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Detection primers, detection kit and detection method for chicken-derived components in mutton, and application of detection primers

A technology for detecting kits and chicken-derived ingredients, which is applied in the field of biological and biological inspection and quarantine, can solve the problems of being unable to effectively distinguish between intentional adulteration and unintentional pollution, and cannot effectively ensure the correct law enforcement of regulatory authorities, achieving good practical application value, good specific effect

Inactive Publication Date: 2018-12-14
SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECHN CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to literature reports, from 2013 to 2015, mutton products in Beijing, Guangdong, Shandong, Tianjin, Shanghai and other regions in China were found to contain chicken-derived components. Effectively distinguish between intentional adulteration and unintentional pollution, and cannot effectively guarantee the correct enforcement of regulatory authorities

Method used

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  • Detection primers, detection kit and detection method for chicken-derived components in mutton, and application of detection primers
  • Detection primers, detection kit and detection method for chicken-derived components in mutton, and application of detection primers
  • Detection primers, detection kit and detection method for chicken-derived components in mutton, and application of detection primers

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Experimental program
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Effect test

Embodiment 1

[0062] This embodiment provides detection primers for chicken-derived components in mutton. The detection primers include a first primer pair and a first probe for detecting mutton components, and a second primer pair and a second probe for detecting chicken-derived components; the first primer pair The base sequence of the first probe is shown in SEQ ID NO.1-2, the base sequence of the first probe is shown in SEQ ID NO.3, and the base sequence of the second primer pair is shown in SEQ ID NO.4-5 , the base sequence of the first probe is shown in SEQ ID NO.6.

[0063] The 5' end of the first probe and the second probe includes a first fluorescent group, and the 3' end includes a second fluorescent group, the first fluorescent group and the second fluorescent group are different, and the first fluorescent group is One of FAM, MGB, VIC, TET, BHQ1, JOE, HEX, CY3, CY5, ROX, RED610, Texas Red, RED670 or NED, the second fluorescent group is FAM, MGB, VIC, TET, BHQ1, JOE , HEX, CY3, ...

Embodiment 2

[0080] This embodiment provides a detection kit for chicken-derived components in mutton, which includes the detection primers for chicken-derived components in mutton provided in Example 1; the detection kit includes genomic DNA extraction reagents, ddPCR reaction reagents and fluorescence quantitative PCR reaction reagents.

[0081] Genomic DNA extraction reagent is at least one of PBS buffer, proteinase K, tissue digestion solution GHA and TB solution.

[0082] PCR reaction reagents include Taq DNA polymerase, dNTPs, Mg 2+ at least one of the

[0083] ddPCR reaction reagents include TM Supermix for probes reagents.

Embodiment 3

[0085] The present embodiment has provided a kind of detection method of chicken source component in mutton, and detection method comprises the following steps:

[0086] Genomic DNA was extracted from chicken and mutton samples, and ddPCR and fluorescent quantitative PCR were performed to determine the linear relationship between the gene copy number and Ct value of chicken and mutton samples;

[0087] Quantitative sampling of chicken samples and mutton samples, and genomic DNA extracted respectively, for ddPCR test, to obtain the linear relationship between the quality of chicken samples and mutton samples and the gene copy number;

[0088] Through the linear relationship between the gene copy number and the Ct value and the linear relationship between the quality and the gene copy number, the relationship between the mass of the chicken sample and the mutton sample and the Ct value is obtained, and the relationship between the mass percentage of chicken and mutton is obtained...

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Abstract

The invention provides detection primers, a detection kit and a detection method for chicken-derived components in mutton, and application of the detection primers, belonging to the field of biological inspection and quarantine. The detection primers for the chicken-derived components in mutton have good specificity and can specifically detect chicken components. As the detection primers for the chicken-derived components in mutton are applied to the detection kit, the implementation of detection is better facilitated. The detection method for the chicken-derived components in mutton can detect and calculate the contents of chicken components in a sample, is a stable, accurate, effective and quantitative detection method, and has good practical application value.

Description

technical field [0001] The invention relates to the field of biological and biological inspection and quarantine, in particular to detection primers, detection kits and application and detection methods of chicken-derived components in mutton. Background technique [0002] With the continuous improvement of people's living standards, mutton, as a low-fat, low-cholesterol and high-protein health meat food, is more and more popular among consumers. According to statistics, the output of the mutton industry reached 4.54 million tons in 2016, and it is predicted that its output will rise to 5.33 million tons in 2022 [1]. The huge mutton consumption market has led to frequent occurrence of adulteration incidents of mutton products, which has aroused widespread concern from the whole society. In order to make huge profits, many unscrupulous traders use low-priced meat to partially fill or completely replace high-priced meat, such as duck, fox meat, pork, horse meat, chicken, rat ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 陈世界林华张婧杨苗安微唐廷廷
Owner SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECHN CENT
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