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Short-tandem-repeats (STR) primer combination for source-traced identification on pork product and detection kit

A primer set and pork technology, applied in the field of traceability detection, can solve problems such as disrupting market order, many circulation links, and counterfeit labels

Active Publication Date: 2016-07-20
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Driven by economic interests in recent years, low-value products have often been used to pass off high-value products, counterfeit brands, and counterfeit origins in the market, which not only disrupts market order and violates consumer rights, but also may cause consumers to suffer from allergies and other problems. health risks
Although my country has begun to implement the traceability management and application of meat products, electronic ear tags are mainly used as the main identification. According to the survey, the probability of ear tag falling off is about 10% on average, and the ear tag falling off rate of individual farmers is as high as 50%. %, and due to the many circulation links and easy label counterfeiting, it is impossible to achieve true traceability

Method used

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  • Short-tandem-repeats (STR) primer combination for source-traced identification on pork product and detection kit
  • Short-tandem-repeats (STR) primer combination for source-traced identification on pork product and detection kit
  • Short-tandem-repeats (STR) primer combination for source-traced identification on pork product and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1. Primer pair and kit for identifying pork breeds

[0089] 1. Design and synthesis of primer pairs for identification of pork breeds

[0090] The following 23 porcine microsatellite primer pairs were designed and synthesized. The sequences and primer names are shown in Table 1:

[0091] Table 1 is 23 porcine microsatellite primer pairs

[0092]

[0093]

[0094] 2. Composite primer set and its reaction system for identifying pork breeds

[0095] In order to improve the efficiency of PCR amplification, after repeated tests, 3 composite primer sets (Table 2) were obtained by rationally grouping 23 pairs of primers, and the optimal PCR composition system and amplification conditions (Table 3) were obtained by screening. Optimal multiplex amplification method for labeled loci. All target fragments can be obtained by only 3 PCR reactions for the 23 marker loci.

[0096] Table 2 is the composite primer set used to identify pork breeds

[0097]

[0098] ...

Embodiment 2、23

[0113] Example 2, 23 primer pairs to identify pork individuals

[0114] 1. Genomic DNA extraction

[0115] The following pork samples were collected respectively, 8 Northeast Min pigs (numbered M1-M8), 4 Duroc pigs (numbered D1-D4), 3 good and miscellaneous pigs (numbered L1-L3), and 13 Sanyuan pigs (numbered S1-S13), 4 Yimeng black pigs (numbered Y1-Y4), 13 Taihu pigs (numbered T1-T13), and 25 Beijing black pigs (numbered H1-H25), a total of 70 Pork samples.

[0116] Genomic DNA was extracted from 70 pork samples respectively.

[0117] 2. Multiplex PCR amplification

[0118] Using the genomic DNAs of 70 pork samples as templates, PCR amplification was carried out with the three reaction systems in Table 3 in Example 1, and the amplification procedures were shown in Table 4.

[0119] 3. On-board detection

[0120]The three PCR amplification products of all pork samples were detected on the machine, and the 3730XL sequencer was used for detection. Add 9 μl of molecular we...

Embodiment 3

[0129] Example 3, 23 primer pairs are used for the identification of different breeds of pork

[0130] 1. Genomic DNA extraction

[0131] Same as 1 of Example 2;

[0132] 2. Multiplex PCR amplification

[0133] Same as 2 of Example 2;

[0134] 3. On-board detection

[0135] Same as 3 of Example 2;

[0136] 4. Analysis of results

[0137] The amplification results of 23 primer combinations of 70 pigs of all 7 breeds were analyzed by partial least squares (OPLS-DA) discriminant method. The result is as figure 2 As shown, it can be seen that the Beijing black pig ( blue), northeast pig ( yellow), Duroc pig ( green), ternary pig ( light blue), Taihu pig ( Purple) 5 pigs for very good breed identification.

[0138] Therefore, some pork breeds can also be identified by using the 23 primer combinations of the present invention.

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Abstract

The invention discloses a short-tandem-repeats (STR) primer combination for source-traced identification on a pork product and a detection kit. The invention provides a primer kit for identifying pork. The primer kit is (1) or (2), wherein the (1) consists of primer pairs Sw632, S0155, Sw936, S0005, Sw830, S0225, S0355, Swr1941, Sw2406, S0097, Sw857, S0226, Sw72, Sw122, S0090, IGF1, Sw951, Sw787, Sw2525, S0107, Sw2448, Sw911 and CGA; the (2) consists of a primer group A, a primer group B and a primer group C. According to the method provided by the invention, the selected 23 STR markers have the advantages that the polymorphism is high, the markers are not in linkage, amplified fragments have polymorphism, and genotyping is facilitated; grouped multiplex PCR (Polymerase Chain Reaction) detection can be achieved, the cost is reduced, the time is shortened, and the efficiency is increased.

Description

technical field [0001] The invention relates to the field of traceability detection, in particular to a microsatellite primer combination for traceability identification of pork products and a detection kit thereof. Background technique [0002] In recent years, driven by economic interests, the phenomenon of using low-value products to pretend to be high-value products, counterfeit brands, and counterfeit places of origin has often occurred in the market, which not only disturbs the market order and infringes on the rights and interests of consumers, but also may cause problems for consumers due to allergies and other issues. health risks. Although my country has begun to implement the traceability management and application of meat products, electronic ear tags are mainly used as the main identification. The survey shows that the probability of ear tags falling off is about 10% on average, and the ear tag falling rate of individual farmers is as high as 50%. %, and because...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 陈爱亮杨曙明赵杰朱超蒋小玲
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS