Primer system, kit and method for simultaneously detecting various derived components in food

Abstract

The invention discloses a primer system for simultaneously detecting various source components in food. The primer system comprises two or more of a soybean primer group A, a common livestock and poultry primer group B, a common livestock and poultry primer group C, a high-value livestock primer group D, a mean-price livestock primer group E and a common poultry primer group F. The invention belongs to the technical field of biological detection, provides a primer system, a kit and a method for simultaneously detecting various source components in food, solves the problem of relatively single source in the existing rapid detection technology, can meet the application scene detection of common livestock and poultry related food, has the advantages of low hardware environment requirement, convenience in implementation, low detection cost and the like, and has a wide application prospect. The method is especially suitable for on-site fast detection and base-layer fast detection.

Description

Technical Field

[0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer system, a kit and a method for simultaneously detecting multiple derived components in food. Background Art

[0002] Adulteration / counterfeiting of meat products cannot be detected by sensory evaluation alone without professional experimental techniques, thus endangering food safety, disrupting industry order, and causing a series of social and economic problems.

[0003] Meat product authenticity testing can be simply divided into two categories in terms of application scenarios: accurate testing and rapid screening. Taking multiplex fluorescence PCR technologies such as Chinese patents ZL 201710052775.7 and ZL201711095505.0 as examples, this type of testing technology has good accuracy and high specificity, but the total cost of laboratories and testing equipment is generally in the millions of yuan level, and professional personnel are required to operate. The charges for common 5-7 source component tests are mostly above 2,000 yuan / batch, which is mainly limited to provincial and developed municipal testing institutions, and it is difficult to meet the testing needs of grassroots institutions and small and medium-sized enterprises, small workshops, and individuals. Rapid testing and screening technologies such as LAMP, HDA, HAA / HPA and other constant temperature amplification detection technologies have the core characteristics of a single reaction temperature and are not restricted by thermal cyclers. They do not require expensive qPCR instruments, and the equipment cost is generally between 5,000 yuan and 50,000 yuan. There are no special requirements for the environment, the professional requirements for operators are not high, and the results are visualized.

[0004] At present, most rapid test kits are centered on single-source detection, and even the multi-in-one test kits are prepared with reagents separately; in daily source detection, due to the wide range of food sources and vague raw material characteristics, it is generally necessary to screen sources such as cattle, sheep, pigs, horses, chickens, ducks, and geese to ensure food safety and combat counterfeiting. Therefore, it is of great significance to provide a primer system, test kit, and method that has low hardware environment requirements and is easy to implement for simultaneous detection of multiple source components in food. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the inventors rationally used LAMP (loop-mediated isothermal amplification) detection technology while fully considering the application scenarios. The primer system and kit provided solved the problem of the relatively single source of existing rapid detection technology, and can meet the application scenario detection of common livestock and poultry-related foods. It has the advantages of low hardware environment requirements, easy implementation, and low detection cost, and is particularly suitable for on-site rapid detection and grassroots rapid detection.

[0006] The purpose of the present invention will be further illustrated by the following detailed description.

[0007] The present invention provides a primer system for simultaneously detecting multiple derived components in food, comprising two or more of a soybean primer group A, a common livestock and poultry primer group B, a common livestock primer group C, a high-value livestock primer group D, an affordable livestock primer group E and a common poultry primer group F; the soybean primer group A comprises: a first outer primer F3 as shown in SEQ ID No.1, a first outer primer B3 as shown in SEQ ID No.2, a first inner primer FIP as shown in SEQ ID No.3, and a first inner primer BIP as shown in SEQ ID No.4; the common livestock and poultry primer group B comprises: a second outer primer F3 as shown in SEQ ID No.5, a second outer primer B3 as shown in SEQ ID No.6, a second inner primer FIP as shown in SEQ ID No.7, and a second inner primer BIP as shown in SEQ ID No.8; the common livestock primer group C comprises: a third outer primer F3 as shown in SEQ ID No.9, a third outer primer B3 as shown in SEQ ID No.10, a first inner primer FIP as shown in SEQ ID No.11, and a first inner primer BIP as shown in SEQ ID No.12. The third inner primer FIP shown in No.11, and the third inner primer BIP shown in SEQ ID No.12; the high-value livestock primer group D includes: the fourth outer primer F3 shown in SEQ ID No.13, the fourth outer primer B3 shown in SEQ ID No.14, the fourth inner primer FIP shown in SEQ ID No.15, and the fourth inner primer BIP shown in SEQ ID No.16; the affordable livestock primer group E includes: the fifth outer primer F3 shown in SEQ ID No.17, the fifth outer primer B3 shown in SEQ ID No.18, the fifth inner primer FIP shown in SEQ ID No.19, and the fifth inner primer BIP shown in SEQ ID No.20; the common poultry primer group F includes: the sixth outer primer F3 shown in SEQ ID No.21, the sixth outer primer B3 shown in SEQ ID No.22, the sixth inner primer FIP shown in SEQ ID No.23, and the sixth inner primer BIP shown in SEQ ID No.24.

[0008] Preferably, the common livestock and poultry refer to cattle, sheep, pigs, horses, chickens, ducks and geese; the common livestock refer to cattle, sheep, pigs and horses; the high-value livestock refer to cattle and sheep; the affordable livestock refer to pigs and horses; and the common poultry refer to chickens, ducks and geese.

[0009] Preferably, the primer system for simultaneously detecting multiple derived components in food consists of soybean primer group A and common livestock and poultry primer group B, or consists of high-value livestock primer group D and affordable livestock primer group E, or consists of high-value livestock primer group D, affordable livestock primer group E and common poultry primer group F, or consists of common livestock primer group C and common poultry primer group F.

[0010] With reference to the realities of fresh meat prices and common eating habits in my country in recent years, and in response to the adulteration / counterfeiting of meat products such as soy protein counterfeit meat puree, chicken and / or duck and / or pork impersonating mutton or beef, the above-mentioned technical scheme is adopted to rationally design a primer system for simultaneously detecting multiple source components in food, avoid cross-reaction or interference between different primer groups, solve the problem of relatively single source of existing rapid detection technology, improve detection efficiency and reduce reagent waste, which can meet the rapid initial screening detection of specific application scenarios of common livestock and poultry related foods, and greatly reduce costs while rectifying the market order.

[0011] Correspondingly, the present invention also provides a kit for simultaneously detecting multiple derived components in food, comprising the primer system, LAMP reagent, reaction enzyme, positive control, negative control and blank control.

[0012] Preferably, the components of the LAMP reagent include 18-22mmol / L Tris-HCl, 8-12mmol / L KCl, 8-12mmol / L (NH4)2SO4, 5-7mmol / L MgCl2, 1.8-2.2mmol / L MgSO4, 1.3-1.5mmol / L dNTP, 0.08-0.12% TritonX-100, and 1-1.4mol / L betaine.

[0013] Preferably, the kit for simultaneously detecting multiple derived ingredients in food further comprises a visual agent; the visual agent is a calcein color-changing agent, which is added to the reaction system during detection. If it turns green, it indicates a positive test result; if it remains orange, it indicates a negative test result.

[0014] Preferably, the kit for simultaneously detecting multiple derived ingredients in food further comprises Sybr green dye, which is added to the reaction system during detection. If the fluorescence spectrum obtained by the fluorescence PCR instrument is an S-type amplification curve, the detection result is positive; if the fluorescence spectrum obtained by the fluorescence PCR instrument is not an S-type amplification curve, the detection result is negative.

[0015] In addition, the present invention also provides a method for simultaneously detecting multiple derived components in food, comprising the following steps:

[0016] S1: Rapidly extract the DNA sample liquid of the food to be tested;

[0017] S2: Using the DNA sample solution as a reaction template, performing LAMP amplification and detection using the kit, and determining the results.

[0018] Preferably, the step S1 comprises: placing the food to be tested into a grinding tube filled with stainless steel beads, adding triple distilled water, grinding the food in a portable grinder, centrifuging, and taking the supernatant to obtain a DNA sample solution;

[0019] In the step S2, the steps of performing LAMP amplification and detection include: adding the primer system, DNA sample solution or positive control or negative control or blank control, and reaction enzyme as described in any one of claims 1 to 3 to the LAMP reagent, mixing, and then performing steps S2.1, S2.2 or S2.3; S2.1: placing a drop of calcein color changing agent on the lid of the reaction tube, tightly covering it, placing it in a constant temperature device, and subjecting it to a program of 65°C, 60min and 80°C, 10min, and then instantaneously centrifuging it to observe the color change; S2.2: adding Sybr green dye to the reaction system, tightly covering it, and placing it in a fluorescent PCR, with 63°C, 15s and 63°C, 45s as one cycle, collecting the fluorescent signal at 63°C, 45s, and performing 45 cycles to observe the amplification curve results; S2.3: placing the lid of the reaction tube in a turbidity meter, and subjecting it to a program of 65°C, 60min and 80°C, 10min, and then observing the amplification value results.

[0020] Preferably, in step S2, the step of result judgment includes: 1) quality control judgment: the positive control result is positive, and the negative control and blank control results are negative, indicating that the kit is effective, and entering the sample detection result judgment step; 2) sample detection result judgment: when the soybean primer group A test result of the sample DNA is negative, it indicates that the sample does not contain soybean-derived components; when the soybean primer group A test result of the sample DNA is positive, it indicates that the sample contains soybean-derived components; when the common livestock and poultry primer group B test result of the sample DNA is negative, it indicates that the sample does not contain cattle, sheep, pigs, horses, chickens, ducks, and geese-derived components; when the common livestock and poultry primer group B test result of the sample DNA is positive, it indicates that the sample contains at least one of cattle, sheep, pigs, horses, chickens, ducks, and geese-derived components; when the common livestock primer group C test result of the sample DNA is negative, it indicates that the sample does not contain cattle, sheep, pigs, horses, chickens, ducks, and geese-derived components. points; when the common livestock primer group C test result of the sample DNA is positive, it indicates that the sample contains at least one of cattle, sheep, pig and horse origin components; when the high-value livestock primer group D test result of the sample DNA is negative, it indicates that the sample does not contain cattle and sheep origin components; when the high-value livestock primer group D test result of the sample DNA is positive, it indicates that the sample contains at least one of cattle and sheep origin components; when the affordable livestock primer group E test result of the sample DNA is negative, it indicates that the sample does not contain pig and horse origin components; when the affordable livestock primer group E test result of the sample DNA is positive, it indicates that the sample contains at least one of pig and horse origin components; when the common poultry primer group F test result of the sample DNA is negative, it indicates that the sample does not contain chicken, duck and goose origin components; when the common poultry primer group F test result of the sample DNA is positive, it indicates that the sample contains at least one of chicken, duck and goose origin components.

[0021] Compared with the prior art, the beneficial effects of the present invention include:

[0022] (1) The present invention provides a primer system for simultaneously detecting multiple source components in food, which has good specificity and avoids cross-reaction or interference between different primer sets, thus solving the problem of relatively single source of existing rapid detection technology.

[0023] (2) The kit and method for simultaneously detecting multiple source components in food provided by the present invention have the characteristics of low hardware environment requirements, simpler and faster (when converted to a single source component, the detection efficiency is improved by 1-6 times), and lower detection cost (when converted to a single source component, the detection cost can be as low as 7-25 yuan / batch), etc., which improves the detection efficiency and reduces the waste of reagents, can meet the application scenario detection of common livestock and poultry related foods, and is more suitable for on-site rapid detection and grassroots rapid detection. When using calcein color changing agent, the result can be judged directly by observing the obvious orange / green color change phenomenon with the naked eye, which is conducive to strengthening daily supervision and combating counterfeit and shoddy products, and greatly reducing costs while rectifying the market order. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The fluorescence spectra of the samples detected by the LAMP detection method (using the visual method) of Example 3 of the present invention and the positive, negative and blank controls of the kit are shown.

[0025] Figure 2 The fluorescence spectra of the positive, negative and blank controls of the LAMP detection method (using fluorescence method) detection kit of Example 3 of the present invention are shown.

[0026] Figure 3 This is the fluorescence spectrum of the sample detected by the LAMP detection method (using fluorescence method) of Example 3 of the present invention.

[0027] Figure 4 The turbidity spectra of the positive, negative and blank controls of the LAMP detection method (using turbidity method) detection kit of Example 3 of the present invention are shown.

[0028] Figure 5 This is the turbidity spectrum of the sample detected by the LAMP detection method (using turbidity method) of Example 3 of the present invention. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below with reference to the accompanying drawings and embodiments.

[0030] In the present invention, the reagents and materials involved are conventional commercial products, or can be obtained by conventional technical means in the art. Unless otherwise specified, % refers to volume percentage.

[0031] Example 1 LAMP primer system for simultaneous detection of multiple derived ingredients in food

[0032] When selecting relevant species sequences in each genomic library, the present invention only selects common species with correct names and clear sources to improve accuracy, specifically domestic cattle (Bos taurus domestica), domestic sheep including domesticated goats (Capraaegagrus hircus) and sheep (Ovis aries), domestic pigs (Sus scrofa domestica), domestic horses (Equuscaballus caballus), domestic chickens (Gallusgallus domesticus), domestic ducks (Anasplatyrhynchosdomesticus), domestic geese including Chinese geese (Anser cygnoides domestica) and European geese (Anseranserdomestica).

[0033] The present invention first screens target genes based on mitochondria. As extranuclear genetic material, mitochondria have a higher mutation rate than the nuclear genome, and have extensive polymorphism and high specificity both between and within species, which is more conducive to comparing the differences between species. However, whether it is suitable for range detection and whether it can achieve a balance between breadth and specificity is difficult for technical personnel in this field to predict.

[0034] In this study, more than three genes to be tested were collected from cattle, sheep, pig, horse, chicken, duck and goose in the Genbank of NCBI, and more than 30 data were collected for each gene. The conserved gene fragments within 2 kbp were aligned using the MegAlign software in Lasergene v7.1.0, and the primer sets were generated using the loop-mediated primer online design software LAMP primer designing softwaveprimerexplorerV5.0 of Japan Eigan Co., Ltd., as shown in Table 1.

[0035] Table 1 Primer sets for seven animal-derived specific genes

[0036]

[0037] Example 2 LAMP Primer Screening of Multiple Origin Ingredients in Poultry and Livestock Related Foods

[0038] This example purchased synthesis services from Sangon Biotech (Shanghai) Co., Ltd. 100 μmol / L of FIP, BIP, F3, and B3 independent primer solutions of each primer set were prepared into mixed solutions with final concentrations of 8, 8, 2, and 2 μmol / L.

[0039] Positive controls and negative controls can be obtained by purchasing quality control samples, extracting fresh meat, etc. In this embodiment, nucleic acid quality control samples derived from soybeans, cattle, sheep, pigs, horses, chickens, ducks, and geese were purchased from the mall Beina Chuanglian Biotechnology Co., Ltd.

[0040] The blank control was triple-distilled water.

[0041] Reaction solution: concentration is 20mmol / LTris-HCl, 10mmol / L KCl, 10mmol / L(NH4)2SO4, 6mmol / LMgCl2, 2mmol / LMgSO4, 1.4mmol / L dNTP, 0.1% TritonX-100, and 1.2mol / L betaine.

[0042] Reaction enzyme: Bst polymerase, purchased from Sangon Biotechnology (Shanghai) Co., Ltd. in this example, with an enzyme activity of 8 U / μL.

[0043] Visual agent: calcein indicator, purchased from Guangzhou Diao Biotechnology Co., Ltd. in this example, turns green when the test result is positive, and yellow when the test result is negative.

[0044] Reaction formula: Add 22 μL of the reaction solution, 1 μL of a primer set, 1 μL of Bst polymerase, and 1 μL of the positive, negative, and blank controls corresponding to the primer set to a 100 μL reaction tube, and mix gently. Visual agent usage: Add 1 μL of calcein indicator to the reaction tube cap and cover tightly.

[0045] Reaction conditions: 65°C, 60 min, 80°C, 10 min. The incubation equipment was Shanghai Jingqi Instrument MK-10S dry thermostat, and the reaction was centrifuged at 3krpm immediately after completion.

[0046] Verification method: Determine based on the expected results of positive, negative and blank controls.

[0047] The results of LAMP primer validation are shown in Table 2.

[0048] Table 2 LAMP primer verification results

[0049]

[0050]

[0051]

[0052] Note: “+” indicates positive; “-” indicates negative; “ / ” indicates not tested.

[0053] Example 3: Verification of the LAMP kit assembly for multiple derived ingredients in livestock-related foods

[0054] Some problematic samples accumulated during actual inspections were selected for testing: meat floss, fresh beef, mutton skewers, and bursting meatballs.

[0055] 1. Basic components of the kit

[0056] ① Primer solution: This example purchased synthesis services from Sangon Biotech (Shanghai) Co., Ltd. 100 μmol / L of FIP, BIP, F3, and B3 independent primer solutions in primer sets A, B, C, D, E, and F were respectively prepared into mixed solutions with final concentrations of 8, 8, 2, and 2 μmol / L, with a specification of 100 μL in a 1.5 mL centrifuge tube.

[0057] ② Positive control: Take the primers of primer group A, B, C, D, E, F, F3, B3 independent primer solution, dilute to 10μmol / L respectively. Use the above diluted primers to amplify the target gene in the nucleic acid quality control samples from soybean, cattle, sheep, pig, horse, chicken, duck, goose, and transfer each amplified target band into the pMV-20T vector after gel excision and recovery, replicate with competent E.coli cells JM109, extract the plasmid and dilute to 10 6 Finally, the plasmid was diluted and mixed to 10 5 The positive plasmid control with 100 copies / μL is 100 μL in a 1.5 mL centrifuge tube.

[0058] ③Negative control: DNA extracted from salmon sperm, concentration 10 5 –10 6 copies / μL, the specification is 100μL in a 1.5mL centrifuge tube.

[0059] ④ Blank control: Same as Example 2, specification is 100 μL in a 1.5 mL centrifuge tube.

[0060] ⑤ Reaction enzyme: Same as Example 2, specification is 100 μL in a 1.5 mL centrifuge tube.

[0061] ⑥ Reaction solution: Same as in Example 2, with the specification of 1 mL in a 1.5 mL centrifuge tube.

[0062] 2. Method Validation

[0063] 2.1 Visual verification:

[0064] ① Visual agent: calcein indicator.

[0065] ② Reaction formula: add 22 μL of the reaction solution, 1 μL of a primer set, 1 μL of Bst polymerase, and then add 1 μL of the positive, negative, blank control or sample corresponding to the primer set into a 100 μL reaction tube, and mix gently.

[0066] ③Reaction conditions: same as in Example 2.

[0067] ④ Result determination: After the reaction is completed, it turns green, indicating a positive test result, and turns yellow, indicating a negative test result.

[0068] 2.2 Fluorescence verification:

[0069] ① Fluorescent agent: 100 μL Sybr green solution.

[0070] ② Reaction formula: add 22 μL of the reaction solution, 1 μL of a primer set, 1 μL of Bst polymerase, and 1 μL of the positive, negative, blank control or sample corresponding to the primer set into a 100 μL reaction tube, mix gently, and then add 1 μL of fluorescent agent.

[0071] ③ Reaction conditions: 63℃15s, 63℃45s as one cycle, collecting fluorescence signals at 63℃45s, 45 cycles, fluorescence PCR instrument is ABI Step one plus.

[0072] ④ Result determination: If the fluorescence spectrum is an S-shaped amplification curve, it indicates that the test result is positive; if it is not an S-shaped amplification curve, it indicates that the test result is negative.

[0073] 2.3 Turbidimetric verification:

[0074] ① Reaction formula: add 22 μL of the reaction solution, 1 μL of a primer set, 1 μL of Bst polymerase, and then add 1 μL of the positive, negative, blank control or sample corresponding to the primer set into a 100 μL reaction tube, and mix gently.

[0075] ② Reaction conditions: 65℃, 60min, 80℃, 10min. Turbidimeter is LA320C.

[0076] ③Result determination: If the amplification exceeds the threshold, it indicates that the test result is positive; if it does not exceed the threshold, it indicates that the test result is negative.

[0077] 3. Testing process: First, conduct preliminary research on the sample ingredient list, appearance, etc.

[0078] ① Meat floss: no package, about 250g, the ingredient list of the large package clearly stated that it contained meat floss and soybean powder. Therefore, the kit with a primer system composed of soybean primer group A and common livestock and poultry primer group B was selected for testing.

[0079] ② Fresh beef: agricultural product, about 2kg in total, clearly stated on the bill as beef. Based on the appearance, chicken, duck, and goose meat were ruled out, so a kit consisting of a primer system consisting of high-value livestock primer set D and low-cost livestock primer set E was selected for testing.

[0080] ③ Lamb skewers: catering food, 10 skewers weigh about 500g, and the merchant claims it is lamb. Based on the appearance and general price, a kit with a primer system consisting of high-value livestock primer group D, affordable livestock primer group E, and common poultry primer group F was selected for detection.

[0081] ④ Exploding meatballs: Bulk weighing, 12 pieces are about 250g, and the ingredients of the large package indicate that it contains beef, pork, and chicken. Therefore, the kit with the primer system composed of common livestock primer group C and common poultry primer group F was selected for detection.

[0082] The primer system combinations of some typical cases are shown in Table 3.

[0083] Table 3 Primer system combinations of some typical cases

[0084]

[0085]

[0086] The test results are shown in Table 4, which can quickly identify the problem. The results of visual, fluorescence, and turbidity methods are as follows: Figures 1 to 5 As shown, it can be seen that the positive, negative and blank controls are in line with expectations and the sample test results are normal. Figure 2 In the figure, curves 1-6 are the S-type amplification curves of the positive control, and curves 7-18 are the amplification curves of the negative control and the blank control, respectively. Figure 3 Among them, curves 19-22 are the amplification curves of the detected sources of meat floss, fresh beef, mutton skewers, and bursting meatballs, and curves 23-26 are the amplification curves of the undetected sources of meat floss, fresh beef, mutton skewers, and bursting meatballs. Figure 4 In the figure, curves 1-6 are turbidity detection curves of positive control, and curves 7-18 are turbidity detection curves of negative control and blank control, respectively. Figure 5 Among them, curves 19-22 are the turbidity detection curves of the detected sources of meat floss, fresh beef, mutton skewers, and bursting meatballs, and curves 23-26 are the turbidity detection curves of the undetected sources of meat floss, fresh beef, mutton skewers, and bursting meatballs.

[0087] In-depth confirmation is carried out according to the needs (Table 5). This laboratory adopts relevant standard methods such as GB / T 38164-2019 and SN / T 3730.5-2013 to conduct inspections on cattle, sheep, pigs, horses, chickens, ducks and geese, so as to clean up the consumption environment and support market supervision.

[0088] Table 4 Test results

[0089]

[0090]

[0091] Table 5 Verification results of relevant standard methods

[0092]

[0093] 4. Method comparison: For the common cases mentioned above, single-source and multi-source rapid detection methods were compared with the fluorescent PCR method. As shown in Table 6, it can be seen that the multi-source rapid detection method has the characteristics of low hardware environment requirements, simpler and faster (when converted to a single source component, the detection efficiency is increased by 1-6 times), and lower detection cost (when converted to a single source component, the detection cost can be as low as 7-25 yuan / batch). It is more suitable for on-site rapid detection and grassroots rapid detection, which is conducive to strengthening daily supervision and combating counterfeit products.

[0094] Table 6 Method comparison

[0095]

[0096] 5. Verification in different laboratories: The samples of this embodiment and the kit were sent to two testing units in the same industry in Guizhou Province. Laboratory testing was also carried out in accordance with GB / T 38164-2019 and SN / T 3730.5-2013, and rapid testing was carried out according to the instructions of the kit. The experimental results were consistent.

[0097] In summary, the primer system for simultaneously detecting multiple derived components in food provided by the present invention has good specificity, and there is no cross-reaction or interference between different primer groups. The provided kit and method have the advantages of simplicity, speed, simple operation and low detection cost. It can meet the application scenario detection and rapid screening of common livestock and poultry-related foods and has good practical value.

[0098] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.

Claims

1. A primer system for simultaneously detecting multiple derived components in food, characterized in that: Including two or more of soybean primer group A, common livestock and poultry primer group B, common livestock primer group C, high-value livestock primer group D, affordable livestock primer group E and common poultry primer group F; The soybean primer set A comprises: a first outer primer F3 as shown in SEQ ID No.1, a first outer primer B3 as shown in SEQ ID No.2, a first inner primer FIP as shown in SEQ ID No.3, and a first inner primer BIP as shown in SEQ ID No.4; the common livestock and poultry primer set B comprises: a second outer primer F3 as shown in SEQ ID No.5, a second outer primer B3 as shown in SEQ ID No.6, a second inner primer FIP as shown in SEQ ID No.7, and a second inner primer BIP as shown in SEQ ID No.8; the common livestock primer set C comprises: a third outer primer F3 as shown in SEQ ID No.9, a third outer primer B3 as shown in SEQ ID No.10, a third inner primer FIP as shown in SEQ ID No.11, and a third inner primer BIP as shown in SEQ ID No.12; the high-value livestock primer set D comprises: a fourth outer primer F3 as shown in SEQ ID No.13, a fourth outer primer B3 as shown in SEQ ID No.14, a fourth inner primer FIP as shown in SEQ ID No.15, a fourth inner primer BIP as shown in SEQ ID No.16; The fourth inner primer FIP shown in No.15, and the fourth inner primer BIP shown in SEQ ID No.16; the affordable livestock primer group E includes: the fifth outer primer F3 shown in SEQ ID No.17, the fifth outer primer B3 shown in SEQ ID No.18, the fifth inner primer FIP shown in SEQ ID No.19, and the fifth inner primer BIP shown in SEQ ID No.20; the common poultry primer group F includes: the sixth outer primer F3 shown in SEQ ID No.21, the sixth outer primer B3 shown in SEQ ID No.22, the sixth inner primer FIP shown in SEQ ID No.23, and the sixth inner primer BIP shown in SEQ ID No.

24.

2. The primer system for simultaneously detecting multiple derived components in food according to claim 1, characterized in that: The common livestock and poultry refer to cattle, sheep, pigs, horses, chickens, ducks and geese; the common livestock refer to cattle, sheep, pigs and horses; the high-value livestock refer to cattle and sheep; the affordable livestock refer to pigs and horses; the common poultry refer to chickens, ducks and geese.

3. The primer system for simultaneously detecting multiple derived components in food according to claim 1 or 2, characterized in that: It is composed of soybean primer group A and common livestock and poultry primer group B, or of high-value livestock primer group D and affordable livestock primer group E, or of high-value livestock primer group D, affordable livestock primer group E and common poultry primer group F, or of common livestock primer group C and common poultry primer group F.

4. A kit for simultaneously detecting multiple derived components in food, characterized in that: The method comprises the primer system according to any one of claims 1 to 3, a LAMP reagent, a reaction enzyme, a positive control, a negative control, and a blank control.

5. The kit for simultaneously detecting multiple derived components in food according to claim 4, characterized in that: The components of the LAMP reagent include 18-22mmol / LTris-HCl, 8-12mmol / LKCl, 8-12mmol / L(NH4)2SO4, 5-7mmol / LMgCl2, 1.8-2.2mmol / L MgSO4, 1.3-1.5mmol / L dNTP, 0.08-0.12% TritonX-100, and 1-1.4mol / L betaine.

6. The kit for simultaneously detecting multiple derived components in food according to claim 4 or 5, characterized in that: It also includes a visual agent; the visual agent is a calcein color-changing agent, which is added to the reaction system during detection. If it turns green, it indicates that the test result is positive, and if it remains orange, it indicates that the test result is negative.

7. The kit for simultaneously detecting multiple derived components in food according to claim 4 or 5, characterized in that: It also includes Sybr green dye; when added to the reaction system during detection, if the fluorescence spectrum obtained by the fluorescence PCR instrument is an S-type amplification curve, it indicates that the test result is positive; if the fluorescence spectrum obtained by the fluorescence PCR instrument is not an S-type amplification curve, it indicates that the test result is negative.

8. A method for simultaneously detecting multiple derived components in food, characterized in that: The steps include: S1: Rapidly extract the DNA sample of the food to be tested; S2: Using the DNA sample solution as a reaction template, performing LAMP amplification and detection using a kit as described in any one of claims 4 to 7, and determining the results.

9. The method for simultaneously detecting multiple derived components in food according to claim 8, characterized in that: The step S1 comprises: placing the food to be tested into a grinding tube filled with stainless steel beads, adding triple distilled water, grinding the food in a portable grinder, centrifuging the mixture, and taking the supernatant to obtain a DNA sample solution; In the step S2, the steps of performing LAMP amplification and detection include: adding the primer system, DNA sample solution or positive control or negative control or blank control, and reaction enzyme as described in any one of claims 1 to 3 to the LAMP reagent, mixing, and then performing steps S2.1, S2.2 or S2.3; S2.1: placing a drop of calcein color changing agent on the lid of the reaction tube, tightly covering it, placing it in a constant temperature device, and subjecting it to a program of 65°C, 60min and 80°C, 10min, and then instantaneously centrifuging it to observe the color change; S2.2: adding Sybr green dye to the reaction system, tightly covering it, and placing it in a fluorescent PCR, with 63°C, 15s and 63°C, 45s as one cycle, collecting the fluorescent signal at 63°C, 45s, and performing 45 cycles to observe the amplification curve results; S2.3: placing the lid of the reaction tube in a turbidity meter, and subjecting it to a program of 65°C, 60min and 80°C, 10min, and then observing the amplification value results.

10. The method for simultaneously detecting multiple derived components in food according to claim 8 or 9, characterized in that: In the step S2, the step of result judgment includes: 1) quality control judgment: the positive control result is positive, and the negative control and blank control results are negative, indicating that the kit is effective, and entering the sample test result judgment step; 2) sample test result judgment: when the soybean primer group A test result of the sample DNA is negative, it indicates that the sample does not contain soybean-derived components; when the soybean primer group A test result of the sample DNA is positive, it indicates that the sample contains soybean-derived components; when the common livestock and poultry primer group B test result of the sample DNA is negative, it indicates that the sample does not contain cattle, sheep, pigs, horses, chickens, ducks, and geese-derived components; when the common livestock and poultry primer group B test result of the sample DNA is positive, it indicates that the sample contains at least one of cattle, sheep, pigs, horses, chickens, ducks, and geese-derived components; when the common livestock primer group C test result of the sample DNA is negative, it indicates that the sample does not contain cattle, sheep, pigs, and horses-derived components; When the test result of common livestock primer group C of sample DNA is positive, it indicates that the sample contains at least one of cattle, sheep, pig and horse origin components; when the test result of high-value livestock primer group D of sample DNA is negative, it indicates that the sample does not contain cattle and sheep origin components; when the test result of high-value livestock primer group D of sample DNA is positive, it indicates that the sample contains at least one of cattle and sheep origin components; when the test result of affordable livestock primer group E of sample DNA is negative, it indicates that the sample does not contain pig and horse origin components; when the test result of affordable livestock primer group E of sample DNA is positive, it indicates that the sample contains at least one of pig and horse origin components; when the test result of common poultry primer group F of sample DNA is negative, it indicates that the sample does not contain chicken, duck and goose origin components; when the test result of common poultry primer group F of sample DNA is positive, it indicates that the sample contains at least one of chicken, duck and goose origin components.

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