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Multiplex PCR detection kit for fox origin component identification and identification of fox, rabbit and dog components in animal products

A detection kit and kit technology, applied in the direction of microbial measurement/inspection, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of low accuracy rate, easy misjudgment, large human error, etc., and achieve low cost , wide applicability and strong specificity

Active Publication Date: 2016-02-03
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are also many unscrupulous traders who process fox carcasses, viscera or bones to make or mix them into edible meat products and feed, defrauding consumers and disrupting market order
[0004] Using traditional morphological methods to identify the sources of fur, meat or feed ingredients has large human errors, is prone to misjudgment, and has a low accuracy rate; however, there has not been a simple and quick method in China that can accurately distinguish fox products from other animal products. products

Method used

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  • Multiplex PCR detection kit for fox origin component identification and identification of fox, rabbit and dog components in animal products
  • Multiplex PCR detection kit for fox origin component identification and identification of fox, rabbit and dog components in animal products
  • Multiplex PCR detection kit for fox origin component identification and identification of fox, rabbit and dog components in animal products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Specific detection of fox-derived components in the sample of Example 1

[0058] 1. Sample preparation and storage

[0059] 1.1 Sampling

[0060] Collect 1 g of fox samples and store them at -20°C for future use.

[0061] 1.2 DNA template preparation

[0062] The DNA template was prepared using the commonly used phenol-chloroform crude extraction method (Sambrook J, Fritsch EF, Maniartis T. Molecular Cloning Experiment Guide [M]. 2nd Edition. Jin Dongyan, Li Mengfeng. Beijing: Science Publishing Society, 1999.465-467) or recognized other extraction methods with the same efficacy, these methods are the commonly used methods reported.

[0063] 2. Primer Design

[0064] The primer sequences of this embodiment are shown in Table 3 and SEQ ID NO: 2 and 3 in the sequence listing.

[0065] Table 3 PCR amplification primers designed by the present invention

[0066] Primer name

Primer sequence

VULPES-F

5′-ACAGGGGAGAAAACGCTGTTC-3′

VULPES-R

...

Embodiment 2

[0089] Embodiment 2 sensitivity test

[0090] Using 1ng / μL, 5ng / μL, 10ng / μL, 25ng / μL, 50ng / μL, and 100ng / μL of fox DNA as templates, the nucleotide-specific fragment of SEQ ID NO.1 was amplified according to the conditions of Example 1. The result is as figure 2 As shown, there is amplification at a template concentration of 5 ng / μL, indicating that the fragment amplified by the specific primer has good sensitivity.

Embodiment 3

[0091] Example 3 Detection of primer sets for fox-derived products incorporating non-fox-derived components

[0092] The present invention is for the detection of the source of animal products, through the PCR amplification of the corresponding primers of each species, it is judged whether it is or contains the DNA of a certain species, and then it is judged whether there is any adulteration of animal-derived components of a certain species.

[0093] 1 Extraction of DNA from samples

[0094] The extraction and storage methods of the DNA of each species are the same as those described in Example 1 above.

[0095] 2 Primer design and screening

[0096] In order to find the specific detection sequence of each species, we screen the specific sequence from the aspects of intra-species consistency and stability, inter-species specificity, copy number, etc., by comparing with the nucleic acid sequences of non-native species and different species of this species , and screen the nucle...

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Abstract

Belonging to the technical field of animal origin component molecular detection, the invention provides a multiplex PCR detection kit for fox origin component identification and identification of fox, rabbit and dog components in animal products. The kit provided by the invention includes fox, rabbit, dog and other species-specific primers, a reaction reagent, and positive control and blank control. The fox specific primer is a primer pair able to specifically amplify the nucleotide sequence shown as SEQ ID No.1. The primer provided by the invention is employed to amplify sample DNA, and the PCR amplification product is analyzed to determine whether the sample contains fox species origin ingredients, and at the same time can identify whether rabbit origin or dog origin components are mixed, thus providing means for molecular identification of fur products. In addition, by means of multiplex PCR detection, fox origin, rabbit origin and dog origin components can be identified at one time. The invention establishes an effective method for identification of meat adulteration, fur and feed components. The method and kit have the advantages of simplicity, fastness, comprehensiveness, strong specificity, low cost, and wide applicability.

Description

technical field [0001] The invention relates to the field of animal molecular biology, in particular to the identification of fox-derived components and a multiplex PCR detection kit for fox, rabbit and dog components in animal products. Background technique [0002] my country is a big country of fur production and consumption. Fur usually comes from foxes, mink, sheep, rabbits, dogs, etc. The more expensive sources of fur are foxes, sea otters, mink, nutria, seals, etc. With the development of the economy and the progress of society, although fur is no longer the exclusive product of the upper class, it is still a luxury for mass consumers due to its high price. In order to make huge profits, some unscrupulous operators use artificial fur instead of natural animal fur. Rex rabbit fur, raccoon fur, etc. have also become substitutes for mink and fox fur. [0003] In terms of animal taxonomy, foxes belong to Mammalia, Carnivora, Canidae, and Fox. The main economic value lies...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/16
Inventor 刘榜王文君向胜男赵金艳甄月然张庆德
Owner HUAZHONG AGRI UNIV
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