bp-F: 5'-GGCCTCGGAGTGTGTATTCAG-3' (SEQ ID NO: 1)

[0046] Bovine-62bp-R: 5'-GCCCCAGAATGAGGTTCACTT-3' (SEQ ID NO: 2)

[0047] Bovine-62bp-P: FAM-AGGTGCACAGTACGTTC-MGB (SEQ ID NO: 3)

[0048] Among them, FAM represents a fluorescent reporter group, and MGB represents a quencher group. The present invention adopts the fluorescent probe method, and its detection principle is to identify templates by using fluorescently labeled specific probes. Compared with the SYBR dye method in the prior art, the fluorescence-labeled specific probe of the present invention has stronge...

Embodiment 2

[0054] The preparation of embodiment 2 test samples

[0055] Animal meat samples were shredded, dried, and ground into powder using a freezer grinder (SPEX 6850). Plant samples and samples for practical validation were directly ground into powder using a cryo-mill (SPEX 6850). Taking corn flour as the base, adding beef powder to prepare the content of 0.1% (W / W), 0.01% (W / W), 0.001% (W / W), 0.0001% (W / W) and 0.00001% (W / W) / W) mixed beef powder samples for sensitivity testing.

Embodiment 3

[0056] The extraction of embodiment 3DNA

[0057] DNA from animal samples was extracted using the Animal Genome Extraction Kit (Tiangen Biochemical Technology Co., Ltd.; catalog number: DP323), and DNA from plant samples was extracted using the Plant Genome Extraction Kit (Tiangen Biochemical Technology Co., Ltd.; catalog number: DP305). Plant Feed Genome Extraction Kit (Tiangen Biochemical Technology Co., Ltd.; catalog number: DP323) was used to extract DNA from mixed samples and actual verification samples. For the extraction method, see the kit operation manual for details. The DNA solution was stored at -20°C for later use.

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 discloses a real-time fluorescence PCR detection method for detecting bovine-derived materials in food and feed by use of a single-copy nuclear gene. The method comprises the following steps: (I) with the DNA of a to-be-detected sample as a template, performing fluorescence quantitative PCR amplification to obtain a PCR amplification product; (II) detecting the fluorescence signal of the amplification product; and (III) judging whether the sample contains bovine-derived materials according to the Ct value of the detection result, wherein the reaction system for PCR amplification contains a specific primer pair for amplifying the bovine-derived materials and a specific probe of the bovine-derived materials. In the invention, the specific primer pair and probe for real-time fluorescence PCR amplification of the bovine-derived materials have good specificity and high sensitivity; a quantitative detection method is provided for quickly and accurately detecting whether the food and feed contain bovine-derived materials; and the method has relatively good application prospects.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a real-time fluorescent PCR detection method for detecting bovine-derived components in food and feed by using a single-copy nuclear gene. Background technique [0002] Since mad cow disease first occurred in the UK in 1987, countries and regions around the world have promulgated laws and regulations to control the use of animal-derived ingredients. In 2004, my country promulgated the "Administrative Measures for the Safety and Hygiene of Animal-derived Feed Products", which prohibits the use of animal-derived feed products other than milk and dairy products in ruminant feed. At the beginning of 2013, Sweden, the United Kingdom and other European countries were involved in the "horse meat scandal" scandal, which aroused the attention of various countries on the adulteration of meat and meat products. [0003] In my country, the issue of "adulteration" has always been th...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2531/113C12Q2563/107C12Q2561/113
Inventor 蔡一村吕蓉潘良文
Owner PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com