CN105886646A - Primer, probe and kit for evaluating sun protection condition of skin - Google Patents
Primer, probe and kit for evaluating sun protection condition of skin Download PDFInfo
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- CN105886646A CN105886646A CN201610367704.1A CN201610367704A CN105886646A CN 105886646 A CN105886646 A CN 105886646A CN 201610367704 A CN201610367704 A CN 201610367704A CN 105886646 A CN105886646 A CN 105886646A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention discloses a primer, probe and kit for evaluating the sun protection condition of skin. The sequences of the primer and the probe are SEQ ID NO:1-4, SED ID NO:5-8, SEQ ID NO:9-12 and SEQ ID NO:13-16, wherein the 5' ends and the 3'ends of SEQ ID NO:3-4, SEQ ID NO:7-8, SEQ ID NO:11-12 and SEQ ID NO:15-16 are connected with different fluorescence markers respectively. The invention further discloses a method for assisting a user in selecting a correct beautifying method through gene detection. The internal cause and the external cause of sunburn, freckles and pigment accumulation of skin are leant, a user-friendly beautifying healthy solution is provided, and the effects of sun protection beautifying products are maximized.
Description
Technical field
The present invention relates to beauty treatment fields, particularly relate to a kind of primer for assessing antitan agent situation and probe and reagent
Box.
Background technology
SNP (single nucleotide polymorphism) refers to the variation of single core thuja acid on genome, including conversion, transversion, disappearance
And insertion, it is a kind of modal genetic marker, the frequency of carrying in crowd is more than 1%, is widely used in human diseases and examines
The fields such as disconnected, medicine individuation use, Basic of Biology research.Along with different SNP site researchs are goed deep into, by multiple
The combine detection of SNP site and follow-up data analysis, can formulate personalized health for the gene information of different crowd
Solution.
The skin appearance of human body is affected by endogenous cause of ill and exopathogenic factor, and exopathogenic factor includes that ultraviolet, environmental pollution, improper beauty treatment are produced
Product use etc., endogenous cause of ill is then embodied in the Biochemical changes relevant to skin, including protein active, hormone level etc., these changes
Mostly determined by gene.Mostly the skin care method of present stage is to take according to exopathogenic factor, as used sunscreen cream, guarantor
Humectant etc., but these have little effect mostly without maintenance method targetedly, cure the symptoms, not the disease.Along with growth in the living standard,
Increasing for improving the demand of skin smooth and tautness, carrying out specific aim beauty and skin care in conjunction with internal and external reasons will ratio tradition side
Method more advantage, the most accurately understands endogenous cause of ill and seems most important.
Skin sunburn and freckle produce by multiple effect genes, and each gene exists again multiple SNP site, these sites
The enzymatic activity of related gene can be affected.The antitan agent energy that can evaluate and test Different Individual is detected by the SNP site of related gene
Power, it is also possible to whether the relevant sunscreen product of assessment is applicable to tester, makes product effect reach to maximize, combines on this basis
Daily life custom adjusts, and makes sun-proof result promote further.For this detection method, choose accurately Sites Combination and
Subsequent data analysis is particularly important.Having patent report both at home and abroad utilizes SNP site genotype to judge that cosmetics are suitable for skin
Type, but there are still susceptible SNP site and choose inaccurate, testing result the most energetic total score analysis problem.Currently have no knot
Close different genes SNP site combine detection and the report of quantization algorithm assessment antitan agent ability.
Chinese utility model patent 201320009798.7 discloses a kind of gene chip, utilizes this genechip detection beautiful
White and aging related gene SNP site, the SNP site of whitening gene includes SLC45A2 (re35391), KITLG
(rs642742)、ASIP(rs1015362)、TYR(rs1042602)、OCA2(rs1800414)、DCT(rs2031526)、MC1R
(rs885479), the SNP of aging gene include MMP1 (rs1799750), TIMP1 (rs4898), SIRT6 (rs107251),
SOD2 (rs4880), NQO1 (rs1800566), TNFA (rs1800629), EPHX (rs1051740), but the base that this patent uses
Because chip method is cumbersome, it is difficult to be suitable for a large amount of detection.Chinese invention patent application 201180056031.3 has screened one group
The gene (LPL, ADIPOQ, PLN etc.) relevant to skin aging, and according to this group genescreen to a kind of aging resistance new material,
Not mentioned gene-correlation SNP site in this patent.Chinese patent application 201080044917.1 provide one utilize natural instead
Justice polynucleotide or compound modulates FLG gene expression, thus reach prevention or treat the disease relevant to FLG.
United States Patent (USP) US200812286033 induces related gene expression to reach to improve wrinkle by Monoterpenes
Effect.United States Patent (USP) US200912512368 induces ELN and COL3A1 gene by the cosmetics containing iso-amylene isoflavone
Express, reached to improve the effect of skin appearance.United States Patent (USP) US201514618080 and US201313932565 are by skin
The sun-proof position of skin uses Carlina acaulis extract and olive oil extract, makes the related gene including ELN and COL3A1 be subject to
Regulation and control up-regulated expression, thus reach to improve effect of sun-protected skin.
Above patent/patent application all utilizes foreign substance to regulate and control the intragentic expression of body, but in the gene of each individuality
But there are differences SNP site, this method is difficult to reach personalized maintenance or treatment.British patent GB20110021917 detects
The target gene SNP site relevant to cosmetics ingredients Biogenic approach, and then judge whether this product is not suitable for use with person,
The SNP site that this patent relates to has MMP-1 (rs1799750), NQO1 (rs1800566), GSTP1 (rs1695), hGPX1
(rs10504)。
Summary of the invention
It is an object of the invention to provide a kind of primer for assessing antitan agent situation and probe.
For achieving the above object, the present invention provides a kind of primer for assessing antitan agent situation and probe, its feature
It is, described sequence such as SEQ ID NO:1-4, and SEQ ID NO:5-8, and SEQ ID NO:9-12, and SEQ ID NO:13-
16；Wherein 5 ' the ends of SEQ ID NO:3-4,7-8,11-12,15-16 connect different fluorescent labelinies respectively with 3 ' ends.
The present invention provides a kind of test kit for assessing antitan agent situation, it is characterised in that containing described primer
And probe.
The present invention provides a kind of method utilizing the gene test auxiliary correct beauty method of customer selecting, it is characterised in that
Use described primer and probe, or the test kit described in use.
Further, step is:
1) use described in primer and probe or described test kit client is carried out SNP site detection；
2) the antitan agent ability value of client is gone out according to susceptible SNP site combination calculation；Described susceptible SNP site is combined as
MC1R rs2228479、MC1R rs885479、ASIP rs1015362、OCA2rs1800414；
3) combine 1) SNP site testing result and 2) antitan agent ability value, carry out result judgement.
Further, described step 1) be,
Extract the DNA sample of individuality to be detected；
Described primer and probe or described test kit is used to carry out PCR amplification；Preferably, the response procedures of PCR amplification is
50 DEG C of 2min, 95 DEG C of 10min, 95 DEG C of 15s, 60 DEG C of 1min circulate 40 times.
Further, described step 2) be,
The pathogenic risk ratio ratio of each loci gene type according to the combination of known susceptible SNP site calculates skin and shines
Speckle, freckle produce individual integrated risk ratio:
Produce average risk ratio odds (D) by known Chinese population different age group antitan agent, calculate detection
Individual antitan agent produces value-at-risk ratio odds (X), and calculating formula is odds (X)=OR (C) × odds (D)；Calculate comprehensive
Conjunction value-at-risk:
According to known normal genotype crowd's value-at-risk F (N), calculate the sun-proof ability value of individual's skin:
Further, described step 3) result be judged as,
1) if the sun-proof ability value of individual's skin is less than 100%, then it represents that the antitan agent ability of this individuality is less than this age
The normal value of Duan Renqun, sun-proof ability, belong to and be difficult to sun-proof body constitution；Being worth the lowest, sun-proof ability is the poorest；
2) if the sun-proof ability value of individual's skin is equal to 100%, then it represents that the antitan agent ability of this individuality and this age bracket
The normal value of crowd is the same, and sun-proof ability is general；
3) if the sun-proof ability value of individual's skin is higher than 100%, then it represents that the antitan agent ability of this individuality is higher than this age
The normal value of Duan Renqun, sun-proof ability is preferable, belongs to the most sun-proof body constitution；Being worth the highest, sun-proof ability is the best；
Simultaneously；
If SNP site rs2228479 of gene M C1R is A/G, showing to carry heterozygous mutant, antitan agent is less able,
It is easily subject to the damage of ultraviolet so that in a large amount of melanin deposition epidermal areas, with the passing of time can accelerate skin aging；If A/A, table
The bright homozygous mutation that carries, antitan agent is less able, is easily subject to the damage of ultraviolet so that a large amount of melanin deposition epidermal areas
In, with the passing of time can accelerate skin aging；If G/G, then it it is normal genotype；
If SNP site rs885479 of gene M C1R is A/G, show to carry heterozygous mutant, produce sunburn and the wind of freckle
Danger is higher；If A/A, showing to carry homozygous mutation, the risk producing sunburn and freckle is higher；If G/G, then it it is normal gene
Type；
If SNP site rs1015362 of Gene A SIP is C/T, show to carry heterozygous mutant, sensitive to sunlight, can increase
Freckle and sunburn risk；If T/T, showing to carry homozygous mutation, the risk producing sunburn and freckle is lower；If C/C, it is then
Normal genotype, but owing to carrying C, sensitive to sunlight, freckle and sunburn risk can be increased；
If SNP site rs1800414 of gene OCA2 is C/T, showing to carry heterozygous mutant, optimum sudden change, melanin amasss
Tired risk is lower；If C/C, showing to carry homozygous mutation, optimum sudden change, melanin accumulation risk is lower；If T/T, it is then
Normal genotype.
The present invention (1) filters out one group and can verify through too much piece document, and molecular mechanism is clear, can be used for accurately commenting comprehensively
Estimate the SNP site combination of antitan agent ability；(2) high specificity, highly sensitive TaqMan-MGB Probe-detection methods are used
Detect, multiple sample can be detected simultaneously；(3) produce risk by algorithm precise quantification sunburn, freckle to prevent with assessment skin
Solarization ability, provides personalized antitan agent scheme for tester.
The technical solution used in the present invention is: binding molecule Mechanism Study, literature search and clinical data screening, obtains one
The susceptible SNP site that group is relevant with skin sunburn, freckle combines, described combination site be respectively MC1R (rs2228479,
Rs885479), ASIP (rs1015362), OCA2 (rs1800414), the site information of described combination is shown in Table 1, this combination site
Can be with the antitan agent ability of accurate evaluation Different Individual.
Table 1 susceptible SNP site information table
| SNP_ID | Gene |
| rs2228479 | MC1R |
| rs885479 | MC1R |
| rs1015362 | ASIP |
| rs1800414 | OCA2 |
In order to detect described site, design the primer needed for detection and probe according to primer and fluorescent probe principle of optimality,
Described primer and fluorescent probe are following (underscore is SNP site):
MC1R (rs2228479) primer and fluorescent probe sequence:
Forward primer CACCATCGCCAAGAACCGGAA SEQ ID NO:1；
Reverse primer AGCTGCAGGTGATCACGTC SEQ ID NO:2；
Fluorescent probe FAM-AGCAACATGCTGGAGACGGCC-MGB SEQ ID NO:3；
Fluorescent probe VIC-AGCAACGTGCTGGAGACGGCC-MGB SEQ ID NO:4；
MC1R (rs885479) primer and fluorescent probe sequence:
Forward primer ACATCTCCATCTTCTACGCACT SEQ ID NO:5；
Reverse primer CCATGAGCACCAGCATAGCC SEQ ID NO:6；
Fluorescent probe FAM-CCTGCCGCGGGCGCGGCAAGCCG-MGB SEQ ID NO:7；
Fluorescent probe VIC-CCTGCCGCGGGCGCGGCGAGCCG-MGB SEQ ID NO:8；
ASIP (rs1015362) primer and fluorescent probe sequence:
Forward primer CAGGTAGAATGTACTAACCCCT SEQ ID NO:9；
Reverse primer TGAACAAATAGTCCCGACCAG SEQ ID NO:10；
Fluorescent probe FAM-CTGAAACAGTATGAGATGTTT-MGB SEQ ID NO:11；
Fluorescent probe VIC-CTGAAACAGTGTGAGATGTTT-MGB SEQ ID NO:12；
OCA2 (rs1800414) primer and fluorescent probe sequence:
Forward primer TTGTAACACAGTACTTTGCCAT SEQ ID NO:13；
Reverse primer CATTTGGCGAGCAGAATCCC SEQ ID NO:14；
Fluorescent probe FAM-CTCTCTTACAGCATAGGATAT-MGB SEQ ID NO:15；
Fluorescent probe VIC-CTCTCTTACAGCGTAGGATAT-MGB SEQ ID NO:16；
Described loci detection method specifically comprises the following steps that
1, human saliva sample collection.Using non-injurious salivary automatically to flow out acquisition mode, before saliva gathers, 30min needs to use
Foreign material in drinking water cleaning oral cavity, gather 2mL saliva and deposit in saliva preservation liquid for detection.
2, human body complete genome DNA extracts.Use potassium iodide method, with reference to concretely comprising the following steps: take 0.1mL saliva in 1.5mL
In centrifuge tube, adding 500 μ L 0.01mol/L PBS solution and repeatedly blow and beat, 10000rpm is centrifuged 5min；Supernatant, precipitation is gone to add
50 μ L 5mol/LKI solution, concussion mixing, add 100 μ L 0.9%NaCl solution, 150 μ L phenol/chloroform (25: 24, V/V)
Mixed liquor, concussion 30s, 10000rpm are centrifuged 5min；Taking 200 μ L of supernatant, add 200 μ L isopropanols, mixing, 10000rpm is centrifuged
5min；Going supernatant, precipitation to add 1mL absolute ethanol washing, 10000rpm is centrifuged 5min；Abandon dehydrated alcohol, be deposited in super-clean bench
In dry up, add 50 μ L TE buffer solution ,-20 DEG C save backup.
3, PCR reaction system and response procedures.10 μ L reaction systems containUniversal PCR Master Mix
(Applied Biosystems, Applied Biotechnology company limited of the U.S.) 5 μ L, DNA profiling 1 μ L, fluorescent probe (each SNP
The detection correspondence probe in site mixes) (2.5 μMs) (the corresponding primer of detection of each SNP site mixes with primer mixed liquor
It is combined) 1.5 μ L, ddH2O 2.5μL.Response procedures is 50 DEG C of 2min, 95 DEG C of 10min, 95 DEG C of 15s, 60 DEG C of 1min circulations
40 times.Detecting instrument is StepOnePlus real-time fluorescence quantitative PCR instrument (Applied Biosystems, the biological skill of U.S.'s application
Art company limited).
4, interpretation.Use StepOnePlusTMSoftware v2.3 (Applied Biosystems, the U.S.
Applied Biotechnology company limited) it is analyzed, according to different fluorescence signals, draw the genotype of corresponding SNP site.
Antitan agent ability quantization method is as follows:
1, skin sunburn, freckle produce individual integrated risk ratio ratio and calculate.Each locus gene of known detection gained
The sunburn of type, freckle risk ratio are than being respectively OR1、OR2…ORm(literature query is learnt, the results are shown in Table 3), can calculate
Go out skin sunburn, freckle produces individual integrated risk ratio than OR (C), and calculation equation is as follows:
2, the skin sunburn that calculated by step 1, freckle produce individual integrated risk ratio than OR (C), pass through document
Or other statistical approach can obtain Chinese population different age group skin sunburn, freckle produces average risk ratio odds (D)
(known Chinese population different age group antitan agent produce average risk ratio odds (D) be respectively as follows: 20-35 year: 0.06,36-
45 years old: 0.15,46-55 years old: 0.47), can calculate the individual skin sunburn of detection by odds ratio computing formula, freckle is produced
Raw value-at-risk ratio odds (X), calculation equation is as follows:
3, the individual's skin sunburn, the freckle that are calculated by step 2 produce risk ratio odds (X), i.e. can pass through wind
Danger value prediction equation can calculate integrated risk value F (X), and calculation equation is as follows:
4, the antitan agent ability of Different Individual can be assessed by the calculated integrated risk value of step 3, it is known that just
Often genotype crowd's value-at-risk F (N) (known normal genotype crowd's value-at-risk F (N) be respectively as follows: 20-35 year: 5.67%, 36-
45 years old: 13.04%, 46-55 year: 31.97%), individual's skin sun-proof ability value V can be calculated, calculation equation is as follows:
Testing result personality analysis and solution:
According to sun-proof ability quantum chemical method result, in conjunction with function and the molecular mechanism of related locus, it is judged that skin problem
Probability, assessment tester can use the cosmetics containing which kind of composition, and adjust the daily life being conducive to skin protection for it
Dietary habit alive.The molecular mechanism of related gene is as follows: if MC1R carries sudden change, freckle and sunburn produces risk reduction；
ASIP carries sudden change, freckle and sunburn and produces risk reduction；OCA2 carries sudden change, is easily generated melanin.
Beneficial effect:
Skin sunburn, freckle and the endogenous cause of ill of pigment accumulation and exopathogenic factor can be fully understood by, provide the user personalized beauty care
Healthy solution, makes sun-rays proof beauty-care product using effect maximize.Skin protection is conducive to further, it is also possible to adjust for tester
Daily life dietary habit.
Detailed description of the invention
Embodiments of the invention are described below in detail, and the example of described embodiment is being intended to for explaining the present invention, and not
It is understood that as limitation of the present invention.In embodiment, unreceipted concrete technology or condition person, retouched according to the document in this area
The technology stated or condition or carry out according to product description.Agents useful for same or instrument unreceipted production firm person, be permissible
By city available from conventional products.
Embodiment 1: primer and probe design
1, design and synthesize following primer and probe, synthesize unit: Applied Biosystems, the U.S. applies biological skill
Art company limited.
MC1R (rs2228479) primer and fluorescent probe sequence:
Forward primer CACCATCGCCAAGAACCGGAA SEQ ID NO:1；
Reverse primer AGCTGCAGGTGATCACGTC SEQ ID NO:2；
Fluorescent probe FAM-AGCAACATGCTGGAGACGGCC-MGB SEQ ID NO:3；
Fluorescent probe VIC-AGCAACGTGCTGGAGACGGCC-MGB SEQ ID NO:4；
MC1R (rs885479) primer and fluorescent probe sequence:
Forward primer ACATCTCCATCTTCTACGCACT SEQ ID NO:5；
Reverse primer CCATGAGCACCAGCATAGCC SEQ ID NO:6；
Fluorescent probe FAM-CCTGCCGCGGGCGCGGCAAGCCG-MGB SEQ ID NO:7；
Fluorescent probe VIC-CCTGCCGCGGGCGCGGCGAGCCG-MGB SEQ ID NO:8；
ASIP (rs1015362) primer and fluorescent probe sequence:
Forward primer CAGGTAGAATGTACTAACCCCT SEQ ID NO:9；
Reverse primer TGAACAAATAGTCCCGACCAG SEQ ID NO:10；
Fluorescent probe FAM-CTGAAACAGTATGAGATGTTT-MGB SEQ ID NO:11；
Fluorescent probe VIC-CTGAAACAGTGTGAGATGTTT-MGB SEQ ID NO:12；
OCA2 (rs1800414) primer and fluorescent probe sequence:
Forward primer TTGTAACACAGTACTTTGCCAT SEQ ID NO:13；
Reverse primer CATTTGGCGAGCAGAATCCC SEQ ID NO:14；
Fluorescent probe FAM-CTCTCTTACAGCATAGGATAT-MGB SEQ ID NO:15；
Fluorescent probe VIC-CTCTCTTACAGCGTAGGATAT-MGB SEQ ID NO:16；
2, SNP site detection
1), human saliva sample collection.Using non-injurious salivary automatically to flow out acquisition mode, before saliva gathers, 30min needs
With foreign material in drinking water cleaning oral cavity, gather 2mL saliva and deposit in saliva preservation liquid for detection.
2), human body complete genome DNA extracts.Use potassium iodide method, with reference to concretely comprising the following steps: take 0.1mL saliva in 1.5mL
In centrifuge tube, adding 500 μ L 0.01mol/L PBS solution and repeatedly blow and beat, 10000rpm is centrifuged 5min；Supernatant, precipitation is gone to add
50 μ L 5mol/LKI solution, concussion mixing, add 100 μ L 0.9%NaCl solution, 150 μ L phenol/chloroform (25: 24, V/V)
Mixed liquor, concussion 30s, 10000rpm are centrifuged 5min；Taking 200 μ L of supernatant, add 200 μ L isopropanols, mixing, 10000rpm is centrifuged
5min；Going supernatant, precipitation to add 1mL absolute ethanol washing, 10000rpm is centrifuged 5min；Abandon dehydrated alcohol, be deposited in super-clean bench
In dry up, add 50 μ L TE buffer solution ,-20 DEG C save backup.
3), PCR reaction system and response procedures.10 μ L reaction systems containUniversal PCR Master
Mix (Applied Biosystems, Applied Biotechnology company limited of the U.S.) 5 μ L, DNA profiling 1 μ L, fluorescent probe are (each
The detection correspondence probe of SNP site mixes) (2.5 μMs) and primer mixed liquor (the corresponding primer of detection of each SNP site
Mix) 1.5 μ L, ddH2O 2.5μL.Response procedures is 50 DEG C of 2min, 95 DEG C of 10min, and 95 DEG C of 15s, 60 DEG C of 1min follow
Ring 40 times.Detecting instrument is StepOnePlus real-time fluorescence quantitative PCR instrument (Applied Biosystems, U.S.'s application biology
Technology Co., Ltd.).
4), detection program end of run after, use StepOnePlusTMSoftwarev2.3 (AppliedBiosystems,
Applied Biotechnology company limited of the U.S.) testing result is analyzed, draw the genotype (being shown in Table 2) of corresponding SNP.
Table 2 SNP site testing result table
| Detection gene | SNP_ID | Genotype |
| MC1R | rs2228479 | G/G |
| MC1R | rs885479 | A/G |
| ASIP | rs1015362 | T/T |
| OCA2 | rs1800414 | C/C |
3, sun-proof ability quantitative evaluation and personalized solution
1), sunburn, freckle produce integrated risk value and calculate.Learnt by literature query, the ratio of different loci such as table 3
Shown in.
Risk ratio ratio (OR) of table 3 related locus genotype
(1) skin sunburn, freckle produce individual integrated risk ratio ratio and calculate.The OR value of different loci is as shown in table 3, then
(2) calculate skin sunburn by step (1), freckle produces individual integrated risk ratio than OR (C), comprehensive literature
Learn that the ratio odds (D) that Chinese population women 20-35 age bracket skin freckle and sunburn produce is about 0.06 with survey data
(the detection sample of the present embodiment belongs to this age bracket, therefore selects the data of this age bracket), permissible by odds ratio computing formula
Calculate the individual skin freckle of detection and sunburn produce value-at-risk ratio odds (X)=OR (C) × odds (D)=1.54 ×
0.06=0.092.
(3) detection individual risk ratio odds (X) calculated by step (2), integrated risk value
(4) known normal genotype crowd's value-at-risk F (N) is 5.67%, can calculate antitan agent ability value
2), testing result is comprehensively analyzed.The antitan agent ability value of this tester is 67%, the antitan agent ability of heredity
Poor.In combination with the molecular mechanism of related gene, ASIP, OCA2 in this detection individuality carry susceptibility loci as shown in Table 2
Useful homozygous mutation, can reduce sunburn, freckle generation risk；MC1R carries susceptibility loci heterozygous mutant, can increase sunburn, freckle
Produce risk.Therefore, MC1R heterozygous mutant the adverse effect the brought beneficial mutation more than ASIP and OCA2, in daily skin protection
In it is noted that use and can offset the skin care method of MC1R adverse effect.
3), personalized solution: these individual sun-proof ability relatively common people are poor, and the sudden change of MC1R makes skin more be subject to
The infringement of ultraviolet, it is important to note that the precipitation of dermal melanin.In order to offset the adverse effect that gene bring, should be special
Note sun-proof, select the sun-proof articles of oneself SPF (sun protection factor) suitable.Eat more fish can improve antitan agent ability (in fish rich in
Omega-3 unsaturated fatty acid can protect skin DNA pathological changes to be attacked)；Vitamin C has whitening effect, can have
Prevent skin from aoxidizing to effect, help skin to resist ultraviolet damage, it is to avoid black speck, freckle produce；Strengthening skin reply daylight wound
The ability of evil, can arrange in pairs or groups vitamin C and vitamin E.
Although above it has been shown and described that embodiments of the invention, it is to be understood that above-described embodiment is example
Property, it is impossible to be interpreted as limitation of the present invention, those of ordinary skill in the art is without departing from the principle of the present invention and objective
In the case of above-described embodiment can be changed within the scope of the invention, revise, replace and modification.
Claims (7)
1. primer and the probe being used for assessing antitan agent situation, it is characterised in that described sequence such as SEQ ID NO:1-
4, and SEQ ID NO:5-8, and SEQ ID NO:9-12, and SEQ ID NO:13-16；Wherein SEQ ID NO:3-4,7-8,
5 ' the ends of 11-12,15-16 connect different fluorescent labelinies respectively with 3 ' ends.
2. the test kit being used for assessing antitan agent situation, it is characterised in that containing the primer described in claim 1 and spy
Pin.
3. the method utilizing the gene test auxiliary correct beauty method of customer selecting, it is characterised in that use claim 1
Primer and probe, or use claim 2 test kit.
4. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 3, it is characterised in that step
For:
1) use the primer of claim 1 and the test kit of probe or claim 2 that client is carried out SNP site detection；
2) the antitan agent ability value of client is gone out according to susceptible SNP site combination calculation；Described susceptible SNP site is combined as MC1R
rs2228479、MC1R rs885479、ASIP rs1015362、OCA2rs1800414；
3) combine 1) SNP site testing result and 2) antitan agent ability value, carry out result judgement.
5. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 4, it is characterised in that described
Step 1) be,
Extract the DNA sample of client；
Test kit described in primer described in claim 1 and probe or claim 2 is used to carry out PCR amplification；Preferably, PCR expands
The response procedures increased is 50 DEG C of 2min, 95 DEG C of 10min, and 95 DEG C of 15s, 60 DEG C of 1min circulate 40 times.
6. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 4, it is characterised in that described
Step 2) be,
The pathogenic risk ratio ratio of each loci gene type according to the combination of known susceptible SNP site calculates skin sunburn, passeris montani saturati
Speckle produces individual integrated risk ratio:
Produce average risk ratio odds (D) by known Chinese population different age group antitan agent, calculate detection individuality
Antitan agent produce value-at-risk ratio odds (X), calculating formula is odds (X)=OR (C) × odds (D)；Calculate comprehensive wind
Danger value:
According to known normal genotype crowd's value-at-risk F (N), calculate the sun-proof ability value of individual's skin:
7. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 4, it is characterised in that described
Step 3) result be judged as,
1) if the sun-proof ability value of individual's skin is less than 100%, then it represents that the antitan agent ability of this individuality is less than this age bracket people
The normal value of group, sun-proof ability, belong to and be difficult to sun-proof body constitution；Being worth the lowest, sun-proof ability is the poorest；
2) if the sun-proof ability value of individual's skin is equal to 100%, then it represents that the antitan agent ability of this individuality and this age bracket crowd
Normal value the same, sun-proof ability is general；
3) if the sun-proof ability value of individual's skin is higher than 100%, then it represents that the antitan agent ability of this individuality is higher than this age bracket people
The normal value of group, sun-proof ability is preferable, belongs to the most sun-proof body constitution；Being worth the highest, sun-proof ability is the best；
Simultaneously；
If SNP site rs2228479 of gene M C1R is A/G, showing to carry heterozygous mutant, antitan agent is less able, easily
Damaged by ultraviolet so that in a large amount of melanin deposition epidermal areas, with the passing of time can accelerate skin aging；If A/A, show to take
Band homozygous mutation, antitan agent is less able, is easily subject to the damage of ultraviolet so that in a large amount of melanin deposition epidermal areas,
With the passing of time skin aging can be accelerated；If G/G, then it it is normal genotype；
If SNP site rs885479 of gene M C1R is A/G, show to carry heterozygous mutant, produce the risk of sunburn and freckle more
High；If A/A, showing to carry homozygous mutation, the risk producing sunburn and freckle is higher；If G/G, then it it is normal genotype；
If SNP site rs1015362 of Gene A SIP is C/T, show to carry heterozygous mutant, sensitive to sunlight, freckle can be increased
With sunburn risk；If T/T, showing to carry homozygous mutation, the risk producing sunburn and freckle is lower；If C/C, then it is normal
Genotype, but owing to carrying C, sensitive to sunlight, freckle and sunburn risk can be increased；
If SNP site rs1800414 of gene OCA2 is C/T, show to carry heterozygous mutant, optimum sudden change, melanin accumulation wind
Danger is lower；If C/C, showing to carry homozygous mutation, optimum sudden change, melanin accumulation risk is lower；If T/T, then it is normal
Genotype.
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| CN106755390A (en) * | 2016-12-15 | 2017-05-31 | 上海东方杰玛基因生物科技有限公司 | A kind of Primer composition and detection method for skin anti-aging ability genetic test |
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| CN106755390B (en) * | 2016-12-15 | 2020-12-04 | 上海东方杰玛基因生物科技有限公司 | Primer composition for detecting skin anti-aging capability gene and detection method |
| KR20180129695A (en) * | 2017-05-26 | 2018-12-05 | (주)아모레퍼시픽 | Genetic Polymorphic marker for predicting skin-temperature and use thereof |
| KR102610856B1 (en) | 2017-05-26 | 2023-12-06 | (주)아모레퍼시픽 | Genetic Polymorphic marker for predicting skin-temperature and use thereof |
| CN107944224A (en) * | 2017-12-06 | 2018-04-20 | 懿奈(上海)生物科技有限公司 | Build method and the application of skin-related gene standard type database |
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