CN1721549A - The test kit and the primer of forecast China people's diabetes B susceptibility - Google Patents

Abstract

The present invention relates to be used to predict the test kit and the primer of diabetes B susceptibility, more specifically, the present invention relates to utilize the test kit and the primer of prediction diabetes B susceptibility of a single nucleotide polymorphism (SNP) site design of the diabetes B tumor susceptibility gene KCNJ11 of new discriminating.The invention still further relates to the application of KCNJ11 gene in the diagnostic reagent of preparation prediction diabetes B susceptibility.

Description

The test kit and the primer of forecast China people's diabetes B susceptibility

Technical field

The present invention relates to be used to predict the test kit and the primer of diabetes B susceptibility, more specifically, the present invention relates to utilize diabetes B tumor susceptibility gene KCNJ11 (the potassium inwardly-rectifying channel of new discriminating, subfamily J, member 11) the test kit and the primer of prediction diabetes B susceptibility of a single nucleotide polymorphism (SNP) site design.The invention still further relates to the application of KCNJ11 gene in the diagnostic reagent of preparation prediction diabetes B susceptibility.

Background technology

The massive epidemiology data shows that diabetes are a kind of disease of multifactorial inheritance with complexity of obvious genetic predisposition.Inherited genetic factors can obviously influence the susceptibility of body to Regular Insulin.At present diabetes Studies on Susceptibility Genes strategy is mainly comprised positional cloning method, candidate gene method and the positional candidate cloning method that the two combines

In recent years, most researchers is devoted to seek the diabetes genes involved by the positional cloning method, promptly seeks interregional chain of the chromosomal DNA shared between ill family member.Found some the relative more rare single-gene forms in the diabetes B, (maturity-onset diabetes of the young MODY), is because sudden change has caused the β cell function low to be called young late hair style diabetes.These patients fall ill to the adult is early stage in pubescence usually, show as the glucose reactivity insulin hyposecretion.6 glucokinases of encoding respectively, transcription factor HNF1 α, 1 β, 4 α and insulin promoter factor (IPF1) are arranged, neurocyte differential protein (NeuroD1) in these Disease-causing genes.These transgenations finally cause the hyperglycemia mass formed by blood stasis.The sickness rate of these single-gene diabetes is also imprecise, estimates to account for 5% of all diabetes Bs.But, utilize the strategy of positional cloning to determine that the successful report of disease of multifactorial inheritance is still rare merely.

Except that the positional cloning method, the genes involved research to disease of multifactorial inheritance also has a kind of method commonly used-candidate gene method at present, i.e. the variation of direct study of disease candidate gene and the relation between the disease phenotype.This method is mainly based on the case-contrast association analysis to family or crowd.Association analysis does not need big family research, but relatively certain or a certain cover are marked at the distributed degrees of patient and normal individual.If it is fairly obvious that certain mark distributes, so just can think that this mark is associated with disease phenotype in diseased individuals.

But, because the Disease-causing gene position is not enough understood,, be primarily aimed at some metabolic pathways relevant and study in the candidate gene method with disease development selecting there is bigger blindness on the candidate gene.The positional candidate cloning method that above-mentioned two kinds of methods are combined has remedied the limitation of any single a kind of method to a certain extent, is the general strategy of employing of institute in the disease of multifactorial inheritance research at present.

The selection of diabetes candidate gene at present mainly contains two sources, and the one, some metabolic pathways relevant with disease development, the one, by the determined chromosomal region of positional cloning method.Diabetes B is a kind of disease of multifactorial inheritance, its susceptibility is the result of a plurality of gene interactions, thereby the inventor selects candidate gene with a kind of new thinking, promptly the Chinese han population diabetes B tumor susceptibility gene that is obtained with this group former studies is a target gene, in its co-expression gene and residing metabolic pathway, seek candidate gene, in the hope of making up diabetes B tumor susceptibility gene network chart.

Corresponding with the inherited disease research strategy, its research tool-polymorphism mark is also from restriction fragment length polymorphism (the Restrictive Fragment Length Polymorphism of the first-generation, RFLP), the microsatellite marker of the s-generation (Microsatellite Marker) develop into the third generation single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP).Especially disease of multifactorial inheritance is being carried out need utilize a large amount of highdensity markers in case-contrast association analysis, preceding two generation polymorphism mark can not satisfy this requirement.Therefore, in recent years, SNP more and more causes people's extensive concern as third generation construction of genetic atlas person.

Single nucleotide polymorphism (SNP) mainly is meant on the genomic level by the caused dna sequence polymorphism of the variation of mononucleotide.It is modal a kind of in human heritable variation, accounts for more than 90% of all known polymorphisms.SNP extensively distributes in human genome, just has 1 in average every 500-1000 base, and its Estimate of Total Number can reach 3,000,000 even more.In recent years, along with the progress of human genome and genetics research, SNP also more and more demonstrates its significance in depth understanding between individuality and colony genomic variation or aspects such as polymorphism and disease research comprehensively.It is exactly its widely distributed property that SNP is used for the localized biggest advantage of diseases predisposing gene.Next is that it has the stability higher than little satellite, can reduce to greatest extent in the genetic process because the information loss that sequence variations caused.In addition, SNP is easy to realize high-throughout somatotype, can produce a large amount of results in the short period of time, and this point is that little satellite is incomparable.The These characteristics of SNP has determined it to be very suitable for multigenic disease, the especially Position Research of the disease that a plurality of minor gene caused as diabetes.Selecting appropriate cases and normal control, take the research method of association analysis, for SNP, is fit closely.

The inventor was engaged in Han People of North China diabetes B (non insulin dependent diabetes, NIDDM) tumor susceptibility gene Position Research in recent years always.Formerly in candidate's positional cloning research that this group is carried out, 6 Chinese han population diabetes B tumor susceptibility gene (PRKCZ have been found, UTS2, CASP9, CDC212, PGD, SAC), the inventor is with a kind of new thinking, trial from said gene path of living in the function genes involved and co-expression gene in select suitable candidate gene, utilize SNP to carry out case-contrast association analysis, finally identified a Chinese han population diabetes B tumor susceptibility gene (being the KCNJ11 gene), and a SNP site relevant especially with the Chinese han population diabetes B on this gene.Finished the present invention through further investigation thus.

Summary of the invention

According to an aspect of the present invention, a kind of test kit of predicting the diabetes B susceptibility is provided, and described test kit comprises based on the special primer that is used for pcr amplification of the sequences Design of Figure 1A or Figure 1B and the contained conventional assembly of test kit that detects by nucleic acid amplification.Described primer designs at the SNP site shown in the figure, and length is 18bp-28bp.Preferably, described primer is selected from a group that SEQ ID NO:1-25 forms, and more preferably, described primer is amplification with sequence shown in SEQ ID NO:26-29 sudden change system (ARMS) primer that is obstructed.

According to a further aspect in the invention, the primer that is used to predict the diabetes B susceptibility is provided, described primer is that described primer designs at the SNP site shown in the figure based on the special primer that is used for pcr amplification of the sequences Design of Figure 1A or Figure 1B, and length is 18bp-28bp.Preferably, described primer is selected from one group that SEQ ID NO:1-25 forms.According to a preferred embodiment of the present invention, primer of the present invention is SEQ ID NO:26-29, shown amplification sudden change system (ARMS) primer that is obstructed.

In accordance with a further aspect of the present invention, KCNJ11 gene (inward rectifier potassium channels is provided, the J subunit, No. 11) be used for the application of the diagnostic reagent of diabetes diagnosis as the diabetes tumor susceptibility gene in preparation, the nucleotide sequence of KCNJ11 gene is shown in SEQ ID NO:30.

Description of drawings

Figure 1A illustrates the nucleotide sequence of the KCNJ11 gene region at place, SNP of the present invention site (R).

Figure 1B is the complementary sequence of Figure 1A sequence

Definition:

Single nucleotide polymorphism (SNP): mainly be meant on the genomic level by the caused dna sequence polymorphism of the variation of single Nucleotide.It is modal a kind of in human heritable variation, accounts for more than 90% of all known polymorphisms.

Gene type and gene frequency distribute: in the same population biology, the sequence analysis of phase isoallele nucleotide sequence is represented determining of the moiety of identical SNP site in Different Individual at this, is gene type.Gene frequency is meant in the same kind allelotrope of being studied, and the allelic proportion that changes just appears in the ratio that the allelotrope of desired appearance accounts for.

Disease susceptibility (diabetes susceptibility): the degree that the crowd experiences easily to disease.

Sudden change system (the amplification refractory mutation system that is obstructed increases, ARMS): claim allele-specific PCR (ASPCR) or specific alleles pcr amplification (PASA) again, this method lacks the characteristics of 3 ' → 5 ' circumscribed proofreading activity based on heat-resisting Taq archaeal dna polymerase, the special base of design 3 ' end is complementary to the primer of the corresponding base of wild-type and mutant allele respectively, can detect a kind of novel method of known point sudden change in the dna sequence dna by PCR reaction and gel electrophoresis.Its characteristics are: fast, repeatable high, cost is low and be convenient to automatization.

Case-contrast association analysis: be a kind of a priori retrospective study.It is earlier to determine the respondent by morbid state, is divided into case and control group, and the difference by the genetic marker frequency of occurrences studied between case group relatively and control group obtains information related between this sign and disease.

Parameter linkage analysis and Non-Parametric Linkage Analysis Methods: calculate two chromosome maps between the gene apart from being linkage analysis according to the recombination fraction of gene, analyze with classified variable (discrete variable), measurement index is sorted out by a certain standard, counting, analyze then and be the parameter analysis, the analysis of carrying out with the result after the actual measurement is a Non-Parametric Linkage Analysis Methods.

K _ATP(ATP sensitive potassium channel): be a kind of inward rectifier potassium channels that is subjected to ATP concentration regulation and control in the cell, by with cellular metabolism and bioelectric coupling connection mutually, performance important physical and physiopathology function in secretion and Muscle contraction process.By to clonal expression and natural type K _ATPThe comparative study of characteristic shows, K _ATPCan be divided into following 5 types: 1. pancreas type, Kir6.2/SUR1 is cardiac muscle and skeletal muscle type 2., and Kir6.2/SUR2A is the vascular smooth muscle type 3., and Kir6.2/SUR2B is the kidney type 4., and Kir1.1/CFTR is the plastosome type 5., Kir6.1/?

The KCNJ11 that this patent relates to (member 11 for potassium inwardly-rectifying channel, subfamily J) gene is called BIR or Kir6.2 again (Chandy and Gutman (1993)In the nomenclature), its proteins encoded is a kind of inward rectifier potassium channels, expresses at the pancreas height, forms the interior K of above-mentioned pancreatic beta cell jointly with sulfonylureas receptor SUR1 (a kind of action target spot of ofhypoglycemic medicine of widespread use) _ATP, be subjected to the G protein regulation and in the dispose procedure of Regular Insulin, play important regulatory role.The proteins encoded of KCNJ11 contains 390 amino acid, form (between the 119th～135 amino-acid residue) by 2 hydrophobic TMDs (laying respectively between the 74th～95 and the 145th～166 amino-acid residue) and H5 district, the latter also is made up of glycine-phenylalanine-glycine.C-terminal has the phosphorylation site (being respectively the 224th Threonine and 363 Serine) that 2 PKA rely in its born of the same parents, the phosphorylation site that interior amino of its born of the same parents and C-terminal have 5 PKC dependences (is respectively the 3rd, 37,36 Serine and 335,344 s' Threonine).It is expressed at the pancreas height, relative low expression the with skeletal muscle of heart.

The long 4.5kb of the encoding gene of KCNJ11, Locus ID3767, merge with the encoding gene of sulfonylureas receptor SUR1, be positioned altogether on the karyomit(e) 11p15.1, the sudden change that has proved the KCNJ11 gene is the important cause of disease of the low high glucose mass formed by blood stasis of Regular Insulin (PHHI) of familial inheritance baby

(http：//www.ncbi.nlm.nih.gov/LocusLink/LocRpt.cgi？l＝3767)。Find that in research the E23K sudden change of KCNJ11 is relevant with diabetes B to European white man.

Embodiment

Have a plurality of public databases to store millions of SNP information at present, people can be in several ways, as with gene name, registration number, karyomit(e) number, functional class etc., searches.But, the SNP in these databases most just " candidates ", but not real SNP through verifying.They utilize computer to excavate out when a plurality of cloned sequences are compared, and wherein certainly exist a part of false positive SNP.In addition, these SNP great majority are the SNP that utilize Caucasian and Black people to check order and seek, and some is not necessarily identical for the distribution in Chinese population, and we utilize the diabetic subject and the normal control of Chinese han population, and SNP is sought in order-checking again.And utilize the SNP of the high frequency that we find in the China Han diabetic population to carry out gene type.

For seeking the diabetes B tumor susceptibility gene, the information that the inventor announces according to American National information biology (National Center for Biotechnology Information) center, trial is from previous 6 the Chinese han population diabetes B tumor susceptibility gene (PRKCZ that find of this group, UTS2, CASP9, CDC212, PGD, SAC) 18 Chinese han population diabetes B candidate genes have been selected in function genes involved in the path of living in and the co-expression gene, exon to these genes, Promotor district 2kb, and check order SNP in the examination Han People of North China of the zone of the intron that closes on.And the SNP of part high frequency wherein carried out gene type, by methods such as ARMS reaction, sequencings it is carried out somatotype and carry out case-contrast association analysis in case group and control group.

Go through as following examples, in the SNP site of selected 72 high frequencies (gene frequency 〉=15%), choose 7 sites in the KCNJ11 gene and carried out gene type, find that this site is relevant with diabetes, it is positioned at a SNP site (R at 990 places of No. second exon, be G/A), numbering among the NCBI is rs5213, the difference that its gene frequency is distributed in case group and the control group has statistical significance (P＜0.05), point out this SNP site relevant with diabetes B, further reach a conclusion, its place gene KCNJ11 gene is the tumor susceptibility gene of Chinese han population diabetes B.

Therefore, one aspect of the present invention relates to the application of diabetes tumor susceptibility gene KCNJ11 gene in the prediction diabetes susceptibility.Based on the KCNJ11 gene, can obtain various diagnostic reagents and test kit to be used to predict diabetes susceptibility.

KCNJ11 place genome sequence is shown in SEQ ID NO:30, total length is 6,458bp, disclosed SNP site (R) is positioned at 4 of this sequence, 107-4, between 976, this regional nucleotide sequence is shown in Figure 1A, and R is the SNP site among the figure, and it represents bases G/A polymorphism, promptly this site can be G, also can be A.This regional complementary sequence is shown in Figure 1B, and R represents the complementary base in SNP of the present invention site among the figure, and it represents base C/T.

The invention still further relates to the primer that is used to predict diabetes susceptibility.Preferably, described primer designs based on the sequence of Figure 1A, and thus, the product order-checking behind the pcr amplification can directly record the SNP site.Certainly, primer of the present invention also can be based on the sequences Design of Figure 1B, and the order-checking of the product behind the pcr amplification obtains the complementary base of SNP like this.Preferably, primer of the present invention has the sequence shown in the SEQ ID NO:1-25, particularly preferably is the ARMS-PCR primer shown in the SEQ ID NO:26-29.Following table 1 shows some combinations of primer of the present invention, the product size that the melting temperature(Tm) of primer (Tm) and these combinations obtain for example.It will be appreciated by those skilled in the art that primer of the present invention is not limited to these primer of listing and combinations thereof.

Primer sequence shown in the table 1 SEQ ID NO:1-25

SEQ?ID?NO：	Primer sequence	Tm	Product length
				1 2	Adopted CAGCCTACTGGAAGCTCTGAC antisense CTTAGCCAGGTGCTTTTTCAG is arranged	57 58	563
3 4	Adopted GCTTGATGAGGACCACAGCCTACT antisense AACACCCTCTCTCATCAACTGCCCT is arranged	59 62	623
				3 5	Adopted GCTTGATGAGGACCACAGCCTACT antisense AACACCCTCTCTCATCAACTGCC is arranged	59 58	623
3 6	Adopted GCTTGATGAGGACCACAGCCTACT antisense ACACCCTCTCTCATCAACTGCCCT is arranged	59 61	622
				7 4	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCCCT is arranged	62 62	624
3 8	Adopted GCTTGATGAGGACCACAGCCTACT antisense AACACCCTCTCTCATCAACTGCCC is arranged	59 61	623
				7 5	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCC is arranged	62 58	624
7 6	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense ACACCCTCTCTCATCAACTGCCCT is arranged	62 61	623
				7 8	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCCC is arranged	62 61	624
3 9	Adopted GCTTGATGAGGACCACAGCCTACT antisense TCTCTCATCAACTGCCCTCCCT is arranged	59 58	616
				10 4	Adopted GCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCCCT is arranged	62 62	623
3 11	Adopted GCTTGATGAGGACCACAGCCTACT antisense GGAACACCCTCTCTCATCAACTGC is arranged	59 60	625
				12 4	Adopted AGGACCACAGCCTACTGGAAGCT antisense AACACCCTCTCTCATCAACTGCCCT is arranged	59 62	615
10 5	Adopted GCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCC is arranged	62 58	623
				7 13	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense GAACACCCTCTCTCATCAACTGCC is arranged	62 60	625
10 6	Adopted GCTTGATGAGGACCACAGCCTACTG antisense ACACCCTCTCTCATCAACTGCCCT is arranged	62 61	622

3 14	Adopted GCTTGATGAGGACCACAGCCTACT antisense ACCCTCTCTCATCAACTGCCCT is arranged	59 57	620
				7 9	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense TCTCTCATCAACTGCCCTCCCT is arranged	62 58	617
3 15	Adopted GCTTGATGAGGACCACAGCCTACT antisense ACACCCTCTCTCATCAACTGCCC is arranged	59 60	622
				12 5	Adopted AGGACCACAGCCTACTGGAAGCT antisense AACACCCTCTCTCATCAACTGCC is arranged	59 58	615
12 6	Adopted AGGACCACAGCCTACTGGAAGCT antisense ACACCCTCTCTCATCAACTGCCCT is arranged	59 61	614
				10 8	Adopted GCTTGATGAGGACCACAGCCTACTG antisense AACACCCTCTCTCATCAACTGCCC is arranged	62 61	623
7 11	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense GGAACACCCTCTCTCATCAACTGC is arranged	62 60	626
				12 8	Adopted AGGACCACAGCCTACTGGAAGCT antisense AACACCCTCTCTCATCAACTGCCC is arranged	59 61	615
16 4	Adopted GCTTGATGAGGACCACAGCCTAC antisense AACACCCTCTCTCATCAACTGCCCT is arranged	58 62	623
				7 14	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense ACCCTCTCTCATCAACTGCCCT is arranged	62 57	621
10 13	Adopted GCTTGATGAGGACCACAGCCTACTG antisense GAACACCCTCTCTCATCAACTGCC is arranged	62 60	624
				7 15	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense ACACCCTCTCTCATCAACTGCCC is arranged	62 60	623
3 17	Adopted GCTTGATGAGGACCACAGCCTACT antisense TCTCTCATCAACTGCCCTCCCTG is arranged	59 61	616
				10 9	Adopted GCTTGATGAGGACCACAGCCTACTG antisense TCTCTCATCAACTGCCCTCCCT is arranged	62 58	616
16 5	Adopted GCTTGATGAGGACCACAGCCTAC antisense AACACCCTCTCTCATCAACTGCC is arranged	58 58	623
				3 18	Adopted GCTTGATGAGGACCACAGCCTACT antisense ACACCCTCTCTCATCAACTGCC is arranged	59 57	622
3 19	Adopted GCTTGATGAGGACCACAGCCTACT antisense CTCTCTCATCAACTGCCCTCCCT is arranged	59 59	617
				12 20	Adopted AGGACCACAGCCTACTGGAAGCT antisense CGGGAACACCCTCTCTCATCAA is arranged	59 60	619
10 11	Adopted GCTTGATGAGGACCACAGCCTACTG antisense GGAACACCCTCTCTCATCAACTGC is arranged	62 60	625
				12 9	Adopted AGGACCACAGCCTACTGGAAGCT antisense TCTCTCATCAACTGCCCTCCCT is arranged	59 58	608
16 6	Adopted GCTTGATGAGGACCACAGCCTAC antisense ACACCCTCTCTCATCAACTGCCCT is arranged	58 61	622
				12 13	Adopted AGGACCACAGCCTACTGGAAGCT antisense GAACACCCTCTCTCATCAACTGCC is arranged	59 60	616

12 21	Adopted AGGACCACAGCCTACTGGAAGCT antisense CGGGAACACCCTCTCTCATCAACT is arranged	59 61	619
				7 22	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense CACCCTCTCTCATCAACTGCCCT is arranged	62 60	622
16 8	Adopted GCTTGATGAGGACCACAGCCTAC antisense AACACCCTCTCTCATCAACTGCCC is arranged	58 61	623
				12 11	Adopted AGGACCACAGCCTACTGGAAGCT antisense GGAACACCCTCTCTCATCAACTGC is arranged	59 60	617
7 17	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense TCTCTCATCAACTGCCCTCCCTG is arranged	62 61	617
				10 14	Adopted GCTTGATGAGGACCACAGCCTACTG antisense ACCCTCTCTCATCAACTGCCCT is arranged	62 57	620
7 18	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense ACACCCTCTCTCATCAACTGCC is arranged	62 57	623
				7 19	Adopted AGCTTGATGAGGACCACAGCCTACTG antisense CTCTCTCATCAACTGCCCTCCCT is arranged	62 59	618
10 15	Adopted GCTTGATGAGGACCACAGCCTACTG antisense ACACCCTCTCTCATCAACTGCCC is arranged	62 60	622
				12 14	Adopted AGGACCACAGCCTACTGGAAGCT antisense ACCCTCTCTCATCAACTGCCCT is arranged	59 57	612
3 22	Adopted GCTTGATGAGGACCACAGCCTACT antisense TCTCTCATCAACTGCCCTCCC is arranged	59 57	616
				3 23	Adopted GCTTGATGAGGACCACAGCCTACT antisense TCTCATCAACTGCCCTCCCTG is arranged	59 58	614
16 13	Adopted GCTTGATGAGGACCACAGCCTAC antisense GAACACCCTCTCTCATCAACTGCC is arranged	58 60	624
				24 4	Adopted AGCTTGATGAGGACCACAGCC antisense AACACCCTCTCTCATCAACTGCCCT is arranged	58 62	624
25 4	Adopted GGACCACAGCCTACTGGAAGCT antisense AACACCCTCTCTCATCAACTGCCCT is arranged	58 62	614

The invention further relates to the test kit that comprises one or more primer of the present invention, this test kit is used to detect the susceptibility of diabetes.Except primer of the present invention, described test kit also comprises the utilization pcr amplification and the conventional assembly of the test kit that detects, reagent, damping fluid etc., and those skilled in the art are familiar with these conventional assembly and detection methods.

Further describe the present invention hereinafter with reference to embodiment.

The correlative study of SNP in the evaluation of embodiment 1, Chinese han population diabetes B tumor susceptibility gene KCNJ11 and this gene

1, the collection of Chinese han population diabetes B patient and normal control sample

Used diabetes B group of the present invention and normal group crowd peripheral blood sample (10ml) are mainly from the outpatient service of Endocrinology Department of BJ Union Hospital.Diabetes cases can be distributes, and also can be member's (but have only in each family this moment a patient selected usually, the member without any genetic connection in the same family can select more than) of diabetic pedigree; Normal control requires the three generations of its family with the interior diabetic subject of not having (comprising type i diabetes) from normal population, and control group and ill group of age and sex are complementary.

The diagnosis of diabetes B is carried out in strict accordance with the standard that WHO issued in 1985.That is: fasting blood glucose concn 〉=7.8mmol/L or 2 hours after the meal blood sugar 〉=11.1mmol/L are diabetes.The judging criterion of OGTT is: back 2 hours blood glucose concentration＜7.8mmol/L of clothes sugar are for normal, and 〉=11.1mmol/L is diabetes, between 7.8-11.1mmol/L be impaired glucose tolerance (impaired glucose tolerance, IGT).

Each selected member record generalized case, comprise projects such as sex, date of birth, native place, height, body weight, heart rate, breathing, blood pressure, the investigation and the special examined of going where necessary are associated with diseases such as hypertension, heart trouble, atherosclerosis with eliminating.Except that above-mentioned project, the diabetic subject is also write down projects such as age of onset, the highest body weight of the past, blood fat, glycolated hemoglobin and diabetic complication, normal control group is also measured fasting plasma glucose and got rid of diabetes family history person.

Guarantee before the blood sample collection that all members understand the blood sampling purpose, right to know is promptly arranged, and go up signature at " Informed Consent Form ".The data of collecting is holded in close confidence, externally do not announce all data such as name, age, disease of blood supply member.

Collect 714 parts of samples altogether, wherein the case group is 371 parts, 343 parts of control groups.Two groups in sex, be complementary on the age, and diabetic subject's mean age is 53.01 years old, and the mean age during the normal control sampling is 49.68 years old.In all members, men and women's ratio is 361: 353, the case group male sex 190 people, women 181 people; The control group male sex 168 people, women 175 people.Two group memberships' age structure situation such as table 2:

Ill group of table 2 and control group member's age composition

Age bracket	The case group	Control group
			＜30	5	7
30-39	17	89
			40-49	73	218
≥50	276	129
			Amount to	371	343

2, be used for the selection in the SNP site of gene type in the definite and candidate gene of candidate gene

Based on previous 6 the Chinese han population diabetes B tumor susceptibility gene (PRKCZ that find of this group, UTS2, CASP9, CDC212, PGD, SAC), the information that the inventor announces according to American National information biology center (National Center for Biotechnology Information), 18 Chinese han population diabetes B candidate genes have been selected at these 6 in based on function genes involved in the related path and co-expression gene, to the exon of these genes, Promotor district 2kb, the zone of the intron that closes on and 3 ' the non-translational region SNP in the examination Han People of North China that checks order.And the SNP of 72 high frequencies (gene frequency 〉=15%) wherein carried out gene type by methods such as ARMS reaction, sequencings.

3, SNP locus gene phenotypic analysis

The main somatotype that adopts 2 kinds of methods to carry out SNP: sudden change system (amplificationrefractory mutation system, ARMS) somatotype and the sequencing and typing of being obstructed increases.

1). the amplification be obstructed the sudden change system (amplification refractory mutation system, ARMS)

1.1) design of primers and synthetic: utilize Tetra-primer ARMS-PCR software to carry out design of primers (http://cedar.genetics.soton.ac.uk/public_html/primerl.html), 4 primers of each SNP site design in 7 SNP sites in the KCNJ11 gene are respectively the inner primer of forward, reverse inner primer, the outside primer of forward, adverse external primer.

ARMS-PCR design of primers principle: primer length is between 18～30base, and the Tm value is between 55～80 ℃.The suitableeest inner PCR product length 350bp, two inner PCR product length ratios are 1.3-2.

1.2) ARMS PCR reaction and product evaluation: carry out Tetra-prmer ARMS-PCR at each the high frequency SNP site in the KCN11 gene, reaction system 15 μ l, each component such as table 3:

Table 3 ARMS PCR reaction system

Title original liquid concentration application of sample amount (μ l)

Q-solution 5× 3

buffer 10× 1.5

primer ^*4 20μM 0.3×4

Mgcl2 25mM 0.9

dNTP 10mM 0.45

HotstarTaq 5units/μl 0.12

template 20ng/μl 1

DdH2O polishing to 15 μ l

Annotate: employed Q-solution, buffer, Mgcl2, HotstarTaq provide by the HotstarTaq test kit of QIAGEN company

Adopt Touch-down PCR, its program is as follows: after 94 ℃, 12 minutes, carry out 94 ℃ of 12 round-robin 30 seconds, (Tm+2 ℃) 60 seconds (each circulating temperature descends 0.5 ℃), 72 ℃ 50 seconds, 30 seconds, 72 ℃ of 94 ℃ of 30 round-robin 30 seconds, (Tm-3 ℃) are 45 seconds subsequently, last 72 ℃ 10 minutes 1 time.Reaction finishes the back to be identified with 2% agarose electrophoresis, observes the product situation under the UV lamp, and takes pictures writing time, site title, sample number etc. with ultraviolet gel imaging instrument.

1 .3) genotype is judged and data preparation: according to band quantity and location determination SNP genotype.With each site somatotype result arrangement is that the Excel form is to carry out statistical treatment.

(2) sequencing and typing: carry out sequencing and typing with above-mentioned ARMS method somatotype or the unfavorable site of somatotype result for being not suitable for.

More than the somatotype in each SNP site all in the normal control individuality of 96 picked at random, carry out earlier.

In case group and control group, the SNP site of above-mentioned 72 somatotypes all meets the Hardy-Weinberg balance.In view of the KCNJ11 gene order lacks 6,458bp, it is carried out full gene sequencing, examination obtains 7 in high frequency SNP site, adopt in above-mentioned 2 method to carry out gene type, analyze through SPSS statistical software 11.0 editions (free download is in http://linkage.rockefeller.edu/soft/), there is 1 SNP site to be positioned at the 989th of second exon of KCNJ11 gene, rs5213 in the snp database of NCBI, gene frequency is distributed in that difference has statistical significance (P＜0.05) in two groups, and is as shown in table 4.

Table 4 case group and control group SNP somatotype SPSS statistic analysis result

Allelotrope

SNP C T A G amounts to the P value

Case group 306 436 742 0.002

Rs203849 control group 339 347 686

Amount to 645 783 1428

There is significant difference in the gene frequency that is arranged in 1 SNP site of second exon of KCNJ11 gene in case group and control group, therefore points out this site relevant with diabetes B.And then prompting KCNJ11 gene is a kind of new diabetes tumor susceptibility gene.

KCNJ11 place genome sequence is shown in SEQ ID NO:30, total length is 6,458bp, disclosed SNP site (R) is positioned at 4 of this sequence, 107-4, between 976, this regional nucleotide sequence is shown in Figure 1A, and R is the SNP site among the figure, and it represents bases G/A polymorphism, promptly this site can be G, also can be A.This regional complementary sequence is shown in Figure 1B, and R represents the complementary base in SNP of the present invention site among the figure, and it represents base C/T.

Based on this SNP site, can design the primer of suitable length, be used for predicting diabetes susceptibility through pcr amplification.

The prediction of embodiment 2 prediction diabetes susceptibility primer design and diabetes susceptibility

1.ARMS-PCR primer

The ARMS-PCR design of primers is followed following principle: primer length is between 18～30base, and the Tm value is between 55～80 ℃.The suitableeest inner PCR product length 350bp, two inner PCR product length ratios are 1.3-2.At 4 primers of this SNP site design in the KCNJ11 gene, as shown in table 5 below:

ARMS-PCR primer shown in the table 5 SEQ ID NO:26-29

SEQ?ID?NO：	Primer title primer sequence	Tm	Product length
				26 27	Inner primer (G allele) CCTCACACCAGGCCCTGTCC of adopted inner primer (A allele) CACGGACCATGTGGTATGTAGCACA antisense is arranged	67 67	172 269
28 29	Outside primer (5 '-the 3 ') GTTGGGAGGAGCAGGGACAAAAATA of adopted outside primer (5 '-3 ') ACTACTCCAAGTTTGGCAACACCGTC antisense is arranged	67 67	396

Organize Tetra-primer ARMS-PCR primer carries out ARMS-PCR reaction detection SNP site to genes of individuals group DNA to be measured sequence with this.

2, regular-PCR amplimer

According to the sequence (Figure 1A and 1B) at place, SNP site, the primer of design shown in SEQ ID NO:1-25.As shown in table 1ly carry out various combinations.With the described primer that is combined as, be that template is carried out pcr amplification with the genomic dna that derives from individual blood sample to be measured, amplified production is checked order detect the single nucleotide polymorphism in described SNP site.

SEQUENCE?LISTING

＜110〉Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences

Sinogenomax Co., Ltd.

＜120〉test kit and the primer of forecast China people diabetes B susceptibility

<130>I20031212CB

<160>30

<170>PatentIn?version?3.1

<210>1

<211>21

<212>DNA

＜213〉artificial sequence

<400>1

cagcctactg?gaagctctga?c 21

<210>2

<211>21

<212>DNA

＜213〉artificial sequence

<400>2

cttagccagg?tgctttttca?g 21

<210>3

<211>24

<212>DNA

＜213〉artificial sequence

<400>3

gcttgatgag?gaccacagcc?tact 24

<210>4

<211>25

<212>DNA

＜213〉artificial sequence

<400>4

aacaccctct?ctcatcaact?gccct 25

<210>5

<211>23

<212>DNA

＜213〉artificial sequence

<400>5

aacaccctct?ctcatcaact?gcc 23

<210>6

<211>24

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<400>6

acaccctctc?tcatcaactg?ccct 24

<210>7

<211>26

<212>DNA

＜213〉artificial sequence

<400>7

agcttgatga?ggaccacagc?ctactg 26

<210>8

<211>24

<212>DNA

＜213〉artificial sequence

<400>8

aacaccctct?ctcatcaact?gccc 24

<210>9

<211>22

<212>DNA

＜213〉artificial sequence

<400>9

tctctcatca?actgccctcc?ct 22

<210>10

<211>25

<212>DNA

＜213〉artificial sequence

<400>10

gcttgatgag?gaccacagcc?tactg 25

<210>11

<211>24

<212>DNA

＜213〉artificial sequence

<400>11

ggaacaccct?ctctcatcaa?ctgc 24

<210>12

<211>23

<212>DNA

＜213〉artificial sequence

<400>12

aggaccacag?cctactggaa?gct 23

<210>13

<211>24

<212>DNA

＜213〉artificial sequence

<400>13

gaacaccctc?tctcatcaac?tgcc 24

<210>14

<211>22

<212>DNA

＜213〉artificial sequence

<400>14

accctctctc?atcaactgcc?ct 22

<210>15

<211>23

<212>DNA

＜213〉artificial sequence

<400>15

acaccctctc?tcatcaactg?ccc 23

<210>16

<211>23

<212>DNA

＜213〉artificial sequence

<400>16

gcttgatgag?gaccacagcc?tac 23

<210>17

<211>23

<212>DNA

＜213〉artificial sequence

<400>17

tctctcatca?actgccctcc?ctg 23

<210>18

<211>22

<212>DNA

＜213〉artificial sequence

<400>18

acaccctctc?tcatcaactg?cc 22

<210>19

<211>23

<212>DNA

＜213〉artificial sequence

<400>19

ctctctcatc?aactgccctc?cct 23

<210>20

<211>22

<212>DNA

＜213〉artificial sequence

<400>20

cgggaacacc?ctctctcatc?aa 22

<210>21

<211>24

<212>DNA

＜213〉artificial sequence

<400>21

cgggaacacc?ctctctcatc?aact 24

<210>22

<211>23

<212>DNA

＜213〉artificial sequence

<400>22

caccctctct?catcaactgc?cct 23

<210>23

<211>21

<212>DNA

＜213〉artificial sequence

<400>23

tctcatcaac?tgccctccct?g 21

<210>24

<211>21

<212>DNA

＜213〉artificial sequence

<400>24

agcttgatga?ggaccacagc?c 21

<210>25

<211>22

<212>DNA

＜213〉artificial sequence

<400>25

ggaccacagc?ctactggaag?ct 22

<210>26

<211>25

<212>DNA

＜213〉artificial sequence

<400>26

cacggaccat?gtggtatgta?gcaca 25

<210>27

<211>20

<212>DNA

＜213〉artificial sequence

<400>27

cctcacacca?ggccctgtcc 20

<210>28

<211>26

<212>DNA

＜213〉artificial sequence

<400>28

actactccaa?gtttggcaac?accgtc 26

<210>29

<211>25

<212>DNA

＜213〉artificial sequence

<400>29

gttgggagga?gcagggacaa?aaata 25

<210>30

<211>6458

<212>DNA

＜213〉artificial sequence

<400>30

tgttaagagt?gttggttgat?cagggaggtg?ggagagagat?tacagcagtg?gggagggcat 60

catatcagag?agccctgaat?gccagagcaa?ggagtgtggg?ctatgcccat?ggacaaaaag 120

ggagaagggt?tttgagcaag?aaagcgactt?gacaagttca?gtatttggtg?gagatgacta 180

taagcaagtg?caggatggag?caaaagggga?ggctgagcct?ttcaggtggg?agactcagcc 240

tctcagaagg?gtgagcagca?gggagggagg?taaggcctca?aggctgcacc?aggaggcaga 300

acccagcctt?tccagctctg?agatttcccc?aggggcccag?cagcttcctc?cactgtgaga 360

tctcccaaga?gcagagccgg?ggaactgaaa?gaggactcca?caagcctacc?actgctccaa 420

agccacccca?cctcaacaga?ttgaagggaa?gtagcagggg?cagggctggg?cccacagtga 480

ggtgttgcag?ggatgcccaa?ggtcatcctg?gcccaaaatc?cctgtataac?atccctcatg 540

gcccaccgac?ttgatttggg?taccccacag?atgtggctca?tctctggtta?ttgttattag 600

aactgactct?tgcatatgtg?aaaccaaagc?ctgcctctgt?tccacttagt?gtcctggttc 660

taccctttgg?gccacaaaaa?actactagtt?catcctcctt?caaggccagg?tgtttctgca 720

atagtgccca?aatccctggg?ggactttcct?tttctgatgg?ctggacagtt?gtagttgcct 780

cagagatttc?cccatgtaac?cctgtctcct?tgagcttccc?actctgctca?tctcatttta 840

ccctcccctt?cctagacact?gcatctttgt?taatatagcc?catgatcaca?cccatccttt 900

gtagcggtgt?tcgctctgcc?tgcaggtggg?tagggagaga?aaggaaaatc?caatataaaa 960

tgcatatctt?gcccccactg?tggcccacag?ccacactgcc?ttttccagac?cctgtctcaa 1020

gcccccactc?caacaaaatg?gccgcagatg?gtcaggggtt?cttccataag?gatctgagcc 1080

acagacagcg?gtaaagccgt?tactgttcca?acaacagaga?tctagcaggc?tttccctccc 1140

agctgagaac?ccgggtccat?gccacagggc?agcctcaggc?tgatgcctga?acagaaacac 1200

caccatgccg?ggaccaagag?cctgggtccc?ccagtgcccg?aggcagccct?aacacactgt 1260

tcaggcctaa?ataccaccca?tgacctaacg?tggaagaagt?caggtgacag?gtgtgagaaa 1320

gtgtaaagca?agtggcctcg?gctcaaacct?tctttaaagt?agggccggaa?tggctgagaa 1380

gatggagccg?cccaggctgc?cccacagcag?tagcaggtgt?taaactgtgc?tcctttcctg 1440

ccgaggcctt?tttttttttt?tttttttttt?ttttttttaa?tgagagggag?tctgagcttc 1500

catcagattg?tcaaaggatt?caagactaag?attaagagca?tcccccaaag?agaagcagct 1560

gcaggttgag?aagtcctcgg?accgcctaat?ttccgccagc?cctgggattg?gtggcgccgc 1620

ctctcagcgc?ctcctttccc?ggggaacctg?caaacctggc?gcggccgaga?ggcggattac 1680

tgacccaaat?acggatgggg?atattggggg?aggggagcca?gggcgacagg?gcggcggccc 1740

cagactggga?gggcgggtca?cagatccctc?ccagcccccc?actttcgtag?agcgtgggca 1800

ggaggaacct?ctgggttccg?cggatcagaa?aaactccaaa?ggccgggttg?tgagtcccgg 1860

gaggggaggt?ggagggcggg?ggcggggagg?ggcgcgggag?gggcgggggc?ttgctccggc 1920

cccgccccct?gcgctccggt?gcaggtcggg?ctccccctgg?cggtccccgg?ccccgttcct 1980

ctcctcgtgc?gcccccctcc?cgccgtccta?gacccctgcc?tagcccaggt?cggtctccgc 2040

ggacccacgg?acggacagac?agacgggagg?acggccagcc?gcgagcgccc?gggcggcggg 2100

agggggcggg?gaggcgacgg?ccgtggcgtg?aggagcagga?gcaggtgcag?cggcggcggc 2160

gggcggggcc?gggacccggc?gcggagcggg?agccgcggcg?cgggcgggcg?gcagggaccg 2220

ggaggccgcg?actcggagtc?agccccgccg?ggtcgcgcgc?aggtccgggg?agccgcggtt 2280

gagccgggtg?gggtggtgac?tccagagaac?gcaggatccc?aaggagacag?agaggacgag 2340

agctggaggg?ggatccggaa?agcggcgggg?gcgctccggg?aggggtggag?taggacatag 2400

ggggcgcacc?tggaggagag?acggggcggg?ggtggccagg?acctgagctg?gagcctggga 2460

gcccgaaggc?cagacaggtg?aggcgggaga?cccggaggtg?ggggtgaggt?ccggttagtg 2520

ggagagatcc?ggaggtgtta?agttctgagc?tgggctggga?aggcaggctg?ggcggggaga 2580

agggctctta?gcgggaggcc?caggggtggt?cagctggtgg?gggaagctgg?gggaggacgc 2640

agggccaggt?ggagagccgg?cagggttggg?ggctccctag?gcgccaggca?ggtgggctca 2700

agggtgaggc?tgtttttttt?gttttgtttt?ttgtttttga?gacggagtct?cgctctgtcg 2760

cccaggctgg?agtgcagtgg?cgtgatcttg?gctcactgca?acctccgcct?ctcgggttca 2820

agcgattctc?ctgcctcagc?ttcctgagta?gctgggatta?caggtgcgca?ccaccatgcc 2880

cggctaactt?ttgtattttt?agtagagatg?gggtttcacc?atgttggtca?ggctggtctc 2940

gaactcctga?cctagtgatc?tgccctcctc?agcctcccaa?cgtactggga?ttacaggcgt 3000

gagccaccgc?gcccggcctg?aggctggtat?taagaagtga?agtgggaccc?aggtggaggt 3060

aaggaagagt?ctggtgggga?gttatctcag?aagtgaggcc?agcacaggct?gagtgcagcc 3120

ccagggtgag?aaggtgccca?ccgagaggac?tctgcagtga?ggccctaggc?cacgtccgag 3180

gggtgcctcc?gatgggggaa?gcccctccct?gggggtcacc?ggagccatgc?tgtcccgcaa 3240

gggcatcatc?cccgaggaat?acgtgctgac?acgcctggca?gaggaccctg?ccaagcccag 3300

gtaccgtgcc?cgccagcgga?gggcccgctt?tgtgtccaag?aaaggcaact?gcaacgtggc 3360

ccacaagaac?atccgggagc?agggccgctt?cctgcaggac?gtgttcacca?cgctggtgga 3420

cctcaagtgg?ccacacacat?tgctcatctt?caccatgtcc?ttcctgtgca?gctggctgct 3480

cttcgccatg?gcctggtggc?tcatcgcctt?cgcccacggt?gacctggccc?ccagcgaggg 3540

cactgctgag?ccctgtgtca?ccagcatcca?ctccttctcg?tctgccttcc?ttttctccat 3600

tgaggtccaa?gtgactattg?gctttggggg?gcgcatggtg?actgaggagt?gcccactggc 3660

catcctgatc?ctcatcgtgc?agaacatcgt?ggggctcatg?atcaacgcca?tcatgcttgg 3720

ctgcatcttc?atgaagactg?cccaagccca?ccgcagggct?gagaccctca?tcttcagcaa 3780

gcatgcggtg?atcgccctgc?gccacggccg?cctctgcttc?atgctacgtg?tgggtgacct 3840

ccgcaagagc?atgatcatca?gcgccaccat?ccacatgcag?gtggtacgca?agaccaccag 3900

ccccgagggc?gaggtggtgc?ccctccacca?ggtggacatc?cccatggaga?acggcgtggg 3960

tggcaacagc?atcttcctgg?tggccccgct?gatcatctac?catgtcattg?atgccaacag 4020

cccactctac?gacctggcac?ccagcgacct?gcaccaccac?caggacctcg?agatcatcgt 4080

catcctggaa?ggcgtggtgg?aaaccacggg?catcaccacc?caggcccgca?cctcctacct 4140

ggccgatgag?atcctgtggg?gccagcgctt?tgtgcccatt?gtagctgagg?aggacggacg 4200

ttactctgtg?gactactcca?agtttggcaa?caccgtcaaa?gtgcccacac?cactctgcac 4260

ggcccgccag?cttgatgagg?accacagcct?actggaagct?ctgaccctcg?cctcagcccg 4320

cgggcccctg?cgcaagcgca?gcgtgcccat?ggccaaggcc?aagcccaagt?tcagcatctc 4380

tccagattcc?ctgtcctgag?ccatggtctc?tcgggccccc?cacacgcgtg?tgtacacacg 4440

gaccatgtgg?tatgtagccc?ggccagggcc?tggtgtgagg?ctgggccagc?ctcagctcag 4500

cctccccctg?ctgctcatcc?agggtgttac?aaggcacttg?tcactatgct?atttctggcc 4560

tcagcaggaa?cctgtactgg?gttatttttg?tccctgctcc?tcccaaccca?attcaggact 4620

ggctcacccc?tctcccccgc?ccaaggctgc?agaggctgtg?ggaggtactg?ggccctagag 4680

ctgtgcgtcc?agccagtcct?gggtccccac?gattgaccag?ccacactctg?ggccggtggc 4740

tggggaagaa?caatccccga?gggctgctgc?tttgcgtctg?tggctccaag?aagtgcctgt 4800

ggtcaggccc?cagctctact?tggtccctga?aaaagcacct?ggctaagggc?tgggcctggc 4860

cagcagggag?ggcagttgat?gagagagggt?gttcccgctg?gagggttggt?gctgtggagc 4920

ctacaccggc?agggacagcc?tggggctgac?agggctcccc?tccgagggcc?agtttcaggt 4980

ctggaagggg?aggaagcagg?ggaaggtgac?ctgaggaggc?tcggctttgt?agagccccgc 5040

tcaggcacag?ggaggaggag?atgccagggc?tcctgccttt?tgccacatcg?gcctcgtgca 5100

gtgagggctc?tgtgggctgg?ggctgctgcc?cctgcctacc?tcctgcctgt?ccccagaggc 5160

tgaggagagg?gggtactgtg?cccaccacac?atgattaggc?ctcagaccca?actctggtcc 5220

tggctccaca?acagtggctg?ccactcactt?tgtccagaag?gtggcttggg?ggtggatatc 5280

tttgggttgc?tggaaaaggt?gtgggaaggt?tcaggatggt?gggagggact?gaggtccctg 5340

aggtgaagag?gcccttggtc?ctgacgggtt?tgacccgtgc?ctggaccctt?ggagcagtgt 5400

tgtgtgaact?tgcctagaac?tctgccttct?ccgttgtcaa?taaagcctcc?ccctcatgac 5460

ctaaactctg?ggcttttctt?gctggggagg?cagcaagcat?gctggtggga?agggaggcag 5520

ggactggcag?ctgccacccc?cttcaagagg?cgccatagac?cctagcgggg?agggcagggg 5580

agggacggaa?ggctggcacc?tcttccacca?gttcaggggg?actttcccct?ctcctgtctc 5640

aggtggccca?gccctgtcag?cctgtctggc?caactcagcc?tttgggcact?caccaggctt 5700

tgcagccctg?ggctctgtct?ctactcccag?ggacctgctg?gaaggctgga?gtgcccaggg 5760

agaggtatag?aggtgtcata?ggcattagtg?tagtaattgg?agcactaact?ctcgagccaa 5820

ctgcctgggt?tcgaatcctg?gctctagctg?tatgactttt?gtcaagtaac?ttagcctctc 5880

tgtgtctcag?ttgcctcttc?tataacatgg?atgctaatag?tacctacctc?atagaattgt 5940

tttggaagta?aatgaaaaat?atgtaaaatg?ctgaagtgcc?tggtctacag?taagtgctca 6000

ataaatgtta?actattgtga?ttgctgctga?atcagctaca?tgctgaggaa?acggccaaac 6060

aagtgttaaa?gagagacggt?gcttttattt?tgcccttggt?catgtttctt?tgccaggtct 6120

ttctttccac?acggtgtccc?ctcctagctg?agaccacgat?tttaggggct?gtggcaatgc 6180

ctgtgacctg?gtccccttgt?ctccatctca?cccactgtag?tccatctgcc?acacagcctc 6240

cagggtggtc?ttagaagggt?gggtcacccc?ccaccctcat?catgaaatac?cttcggtagt 6300

tccccatcac?ctctgggtca?aggctagctt?cttagcagcc?atttcaggct?gtgactgcct 6360

ctccggcctt?atggcccact?gttttcttgc?caggagtctg?cacttgccca?cccacttccc 6420

taggtacctc?gctgtcctat?ccctcctgac?acataatg 6458

Claims (5)

1, be used to predict the primer of diabetes B susceptibility, the special primer that is used for pcr amplification that described primer designs for the sequence based on Figure 1A or Figure 1B, described primer designs at the SNP site R shown in the figure, and length is 18-28 Nucleotide.

2, primer as claimed in claim 1, it comprises one group a kind of sequence that is selected from SEQ ID NO:1-25 composition.

3, primer as claimed in claim 2, it is the ARMS-PCR primer with the sequence shown in the SEQ ID NO:26-29.

4, be used to predict the test kit of diabetes B susceptibility, it comprises each described primer as claim 1-3.

5, the KCNJ11 gene is used for predicting the application of the diagnostic reagent of diabetes B susceptibility in preparation.

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