CN106434980A - Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit - Google Patents
Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit Download PDFInfo
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Abstract
The invention discloses an SNP (single nucleotide polymorphism) marker for detecting esophagus cancer susceptivity. The SNP marker comprises 24 SNP loci. The invention further discloses a PCR (polymerase chain reaction) amplification primer, a single-basic-group extension primer and a kit of the SNP marker. An important basis is provided for illness risk evaluation and diagnosis reference of esophagus cancer.
Description
Technical field
The invention belongs to Genetic Detection field, particularly to a kind of kit for detecting esophageal cancer susceptibility and its
SNP mark.
Background technology
The cancer of the esophagus is the malignant tumour that Esophageal Mucosa epithelium and oesophagus galandular epithelium occur.Or inside and outside substantial amounts of association study table
Bright, the morbidity of the cancer of the esophagus is relevant with inherent cause.
Using SNP (Single Nucleotide Polymorphsm, SNP) as genomic marker
Association analysis method is one of inheritance susceptible gene tester of disease the most frequently used at present.SNP refers in genomic level
The DNA sequence polymorphism being caused by single nucleotide variations, the occurrence frequency in crowd is more than 1%, and it is that the mankind are heritable
Modal one kind in variation.By force it is easy to detect, the SNP positioned at gene internal can directly influence albumen to SNP genetic stability
The structure of matter or expression, and then have influence on tissue, organ or even physiological function.
To common disease, related multiple genetic markers carry out early detection and system evaluation will be helpful to the early stage to disease
Prevention, diagnosis and treatment.At present, research has been found that the related science of heredity mark of a large amount of and various common diseases, however, due to
Lack and have enough sensitivity and multiple disease correlated identities can be carried out with extensively (high flux detection site, high flux detection
Sample) examination and inspection method so that these common diseases science of heredity mark cannot extensively apply.Additionally, it is common at present
Disease genetic mark shortage system, effectively integration, constrain sending out of common disease early prevention, diagnosis and treatment aspect significantly
Exhibition.Existing detection only has the genetic marker detection of single kind of disease or a class disease, and detection range is limited, and some common diseases
The method that there is no early detection.
At present, the detection of some traditional medicine means, such as histocyte has its intrinsic defect, position of drawing materials is improper,
Histocyte sample material is not enough or thinks to lack experience etc. and all will lead to carcinoma of endometrium mistaken diagnosis.Other technologies such as iconography
Although being widely used in inspection and the diagnosis of carcinoma of endometrium, its disease carcinoma of endometrium degree qualitative on still deposit
In significant limitation.
In sum, research and development a kind of for by multiple susceptibility locis detect esophageal cancer susceptibility kit to the cancer of the esophagus
The prediction of onset risk, prevention, diagnosis are provided according to significant.
Content of the invention
In view of the defect that above-mentioned prior art exists, the purpose of the present invention is to propose to one kind is used for detecting esophageal cancer susceptibility
SNP mark, the pcr amplification primer thing of SNP mark and Single base extension primer and its kit, 24 SNP of the present invention
Site is assessed, is diagnosed with reference to offer important evidence for the risk of the cancer of the esophagus.
The purpose of the present invention will be achieved by the following technical programs：
A kind of SNP mark for detecting esophageal cancer susceptibility, described SNP mark includes 24 SNP site, described
24 SNP site are rs10204525, rs1033667, rs1042026, rs1042522, rs10484761, rs11066015,
rs11066280、rs1346044、rs1642764、rs17761864、rs1805034、rs1883965、rs2074356、
rs2239612、rs2274223、rs238406、rs2847281、rs35597309、rs4785204、rs4822983、
Rs63749993, rs671, rs7206735 and rs7447927.
The pcr amplification primer thing of above-mentioned SNP mark and Single base extension primer,
The sequence of the pcr amplification primer thing of described detection rs10204525 is respectively SEQ ID NO：1 and SEQ ID NO：2,
And the sequence of Single base extension primer is SEQ ID NO：3；
The sequence of the pcr amplification primer thing of described detection rs1033667 is respectively SEQ ID NO：4 and SEQ ID NO：5, with
And the sequence of Single base extension primer is SEQ ID NO：6；
The sequence of the pcr amplification primer thing of described detection rs1042026 is respectively SEQ ID NO：7 and SEQ ID NO：8, with
And the sequence of Single base extension primer is SEQ ID NO：9；
The sequence of the pcr amplification primer thing of described detection rs1042522 is respectively SEQ ID NO：10 and SEQ ID NO：11,
And the sequence of Single base extension primer is SEQ ID NO：12；
The sequence of the pcr amplification primer thing of described detection rs10484761 is respectively SEQ ID NO：13 and SEQ ID NO：
14, and the sequence of Single base extension primer be SEQ ID NO：15；
The sequence of the pcr amplification primer thing of described detection rs11066015 is respectively SEQ ID NO：16 and SEQ ID NO：
17, and the sequence of Single base extension primer be SEQ ID NO：18；
The sequence of the pcr amplification primer thing of described detection rs11066280 is respectively SEQ ID NO：19 and SEQ ID NO：
20, and the sequence of Single base extension primer be SEQ ID NO：21；
The sequence of the pcr amplification primer thing of described detection rs1346044 is respectively SEQ ID NO：22 and SEQ ID NO：23,
And the sequence of Single base extension primer is SEQ ID NO：24；
The sequence of the pcr amplification primer thing of described detection rs1642764 is respectively SEQ ID NO：25 and SEQ ID NO：26,
And the sequence of Single base extension primer is SEQ ID NO：27；
The sequence of the pcr amplification primer thing of described detection rs17761864 is respectively SEQ ID NO：28 and SEQ ID NO：
29, and the sequence of Single base extension primer be SEQ ID NO：30；
The sequence of the pcr amplification primer thing of described detection rs1805034 is respectively SEQ ID NO：31 and SEQ ID NO：32,
And the sequence of Single base extension primer is SEQ ID NO：33；
The sequence of the pcr amplification primer thing of described detection rs1883965 is respectively SEQ ID NO：34 and SEQ ID NO：35,
And the sequence of Single base extension primer is SEQ ID NO：36；
The sequence of the pcr amplification primer thing of described detection rs2074356 is respectively SEQ ID NO：37 and SEQ ID NO：38,
And the sequence of Single base extension primer is SEQ ID NO：39；
The sequence of the pcr amplification primer thing of described detection rs2239612 is respectively SEQ ID NO：40 and SEQ ID NO：41,
And the sequence of Single base extension primer is SEQ ID NO：42；
The sequence of the pcr amplification primer thing of described detection rs2274223 is respectively SEQ ID NO：43 and SEQ ID NO：44,
And the sequence of Single base extension primer is SEQ ID NO：45；
The sequence of the pcr amplification primer thing of described detection rs238406 is respectively SEQ ID NO：46 and SEQ ID NO：47,
And the sequence of Single base extension primer is SEQ ID NO：48；
The sequence of the pcr amplification primer thing of described detection rs2847281 is respectively SEQ ID NO：49 and SEQ ID NO：50,
And the sequence of Single base extension primer is SEQ ID NO：51；
The sequence of the pcr amplification primer thing of described detection rs35597309 is respectively SEQ ID NO：52 and SEQ ID NO：
53, and the sequence of Single base extension primer be SEQ ID NO：54；
The sequence of the pcr amplification primer thing of described detection rs4785204 is respectively SEQ ID NO：55 and SEQ ID NO：56,
And the sequence of Single base extension primer is SEQ ID NO：57；
The sequence of the pcr amplification primer thing of described detection rs4822983 is respectively SEQ ID NO：58 and SEQ ID NO：59,
And the sequence of Single base extension primer is SEQ ID NO：60；
The sequence of the pcr amplification primer thing of described detection rs63749993 is respectively SEQ ID NO：61 and SEQ ID NO：
62, and the sequence of Single base extension primer be SEQ ID NO：63；
The sequence of the pcr amplification primer thing of described detection rs671 is respectively SEQ ID NO：64 and SEQ ID NO：65, and
The sequence of Single base extension primer is SEQ ID NO：66；
The sequence of the pcr amplification primer thing of described detection rs7206735 is respectively SEQ ID NO：67 and SEQ ID NO：68,
And the sequence of Single base extension primer is SEQ ID NO：69；
The sequence of the pcr amplification primer thing of described detection rs7447927 is respectively SEQ ID NO：70 and SEQ ID NO：71,
And the sequence of Single base extension primer is SEQ ID NO：72.
A kind of kit for detecting esophageal cancer susceptibility, described kit includes SNP mark described in claim 2
Pcr amplification primer thing and Single base extension primer, for detect in peripheral blood DNA 24 SNP site be rs10204525,
rs1033667、rs1042026、rs1042522、rs10484761、rs11066015、rs11066280、rs1346044、
rs1642764、rs17761864、rs1805034、rs1883965、rs2074356、rs2239612、rs2274223、
rs238406、rs2847281、rs35597309、rs4785204、rs4822983、rs63749993、rs671、rs7206735
And rs7447927.
Above-mentioned a kind of for detection be esophageal cancer susceptibility kit, wherein, described kit also include Taq enzyme,
DNTP mixed liquor, MgCl2Solution, PCR reaction buffer, endonuclease reaction buffer solution, deionized water.
Kit of the present invention is the cancer of the esophagus crowd with the present inventor based on multiple international and national large sample amounts
Based on the early-stage Study of normal population oesophagus cancer susceptibility gene screening results, by the cancer of the esophagus of the present inventor's autonomous Design
SNP screening process, screens qualified SNP site from NCBI-pubmed database：1) search the literary composition related to the cancer of the esophagus
Offer, by factor of influence with deliver the time limit and filtered；2) check the summary of candidate's document, further exclusion and cancer of the esophagus SNP
Study unrelated document；3) read document, find the corresponding SNP site of the cancer of the esophagus；4) with the SNP site and the cancer of the esophagus chosen it is
Keyword consulting literatures again；5) judge whether the SNP site chosen is closely related with the cancer of the esophagus, choose the closeest with the cancer of the esophagus
SNP site, corresponding OR value and mutating alkali yl that cut is closed；6) SNP site that previous step is obtained, public by Sequenom
Department software Typer 4.0 carries out online evaluation, obtains 24 final SNP site lists.
Compared with prior art, the invention provides a kind of detection method for detecting esophageal cancer susceptibility and reagent
Box, has also reached following effect：24 SNP site of the present invention, through mcta analysis, have higher practicality, can be used for eating
The earlier evaluations of pipe cancer and extensive examination, and this 24 sites detection success rate is high, technique reproducible is good, cost performance is high,
For the risk assessment of the cancer of the esophagus, diagnosis with reference to providing important evidence, to realize the earlier evaluations to cancer of the esophagus disease.
Below just in conjunction with the embodiments, the specific embodiment of the present invention is described in further detail, so that technical scheme is more
Should be readily appreciated that, grasp.
Specific embodiment
Below by specific embodiment, the present invention will be described, but the invention is not limited in this.In following embodiments
Described experimental technique, if no special instructions, is conventional method；Described reagent and material, if no special instructions, all can be from business
Approach obtains, and example below simultaneously is not used to limit the scope of the claims of the present invention, all equivalence enforcements done without departing from the present invention
Or change, it is intended to be limited solely by this patent protection domain.
Embodiment 1 is used for detecting the SNP site that esophageal cancer susceptibility is related
Wherein, rs10204525 and rs1033667 is located at gene HEATR3 region, rs1042026 and rs1042522 is located at
Gene C HEK2 region, rs10484761 is located at gene ST6GAL1 region, and rs11066015 is located at gene SMG6 region,
Rs11066280 is located at gene PTPN2 region, and rs1346044 is located at Gene A DH1B region, and rs1642764 is located at Gene A LDH2
Region, rs17761864 is located at gene TMEM173 region, and rs1805034 is located at Gene A TP1B2 region, and rs1883965 is located at
Gene HLA region, rs2074356 is located at gene PLCE1 region, and rs2239612 is located at gene M SH2 region, rs2274223 position
In gene WRN region, rs238406 is located at gene ERCC2 region, and rs2847281 is located at gene PD-1 region, rs35597309
Positioned at Gene A LDH2 region, rs4785204 is located at gene TP53 region, and rs4822983 is located at gene RANK region,
Rs63749993 is located at gene mTOR region, and rs671 is located at gene UNC5CL region, and rs7206735 is located at gene RPL6-
PTPN11 region, rs7447927 is located at gene C 12orf51 region.
Embodiment 2 detects the kit of esophageal cancer susceptibility
First, the preparation of kit：
1st, design and the pcr amplification primer thing and the Single base extension primer that synthesize described 24 SNP site.SNP site to be measured
Pcr amplification primer thing and Single base extension primer refer to table 1
The pcr amplification primer thing of table 1 SNP site to be measured and Single base extension primer
2nd, kit also includes Taq enzyme, dNTP mixed liquor, MgCl2Solution, PCR reaction buffer, endonuclease reaction buffering
Liquid, deionized water.
2nd, the detection method of kit：
1st, the extraction of DNA
1.1 buccal swab DNA are extracted
1) buccal swab is placed in 2ml centrifuge tube, adds 400 μ l PBS.
2) 20 μ l QIAGEN Protease and 400 μ lBuffer AL are added.Vortex 15s mixes immediately.In order to ensure have
The cracking of effect, sample and Buffer AL must immediately and be sufficiently mixed.
3) 56 DEG C of incubation 10min.Of short duration centrifugation.
4) add 400 μ l ethanol (96-100%) in sample, be vortexed and mix.Of short duration centrifugation is so that no liquid in lid.
5) liquid is transferred to centrifugal column, 8000rpm (6000 × g) is centrifuged 1min.
6) new 2ml EP pipe put into by pillar, plus 500 μ l Buffer AW1, and 8000rpm is centrifuged 1min, abandons EP pipe and gives up
Liquid.
7) new 2mlEP pipe put into by pillar, plus 500 μ l Buffer AW2, and 14000rpm (20,000 × g) is centrifuged 3min,
Abandon EP pipe and waste liquid.
8) pillar is put into new 2mlEP pipe, 14000rpm blank pipe is centrifuged 1min, abandons EP pipe.
9) pillar is put into new 1.5mlEP pipe, plus 120 μ l Buffer AE, incubated at room 5min, 8000rpm is centrifuged
1min, -20 DEG C of preservations.
1.2 whole blood DNAs extract
1) 20 μ l QIAGEN Protease are added in 1.5ml EP pipe.
2) to Guan Zhongjia 200 μ l whole blood sample.Note：The purpose being initially charged enzyme is to ensure that the mixing of enzyme and blood.
3) add 200 μ l Buffer AL, be vortexed and mix 15s.
4) 56 DEG C of incubation 10min, of short duration centrifugation.Note：Extending incubation time can not increase yield or improve quality.
5) 200 μ l absolute ethyl alcohols are added, after vortex 15s, of short duration centrifugation is so that no liquid in lid.
6) continue buccal swab 5～10 step.
2nd, PCR amplification
2.1 prepare pcr amplification reaction system in a new 1.5mlEP pipe, are shown in Table 2
Table 2 pcr amplification reaction system
Above reagent adds 2 μ l in every hole after mixing.
2.2 every holes sequentially add DNA 10ng/ul 1 μ l, 0.25uM Primer Mix 2 μ l, cumulative volume 5 μ l.
2.3 set in the PCR instrument of compatible 96 orifice plates PCR reaction condition as：94 DEG C of 4min, 94 DEG C of 20s, 56 DEG C
30min, 72 DEG C of 1min, 45 circulations；72℃3min；4 DEG C of holdings.96 hole PCR reaction plates are positioned in PCR instrument, start PCR
Reaction.
3rd, PCR primer alkaline phosphatase treatment
3.1 after PCR reaction terminates, by PCR primer SAP (shrimp alkaline phosphatase, shrimp alkalescence
Phosphatase) process, with remove system middle reaches from dNTPs.
3.2 preparation alkaline phosphatase treatment reaction systems, are shown in Table 3
Table 3 alkaline phosphatase treatment reaction system
Above reagent adds 2 μ l in every hole after mixing.
3.3 in the PCR instrument of compatible 96 orifice plates, sets PCR reaction condition：37℃40min；85℃5min；4 DEG C of holdings,
Start PCR instrument and carry out alkaline phosphatase treatment.
4 Single base extensions
4.1 after alkaline phosphatase treatment terminates, and carries out single base extension.
4.2 preparation single base extension systems, are shown in Table 4
Table 4 single base extension system
Above reagent adds 1.06 μ l in every hole after mixing
4.3 every holes add iPLEX Extend Primer Mix 0.94 μ l, cumulative volume 9 μ l.
4.4 in the PCR instrument of compatible 96 orifice plates, sets PCR reaction condition：94 DEG C of 30s, 94 DEG C of 5s, 52 DEG C of 5s, 80 DEG C
5s；4 circulations of 52 DEG C of 5s, 80 DEG C of 5s；94 DEG C of 5s, 52 DEG C of 5s, 39 circulations of 80 DEG C of 5s；72℃3min；4 DEG C of maintenances.Start
PCR instrument carries out single base extension.
5th, purifying resin
5.1 Clean Resin resin is tiled to 6mg resin plate in；
5.2 add 16 μ l water to the corresponding in the hole of extension products；
Dried resin is poured in extension products plate by 5.3, sealer, and slow speed vertical rotates 15min, make resin with anti-
Thing is answered to be fully contacted；
5.4 3000rpm 5min centrifugations make resin sink to bottom hole portion.
6th, chip point sample
Start MassARRAY NanodispenserRS1000 point sample instrument, the extension products after purifying resin are moved to 96
On the solid support of hole, produce chip.
7th, Mass Spectrometer Method
Chip is used MALDI-TOF (matrix-assistedlaser desorption/ionization-time
Offlight), matrix solid-dispersion flight time mass spectrum) analysis, testing result is using TYPER4.0 software
(Sequenom) parting output result, analyzes experimenter's cancer of the esophagus occurrence risk, to realize the earlier evaluations to disease.
Embodiment 3 24 documents that SNP site quote closely related with the cancer of the esophagus, are shown in Table 5
Table 5 24 documents that SNP site quote closely related with the cancer of the esophagus
Embodiment 4 draws associating of 24 SNP site and the cancer of the esophagus according to the document quoted in embodiment 3, is shown in Table 6
6 24 SNP site of table and the association analysis result of the cancer of the esophagus
The checking of 5 24 SNP site of embodiment
Mass Spectrometry detection method using kit in above-described embodiment 2 analyzes 24 SNP site, and testing result uses
24 SNP site of table 6 and oesophagus in TYPER4.0 software (Sequenom) parting output result, its OR value and embodiment 4
The association analysis result of cancer is identical, illustrates that this 24 sites have higher practicality, can be used for the cancer of the esophagus earlier evaluations and
Extensive examination, because analytical technique of mass spectrum has, detection success rate is high, technique reproducible is good, cost performance is high, the therefore present invention
Use Mass Spectrometer Method to analyze this 24 sites detection success rate is high, technique reproducible is good, cost performance is high, is the trouble of the cancer of the esophagus
Sick risk assessment, diagnosis reference provide important evidence, to realize the earlier evaluations to cancer of the esophagus disease.
Described above illustrate and describes some preferred embodiments of the present invention, but as previously mentioned it should be understood that the present invention
Be not limited to form disclosed herein, be not to be taken as the exclusion to other embodiment, and can be used for various other combinations,
Modification and environment, and can be in invention contemplated scope described herein, by technology or the knowledge of above-mentioned teaching or association area
It is modified.And the change that those skilled in the art are carried out and change without departing from the spirit and scope of the present invention, then all should be at this
In the protection domain of bright claims.
SEQUENCE LISTING
<110>Nucleon Gene Tech. Company Limited of Shenzhen
<120>A kind of kit for detecting esophageal cancer susceptibility and its SNP mark
<130>
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<170> PatentIn version 3.5
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<213>Artificial sequence
<400> 38
acgttggatg agtccacaca gctggtaaac 30
<210> 39
<211> 17
<212> DNA
<213>Artificial sequence
<400> 39
tggtggttaa cagttca 17
<210> 40
<211> 30
<212> DNA
<213>Artificial sequence
<400> 40
acgttggatg agttcccaga attccagagg 30
<210> 41
<211> 30
<212> DNA
<213>Artificial sequence
<400> 41
acgttggatg gttcagcagc tcatgactcc 30
<210> 42
<211> 21
<212> DNA
<213>Artificial sequence
<400> 42
gtcataaata gaaactccag a 21
<210> 43
<211> 30
<212> DNA
<213>Artificial sequence
<400> 43
acgttggatg agttcccaga attccagagg 30
<210> 44
<211> 30
<212> DNA
<213>Artificial sequence
<400> 44
acgttggatg tccatcgaaa caccctgaac 30
<210> 45
<211> 19
<212> DNA
<213>Artificial sequence
<400> 45
aagatcttcg aagtgaatg 19
<210> 46
<211> 30
<212> DNA
<213>Artificial sequence
<400> 46
acgttggatg agttcccaga attccagagg 30
<210> 47
<211> 30
<212> DNA
<213>Artificial sequence
<400> 47
acgttggatg agtaccagta tgacaccagc 30
<210> 48
<211> 17
<212> DNA
<213>Artificial sequence
<400> 48
ccagtaacct cacagaa 17
<210> 49
<211> 30
<212> DNA
<213>Artificial sequence
<400> 49
acgttggatg agttcccaga attccagagg 30
<210> 50
<211> 30
<212> DNA
<213>Artificial sequence
<400> 50
acgttggatg ctctctgctg agaatgcaag 30
<210> 51
<211> 18
<212> DNA
<213>Artificial sequence
<400> 51
aggcagatca aactcacc 18
<210> 52
<211> 30
<212> DNA
<213>Artificial sequence
<400> 52
acgttggatg agttcccaga attccagagg 30
<210> 53
<211> 30
<212> DNA
<213>Artificial sequence
<400> 53
acgttggatg ccacattaag aagccctcag 30
<210> 54
<211> 14
<212> DNA
<213>Artificial sequence
<400> 54
tgctggccct gcat 14
<210> 55
<211> 30
<212> DNA
<213>Artificial sequence
<400> 55
acgttggatg agttcccaga attccagagg 30
<210> 56
<211> 30
<212> DNA
<213>Artificial sequence
<400> 56
acgttggatg aacctatgag gtaggtgaac 30
<210> 57
<211> 17
<212> DNA
<213>Artificial sequence
<400> 57
cctcactttc ccatttg 17
<210> 58
<211> 30
<212> DNA
<213>Artificial sequence
<400> 58
acgttggatg agttcccaga attccagagg 30
<210> 59
<211> 30
<212> DNA
<213>Artificial sequence
<400> 59
acgttggatg gcgcaacccc aactctaaaa 30
<210> 60
<211> 20
<212> DNA
<213>Artificial sequence
<400> 60
cagcaccatg cacaactaat 20
<210> 61
<211> 30
<212> DNA
<213>Artificial sequence
<400> 61
acgttggatg agttcccaga attccagagg 30
<210> 62
<211> 30
<212> DNA
<213>Artificial sequence
<400> 62
acgttggatg cgcctacttg gtctaagttg 30
<210> 63
<211> 19
<212> DNA
<213>Artificial sequence
<400> 63
actagggtga tagaactca 19
<210> 64
<211> 30
<212> DNA
<213>Artificial sequence
<400> 64
acgttggatg agttcccaga attccagagg 30
<210> 65
<211> 30
<212> DNA
<213>Artificial sequence
<400> 65
acgttggatg tggtggctac aagatgtcag 30
<210> 66
<211> 18
<212> DNA
<213>Artificial sequence
<400> 66
cactcacagt tttcactt 18
<210> 67
<211> 30
<212> DNA
<213>Artificial sequence
<400> 67
acgttggatg agttcccaga attccagagg 30
<210> 68
<211> 30
<212> DNA
<213>Artificial sequence
<400> 68
acgttggatg ccacattctc ttcctcaagc 30
<210> 69
<211> 18
<212> DNA
<213>Artificial sequence
<400> 69
aagaggcact aaagccta 18
<210> 70
<211> 30
<212> DNA
<213>Artificial sequence
<400> 70
acgttggatg agttcccaga attccagagg 30
<210> 71
<211> 30
<212> DNA
<213>Artificial sequence
<400> 71
acgttggatg taggagagcc accagagcac 30
<210> 72
<211> 15
<212> DNA
<213>Artificial sequence
<400> 72
ggaggctagg tggag 15
Claims (4)
1. a kind of SNP mark for detecting esophageal cancer susceptibility is it is characterised in that described SNP mark includes 24 SNP
Site, described 24 SNP site be rs10204525, rs1033667, rs1042026, rs1042522, rs10484761,
rs11066015、rs11066280、rs1346044、rs1642764、rs17761864、rs1805034、rs1883965、
rs2074356、rs2239612、rs2274223、rs238406、rs2847281、rs35597309、rs4785204、
Rs4822983, rs63749993, rs671, rs7206735 and rs7447927.
2. the pcr amplification primer thing of SNP mark according to claim 1 and Single base extension primer it is characterised in that
The sequence of the pcr amplification primer thing of described detection rs10204525 is respectively SEQ ID NO：1 and SEQ ID NO：2, and
The sequence of Single base extension primer is SEQ ID NO：3；
The sequence of the pcr amplification primer thing of described detection rs1033667 is respectively SEQ ID NO：4 and SEQ ID NO：5, Yi Jidan
The sequence of base extension primer is SEQ ID NO：6；
The sequence of the pcr amplification primer thing of described detection rs1042026 is respectively SEQ ID NO：7 and SEQ ID NO：8, Yi Jidan
The sequence of base extension primer is SEQ ID NO：9；
The sequence of the pcr amplification primer thing of described detection rs1042522 is respectively SEQ ID NO：10 and SEQ ID NO：11, and
The sequence of Single base extension primer is SEQ ID NO：12；
The sequence of the pcr amplification primer thing of described detection rs10484761 is respectively SEQ ID NO：13 and SEQ ID NO：14, with
And the sequence of Single base extension primer is SEQ ID NO：15；
The sequence of the pcr amplification primer thing of described detection rs11066015 is respectively SEQ ID NO：16 and SEQ ID NO：17, with
And the sequence of Single base extension primer is SEQ ID NO：18；
The sequence of the pcr amplification primer thing of described detection rs11066280 is respectively SEQ ID NO：19 and SEQ ID NO：20, with
And the sequence of Single base extension primer is SEQ ID NO：21；
The sequence of the pcr amplification primer thing of described detection rs1346044 is respectively SEQ ID NO：22 and SEQ ID NO：23, and
The sequence of Single base extension primer is SEQ ID NO：24；
The sequence of the pcr amplification primer thing of described detection rs1642764 is respectively SEQ ID NO：25 and SEQ ID NO：26, and
The sequence of Single base extension primer is SEQ ID NO：27；
The sequence of the pcr amplification primer thing of described detection rs17761864 is respectively SEQ ID NO：28 and SEQ ID NO：29, with
And the sequence of Single base extension primer is SEQ ID NO：30；
The sequence of the pcr amplification primer thing of described detection rs1805034 is respectively SEQ ID NO：31 and SEQ ID NO：32, and
The sequence of Single base extension primer is SEQ ID NO：33；
The sequence of the pcr amplification primer thing of described detection rs1883965 is respectively SEQ ID NO：34 and SEQ ID NO：35, and
The sequence of Single base extension primer is SEQ ID NO：36；
The sequence of the pcr amplification primer thing of described detection rs2074356 is respectively SEQ ID NO：37 and SEQ ID NO：38, and
The sequence of Single base extension primer is SEQ ID NO：39；
The sequence of the pcr amplification primer thing of described detection rs2239612 is respectively SEQ ID NO：40 and SEQ ID NO：41, and
The sequence of Single base extension primer is SEQ ID NO：42；
The sequence of the pcr amplification primer thing of described detection rs2274223 is respectively SEQ ID NO：43 and SEQ ID NO：44, and
The sequence of Single base extension primer is SEQ ID NO：45；
The sequence of the pcr amplification primer thing of described detection rs238406 is respectively SEQ ID NO：46 and SEQ ID NO：47, and
The sequence of Single base extension primer is SEQ ID NO：48；
The sequence of the pcr amplification primer thing of described detection rs2847281 is respectively SEQ ID NO：49 and SEQ ID NO：50, and
The sequence of Single base extension primer is SEQ ID NO：51；
The sequence of the pcr amplification primer thing of described detection rs35597309 is respectively SEQ ID NO：52 and SEQ ID NO：53, with
And the sequence of Single base extension primer is SEQ ID NO：54；
The sequence of the pcr amplification primer thing of described detection rs4785204 is respectively SEQ ID NO：55 and SEQ ID NO：56, and
The sequence of Single base extension primer is SEQ ID NO：57；
The sequence of the pcr amplification primer thing of described detection rs4822983 is respectively SEQ ID NO：58 and SEQ ID NO：59, and
The sequence of Single base extension primer is SEQ ID NO：60；
The sequence of the pcr amplification primer thing of described detection rs63749993 is respectively SEQ ID NO：61 and SEQ ID NO：62, with
And the sequence of Single base extension primer is SEQ ID NO：63；
The sequence of the pcr amplification primer thing of described detection rs671 is respectively SEQ ID NO：64 and SEQ ID NO：65, and single alkali
The sequence of base extension primer is SEQ ID NO：66；
The sequence of the pcr amplification primer thing of described detection rs7206735 is respectively SEQ ID NO：67 and SEQ ID NO：68, and
The sequence of Single base extension primer is SEQ ID NO：69；
The sequence of the pcr amplification primer thing of described detection rs7447927 is respectively SEQ ID NO：70 and SEQ ID NO：71, and
The sequence of Single base extension primer is SEQ ID NO：72.
3. a kind of kit for detecting esophageal cancer susceptibility is it is characterised in that described kit is included described in claim 2
The pcr amplification primer thing of SNP mark and Single base extension primer, for detecting that in peripheral blood DNA, 24 SNP site are
rs10204525、rs1033667、rs1042026、rs1042522、rs10484761、rs11066015、rs11066280、
rs1346044、rs1642764、rs17761864、rs1805034、rs1883965、rs2074356、rs2239612、
rs2274223、rs238406、rs2847281、rs35597309、rs4785204、rs4822983、rs63749993、rs671、
Rs7206735 and rs7447927.
4. according to claim 3 a kind of be the kit of esophageal cancer susceptibility it is characterised in that described examination for detection
Agent box also includes Taq enzyme, dNTP mixed liquor, MgCl2Solution, PCR reaction buffer, endonuclease reaction buffer solution, deionized water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201611049684.XA CN106434980A (en) | 2016-11-24 | 2016-11-24 | Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611049684.XA CN106434980A (en) | 2016-11-24 | 2016-11-24 | Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110343765A (en) * | 2019-08-09 | 2019-10-18 | 郑州大学第一附属医院 | A kind of SNP marker and kit for cardia cancer people at highest risk's screening |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101608218A (en) * | 2008-06-20 | 2009-12-23 | 上海主健生物工程有限公司 | Esophageal cancer genetic test kit |
| CN103898197A (en) * | 2012-12-25 | 2014-07-02 | 泰州医药城博奥邦科生物科技有限公司 | Common malignant tumor susceptibility gene detection chip |
| CN104593499A (en) * | 2015-01-20 | 2015-05-06 | 汕头大学医学院 | Detection kit for carrying out efficient molecular subtyping on esophageal cancer susceptible gene loci |
-
2016
- 2016-11-24 CN CN201611049684.XA patent/CN106434980A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101608218A (en) * | 2008-06-20 | 2009-12-23 | 上海主健生物工程有限公司 | Esophageal cancer genetic test kit |
| CN103898197A (en) * | 2012-12-25 | 2014-07-02 | 泰州医药城博奥邦科生物科技有限公司 | Common malignant tumor susceptibility gene detection chip |
| CN104593499A (en) * | 2015-01-20 | 2015-05-06 | 汕头大学医学院 | Detection kit for carrying out efficient molecular subtyping on esophageal cancer susceptible gene loci |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110343765A (en) * | 2019-08-09 | 2019-10-18 | 郑州大学第一附属医院 | A kind of SNP marker and kit for cardia cancer people at highest risk's screening |
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