CN103074414A - Diabetes type II gene susceptivity and pre-warning detection kit - Google Patents
Diabetes type II gene susceptivity and pre-warning detection kit Download PDFInfo
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Abstract
The present invention provides a diabetes type II gene susceptivity and pre-warning detection kit, which is characterized in that seven nucleic acid (DNA and RNA) molecule markers are combined, wherein the seven nucleic acid (DNA and RNA) molecule markers comprise APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146) , CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (FokI) and SLC30A8 (rs13266634). The kit comprises: specific amplification primer pairs and DNA sequencing primer pairs of the gene polymorphism loci, PCR reaction liquids, PCR product enzymolysis liquids and DNA sequencing reaction liquids. According to the present invention, detection is performed by using the kit, gene information is processed and analyzed, diabetes type II susceptivity of a healthy subject is evaluated, and guiding significance is provided for personalized treatment and health care of diabetes type II people.
Description
Technical field
The present invention relates to genetics, molecular biology, clinical medicine and preventive medicine technical field, specifically, the present invention relates to a kind of type ii diabetes Gene Susceptibility and early warning detection kit, it is characterized in that a plurality of nucleic acid of joint-detection (DNA and RNA) molecule marker, comprise: APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), the upper gene polymorphism sites of SLC30A8 (rs13266634), in the computational analysis through risk evaluation model, can assess and screen person under inspection's II diabetes susceptibility (susceptibility).
Background technology
In the whole world in 2000 diabetic subject 1.51 hundred million is arranged, and there is diabetic subject 2.85 hundred million in the whole world at present, by present rate of rise, estimate that the year two thousand thirty whole world diabetes population will rise to 500,000,000, wherein developed country will rise to 7,200 ten thousand by 5,100 ten thousand, rate of increase is 42%, and developing country will rise to 200,000,000 3,000 ten thousand by 8,400 ten thousand, and rate of increase has reached 170%.In recent years China's onset diabetes rate continue to raise, by 1980 1% rise to 2008 9.7%.The diabetic subject breaks through 9,800 ten thousand people at present, wherein is type ii diabetes more than 90%, so also be modal diabetes.Be characterized in insulin resistant, namely in-vivo tissue is insensitive to the effect of Regular Insulin, and the Regular Insulin of normal amount does not have normal hypoglycemic activity, or hypoinsulinism.Comprise that Chinese's the type ii diabetes patient of Asia ethnic group is often and without features such as obesities.Studies show that, in following 20 years, the diabetes mellitus in China crowd is to be higher than 2 times speed increment, and the Asia will become the most serious diseased region.Diabetes and chronic complicating diseases serious threat human health and life.
Studies show that in a large number, type ii diabetes (T2DM) is a kind of disease of multifactorial inheritance, is caused the disease take insulin resistant and islet beta cell function obstacle as characteristics by the h and E factor interaction.Inherited genetic factors has increased the danger of body generation type ii diabetes.Wherein major gene plays a major role, and inferior effect gene plays less effect, also may have interaction between the different genes.
Adiponectin (Adiponectin) is to secrete a kind of collagen like cell factor by adipocyte-specific, is present in a kind of neurophysin in the circulation of blood.Adiponectin is comprised of 244 amino acid, and adiponectin is rich content in blood plasma.There are some researches show the biological action such as Plasma adiponectin has atherosclerosis, improves insulin resistant, hypoglycemic, anti-inflammatory.Low adiponectin concentration can increase the risk of metabolism syndrome and type ii diabetes.Human adiponectin (Adiponectin gene or APM1) is single copy gene, is positioned karyomit(e) 3q27, and size is 17kb, is comprised of 3 exons and 2 introns.Full genome scanning shows that there is the susceptibility loci of type ii diabetes (T2DM) and metabolism syndrome in this zone.
CDK5 (Cyclin Dependent Kinase 5, cyclin-dependent kinase 5) can cause the β cytopathy under the effect of the sugared toxicity of height.CDKAL1 (CDK5 regulator subunit associated protein 1 analogue 1, the cyclin-dependent kinase 5 regulatory subunit associated protein 1-like 1) protein of genes encoding can suppress the activation of CDK5.If the CDKAL1 gene morphs, then CDK5 is lost restraining effect, can cause the β cell function impaired, insulin secretion reduces.As seen, the CDKAL1 gene locus morphs, and lacks relevant with insulin secretion among the crowd.Rs57754840 is positioned at the 5th intron of CDKAL1 gene, is and the closely-related SNP of type ii diabetes genetic predisposition.CDKAL1 gene rs7754840 has G, two kinds of allelotrope of C.G is major gene, and C is minor gene, the G-C variation occurs, and then CDKAL1 weakens the restraining effect of CDK5, can cause the β cell function impaired, so C is the excessive risk allelotrope that causes diabetes to occur.The occurrence frequency of C% is about 40% in the normal population.
TCF7L2 (transcription factor 7 analogues 2, transcription factor 7 like2) gene may be one of main Disease-causing gene of T2DM.The TCF7L2 gene is positioned at karyomit(e) 10q25, total length 217,226bp.Formed by 14 exons and 13 introns.619 amino acid of encoding altogether.Its expression product is one group of transcription factor with variability, and these transcription factors play an important role in keeping the stable state of plasma glucose.Florez in 2006 etc. find that by the perspective study in 3 years two sites of rs12255372 and rs7903146 are relevant with the progress of the diabetes state of an illness, and they can increase the risk that those Patients with Impaired Glucose Tolerances are suffered from diabetes.The discovery site rs7903146 such as Sale are relevant with the African diabetic nephropathy of living in the U.S. with rs7901695.Our research discovery, the genovariation type of rs7903146 is the gene the strongest with type ii diabetes onset risk dependency.TCF7L2 be find so far with one of the closest tumor susceptibility gene of the onset relation of type ii diabetes, and in not agnate crowd, good repeatability is arranged.TCF7L2 genovariation increases the onset risk of type ii diabetes, may be by its cross expressing in diabetic subject's beta Cell of islet, the unusual and beta Cell of islet of Wnt signal path have finally caused insulin secretion obstacle and insulin action to descend to approach such as proinsulin processing obstacles.
CDKN2A and CDKN2B are positioned on the Chromosome 9 9P21, and CDKN2A/CDKN2B encodes respectively and presses down cancer factor p15INK4a and p16INK4b.The latter can suppress the CDK4 activity.And CDK4 (Cyclin Dependent Kinase 4) can effectively regulate pancreatic Beta cell proliferation.If near the rs10811661 the CDKN2A/2B gene morphs, so that the overexpression of CDKN2A/2B gene product will significantly suppress the CDK4 activity, further can affect the secreting function of β cell, insulin secretion is reduced, cause the occurrence risk of type ii diabetes to raise.Have clinical study to show, CDKN2A/2B gene rs10811661 has T/T, T/C, three kinds of genotype of C/C, and the dangerous allelotrope T/T of type ii diabetes group homozygote frequency is 37.5%, and dangerous allelotrope T% is 59.5%, all is significantly higher than Normal group.The risk that T allelotrope carrier suffers from diabetes is 1.38 times of C allelotrope carrier.
The KCNQ1 assignment of genes gene mapping is in o.11 karyomit(e) 11p5, the albumen that 676 amino acid of encoding form, and its physiological action is one of member of Voltage Dependent K Channels family.With the Japan internationality medical centre research group headed by the Yasuda etc. relatively type ii diabetes patient and the general population, find to be positioned at the variation of the rs2237892 of KCNQ1 intron, make the onset risk of type ii diabetes will increase 1.3-1.4 times.The people such as Li Huaixing and Lin Xu with 3210 Donors as the study population, systematically analyzed the rs2074196 on the KCNQ1 gene, rs2237892, rs52237895 and rs522375973 pleomorphism site and type ii diabetes and relevant phenotypic incidence relation thereof.Find: rs2237892, rs52237895 and rs2237s973 site all exist significant incidence relation with the ill risk of type ii diabetes, and it increases ill risk respectively up to 32.5%, 18.8% and 35.8%.These sites all are associated with the β cell function is impaired.The haplotype that is comprised of the dangerous allelotrope of these pleomorphism sites also and between the ill risk of type ii diabetes and the β cell function index exists very strong incidence relation.The result of study prompting, the KCNQ1 gene is the main type ii diabetes gene of of Chinese han population.7 pieces of papers are included in the researchs such as Zhang Liuwei altogether in, obtain 7764 routine type ii diabetes patients and 8937 example contrasts, show through the Meta analytical results, KCNQ1 gene rs2237892 site C allelotrope can increase the onset risk (OR=1.300 of T2DM, 95%CI:1.234～1.366), the CC/TC genotype is the risk genotypes (OR=1.453,95%CI:1.279～1.651) of type ii diabetes.
People VDR full length gene is 4605 base pairs approximately, comprise one section noncoding leader sequence of 115bp, the 3 end non-coding sequences of the encoding sequence of one section 1281bp and one section 3209bp.The VDR gene is positioned at karyomit(e) 12q13, is comprised of 9 exons and 8 introns.Vitamins D is by being combined its biological action of performance with VDR, for example beta Cell of islet is expressed Vitamin D Receptor and vitamin D-dependent calcium-binding protein (calbindinD28k, CB), this prompting vitamins D can affect insulin secretion and control β Growth of Cells and differentiation.Studies show that, insulin secretion increases or reduces relevant behind VDR gene polymorphism sites (Fok I) and the oral glucose tolerance.VDR gene locus single nucleotide polymorphism can affect the variation of VDR transcriptional activity.Therefore, VDR (Fok I) gene is as the candidate gene of type ii diabetes.
SLC30A8 (solute carrier family 30, member 8, Zinc transporter-8) gene nucleotide series total length 41617bp contains 8 exons, is positioned at 8q24.11 at karyomit(e).369 amino acid whose protein of SLC30A8 genes encoding.What studies show that the SLC30A8 coding is a kind of zinc ion transport albumen in the islet cells great expression, be a kind of repeatedly transmembrane protein, and albumen has passed through the modification of various ways.Its major function be with intracytoplasmic zinc ion transport in the insulin secretion vesica, participate in the critical function of insulin synthesis, storage and secretion.Multinomial GWAS research discovery, the C allelotrope of SLC30A8 gene rs13266634 pleomorphism site is the risk allelotrope of Chinese's type ii diabetes.
Summary of the invention
Fundamental principle based on ebm, the method that adopts case-control study and cohort study to combine, and retrospective study combined with perspective study, by literature search, evaluation of evidential matter, information extraction, the check of H-W genetic equilibrium, homogeneity test, a series of strict screenings and the experimental verification processes such as large sample Meta analysis, consider and analyze the relational degree (OR value) of each gene locus and II diabetes, genotype occurrence frequency among the crowd, pathogeny, signal paths etc. are determined the susceptible gene pleomorphism site that seven II diabetes are relevant at last.
The invention provides a kind of II diabetes Gene Susceptibility and early warning detection kit, by a plurality of nucleic acid of joint-detection (DNA and RNA) molecule marker, comprise: APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), the upper gene polymorphism sites of SLC30A8 (rs13266634), assess and screen the genetic predisposition of II diabetes.
The feature of this test kit is that the specificity amplification primer in said gene site comprises reaching the dna sequencing primer pair:
For detection of on the APMI gene+Auele Specific Primer of 45 gene polymorphism sites to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of rs10946398 gene polymorphism sites on the CDKAL1 gene to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of rs7903146 polygene pleomorphism site on the TCF7L2 gene to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of rs10811661 gene polymorphism sites on the CDKN2A/2B gene to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of rs2237892 gene polymorphism sites on the KCNQ1 gene to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of Fok I gene polymorphism sites on the VDR gene to and the dna sequencing primer pair;
For detection of the Auele Specific Primer of rs13266634 gene polymorphism sites on the SLC30A8 gene to and the dna sequencing primer pair;
PCR reaction test solution (comprising: damping fluid, MgCl
2Solution, UdNTPs, dNTPs, Auele Specific Primer pair, Taq archaeal dna polymerase, UNG enzyme, template DNA, pure water etc.);
PCR product enzymolysis solution (comprising: SAP enzyme, ExoL enzyme MIX, pure water etc.);
Dna sequencing reaction liquid component (comprising: EDTA, dehydrated alcohol, 70% pre-cooled ethanol, deionized formamide (Hi-Di Formamide), PCR enzymolysis product, BigDye Mix, dna sequencing primer, pure water etc.).
The described feature of this test kit is that APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), upper seven gene polymorphism sites of SLC30A8 (rs13266634) carry out joint-detection simultaneously.
The described feature of this test kit is seven specificity amplification primers to respectively for APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), the upper gene polymorphism sites of SLC30A8 (rs13266634), can go out dna fragmentation by specific amplification.Seven Auele Specific Primers of the present invention can synthesize with the synthetic method of routine, and these primers those skilled in the art can design easily, synthetic and use etc.
The described feature of this test kit is that seven dna sequencing primer pairs are gone up gene polymorphism sites for APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), SLC30A8 (rs13266634) respectively, and the energy specificity is carried out the sequential detection of dna fragmentation.Seven dna sequencing primer pairs of the present invention can synthesize with the synthetic method of routine, and these primers those skilled in the art can design easily, synthetic and use etc.
The described feature of this test kit is that composition and the content of described test kit comprises:
(1) PCR reaction test solution: single PCR reaction system comprises: 10 * PCR damping fluid, 2.5 μ l, 25mM MgCl
2Solution 2.5 μ l, 25mM UdNTPs 0.2 μ l, 25mM dNTPs 0.2 μ l, the pair of primers 0.5 μ l of 20 μ M Auele Specific Primer centerings, 5U/ μ l mixed enzyme (Taq archaeal dna polymerase and UNG enzyme) 1 μ l, template DNA 3 μ l (50ng) add pure water to 25 μ l.Wherein, use the anti-pollution system of UdNTPs-uridine enzyme (UNG), through heating the U-DNA that optionally to degrade, to prevent the pollution of pcr amplification product.
(2) PCR product enzymolysis solution: 2 μ l SAP enzyme+ExoL enzyme MIX.
(3) dna sequencing reaction liquid: formed by two portions: sequencing reaction system and order-checking product purification.Single DNA sequencing reaction system comprises: 3 μ l PCR enzymolysis products, 1 μ l sequencing reagent (25%BigDye Mix), 1 μ l dna sequencing primer pair (only using one of them primer), 1 μ l pure water.The order-checking product purification comprises: 100mM EDTA 2 μ l, 16 μ l dehydrated alcohols, 70% pre-cooled ethanol, 70 μ l * 2,10 μ l deionized formamides (Hi-Di Formamide) etc.
The storage temperature of above-mentioned three components is-20 ℃.
Embodiment
Further set forth the present invention below in conjunction with specific embodiment, embodiment only is used for illustration purpose, and is not used in the restriction scope of the invention.Do not elaborate in the following example and specifically described experiment condition and experimental technique, should be normal experiment condition and method, or undertaken by operation steps and method that producer provides.
The use of this test kit of embodiment
1, obtaining of sample genomic dna: gargle to secondary with clear water, remove the swill in the oral cavity, and note remaining clear water in the most oral cavity of pharynx.With cotton swab scraping oral mucosa cast-off cells and put it in the cell stationary liquid of Eppendorf pipe, tighten bottle cap for subsequent use.
2, DNA extracting and purifying: the DNA with in the extraction of lysis method is suspended in the 1ml physiological saline the centrifugal 10min of 2000 * g; Abandon supernatant, every pipe adds 1ml physiological saline repeated washing once, the more centrifugal 10min of 2000 * g; Abandon supernatant, every pipe add 400 μ l cell lysis buffer solution (10mmol/L Tris-HCl, PH 8.0; 0.1mol EDTA, PH 8.0; 0.5%SDS), be placed in 50 ℃ of water-baths incubation 30 minutes; Then be cooled to room temperature, add isopyknic phenol one chloroform one primary isoamyl alcohol (volume ratio 25: 24: 1), mixing, the centrifugal 10min of 5000 * g; Shift the upper strata water in another Eppendorf pipe, repeat extracting once; Shift again the upper strata water in another Eppendorf pipe, add the 3M NaAc (pH5.2) of 1/10 volume, mixing; The 95% cold ethanol that adds 2.5 times of volumes, mixing ,-20 ℃ precipitate DNA 30 minutes; Centrifugal 15 minutes of 10000 * g abandons supernatant; Add 1ml 70% cold ethanol washing and precipitating, centrifugal minute of 10000 * g abandons supernatant; The oven dry in 30 minutes of 37 ℃ of incubators; The DNA precipitation is dissolved in the 20 μ l TE damping fluids, adds 20 μ g/ml Pancreatic RNases, 1 μ l, 37 ℃ of temperature were bathed 30 minutes, placed-20 ℃ of preservations.
3, adopt PCR method amplification target dna fragment:
(1) characteristics of this test kit are to use the anti-pollution system of UdNTPs-uridine enzyme (UNG), through heating the U-DNA that can optionally degrade, to prevent the pollution of pcr amplification product.
(2) the PCR reaction reagent component of this test kit is by PCR reaction solution volume 21 μ l, 5U/ μ l mixed enzyme (Taq archaeal dna polymerase and UNG enzyme) 1 μ l, and template DNA 3 μ l (approximately 50ng) form.The PCR reaction solution comprises: 10 * PCR damping fluid, 2.5 μ l, 25mMMgCl
2Solution 2.5 μ l, 25mM UdNTPs 0.2 μ l, 25mM dNTPs 0.2 μ l.
(3) PCR reaction reagent component is prepared: rear abundant mixing and of short duration centrifugal fully please thaws before the reagent open pipe.Suggestion is the preparation mixed reaction solution after DNA extraction finishes, and application of sample immediately.Such as Special Circumstances, the mixed reaction solution short-term that installed in following minute can be stored in 4 ℃, and in 3 hours, use.
(4) PCR reaction reagent component preparation: by each PCR reaction solution volume 21 μ l, mixed enzyme 1 μ l, template DNA 3 μ l multiply by required reaction tubes number, calculate the amount of reagent, are added in the aseptic centrifuge tube vibration mixing.The mixed reaction solution that in each PCR reaction tubes, adds again the above-mentioned mixing of 25 μ l with micro sample adding appliance.
(5) PCR reaction reagent component application of sample: in the PCR reaction tubes that mixed reaction solution is housed that installed in above-mentioned minute, add respectively each 3 μ l of template DNA and negative control.That is: single PCR reaction system component sees Table 1:
The single PCR reaction reagent of table 1 component
Attention: if template DNA less than 3 μ l can supply with pure water.
(6) PCR reaction conditions: circulation 1:94 ℃, 3 minutes; 45 seconds, 72 ℃ extensions of 45 seconds, 52 ℃ annealing of circulation 2-31:94 ℃ sex change 1 minute; Totally 30 circulations; Last 72 ℃ were extended 10 minutes.
4, PCR product purification
(1) electrophoresis: get 5 μ l amplified productions, the sepharose with 2.0% carries out electrophoresis detection, contrasts as molecular size range with suitable DNA Marker, uses gel imaging system to observe electrophoresis result.All the other PCR products can 4 ℃ of short-term preservations (be no more than 2 days, do not use if surpass 2 days, please place-20 ℃ of preservations).
(2) observe electrophoresis result: (A) negative control should be without amplified production, otherwise should all again increase; (B) band is clear, and size meets target product, then can be used for follow-up sequencing reaction; Driftlessness band or band can't clear identification sample, need re-start the amplified reaction in this site; Again after the amplification still without purpose band or band can't clear identification sample, suggestion re-starts the target dna extracting.
(3) PCR enzymolysis product: choose PCR product 3 μ l, add again the purified reagent component: 2 μ l SAP MIX, mixing.Carry out following reaction: 37 ℃ 60 minutes, 80 ℃ 15 minutes.Product can 4 ℃ of short-term preservations (be no more than 2 days, do not use if surpass 2 days, please place-20 ℃ of preservations).
5, sequencing reaction mixes each component of table 2 and carries out sequencing reaction, and sequencing reaction reagent components and continent order reaction conditions see Table 2 and table 3:
Table 2DNA sequencing reaction reagent components
Table 3 sequencing reaction condition
6, order-checking product purification, take 96 orifice plate purification process as example:
(A) in every hole, add 2 μ l EDTA (100mM), 16 μ l dehydrated alcohols, thermal agitation, lucifuge left standstill 15 minutes.More than the 3500g, 4 ℃ centrifugal 30 minutes.Be inverted 96 orifice plates on thieving paper, 500g is of short duration centrifugal.
(B) add 70 μ l, 70% pre-cooled ethanol, thermal agitation, 3500g above 4 ℃ centrifugal 15 minutes.Be inverted 96 orifice plates on thieving paper, 500g is of short duration centrifugal.
(C) add 70 μ l, 70% ethanol, gentleness is put upside down for several times mixing (not thermal agitation), 3500g above 4 ℃ centrifugal 5 minutes.Be inverted 96 orifice plates on thieving paper, 500g is of short duration centrifugal.
(D) make ethanol clean in the room temperature volatilization, add 10 μ l deionized formamide dissolving DNAs.
7, dna sequencing reaction: sex change on the PCR instrument: 95 ℃, 4 minutes; 4 ℃, 4 minutes, upper 3730 type DNA sequencers checked order.
8, as a result interpretation: use the somatotype result of sequencing analysis software to carry out interpretation, data processing and analysis.Those skilled in the art can finish sequencing reaction and as a result interpretation etc. by the operation steps of DNA sequencer easily.
9, risk assessment: calculate first 7 gene locuss OR value separately, again according to risk evaluation model, carry out the genetic risk assessment of the individual type ii diabetes of person under inspection.Be conducive to expert and person under inspection according to the result of gene type, carry out Extraordinary health control, personalized treatment and intervene guidance.
Claims (8)
1. the invention provides a kind of type ii diabetes Gene Susceptibility and early warning detection kit, it is characterized in that may further comprise the steps:
Obtain person under inspection's body fluid, hair, blood or cell tissue equal samples;
Extract the nucleic acid (DNA and RNA) in the sample;
Obtain person under inspection's genetic information by the method for a plurality of nucleic acid of joint-detection (DNA and RNA) molecule marker;
Relevant bioinformation is carried out data processing and statistical study, thereby assess the II diabetes susceptibility of this healthy subject, and personalized treatment and the personalized health care of suffering from the type ii diabetes crowd also had directive significance.
2. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit, it is characterized in that a plurality of nucleic acid of joint-detection (DNA and RNA) molecule marker, comprising: APM1 (+45), CDKAL1 (rs10946398), TCF7L2 (rs7903146), CDKN2A/2B (rs10811661), KCNQ1 (rs2237892), VDR (Fok I), SLC30A8 (rs13266634).
3. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit is characterized in that the detection method of described nucleic acid (DNA and RNA) molecule marker comprises:
The method of various dna sequencings;
Various fluorescence quantifying PCR methods;
Various biochip test methods;
Detect the method for nucleic acid (DNA and RNA) molecule marker by various fluorescent marks and hybridization technique.
4. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit is characterized in that described nucleic acid (DNA and RNA) molecule marker comprises:
Single nucleotide polymorphism (single nucleotide polymorphism, SNP);
Copy number polymorphism (copy number polymorphism, CNP);
Microsatellite DNA (short tandem repeat, STR);
Restriction fragment length polymorphism (restriction fragment length polymorphism, RFLP);
Randomly amplified polymorphic DNA (random amplified polymorphic DNA, RAPD);
Particular sequence site (sequence-tagged site, STS);
Amplified fragment length polymorphism (Amplified Fragment Length Polymorphism, AFLP).
5. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit is characterized in that the specificity amplification primer of described seven nucleic acid (DNA and RNA) molecule marker is to reaching the dna sequencing primer pair.
6. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit is characterized in that the composition of described test kit and content comprise: (1) PCR reaction reagent component; (2) PCR product enzymolysis reagent components; (3) dna sequencing reaction reagent components.
7. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit, the individual fat genetic risk of healthy subject that can be applicable to that it is characterized in that described test kit is assessed, be conducive to expert and person under inspection according to the result of gene type, carry out the Extraordinary health control and intervene guidance.
8. a kind of type ii diabetes Gene Susceptibility according to claim 1 and early warning detection kit, it is characterized in that the detection that can be applicable to suffer from the type ii diabetes crowd of described test kit, be conducive to expert and person under inspection and realize personalized treatment and personalized health care according to the result of gene type.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108384844A (en) * | 2018-02-07 | 2018-08-10 | 深圳鼎新融合科技有限公司 | Detect primer pair, probe and the kit of mankind's VDR, GC, LRP5, SLC30A8 gene pleiomorphism |
| CN108949940A (en) * | 2017-07-20 | 2018-12-07 | 苏州大学附属第二医院 | A kind of pyrosequencing detection secretin acts on the kit and method of relevant single nucleotide polymorphism |
| CN110117649A (en) * | 2019-05-21 | 2019-08-13 | 凯杰（苏州）转化医学研究有限公司 | A kind of Primer composition and its application |
| CN110157798A (en) * | 2019-06-27 | 2019-08-23 | 中南大学湘雅医院 | A kind of primer combination of probe instructing diabetic individual medication gene loci polymorphism and its application and kit |
-
2012
- 2012-04-27 CN CN 201210128332 patent/CN103074414A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949940A (en) * | 2017-07-20 | 2018-12-07 | 苏州大学附属第二医院 | A kind of pyrosequencing detection secretin acts on the kit and method of relevant single nucleotide polymorphism |
| CN108384844A (en) * | 2018-02-07 | 2018-08-10 | 深圳鼎新融合科技有限公司 | Detect primer pair, probe and the kit of mankind's VDR, GC, LRP5, SLC30A8 gene pleiomorphism |
| CN110117649A (en) * | 2019-05-21 | 2019-08-13 | 凯杰（苏州）转化医学研究有限公司 | A kind of Primer composition and its application |
| CN110157798A (en) * | 2019-06-27 | 2019-08-23 | 中南大学湘雅医院 | A kind of primer combination of probe instructing diabetic individual medication gene loci polymorphism and its application and kit |
| CN110157798B (en) * | 2019-06-27 | 2023-04-18 | 中南大学湘雅医院 | Primer probe combination for guiding polymorphism of gene locus of diabetes personalized medicine, application and kit thereof |
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