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Preparation method of kit for fast detecting fox ingredients in food and feed and detection method of fox ingredients in food and feed

A detection kit and kit technology, applied in biochemical equipment and methods, material stimulation analysis, microbial measurement/inspection, etc., can solve the problems of reducing the amount of raw materials, the real attributes of products do not match the labels, and cannot be identified

Active Publication Date: 2013-05-08
山东众合天成检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After various raw materials are processed into food and feed products, it is no longer possible to identify their composition from their appearance
Producers are motivated by profit, change the composition of the product without authorization, or mix in cheap substitutes, or directly reduce the amount of raw materials, resulting in the true attributes of the product not matching the label

Method used

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  • Preparation method of kit for fast detecting fox ingredients in food and feed and detection method of fox ingredients in food and feed
  • Preparation method of kit for fast detecting fox ingredients in food and feed and detection method of fox ingredients in food and feed
  • Preparation method of kit for fast detecting fox ingredients in food and feed and detection method of fox ingredients in food and feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Make the real-time fluorescent PCR detection kit of fox component by following formula, and reagent wherein comprises as follows:

[0034] (1) 2×PCR MasterMix

[0035] Including 0.05u / μL Taq DNA polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);

[0036] Wherein the reaction buffer contains 20mmol / L tris-hydrochloric acid of pH8.8, 100mmol / L potassium chloride and 2% Triton X-100;

[0037] (2) Upstream primer: 10 μmol / L, the sequence is: 5′-TGGAGCATCAGTAGACCTTACAATTT-3′;

[0038] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-GGCGGGAGGTTTTATATTGATAATAG-3′;

[0039] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.

[0040] Follow the procedure below for testing:

[0041] (1) Extraction of DNA from the minced meat sample to be tested

[0042] A. Weigh 0.3g minced meat...

Embodiment 2

[0059] Make the real-time fluorescent PCR detection kit of fox component by following formula, and reagent wherein comprises as follows:

[0060] (1) 2×PCR MasterMix

[0061] Including 0.05u / μL Taq DNA polymerase Polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);

[0062] Wherein the reaction buffer contains 20mmol / L tris-hydrochloric acid of pH8.8, 100mmol / L potassium chloride and 2% Triton X-100;

[0063] (2) Upstream primer: 10 μmol / L, the sequence is: 5′-TGGAGCATCAGTAGACCTTACAATTT-3′;

[0064] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-GGCGGGAGGTTTTATATTGATAATAG-3′;

[0065] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.

[0066] Follow the procedure below for testing:

[0067] (1) DNA extraction of the feed sample to be tested

[0068] A. Weigh 0.3g feed sample,...

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Abstract

The invention belongs to a qualitative detection technology of animal origin ingredients in food and feed, and in particular relates to a kit and a method for detecting the fox ingredients in the food and the feed by using the real-time PCR (polymerase chain reaction) technology. The kit comprises 2*PCRMaster Mix, an upstream primer, a downstream primer and a Taqman probe, wherein 2*PCRMaster Mix contains Taq DNA (deoxyribonucleic acid) polymerase, a reaction buffer, a magnesium chloride and dNTP (diethyl-nitrophenyl thiophosphate), an upstream primer sequence is 5'-TGGAGCATCAGTAGACCTTACAATTT-3 ', a downstream primer sequence is 5'-GGCGGGAGGTTTTATATTGATAATAG-3 ', and a TaqMan probe sequence is 5'-FAM-CCCTGCACCTGGCCGGAGTC-TRAMA-3 '. The method for fast detecting the fox ingredients comprises DNA extraction of the food and the feed, a real-time PCR amplification and a result determination of the fox ingredients. The detection method is fast, strong in specificity, high in sensitivity, easy in operation and good in repeatability.

Description

technical field [0001] The invention relates to a method for rapid detection of animal-derived components using nucleic acid amplification technology, in particular to a real-time fluorescent PCR detection method for fox components. Background technique [0002] The continuous expansion of global economic integration and trade internationalization is both an opportunity and a challenge for developing China. In recent years, my country's foreign trade has shown a sustained and steady growth trend. Statistics show that since 2009, my country has been the second largest foreign trade country in the world for three consecutive years, ranking first in export volume and second in import volume in the world. How to strengthen supervision and supervision, help and guide enterprises to improve product quality, overcome technical barriers in developed countries and regions such as the United States, the European Union, and Japan, and further expand the export of Chinese products; at ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 孙敏高宏伟刘彩霞龚方
Owner 山东众合天成检验有限公司
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