CN105861719A - Primer, probe and kit for evaluating skin anti-aging condition - Google Patents
Primer, probe and kit for evaluating skin anti-aging condition Download PDFInfo
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- CN105861719A CN105861719A CN201610365084.8A CN201610365084A CN105861719A CN 105861719 A CN105861719 A CN 105861719A CN 201610365084 A CN201610365084 A CN 201610365084A CN 105861719 A CN105861719 A CN 105861719A
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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, a probe and a kit for evaluating the skin anti-aging condition. The sequences of the primer and the probe are shown as SEQ ID NO:1-4, SEQ ID NO:5-8, SEQ ID NO:9-12, SEQ ID NO:13-16, SEQ ID NO:17-20, SEQ ID NO:21-22 and SEQ ID NO:23-24, wherein the ends 5' and the ends 3' of SEQ ID NO:3-4, 7-8, 11-12, 15-16 and 19-20 are connected with different fluorescent marks respectively. The invention further discloses a method for assisting a client in selecting a correct beautifying method through gene detection. Internal causes and external causes generated by skin aging of the client are known, an individual beauty and health solution is provided for the client, and the effect of an anti-aging beautifying product is maximal.
Description
Technical field
The present invention relates to beauty treatment fields, particularly relate to a kind of primer for assessing skin anti-aging situation and probe and test kit.
Background technology
SNP (single nucleotide polymorphism) refers to the variation of single core thuja acid on genome, including conversion, transversion, disappearance and
Insert, be a kind of modal genetic marker, in crowd, carry frequency more than 1%, be widely used in diagnosing human disease,
The fields such as the use of medicine individuation, Basic of Biology research.Along with different SNP site researchs are goed deep into, by multiple SNP
The combine detection in site and follow-up data analysis, can formulate personalized health for the gene information of different crowd and solve
Scheme.
The skin appearance of human body is affected by endogenous cause of ill and exopathogenic factor, and exopathogenic factor includes ultraviolet, environmental pollution, improper cosmetics
Using, endogenous cause of ill is then embodied in the Biochemical changes relevant to skin, including protein active, hormone level etc., and these changes
Mostly determined by gene.Mostly the skin care method of present stage is to take according to exopathogenic factor, as use sunscreen cream,
Wetting agent etc., but these have little effect mostly without maintenance method targetedly, cure the symptoms, not the disease.Along with carrying of living standard
Height, 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 be than biography
System method more advantage, the most accurately understands endogenous cause of ill and seems most important.
Be there is again multiple SNP site by multiple effect genes, each gene in skin aging process, these sites can affect phase
The enzymatic activity of correlation gene.The skin anti-aging ability that can evaluate and test Different Individual is detected, also by the SNP site of related gene
Can assess whether relevant anti-aging beauty-care product is applicable to tester, make product effect reach to maximize, tie on this basis
Close daily life custom to adjust, make cosmetic result promote further.For this detection method, choose Sites Combination accurately
And subsequent data analysis is particularly important.Having patent/patent applications report both at home and abroad utilizes SNP site genotype to judge beauty treatment
Product is suitable for skin type, but there are still susceptible SNP site and choose inaccurate, testing result the most energetic total score analysis and ask
Topic.Currently have no and combine different genes SNP site combine detection and the report of quantization algorithm assessment skin anti-aging ability.
Chinese utility model patent 201320009798.7 discloses a kind of gene chip, utilize this genechip detection whitening and
Aging related gene SNP site, the SNP site of whitening gene include 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 gene chip side that this patent uses
Method is cumbersome, it is difficult to be suitable for a large amount of detection.Chinese invention patent application 201180056031.3 screened one group old with skin
Change relevant gene (LPL, ADIPOQ, PLN etc.), and according to this group genescreen to a kind of aging resistance new material, this is special
Not mentioned gene-correlation SNP site in profit application.Chinese patent application 201080044917.1 provide one utilize natural instead
Justice polynucleotide or compound are used for regulating and controlling 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 the effect of wrinkle by Monoterpenes.
United States Patent (USP) US200912512368 induces the expression of ELN and COL3A1 gene by the cosmetics containing iso-amylene isoflavone,
Thus reach to improve the effect of skin appearance.United States Patent (USP) US201514618080 and US201313932565 are by skin
Old and feeble position uses Carlina acaulis extract and olive oil extract, makes the related gene including ELN and COL3A1 be adjusted
Control up-regulated expression, thus reach to improve effect of ageing skin.
Above patent/patent application all utilizes foreign substance to regulate and control the intragentic expression of body, but deposits in the gene of each individuality
In difference SNP site, this method is difficult to reach personalized maintenance or treatment.British patent GB20110021917 have detected
The target gene SNP site relevant to cosmetics ingredients Biogenic approach, and then judge that 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 skin anti-aging situation and probe.
For achieving the above object, the present invention provides a kind of primer for assessing skin anti-aging situation and probe, and its feature exists
In, 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,
With SEQ ID NO:17-20, and SEQ ID NO:21-22, and SEQ ID NO:23-24；Wherein SEQ ID
5 ' the ends of NO:3-4,7-8,11-12,15-16,19-20 connect different fluorescent labelinies respectively with 3 ' ends.
The present invention also provides for a kind of test kit for assessing skin anti-aging situation, it is characterised in that containing described primer
And probe.
The present invention also provides for a kind of method utilizing the gene test auxiliary correct beauty method of customer selecting, it is characterised in that make
With described primer and probe, or gene test is utilized to assist the correct beauty method of customer selecting described in the test kit described in using
Method, it is characterised in that step is:
1) use described in primer and probe or described test kit client is carried out SNP site detection；
2) the skin anti-aging ability value of client is gone out according to susceptible SNP site combination calculation；Described susceptible SNP site is combined as
SOD2rs4880、SOD2rs5746136、NQO1rs1800566、GPX3rs3828599、GPX1rs1050450、
GSTM1GSTM1-null、GSTT1GSTT1-null；
3) combine 1) SNP site testing result and 2) skin anti-aging ability value, carry out result judgement.
Further, described step 1) be,
Extract the DNA sample of client；
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
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 integrated risk ratio
Ratio OR (C), calculation equation is:
Produce average risk ratio odds (D) by known Chinese population different age group skin aging, calculate detection individuality
Skin aging produce value-at-risk ratio odds (X), calculating formula is odds (X)=OR (C) × odds (D)；
Calculate integrated risk value:
According to known normal genotype crowd's value-at-risk F (N), calculate individual's skin defying age ability value:
Further, described step 3) result be judged as,
1) if individual's skin defying age ability value is less than 100%, then it represents that the skin anti-aging ability of this individuality is less than this age
The normal value of Duan Renqun, defying age ability, belong to the oldest and the most feeble body constitution；Ability value is the lowest, and defying age ability is the most weak；
2) if individual's skin defying age ability value is equal to 100%, then it represents that the skin anti-aging ability of this individuality and this age bracket
The normal value of crowd is the same, and defying age ability is general；
3) if individual's skin defying age ability value is higher than 100%, then it represents that the skin anti-aging ability of this individuality is higher than this age
The normal value of Duan Renqun, defying age ability is preferable, belongs to the body constitution being difficult to aging；Ability value is the highest, and defying age ability is the strongest；
Simultaneously；
If SNP site rs4880 of gene SOD2 is A/G, show to carry heterozygous mutant, activity of SOD
Reducing, skin oxidation resistance declines, and easily produces macule, acne etc., and skin becomes obscure tarnish, the most aging；
If G/G, showing to carry homozygous mutation, activity of SOD reduces, and skin oxidation resistance declines, and easily produces
Raw macule, acne etc., skin becomes obscure tarnish, the most aging；If A/A, then it it is normal genotype；
If SNP site rs5746136 of gene SOD2 is C/T, showing to carry heterozygous mutant, activity of SOD reduces,
Skin oxidation resistance declines, easily old and feeble；If T/T, showing to carry homozygous mutation, activity of SOD reduces, skin
Oxidation resistance declines, easily old and feeble；If C/C, then it it is normal genotype；
If SNP site rs1800566 of Gene NQO1 is A/A, showing to carry homozygous mutation, skin oxidation resistance declines,
Easily producing macule, acne etc., skin becomes obscure tarnish, the most aging；If A/G, show to carry heterozygous mutant, skin
Oxidation resistance declines, and easily produces macule, acne etc., and skin becomes obscure tarnish, the most aging；If G/G, then just it is
Often genotype；
If SNP site rs3828599 of gene GPX3 is A/G, showing to carry heterozygous mutant, on accelerating, skin aging impact is little；
If G/G, showing to carry homozygous mutation, enzymatic activity reduces, and skin toxin expelling ability reduces, and is easily subject to external environment and free radical
Destroy, accelerate skin aging；If A/A, then it it is normal genotype；
If SNP site rs1050450 of gene GPX1 is A/A, showing to carry homozygous mutation, enzymatic activity reduces, skin toxin expelling energy
Power reduces, and is easily subject to the destruction of external environment and free radical, accelerates skin aging；If A/G, show to carry homozygous mutation, enzyme
Activity reduces, and skin toxin expelling ability reduces, and is easily subject to the destruction of external environment and free radical, accelerates skin aging；If G/G, then
For normal genotype；
If gene GSTM1 is GSTM1-null, showing gene delection, internal lack this enzyme, total antioxidant status weakens, health toxin expelling
Ability is poor, the most aging；If GSTM1-present, then it it is normal genotype；
If gene GSTT1 is GSTT1-null, showing gene delection, internal lack this enzyme, total antioxidant status weakens, health toxin expelling
Ability is poor, the most aging；If GSTT1-present, then it it is 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 comprehensive accurate evaluation
The SNP site combination of skin anti-aging ability；(2) high specificity, highly sensitive TaqMan-MGB probe in detecting side are used
Method detects, and can detect multiple sample simultaneously；(3) anti-ageing with assessment skin by algorithm precise quantification skin aging risk
Old ability, provides personalized skin anti-aging scheme for tester.
The technical solution used in the present invention is: binding molecule Mechanism Study, literature search and clinical data screening, obtains one group
The susceptible SNP site combination that skin aging is relevant, described combination site is respectively SOD2 (rs4880, rs5746136), NQO1
(rs1800566)、GPX3(rs3828599)、GPX1(rs1050450)、GSTM1(GSTM1-null)、GSTT1
(GSTT1-null), the site information of described combination is shown in Table 1, and this combination site can resist with the skin of accurate evaluation Different Individual
Old and feeble ability.
Table 1 susceptible SNP site information table
In order to detect described site, applicant devises the primer needed for detection and probe, and described primer and fluorescent probe are as follows
(underscore is SNP site):
SOD2 (rs4880) primer and fluorescent probe sequence:
SOD2 (rs5746136) primer and fluorescent probe sequence:
NQO1 (rs1800566) primer and fluorescent probe sequence:
GPX3 (rs3828599) primer and fluorescent probe sequence:
GPX1 (rs1050450) primer and fluorescent probe sequence:
GSTM1 (GSTM1-null) primer sequence:
Forward primer GAACTCCCTGAAAAGCTAAAGC SEQ ID NO:21
Reverse primer GTTGGGCTCAAATATACGGTGG SEQ ID NO:22
GSTT1 (GSTT1-null) primer sequence:
Forward primer TTCCTTACTGGTCCTCACATCTC SEQ ID NO:23
Reverse primer TCACCGGATCATGGCCAGCA SEQ ID NO:24；
Described loci detection method specifically comprises the following steps that
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 need to be with drink
With foreign material in 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；Remove supernatant,
Precipitation adds 50 μ L5mol/L KI solution, concussion mixing, adds 100 μ L 0.9%NaCl solution, 150 μ L phenol
/ chloroform (25: 24, V/V) mixed liquor, concussion 30s, 10000rpm are centrifuged 5min；Take 200 μ L of supernatant, add 200 μ L
Isopropanol, mixing, 10000rpm is centrifuged 5min；Removing supernatant, precipitation adds 1mL absolute ethanol washing, 10000rpm from
Heart 5min；Abandoning dehydrated alcohol, be deposited in super-clean bench and 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 (every
The detection correspondence probe of individual SNP site mixes) (detection of each SNP site is right for (2.5 μMs) and primer mixed liquor
Primer is answered to mix) 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 circulate 40 times.Detecting instrument be StepOnePlus real-time fluorescence quantitative PCR instrument (Applied Biosystems,
Applied Biotechnology company limited of the U.S.).
4, interpretation.Use StepOnePlusTM(Applied Biosystems, the U.S. should for Software v2.3
With Bioisystech Co., Ltd) it is analyzed, according to different fluorescence signals, draw the genotype of corresponding SNP site.
Skin anti-aging ability quantization method is as follows:
1, skin aging integrated risk ratio is than calculating.The risk ratio of each loci gene type of known detection gained is than respectively
For OR1、OR2…ORm(literature query is learnt, the results are shown in Table 3), can calculate Individual senescence integrated risk ratio and compare OR
(C), calculation equation is as follows:
2, the Individual senescence integrated risk ratio calculated by step 1 is than OR (C), by document or other statistical approach
Chinese population different age group skin aging average risk ratio odds (D) (known Chinese population different age group can be obtained
Skin aging produce average risk ratio odds (D) be respectively as follows: 20-35 year: 0.09,36-45 years old: 0.21,46-55
Year: 0.48), can calculate, by odds ratio computing formula, skin aging value-at-risk ratio odds (X) that detection is individual,
Calculation equation is as follows:
3, the individual's skin aging calculated by step 2 produces risk ratio odds (X), i.e. can be pushed away by value-at-risk
Calculating formula and can calculate integrated risk value F (X), calculation equation is as follows:
4, the skin anti-aging ability of Different Individual can be assessed by the calculated integrated risk value of step 3, it is known that normal
Genotype crowd's value-at-risk F (N) (known normal genotype crowd's value-at-risk F (N) is 8.26% to be respectively as follows: 20-35 year:
8.26%, 36-45 year: 17.35%, 46-55 year: 32.43%), individual's skin defying age ability value V can be calculated, meter
Calculation equation is as follows:
Testing result personality analysis and solution:
According to defying age ability quantum chemical method result, in conjunction with function and the molecular mechanism of related locus, it is judged that skin problem can
Energy property, 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.The molecular mechanism of related gene is as follows: if SOD2 carries sudden change, can accelerate skin aging；NQO1 carries
Sudden change, skin oxidation resistance declines, easily old and feeble；GPX3 and GPX1 carries sudden change, and skin oxidation resistance declines,
Skin easy damaged；GSTM1 and GSTT1 carries sudden change, and noxious substance easily accumulates at skin, toxin expelling ability, easily old and feeble.
Beneficial effect:
Endogenous cause of ill and the exopathogenic factor of skin aging can be fully understood by, provide the user personalized beauty care health solution, make anti-ageing
Old cosmetics using effect maximizes.Further, it is also possible to adjust the daily life dietary habit being conducive to skin protection for tester.
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 can not manage
Solve as limitation of the present invention.Unreceipted concrete technology or condition person in embodiment, described by the document in this area
Technology or condition or carry out according to product description.Agents useful for same or instrument unreceipted production firm person, be and can pass through
City available from conventional products.
Embodiment 1: primer and probe design
1, design and synthesize following primer and probe, synthesize unit: Applied Biosystems, U.S.'s Applied Biotechnology is limited
Company.
SOD2 (rs4880) primer and fluorescent probe sequence:
SOD2 (rs5746136) primer and fluorescent probe sequence:
NQO1 (rs1800566) primer and fluorescent probe sequence:
GPX3 (rs3828599) primer and fluorescent probe sequence:
GPX1 (rs1050450) primer and fluorescent probe sequence:
GSTM1 (GSTM1-null) primer sequence:
Forward primer GAACTCCCTGAAAAGCTAAAGC SEQ ID NO:21
Reverse primer GTTGGGCTCAAATATACGGTGG SEQ ID NO:22
GSTT1 (GSTT1-null) primer sequence:
Forward primer TTCCTTACTGGTCCTCACATCTC SEQ ID NO:23
Reverse primer TCACCGGATCATGGCCAGCA SEQ ID NO:24
2, SNP site detection
1), sample collection.Detection sample in the present embodiment uses human saliva, and before saliva gathers, 30min need to be with drinking
Foreign material in water cleaning oral cavity, gather 2mL saliva and deposit in saliva preservation liquid (purchased from the kind limited public affairs of biotechnology share of Xiamen cause
Department) 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 1.5
In mL centrifuge tube, adding 500 μ L 0.01mol/L PBS solution and repeatedly blow and beat, 10000rpm is centrifuged 5min；Remove supernatant,
Precipitation adds 50 μ L5mol/L KI solution, concussion mixing, adds 100 μ L 0.9%NaCl solution, 150 μ L phenol
/ chloroform (25: 24, V/V) mixed liquor, concussion 30s, 10000rpm are centrifuged 5min；Take 200 μ L of supernatant, add 200 μ L
Isopropanol, mixing, 10000rpm is centrifuged 5min；Removing supernatant, precipitation adds 1mL absolute ethanol washing, 10000rpm from
Heart 5min；Abandoning dehydrated alcohol, be deposited in super-clean bench and dry up, add 50 μ L TE buffer solution ,-20 DEG C save backup.
3), reaction system and response procedures 10 μ L reaction system 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 (every
The detection correspondence probe of individual SNP site mixes) (detection of each SNP site is right for (2.5 μMs) and primer mixed liquor
Primer is answered to mix) 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 circulate 40 times.Detecting instrument be StepOnePlus real-time fluorescence quantitative PCR instrument (Applied Biosystems,
Applied Biotechnology company limited of the U.S.).
4), detection program end of run after, use StepOnePlusTMSoftware v2.3 (Applied Biosystems,
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
3, defying age ability quantitative evaluation and personalized solution
1), skin aging integrated risk value calculates.Learnt by literature query, the old and feeble risk ratio such as table of different loci
Shown in 3.
Risk ratio ratio (OR) table of table 3 related locus genotype
(1) Individual senescence integrated risk ratio is than calculating.The OR value of different loci is as shown in table 3, then
(2) calculate Individual senescence integrated risk ratio by step (1) to obtain than OR (C), comprehensive literature and survey data
Know that the ratio odds (D) of Chinese population 20-35 age bracket skin aging is about that 0.09 (the detection sample of the present embodiment belongs to
This age bracket, therefore select the data of this age bracket), can calculate, by odds ratio computing formula, the skin aging that detection is individual
Value-at-risk ratio odds (X)=OR (C) × odds (D)=2.25 × 0.09=0.20.
(3) detection Individual senescence risk ratio odds (X) calculated by step (2), integrated risk value
(4) known normal genotype crowd's allergy value-at-risk F (N) is 8.26%, can calculate resisting skin allergy ability value
2), testing result is comprehensively analyzed.This individual's skin defying age ability value is 49.55%, and skin anti-aging ability is poor,
The oldest and the most feeble.In combination with the molecular mechanism of related gene, the GPX3 in this detection individuality carries susceptibility loci as shown in Table 2
Homozygous mutation, NQO1 carries susceptibility loci heterozygous mutant, and these sudden changes can affect glutathion peroxidase 3 and quinone oxidation
Reductase 1 activity, affects skin oxidation resistance, skin easy damaged, thus causes aging.
3), personalized solution: this individuality belongs to skin easily old and feeble skin quality, and the sudden change of GPX3, NQO1 gene makes body
The oxidation resistance of heredity is more weak, easily old and feeble.Daily life should be noted the selection of cosmetics, should select containing tocopheryl acetate
The cosmetic product that the antioxidant compositions such as ester, grape polyphenols, co-ferment Q10 are high.Reduce contact ultraviolet, X-ray, wine
Essence, some medicine and pollutant etc. may result in the factor that free radical produces；How edible containing three big antioxidant (vitamin
E, vitamin C, beta-carotene) food, such as Garcinia mangostana, Radix Dauci Sativae, Punica granatum L. etc.；Reduce erratic living habit etc.
The factor impact on skin.
Although above it has been shown and described that embodiments of the invention, it is to be understood that above-described embodiment is exemplary,
Being not considered as limiting the invention, those of ordinary skill in the art is without departing from the principle of the present invention and the situation of objective
Under 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 skin anti-aging 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, and SEQ ID NO:17-20,
With SEQ ID NO:21-22, and SEQ ID NO:23-24；Wherein SEQ ID NO:3-4,7-8,11-12,15-16,19-20
5 ' end and 3 ' end respectively connection different fluorescent labelinies.
2. the test kit being used for assessing skin anti-aging situation, it is characterised in that containing the primer described in claim 1
And probe.
3. the method utilizing the gene test auxiliary correct beauty method of customer selecting, it is characterised in that use claim
The primer of 1 and probe, or use the test kit of claim 2.
4. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 3, it is characterised in that step
Suddenly it is:
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 skin anti-aging ability value of client is gone out according to susceptible SNP site combination calculation；Described susceptible SNP site is combined as
SOD2rs4880、SOD2rs5746136、NQO1rs1800566、GPX3rs3828599、GPX1rs1050450、
GSTM1GSTM1-null、GSTT1GSTT1-null；
3) combine 1) SNP site testing result and 2) skin anti-aging 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 institute
State 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
The response procedures of amplification 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 institute
State step 2) be,
The pathogenic risk ratio ratio of each loci gene type according to the combination of known susceptible SNP site calculates integrated risk ratio
Ratio OR (C), calculation equation is:
Produce average risk ratio odds (D) by known Chinese population different age group skin aging, calculate detection individuality
Skin aging produce value-at-risk ratio odds (X), calculating formula is odds (X)=OR (C) × odds (D)；
Calculate integrated risk value:
According to known normal genotype crowd's value-at-risk F (N), calculate individual's skin defying age ability value；
7. the method utilizing the gene test auxiliary correct beauty method of customer selecting described in claim 4, it is characterised in that institute
State step 3) result be judged as,
1) if individual's skin defying age ability value is less than 100%, then it represents that the skin anti-aging ability of this individuality is less than this age
The normal value of Duan Renqun, defying age ability, belong to the oldest and the most feeble body constitution；Ability value is the lowest, and defying age ability is the most weak；
2) if individual's skin defying age ability value is equal to 100%, then it represents that the skin anti-aging ability of this individuality and this age bracket
The normal value of crowd is the same, and defying age ability is general；
3) if individual's skin defying age ability value is higher than 100%, then it represents that the skin anti-aging ability of this individuality is higher than this age
The normal value of Duan Renqun, defying age ability is preferable, belongs to the body constitution being difficult to aging；Ability value is the highest, and defying age ability is the strongest；
Simultaneously；
If SNP site rs4880 of gene SOD2 is A/G, show to carry heterozygous mutant, activity of SOD
Reducing, skin oxidation resistance declines, and easily produces macule, acne etc., and skin becomes obscure tarnish, the most aging；
If G/G, showing to carry homozygous mutation, activity of SOD reduces, and skin oxidation resistance declines, and easily produces
Raw macule, acne etc., skin becomes obscure tarnish, the most aging；If A/A, then it it is normal genotype；
If SNP site rs5746136 of gene SOD2 is C/T, showing to carry heterozygous mutant, activity of SOD reduces,
Skin oxidation resistance declines, easily old and feeble；If T/T, showing to carry homozygous mutation, activity of SOD reduces, skin
Oxidation resistance declines, easily old and feeble；If C/C, then it it is normal genotype；
If SNP site rs1800566 of Gene NQO1 is A/A, showing to carry homozygous mutation, skin oxidation resistance declines,
Easily producing macule, acne etc., skin becomes obscure tarnish, the most aging；If A/G, show to carry heterozygous mutant, skin
Oxidation resistance declines, and easily produces macule, acne etc., and skin becomes obscure tarnish, the most aging；If G/G, then just it is
Often genotype；
If SNP site rs3828599 of gene GPX3 is A/G, showing to carry heterozygous mutant, on accelerating, skin aging impact is little；
If G/G, showing to carry homozygous mutation, enzymatic activity reduces, and skin toxin expelling ability reduces, and is easily subject to external environment and free radical
Destroy, accelerate skin aging；If A/A, then it it is normal genotype；
If SNP site rs1050450 of gene GPX1 is A/A, showing to carry homozygous mutation, enzymatic activity reduces, skin toxin expelling energy
Power reduces, and is easily subject to the destruction of external environment and free radical, accelerates skin aging；If A/G, show to carry homozygous mutation, enzyme
Activity reduces, and skin toxin expelling ability reduces, and is easily subject to the destruction of external environment and free radical, accelerates skin aging；If G/G, then
For normal genotype；
If gene GSTM1 is GSTM1-null, showing gene delection, internal lack this enzyme, total antioxidant status weakens, health toxin expelling
Ability is poor, the most aging；If GSTM1-present, then it it is normal genotype；
If gene GSTT1 is GSTT1-null, showing gene delection, internal lack this enzyme, total antioxidant status weakens, health toxin expelling
Ability is poor, the most aging；If GSTT1-present, then it it is normal genotype.
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| CN106148545A (en) * | 2016-08-29 | 2016-11-23 | 广州肌因港生物科技有限公司 | One group of Primer composition for skin-related gene detection and detection method |
| 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 |
| CN108251521A (en) * | 2018-03-15 | 2018-07-06 | 北京天平永达科技发展有限公司 | For detecting the primer set of tumor susceptibility gene SNP related to skin aging and its application |
| CN108950013A (en) * | 2018-07-27 | 2018-12-07 | 江颖纯 | A kind of skin-related gene site library and its construction method and application |
| CN109207604A (en) * | 2018-09-10 | 2019-01-15 | 广州益养生物科技有限公司 | A kind of method of quick detection heredity skin antioxidant genes |
| CN110819723A (en) * | 2019-11-27 | 2020-02-21 | 嘉文丽（福建）化妆品有限公司 | Detection method for five gene combinations for assessing congenital skin quality and personalized skin care kit formulated according to detection results |
| CN111455035A (en) * | 2020-01-22 | 2020-07-28 | 广州市普森生物科技有限公司 | Primer combination and kit for detecting skin anti-aging capability gene and application of primer combination and kit |
| CN112725437A (en) * | 2021-02-02 | 2021-04-30 | 暨南大学 | Application of rhythm gene protein expression and RNA methylation modification in preparation of aging detection kit |
| CN112980963A (en) * | 2019-12-13 | 2021-06-18 | 宁波海尔施基因科技有限公司 | Kit for detecting individual skin genes and use method thereof |
| WO2021235551A1 (en) * | 2020-05-22 | 2021-11-25 | 株式会社 資生堂 | Method for determining resistance against skin aging using genetic polymorphism |
| CN114686578A (en) * | 2020-12-30 | 2022-07-01 | 北京本真工坊生物科技有限公司 | Skin type assessment method, application and system |
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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 |
| 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 |
| CN108251521A (en) * | 2018-03-15 | 2018-07-06 | 北京天平永达科技发展有限公司 | For detecting the primer set of tumor susceptibility gene SNP related to skin aging and its application |
| CN108251521B (en) * | 2018-03-15 | 2021-08-13 | 北京天平永达科技发展有限公司 | Complete set of primers for detecting susceptibility gene SNP related to skin aging and application thereof |
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| CN112980963A (en) * | 2019-12-13 | 2021-06-18 | 宁波海尔施基因科技有限公司 | Kit for detecting individual skin genes and use method thereof |
| CN111455035A (en) * | 2020-01-22 | 2020-07-28 | 广州市普森生物科技有限公司 | Primer combination and kit for detecting skin anti-aging capability gene and application of primer combination and kit |
| WO2021235551A1 (en) * | 2020-05-22 | 2021-11-25 | 株式会社 資生堂 | Method for determining resistance against skin aging using genetic polymorphism |
| CN114686578A (en) * | 2020-12-30 | 2022-07-01 | 北京本真工坊生物科技有限公司 | Skin type assessment method, application and system |
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