CN111719001A - Primer set, application, kit and method for detecting human orange preference-related SNP loci - Google Patents

Abstract

The invention relates to a primer set, application, kit and method for detecting SNP sites related to human orange preference, and belongs to the field of biotechnology. The invention discloses a series of single nucleotide polymorphism sites related to orange preference, and uses these sites to predict individual orange preference. The present invention provides primer pairs for amplifying these SNP loci, as well as a convenient method for detecting their alleles. The SNP sites disclosed in the present invention have important reference value for detecting the difference in orange preference among individuals in the Chinese population, and also have broad application prospects in the process of developing products related to the taste of oranges in the future.

Description

Primer set, application and kit for detecting SNP loci related to human orange preference and method

technical field

The invention belongs to the field of biotechnology, and in particular relates to a primer set, application, kit and method for detecting SNP sites related to human orange preference.

Background technique

As an important part of the cultures of all countries in the world, diet is the basic guarantee for human survival and development, and it is also an important behavior that affects human life and health. Around the topic of diet, a series of studies have been carried out in many disciplines. Dietary preferences are one of the important topics in dietary research. By revealing the eating habits of different groups of users, it can not only reflect the differences in food culture of different groups, but also provide relevant suggestions for the catering industry and other enterprises.

For the food, beverage, pharmaceutical and other industries, how to discover and satisfy consumers' taste preferences is the core issue to be considered in product design. If the taste of the product is not recognized by consumers, the fate of the product is bound to fail. The results of previous genetic studies have shown that some people like sweet, some like sour, and some people don't like spicy... This is true for both adults and children. The reason for the difference may be hidden in the most mysterious genetic "code" of the body. Different from the traditional taste test model, we can deeply understand the differences of consumers' taste preferences from the level of genetic background. Through scientific qualitative and quantitative research, we can understand consumers' true feelings and preferences for the product itself, provide enterprises with an accurate grasp of the product's market prospects, and provide ideas for product improvement and innovation. However, there are few related studies on genes and specific taste preferences (such as watermelon, orange), and there is not enough evidence to support the relationship between gene variation and orange taste, and it is not enough to guide the development and production of orange taste-related products. Therefore, how to overcome the deficiencies of the prior art is an urgent problem to be solved in the field of biotechnology.

SUMMARY OF THE INVENTION

The purpose of the present invention is to solve the deficiencies of the prior art, and provide a primer set, application, kit and method for detecting SNP sites related to human orange preference.

For achieving the above object, the technical scheme adopted in the present invention is as follows:

A first aspect of the present invention provides a primer set for detecting human orange preference-related SNP sites, the SNP sites corresponding to the primer set are rs1494313, rs72700254, rs17719312, rs1419110, rs10888352, rs4575112, rs4534422, rs6543592, rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、 rs149635739, rs28515851, rs36062241, rs17227288, rs118074899, rs28729663;

The primer set includes 39 pairs of primers, and the primer pairs have the nucleotide sequences shown in SEQ ID NO. 1-78.

The second aspect of the present invention provides the application of the above-mentioned primer set for detecting SNP sites related to human orange preference in preparing a kit for predicting human orange preference.

A third aspect of the present invention provides the above-mentioned kit for predicting human orange preference, the kit comprising the above-mentioned primer set.

Further, preferably, the kit further includes a reagent for detecting the SNP site of claim 1 .

A fourth aspect of the present invention provides the use of a reagent for detecting a plurality of single nucleotide polymorphism sites in the preparation of a detection preparation or a detection device for predicting human orange preference, the plurality of single nucleotide polymorphism sites Spots include the following 39 SNPs:

rs1494313、rs72700254、rs17719312、rs1419110、rs10888352、rs4575112、rs4534422、rs6543592、rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、 rs78404451, rs10732579, rs78025492, rs75062957, rs1038138, rs1761031, rs4900367, rs148847709, rs149635739, rs28515851, rs36062241, rs172297288, rs11722999, rs11963074.

Further, preferably, the rs1494313 allele is A, the rs72700254 allele is G, the rs17719312 allele is T, the rs1419110 allele is T, the rs10888352 allele is A, the rs4575112 allele is G, rs4534422, etc. Allele is G, rs6543592 allele is G, rs72928646 allele is T, rs144657377 allele is G, rs10153756 allele is G, rs74171294 allele is G, rs78301347 allele is G, rs16828741 allele The gene is G, the rs75622246 allele is T, the rs57611848 allele is C, the rs73874387 allele is T, the rs73874391 allele is T, the rs62403686 allele is T, the rs1340612 allele is C, and the rs77159685 allele is Because A, rs116607654 allele is T, rs1441016 allele is A, rs2276320 allele is G, rs117094057 allele is T, rs78404451 allele is C, rs10732579 allele is A, and rs78025492 allele is T, rs75062957 allele is T, rs1038138 allele is C, rs1761031 allele is T, rs4900367 allele is C, rs148847709 allele is G, rs149635739 allele is T, rs28515851 allele is G , rs36062241 allele is T, rs17227288 allele is T, rs118074899 allele is T, rs28729663 allele is A.

Further, preferably, the sample to be tested is from blood, urine, saliva, gastric juice, hair or biopsy of the individual to be tested.

A fifth aspect of the present invention provides a method for detecting the genotype of the SNP locus related to the preference of human oranges, comprising the following steps:

(1) The whole genome DNA of the human to be detected is used as a template, and 38 pairs of primer pairs shown in SEQ ID NO.1-78 are respectively used for PCR amplification to obtain amplified fragments;

(2) The amplified fragment was subjected to agarose gel electrophoresis to separate the target band, the gel was cut to recover the product, and then sanger sequencing was used to determine the genotype of the SNP locus related to human orange preference.

Further, preferably, the PCR amplification system is:

Taq DNA polymerase 1μL, forward primer 10pmol/μL 0.6μL, reverse primer 10pmol/μL 0.6μL, template DNA 2μL, 10x buffer 2μL, dNTP 1.6μL (2.5mM), ddH ₂ O 12.2μL, total 20μL;

The PCR amplification procedure is: pre-heating at 94°C for 3 min to fully denature the template DNA, and then enter the amplification cycle; in each cycle, keep at 94°C for 45s, and then keep at 50-61°C for 45s to make the primer and template Fully annealed and kept at 72°C for 45s to extend the primers on the template to synthesize DNA to complete one cycle; repeat this cycle 30 times; finally, keep at 72°C for 5min to complete the extension of the product and store at 4°C.

A sixth aspect of the present invention provides a detection device for predicting people's preference for oranges, comprising a detection unit and a data analysis unit;

The detection unit includes a detection unit for detecting the condition of the allele of a plurality of single nucleotide polymorphism sites carried by the individual to be tested, and obtains a detection result; wherein, the plurality of single nucleotide polymorphism sites点包括以下39个单核苷酸多态性位点：rs1494313、rs72700254、rs17719312、rs1419110、rs10888352、rs4575112、rs4534422、rs6543592、rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391 、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、rs149635739、rs28515851、rs36062241、rs17227288、rs118074899、rs28729663；

The data analysis unit includes a processing unit for analyzing and processing the detection result of the detection unit, so as to obtain a prediction of personal orange preference.

In the present invention, the data analysis unit preferably uses the constructed random forest model for prediction, and the final prediction score is lower than 1.556 as a favorite, and higher than 1.556 as a dislike.

Among them, the construction method of the random forest model is to use the collected data of 39 loci genotypes and watermelon preferences of people, use randomForest in the R package to simulate, pROC to calculate the efficacy of the model, mtry is 1-4, ntree The value is 500-1000. After evaluation, in the case of mtry=2, ntree=500, the highest AUC value is 0.752, and the random forest model obtained by this is used for prediction.

The present invention provides a series of single nucleotide polymorphism sites (SNPs) associated with orange preference, and uses these sites to predict individual orange preference. The present invention conducts genotyping and phenotypic correlation analysis in more than 1400 Chinese population samples, and obtains 39 SNPs related to the Chinese Han population's orange preference. Based on the 39-locus genotype and orange preference data of 692 people, the construction of The calculation model based on random forest was verified in another 461 people and the ROC curve was drawn. The AUC area was 0.752, and a good prediction effect was obtained.

The physical location of the SNP site of the present invention is determined according to the human reference genome GRCh37/hg19, and the source of the SNP number is dbSNP 151.所述的多个单核苷酸多态性位点包括：rs1494313、rs72700254、rs17719312、rs1419110、rs10888352、rs4575112、rs4534422、rs6543592、rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、 rs73874391、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、rs149635739、rs28515851、rs36062241、rs17227288、rs118074899、rs28729663。 Through these single nucleotide polymorphism sites, the association analysis of orange preference can be carried out.

At the same time, the study found that the related genotypes of these loci are: rs1494313 allele is A, rs72700254 allele is G, rs17719312 allele is T, rs1419110 allele is T, rs10888352 allele is A, rs4575112 Alleles are G, rs4534422 alleles are G, rs6543592 alleles are G, rs72928646 alleles are T, rs144657377 alleles are G, rs10153756 alleles are G, rs74171294 alleles are G, rs78301347, etc. Allele is G, rs16828741 allele is G, rs75622246 allele is T, rs57611848 allele is C, rs73874387 allele is T, rs73874391 allele is T, rs62403686 allele is T, rs1340612 allele The gene is C, the rs77159685 allele is A, the rs116607654 allele is T, the rs1441016 allele is A, the rs2276320 allele is G, the rs117094057 allele is T, the rs78404451 allele is C, and the rs10732579 allele is Because A, rs78025492 allele is T, rs75062957 allele is T, rs1038138 allele is C, rs1761031 allele is T, rs4900367 allele is C, rs148847709 allele is G, and rs149635739 allele is T, rs28515851 allele is G, rs36062241 allele is T, rs17227288 allele is T, rs118074899 allele is T, and rs28729663 allele is A.

The present invention relates to providing primer pairs for amplifying the genotypes of the above loci, as shown in Table 1.

Table 1

Compared with the prior art, the present invention has the following beneficial effects:

(1) The present invention predicts personal orange preference from a genetic perspective, and can easily, quickly and in batches evaluate the orange preference degree in the crowd;

(2) Provided 39 loci related to orange preference in Chinese population;

(3) A primer pair for amplifying the above-mentioned 39 SNP loci was developed, which can be widely used in the identification of the above-mentioned SNP loci in Chinese individuals;

(4) A method for identifying the allele type of the SNP site is provided, which is easy to operate and has accurate and reliable results.

Description of drawings

Figure 1 shows the ROC curve for predicting people's orange preference (Y~X1+X2+...+X39, mtry=2) based on the random forest model, and the AUC area of the prediction model is 0.752.

Detailed ways

The present invention will be further described in detail below in conjunction with the embodiments.

Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. If no specific technology or condition is indicated in the examples, the technology or condition described in the literature in the field or the product specification is used. If the materials or equipment used are not marked with the manufacturer, they are all conventional products that can be obtained through purchase.

Example 1

A primer set for detecting SNP sites related to human orange preference, the SNP sites corresponding to the primer set are rs1494313, rs72700254, rs17719312, rs1419110, rs10888352, rs4575112, rs4534422, rs6543572, rs72928646, rs1437567377, rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、rs149635739、rs28515851、rs36062241、rs17227288、 rs118074899, rs28729663;

The primer set includes 39 pairs of primers, and the primer pairs have the nucleotide sequences shown in SEQ ID NO. 1-78.

Example 2

A kit for predicting human orange preference, the kit includes the primer set described in Example 1.

Example 3

A kit for predicting human orange preference, the kit includes the primer set described in Example 1, and also includes Taq DNA polymerase, 10x buffer, and dNTP.

Example 4

A detection device for predicting people's preference for oranges, comprising a detection unit and a data analysis unit;

The detection unit includes a detection unit for detecting the condition of the allele of a plurality of single nucleotide polymorphism sites carried by the individual to be tested, and obtains a detection result; wherein, the plurality of single nucleotide polymorphism sites点包括以下39个单核苷酸多态性位点：rs1494313、rs72700254、rs17719312、rs1419110、rs10888352、rs4575112、rs4534422、rs6543592、rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391 、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、rs149635739、rs28515851、rs36062241、rs17227288、rs118074899、rs28729663；

The data analysis unit includes a processing unit for analyzing and processing the detection result of the detection unit, so as to obtain a prediction of personal orange preference. Specifically, a random forest model can be constructed based on the existing collected data, and then the random forest model is used for prediction, and it is determined that the final prediction score is lower than 1.556 as a favorite, and higher than 1.556 as a dislike.

Example 5

1. Peripheral blood collection and DNA extraction

Peripheral blood is generally collected by vacuum blood collection tube. The collected whole blood should be quick-frozen with liquid nitrogen within 5 minutes and stored in a -80°C refrigerator. Fresh blood after adding EDTA anticoagulant can generally be placed at 4°C for 12-24 hours. It is important to ensure that the quick-frozen samples are stored at low temperature, and repeated freezing and thawing must be avoided.

The DNA extraction of peripheral blood in the present invention includes the following steps: first, add 5 times the volume of 1× red blood cell lysate to 2 mL of whole blood, vortex and discard the supernatant; then add 0.5 mL of leukocyte lysate and 20 μL of 20 mg/ml proteinase K, Lyse the leukocytes in a water bath at 37°C until the precipitate dissolves; add an equal volume of chloroform to separate layers, transfer the supernatant to a new EP tube, add 1/10 volume of 3M ammonium acetate and an equal volume of ice ethanol to precipitate DNA, and use a volume of 1 mL After washing with 75% ethanol, it was naturally dried and dissolved in TE to obtain DNA. In order to ensure the quality of nucleic acid extraction from subsequent blood samples, more than 1 μg of whole genome DNA should be extracted from each sample, with a concentration of >12.5 ng/μL, and an OD260/280 between 1.8 and 2.0. DNA quality testing, including integrity, total, and concentration testing, can be accomplished with Qubit or AGE instruments. DNA sample content can be detected by one of the following two methods: fluorescence quantification or agarose gel electrophoresis detection. Fluorescence quantification mainly quantifies the sample concentration accurately (such as Qubit Fluorometer), and agarose gel electrophoresis uses 1% agarose gel electrophoresis at 150V for about 40min (buffer is 1x TAE) to test the integrity of the sample and the presence of RNA, protein and secondary metabolite contamination. Blood samples were graded based on the total amount and concentration of DNA. Excellent: The total amount of DNA is ≥2μg, the concentration is ≥12.5ng/μL, and the sample is not degraded or slightly degraded. Good: The total amount of DNA is between 1 and 2 μg, the concentration is greater than or equal to 12.5 ng/μL, and the sample is not degraded or slightly degraded. Try to select DNA samples with excellent quality detection for follow-up work.

2. Single nucleotide polymorphism detection

Using the primer pair designed according to the characteristics of the gene polymorphic region in the present invention, PCR amplification is performed using the above DNA sample as a template to obtain a DNA fragment containing the polymorphic region. details as follows:

Taq DNA polymerase 1μL, forward primer 10pmol/μL 0.6μL, reverse primer 10pmol/μL 0.6μL, template DNA 2μL, 10x buffer 2μL, dNTP 1.6μL (2.5mM), ddH ₂ O 12.2μL, total 20μL;

The PCR amplification procedure is: pre-heating at 94°C for 3 min to fully denature the template DNA, and then enter the amplification cycle; in each cycle, keep at 94°C for 45s, and then keep the annealing temperature shown in Table 1 for 45s to make the primers Fully annealed with the template, kept at 72°C for 45s, extended the primer on the template, synthesized DNA, and completed a cycle; repeated this cycle 30 times; finally kept at 72°C for 5min to make the product extension complete, at 4°C save.

After PCR amplification is completed, agarose gel electrophoresis is used for detection, and the product is recovered by cutting the gel, and then sanger sequencing is used to obtain the genotype of the single nucleotide polymorphism site in the sample.

3. The correlation verification between the single nucleotide polymorphism site involved in the present invention and orange preference

In order to determine that the single nucleotide polymorphisms in the present invention are related to the taste of oranges, 738 cases of people who did not like the taste of oranges were selected and 416 cases of controls were selected for verification. PLINK (v.1.9) was used for single-point association analysis, and the model used was the additive genetic model. The results of the above experiments are summarized in Table 2. The significance of rs75062957, rs78404451, rs148847709, rs1419110 and rs1441016 all reached 10 ^-6 .

Table 2

In order to further verify the correlation between the above loci and orange preference, using the genotype data of 692 people and the results of the orange preference survey, the R package randomForest was used to predict the orange preference based on a random forest model (Y～X1+X2+...+X39 , mtry=2, ntree=500). Validation of the established model in an additional 461 individuals yields an accuracy of 0.706 and a sensitivity of 0.730 when the threshold is set to 1.556. And use the R package pROC to draw the ROC curve, the AUC area of the prediction model is 0.752, reflecting a high prediction effect (Figure 1).

The foregoing has shown and described the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above-mentioned embodiments. The above-mentioned embodiments and descriptions only illustrate the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention will also have Various changes and modifications fall within the scope of the claimed invention. The claimed scope of the present invention is defined by the appended claims and their equivalents.

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<212> DNA

<213> artificial sequence()

<400> 52

acgttggatg ctcaggtcaa gcgtttt 27

<210> 53

<211> 27

<212> DNA

<213> artificial sequence()

<400> 53

acgttggatg ctggctaaca cggtgaa 27

<210> 54

<211> 27

<212> DNA

<213> artificial sequence()

<400> 54

acgttggatg ctttggagac agaatct 27

<210> 55

<211> 27

<212> DNA

<213> artificial sequence()

<400> 55

acgttggatg acaggtgctg gcaagac 27

<210> 56

<211> 27

<212> DNA

<213> artificial sequence()

<400> 56

acgttggatg ataggcatgc atgtgtc 27

<210> 57

<211> 27

<212> DNA

<213> artificial sequence()

<400> 57

acgttggatg aacccgtgtg actttga 27

<210> 58

<211> 27

<212> DNA

<213> artificial sequence()

<400> 58

acgttggatg tagactcgat gatggtt 27

<210> 59

<211> 27

<212> DNA

<213> artificial sequence()

<400> 59

acgttggatg ttgctttggg tagcatg 27

<210> 60

<211> 27

<212> DNA

<213> artificial sequence()

<400> 60

acgttggatg cctcatcgct tatagcc 27

<210> 61

<211> 27

<212> DNA

<213> artificial sequence()

<400> 61

acgttggatg gctctgaaat ccatcac 27

<210> 62

<211> 27

<212> DNA

<213> artificial sequence()

<400> 62

acgttggatg aagaggtcac aaccctg 27

<210> 63

<211> 27

<212> DNA

<213> artificial sequence()

<400> 63

acgttggatg aaatgagctg ggcttct 27

<210> 64

<211> 27

<212> DNA

<213> artificial sequence()

<400> 64

acgttggatg cagcggacac tgctaca 27

<210> 65

<211> 27

<212> DNA

<213> artificial sequence()

<400> 65

acgttggatg atttggacat tgtggcg 27

<210> 66

<211> 27

<212> DNA

<213> artificial sequence()

<400> 66

acgttggatg tgagctcatg ttttgac 27

<210> 67

<211> 27

<212> DNA

<213> artificial sequence()

<400> 67

acgttggatg acctggtata ttcgcac 27

<210> 68

<211> 27

<212> DNA

<213> artificial sequence()

<400> 68

acgttggatg gtttgacatg cagtgca 27

<210> 69

<211> 27

<212> DNA

<213> artificial sequence()

<400> 69

acgttggatg gatacatgca gcattta 27

<210> 70

<211> 27

<212> DNA

<213> artificial sequence()

<400> 70

acgttggatg gtgtctcctt tccaaca 27

<210> 71

<211> 27

<212> DNA

<213> artificial sequence()

<400> 71

acgttggatg acatcccccc gttttat 27

<210> 72

<211> 27

<212> DNA

<213> artificial sequence()

<400> 72

acgttggatg ttctgggagt tgtagtc 27

<210> 73

<211> 27

<212> DNA

<213> artificial sequence()

<400> 73

acgttggatg actgactaca ctgtttc 27

<210> 74

<211> 27

<212> DNA

<213> artificial sequence()

<400> 74

acgttggatg ggaactagtg ggaatca 27

<210> 75

<211> 27

<212> DNA

<213> artificial sequence()

<400> 75

acgttggatg gggtgacggt tacataa 27

<210> 76

<211> 27

<212> DNA

<213> artificial sequence()

<400> 76

acgttggatg cagcttgtcc caacgat 27

<210> 77

<211> 27

<212> DNA

<213> artificial sequence()

<400> 77

acgttggatg ctgtttcttg ttttgga 27

<210> 78

<211> 27

<212> DNA

<213> artificial sequence()

<400> 78

acgttggatg catccatgtc tttcgtc 27

Claims (10)

1. A primer group for detecting SNP sites related to preference of human oranges, which is characterized in that the SNP sites corresponding to the primer group are rs1494313, rs72700254, rs17719312, rs1419110, rs10888352, rs4575112, rs4534422, rs6543592, rs72928646, rs144657377, rs10153756, rs74171294, rs78301347, rs16828741, rs75622246, rs57611848, rs 73874387874, rs73874391, rs62403686, rs1340612, rs 13459659659685, rs116607654, rs1441016, rs2276320, rs117094057, rs 407840404451, rs 107579, rs 02545492, rs75062957, rs1038138, rs4900367, rs 1031847709, rs 14863639, rs 512857241, rs 62281724863, rs 7572969663 and rs 07729663;

the primer group comprises 39 pairs of primer pairs, and the primer pairs have nucleotide sequences shown in SEQ ID NO. 1-78.

2. Use of the primer group for detecting the SNP sites related to the preference of the human oranges as claimed in claim 1 in the preparation of a kit for predicting the preference of the human oranges.

3. A kit for predicting human orange preference, the kit comprising the primer set of claim 1.

4. The kit for predicting human orange preference according to claim 3, further comprising a reagent for detecting the SNP site according to claim 1.

5. The application of a reagent for detecting a plurality of single nucleotide polymorphism sites in preparing a detection preparation or a detection device for predicting the preference of human oranges is characterized in that: the plurality of single nucleotide polymorphic sites comprises the following 39 single nucleotide polymorphic sites:

rs1494313、rs72700254、rs17719312、rs1419110、rs10888352、rs4575112、rs4534422、rs6543592、rs72928646、rs144657377、rs10153756、rs74171294、rs78301347、rs16828741、rs75622246、rs57611848、rs73874387、rs73874391、rs62403686、rs1340612、rs77159685、rs116607654、rs1441016、rs2276320、rs117094057、rs78404451、rs10732579、rs78025492、rs75062957、rs1038138、rs1761031、rs4900367、rs148847709、rs149635739、rs28515851、rs36062241、rs17227288、rs118074899、rs28729663。

6. use of the reagent for detecting multiple single nucleotide polymorphism sites according to claim 5 for preparing a detection preparation or a detection device for predicting preference of human orange, characterized in that:

the allele A, rs72700254 is the allele T, rs1419110 is the allele T, rs10888352 is the allele T, rs4575112 is the allele T, rs4534422 is the allele T, rs72928646 is the allele T, rs144657377 is the allele T, rs 1015353294 is the allele T, rs78301347 is the allele T, rs 168622246 is the allele T, rs 5761618 is the allele T, rs 73387 874387 is the allele T, rs 60301416441644164418180 is the allele T, rs 3641563757 is the allele T, rs 36413741644164416441563757 is the allele T, rs 3641374141413757 is the allele T, rs 364137413741374137413757 is the allele T, rs 3641414141414137413757 364141414141644164416472 is the allele T, rs is the allele 36413641364136413672 is the allele 364141414177413641364141415632413678T, rs is the allele 3641364136413678T, rs is the allele 364136413641364136414136414141413672 is the allele 3641414174 is the allele 36413641364136413641364174 is the allele 40equivalent 36, The rs149635739 allele is T, rs28515851 allele is G, rs36062241 allele is T, rs17227288 allele is T, rs118074899 allele is T, rs28729663 allele is A.

7. Use of the reagent for detecting multiple single nucleotide polymorphism sites according to claim 5 for preparing a detection preparation or a detection device for predicting preference of human orange, characterized in that: the sample to be tested is derived from blood, urine, saliva, gastric juice, hair or biopsy of the subject to be tested.

8. A method for detecting the genotype of SNP loci related to preference of human oranges is characterized by comprising the following steps: the method comprises the following steps:

(1) using human whole genome DNA to be detected as a template, and respectively adopting 38 pairs of primers shown in SEQ ID NO.1-78 to carry out PCR amplification to obtain amplified fragments;

(2) and carrying out agarose gel electrophoresis on the amplified fragment to separate a target band, cutting the gel to recover a product, and then judging the genotype of the SNP locus related to the preference of the human orange by using a sanger sequencing method.

9. The method for detecting the genotype of the SNP site related to orange preference according to claim 8, wherein the PCR amplification system is as follows:

1. mu.L of Taq DNA polymerase, 10 pmol/. mu.L of forward primer 0.6. mu.L of reverse primer, 10 pmol/. mu.L of template DNA 0.6. mu.L, 10 XBuffer 2. mu.L, 1.6. mu.L of dNTP (2.5mM), ddH₂O12.2. mu.L, 20. mu.L in total;

the PCR amplification procedure was: preheating at 94 ℃ for 3min to fully denature the template DNA, and then entering an amplification cycle; in each cycle, maintaining the temperature at 94 ℃ for 45s, then maintaining the temperature at 50-61 ℃ for 45s to ensure that the primer is fully annealed with the template, maintaining the temperature at 72 ℃ for 45s to ensure that the primer is extended on the template to synthesize DNA, and completing one cycle; this cycle was repeated 30 times; finally keeping the temperature at 72 ℃ for 5min to ensure that the product is completely extended and stored at 4 ℃.

10. A detection device for predicting preference of human oranges is characterized by comprising a detection unit and a data analysis unit;

the detection unit comprises a detection unit for detecting the allelic condition of the individual to be detected carrying a plurality of single nucleotide polymorphism sites to obtain a detection result; wherein the plurality of single nucleotide polymorphic sites comprises the following 39 single nucleotide polymorphic sites: rs1494313, rs72700254, rs17719312, rs1419110, rs10888352, rs4575112, rs4534422, rs6543592, rs72928646, rs144657377, rs10153756, rs74171294, rs78301347, rs16828741, rs75622246, rs57611848, rs 73387874, rs73874391, rs62403686, rs 0611342, rs77159685, rs116607654, rs1441016, rs2276320, rs117094057, rs78404451, rs10732579, rs 17278540292, rs75062957, rs 8138, rs 176103110311031, rs 1484900367, rs148847709, rs149635739, rs 51515851, rs36062241, rs 4817288, rs 4811899, rs 287263;

the data analysis unit comprises a processing unit for analyzing and processing the detection result of the detection unit, and the prediction of personal orange preference is obtained.

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