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

A technology for detection kits and source components, applied in the field of multiplex PCR detection kits, can solve the problems of complicated methods, low accuracy, few animal species, etc., and achieve the effects of wide applicability, low cost 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

The collars of mink fur bought in the market may be made of ordinary rabbit fur or dog fur through deep processing, and ordinary consumers cannot tell the difference at all.
In addition, there are also many unscrupulous traders who process or mix mink carcass meat, internal organs or bones into edible meat products and feed, deceiving consumers and disrupting market order
Using traditional morphological methods to identify the source of fur, meat or feed ingredients, human error is large, easy to misjudge, and the accuracy rate is low
At present, in the published patents of our country, there is a method for identifying sable and pine mink (Li Bo, Xu Yanchun. Primers for amplifying the cytochrome b gene of mink

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Specific detection of mink-derived components in the sample of Example 1

[0059] 1. Sample preparation and storage

[0060] 1.1 Sampling

[0061] 1 g of mink samples were collected and stored at -20°C for future use.

[0062] 1.2 DNA template preparation

[0063] 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.

[0064] 2. Primer Design

[0065] The primer sequences of this embodiment are shown in Table 3 and SEQ ID NO: 2 and 3 in the sequence listing.

[0066] Table 3 PCR amplification primers designed by the present invention

[0067] Primer name

Primer sequence

MUSTLA-F

5′-TTCGGCGCTGCCTTAATTGTC-3′

MUSTLA...

Embodiment 2

[0090] Embodiment 2 sensitivity test

[0091] Using 0.1ng / μL, 1ng / μL, 10ng / μL, and 100ng / μL mink 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 1ng / μL, indicating that the fragment amplified by the specific primer has good sensitivity.

Embodiment 3

[0092] Example 3 Detecting the Primer Set of Mink-derived Products Mixed with Non-Mink-derived Products

[0093]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.

[0094] 1 Extraction of DNA from samples

[0095] The extraction and storage methods of the DNA of each species are the same as those described in Example 1 above.

[0096] 2 Primer design and screening

[0097] 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, The nucleotide sequences...

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PUM

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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 mink origin component identification and identification of mink, rabbit and dog components in animal products. The kit provided by the invention includes mink, rabbit, dog and other species-specific primers, a reaction reagent, and positive control and blank control. The mink 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 mink species origin ingredients, and at the same time the kit 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, mink origin, rabbit origin and dog origin components can be identified at one time. The invention establishes an effective method for identification of meat and fur adulteration 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 mink-derived components and a multiplex PCR detection kit for mink, rabbit and dog components in animal products. Background technique [0002] Fur, also known as fur, refers to clothing made of animal fur. It may be one of the earliest clothing materials for humans. With the development of the times, fur has gradually become a symbol of luxury and wealth. Animals commonly used to make fur include mink, fox, rabbit, raccoon dog, beaver, etc., which are expensive and are one of the high-end winter clothing for women. The fur that people often say generally refers to mink fur. [0003] Mink belongs to Mammalia, Carnivora, Mustelidae, Weasel in animal taxonomy, and is one of the small and precious fur animals. Mink fur is one of the three treasures of Northeast China. It is known as "the king of fur" and is called "soft gold" abroad. Mink fur is a ki...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 刘榜王文君吴清清夏瑾华张庆德
Owner HUAZHONG AGRI UNIV
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