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Identification of fox-derived components and multiplex PCR detection kit for 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: 2018-08-03
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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  • Identification of fox-derived components and multiplex PCR detection kit for fox, rabbit and dog components in animal products
  • Identification of fox-derived components and multiplex PCR detection kit for fox, rabbit and dog components in animal products
  • Identification of fox-derived components and multiplex PCR detection kit for fox, rabbit and dog components in animal products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0058] 1. Preparation and storage of samples

[0059] 1.1 Sampling

[0060] Collect 1g fox sample and store it at -20°C for later use.

[0061] 1.2 DNA template preparation

[0062] The DNA template was prepared using the commonly used phenol-chloroform crude extraction method (Sambrook J, Fridge EF, Maniatis T. Molecular cloning experiment guide [M]. 2nd edition. Jin Dongyan, Li Mengfeng. Beijing: Science Publishing Society, 1999.465-467) or other recognized extraction methods with the same effectiveness, these methods are commonly reported methods.

[0063] 2. Primer design

[0064] The primer sequences of this example 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

5′-GGCCTCCCCGAGATGAATC-3′

[0067] The expected amplified f...

Embodiment 2

[0089] Example 2 Sensitivity test

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

Embodiment 3

[0091] Example 3 Detection of primer sets incorporating non-fox-derived ingredients in fox-derived products

[0092] The present invention is to detect the origin of animal products, and determine whether it is or contain DNA of a certain species through PCR amplification with corresponding primers of each species, and then determine whether there is adulteration of animal-derived components of a certain species.

[0093] 1 Extraction of DNA from samples

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

[0095] 2 Design and screening of primers

[0096] In order to find the specific detection sequence of each species, we screened the specific sequence from the aspects of intra-species consistency and stability, inter-species specificity, copy number, etc., through comparison with the nucleic acid sequences of non-this species and different species of this species , Screen the more conservative and inter-species spe...

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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 multiple PCR detection kits for fox, rabbit and dog components in animal products. Background technique [0002] my country is a big country for fur production and consumption. Fur usually comes from foxes, minks, sheep, rabbits, dogs, etc. The more valuable sources of fur are foxes, sea otters, minks, nutrias, seals, etc. With economic development and social progress, although fur is no longer an exclusive product of the upper class, it is expensive and still a luxury item for mass consumers. Some unscrupulous operators used artificial fur instead of natural animal fur to make huge profits. Rex rabbit fur and raccoon fur have also become substitutes for mink and fox fur. [0003] The fox belongs to Mammalia, Carnivora, Canidae, and Fox in the taxonomy of animals. The main economic value lies in the fur. It has good warmth retenti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
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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