CN103898197A - Common malignant tumor susceptibility gene detection chip - Google Patents
Common malignant tumor susceptibility gene detection chip Download PDFInfo
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- CN103898197A CN103898197A CN201210570393.0A CN201210570393A CN103898197A CN 103898197 A CN103898197 A CN 103898197A CN 201210570393 A CN201210570393 A CN 201210570393A CN 103898197 A CN103898197 A CN 103898197A
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Abstract
The present invention discloses a common malignant tumor susceptibility gene detection chip. The detection method comprises: 1) extracting DNA of a subject sample; 2) carrying out target gene PCR amplification, purification, fragmentation and fluorescein labeling on the sample DNA; 3) carrying out gene chip hybridization on the fluorescein-labeled PCR product, and detecting the SNP locus of the target gene; and 4) analyzing susceptibility of the subject on breast cancer, lung cancer, gastric cancer, prostate cancer, pancreatic cancer, bladder cancer, liver cancer, leukemia, esophageal cancer and colorectal cancer according to the genotyping result in the step 3). According to the present invention, with the relevant genotyping, the common malignant tumor susceptibility assessment can be performed on the subject so as to formulate the most scientific personalized care plan, such that occurrence of malignant tumors can be avoided, delayed or even prevented.
Description
Technical field
The present invention relates to a common cancer tumor susceptibility gene detection chip, particularly relate to a kind of malignant tumour susceptibility test and appraisal and cover prescription method.
Background technology
2008, the whole world had 1,270 ten thousand people to suffer from cancer, and death toll is up to 7,600,000.The number of world wide internal cause mortality of malignant tumors, adds up than acquired immune deficiency syndrome (AIDS), malaria and tuberculosis also many.If do not adopted an effective measure, expect the year two thousand thirty, will there are every year 2600 ten thousand newly-increased Malignant Tumor Cases, mortality of malignant tumors number will reach 1,700 ten thousand, and middle and low income country is by " severely afflicated area " that become malignant tumour and wreak havoc.
The M & M of tumour was high in recent years, and with regard to China, malignant tumour has become resident's the second cause of the death, city resident's the first cause of the death.In the world, the Cancer Mortality of China and the U.S., Britain, France approach, higher than most Asian countries.Lung cancer, liver cancer, colorectal cancer, mammary cancer, bladder cancer mortality ratio and formation thereof are obvious ascendant trend, wherein lung cancer and mammary cancer ascensional range maximum, lung cancer has replaced liver cancer to become the first Death Cause for Malignant Tumors of China (account for whole mortality of malignant tumors 22.7%).
The a large amount of knowledge about the intervening measure of the malignant tumour cause of disease and prevention and management malignant tumour are grasped.By implementing taking evidence as basic malignant tumour prevention, early discovery and malignant tumor patient management strategy, can make malignant tumour be reduced and control.
Tobacco using, drink, unsound diet and lack motion be the Major Risk Factors of whole world malignant tumour.Inherited genetic factors is also important risk factor of malignant tumour simultaneously.Because of outside atmosphere, we can control, and internal factor-inherited genetic factors (tumor susceptibility gene) we cannot control.Therefore to fundamentally prevent malignant tumour generation, delay the development of malignant tumour and work out rational disease treatment measure, just must detect the tumor susceptibility gene of these diseases.The positive effect of gene test is embodied in cancer just can find potential risks before occurring, thereby mentions the concern of people to self Health and Disease, and then instructs person under inspection to take effective means to prevent by " personalized health management ".
Summary of the invention
The technical problem to be solved in the present invention is to provide common cancer tumor susceptibility gene detection chip, detect by the key gene genetic site in these genes, judge concrete genotype, can assess individuality and suffer from the risk probability of malignant tumour, thereby play active effect to strengthening prevention and treatment of malignant tumors.
For solving the problems of the technologies described above, common cancer tumor susceptibility gene detection chip of the present invention, comprises step:
1) extract person under inspection's sample DNA;
2) sample DNA is carried out to pcr amplification, purifying, the fragmentation and fluorescein-labelled of heritable variation, described heritable variation comprises: rs505922, rs2294008, rs2279744, rs738409, rs2274223, rs667282, rs6983267 and rs2231142.
3) fluorescein-labeled PCR product carries out gene chip hybridization, detects heritable variation, and described heritable variation comprises: rs505922, rs2294008, rs2279744, rs738409, rs2274223, rs667282, rs6983267 and rs2231142.
4) the gene type result obtaining according to step 3), the curative effect of analysis person under inspection tumour personalized treatment.
Described step 2) in the primer of rs505922 be the primer of sequence shown in SEQ ID NO:1～2, the primer of rs2294008 is the primer of sequence shown in SEQ ID NO:3～4, the primer of rs2279744 is the primer of sequence shown in SEQ ID NO:5～6, the primer of rs738409 is the primer of sequence shown in SEQ ID NO:7～8, the primer of rs2274223 is the primer of sequence shown in SEQ ID NO:9～10, the primer of rs667282 is the primer of sequence shown in SEQ ID NO:11～12, the primer of rs6983267 is the primer of sequence shown in SEQ ID NO:13～14, the primer of rs2231142 is the primer of sequence shown in SEQ ID NO:15～16.
Described step 2) in fragmentation be by the PCR product of purifying after measured after concentration, carry out fragmentation with DNase I; Described fluorescein-labelled be that fragmentation PCR product utilization deoxynucleotidyl transferase is carried out fluorescein-labelled at 3 ' end.
The preparation of described step 3) gene chip is that the probe designing and synthesizing is in advance downloaded to by contact point sample or ink jet type point of sample on the solid phase carrier sheet base of slide or silicon chip material, wherein, probe is the DNA probe of sequence shown in SEQ ID NO:17～SEQ ID NO:32.
The theoretical foundation in the gene in the present invention and SNP site:
Rs505922 is relevant to the abo blood group of human body, and human body abo blood group is relevant to carcinoma of the pancreas.
Rs2294008 is the pleomorphism site on PTSA gene, and PTSA is the significant gene of prostate cancer.The research in this year finds that this gene is also relevant to the generation of other kinds of tumors.
Rs2279744 is a pleomorphism site on MDM2 gene.MDM2 is a member in P53 cancer suppressor gene regulated and control network, and its dysfunction almost occurs to develop relevant to all cancers.
Rs738409 is a pleomorphism site on PNPLA3 gene.PNPLA3 dysfunction is relevant to liver cirrhosis and liver cancer.
Rs2274223 is a polymorphic site on PLCE1 gene.PLCE1 can regulate Growth of Cells, differentiation, and apoptosis and blood vessel occur, and its dysfunction is relevant to kinds of tumors.
Rs667282 is relevant to smoking addiction, and the smoking topmost Hazard Factor that are lung cancer.
Rs6983267 is arranged in MYC enhanser, affects its expression.MYC is oncogene, relevant to the generation development of a lot of tumours.
Rs2231142 is a polymorphic site on ABCG2 gene.ABCG2 is relevant to cancer resistance, but Recent study discovery, this gene also participates in tumour and forms.
Beneficial effect of the present invention:
By genes involved somatotype, can estimate the susceptibility of the common cancers such as person under inspection's mammary cancer, lung cancer, cancer of the stomach, prostate cancer, carcinoma of the pancreas, bladder cancer, liver cancer, leukemia, the esophageal carcinoma, colorectal cancer, thereby work out most scientific individuation health care's scheme, even to avoid, to delay avoiding the generation of malignant tumour.
Brief description of the drawings
Below in conjunction with accompanying drawing and embodiment, the present invention is further detailed explanation:
Fig. 1 is the electrophoresis detection result figure after multiplex PCR amplification;
Fig. 2 is the electrophoresis detection result figure after PCR product fragmentation;
Fig. 3 is detection chip results of hybridization figure;
Embodiment
Embodiment 1: the extraction of sample DNA
Embodiment
Embodiment 1: the extraction of sample DNA
Adopt FlexiGene DNA Kit(QIAGEN, Cat. No. 51206) genomic dna in test kit extracting human peripheral.
Concrete grammar is: in 300 μ l blood samples, add 750 μ l Buffer FG1, turn upside down and 5 times it is mixed.Then centrifugal 1 min under 12,000 rpm rotating speeds.After centrifugal, outwell supernatant liquid, then add 150 μ l Buffer FG2 and 1.5 μ l protein enzyme solutions, vibration immediately, until precipitation is dissolved completely.Next centrifugal 3 ~ 5 s, then 65 DEG C of water-bath 5 min.When solution is from redness becomes olive-green, add 150 μ l 100% Virahols, fully put upside down up and down centrifuge tube, it is mixed, until DNA separates out, be macroscopic wire or bulk.Then centrifugal 3 min under 12,000 rpm rotating speeds.After centrifugal, outwell supernatant liquid, then add 150 μ l 70% ethanol, and 5 s that vibrate.Then centrifugal 3 min under 12,000 rpm rotating speeds again, outwell supernatant liquid after centrifugal, and natural air drying precipitation, until all liquid all evaporates.In the most backward centrifuge tube, add 200 μ l Buffer FG3, vibration 5 s, then 65 DEG C of water-bath 10 min, dissolve DNA.
Use ND-1000 nucleic acid concentration analyser quantitative to the DNA of institute's extracting.DNA working fluid concentration correction to 10 ng/ μ l, is placed in-20 DEG C of Refrigerator stores.
Embodiment 2: chip hybridization detects SNP site
1. the preparation of gene chip
(1) probe dissolves
By every probe TE solution dilution of sequence probe shown in SEQ ID NO:16～SEQ ID NO:32, final concentration is 10 mM.By concentration be the probe of 10 mM and concentration be the PBS solution of 200 mM in the medium volume mixture of 384 orifice plate, seal 384 orifice plates with adhesive sheet, under room temperature, vibrate 2 minutes, centrifugal ,-20 DEG C of preservations, are used in order to point sample.
(2) point sample
The probe designing and synthesizing is in advance downloaded on the solid phase carrier sheet base of the material such as slide, silicon chip by contact point sample or ink jet type point of sample.Sheet base adopts Cell Associates CSS-100 aldehyde radical sheet base, the point sample instrument of Ominigrid 100 models of GeneMachine company, be as the criterion with FullMoon sheet base at humidity: 65-75%(), temperature is point sample under the condition of 25 DEG C, after point sample, place after half an hour, chip is taken out, drying at room temperature is preserved.
2. the processing of testing sample and mark
(1) amplification of goal gene
With the primer (SEQ ID NO:1～2) of rs505922, primer (SEQ ID NO:3～4 of rs2294008, the primer (SEQ ID NO:5～6) of rs2279744, the primer (SEQ ID NO:7～8) of rs738409, the primer (SEQ ID NO:9～10) of rs2274223, the primer (SEQ ID NO:11～12) of rs667282, the primer (SEQ ID NO:13～14) of rs6983267, the primer (SEQ ID NO:15～16) of rs2231142 carries out pcr amplification to sample DNA.Pcr amplification is undertaken by 30 μ l reaction systems, reaction system is 0.3 mM dNTP, 10 mM Tris-HCl, 50 mM KCl, 2 mM MgCl2,20% Q solution(Qiagen), concentration 0.16 μ M, genomic dna 10 ng of upstream and downstream primer, Taq enzyme 0.6 U(Takara).Application Touch-down PCR response procedures: 94 DEG C of sex change 5 min; 94 DEG C of sex change 40 s, 64 DEG C of annealing 1 min, each circulation reduces by 0.5 DEG C, and 72 DEG C are extended 50 s, totally 10 circulations; Then 94 DEG C of sex change 40 s, 59 DEG C of annealing 40 s, 72 DEG C are extended 50 s, totally 30 circulations; Last 72 DEG C are extended 5 min.PCR finishes rear use 1.5% sepharose and detects amplification.
In the time carrying out multi-PRC reaction, the primer of sequence shown in SEQ ID NO:1～SEQ ID NO:16 to be put into a reaction system and increase, system is 50 μ l.The reaction system of multiplex PCR is: every kind of dNTP 0.3 μ mol/L, Tricine-KOH(PH=8.7) 40 mmol/L, KCl 16 mmol/L, MgCl
23.5 mmol/L, BSA 3.75 μ g/ml, every primer 2 μ mol/L, DNA 80 ng and 2.2 × Titanium Taq archaeal dna polymerase (Clontech, USA).Multi-PRC reaction condition: 95 DEG C of sex change 3 min; 95 DEG C of sex change 30s, 66 DEG C of annealing 2 min, 68 DEG C are extended 4 min, totally 40 circulations; Last 68 DEG C extend 10 min.After pcr amplification, get 3 μ l PCR reaction product and do agarose gel electrophoresis, these PCR products can be used for hybridization step below after treatment.
(2) PCR product purification and fragmentation
All PCR products of each sample mix, with QIA quick PCR Purification Kit(Qiagen, Cat. No. 28106) purifying.The PCR product of purifying is after measured after concentration, with DNase I(deoxyribonuclease I) carry out fragmentation.The reaction system of fragmentation comprises: 30 μ l purified pcr products (10 μ g), 10 × DNase I damping fluid of 4 μ l, the DNase I of 0.12 μ l, the ddH of 5.88 μ l
2o.Reaction conditions is that 37 DEG C of temperature are bathed 5 min, then 95 DEG C of 15 min.Product after fragmentation runs 4% sepharose, ensures that most fragments is in 30-200 base pair.
(3) fluorescein-labelled
Utilize deoxynucleotidyl transferase to carry out fluorescein-labelled at 3 ' end, 40 μ l reaction systems of mark comprise: 25 μ l fragmentation PCR products, 5 × deoxynucleotidyl transferase damping fluid of 8 μ l, the CY3-N6-ddCTP(1 mM of 1 μ l), the deoxynucleotidyl transferase of 3 μ l (20 U/ μ l), the ddH of 3 μ l
2o.Reaction conditions is that 37 DEG C of temperature are bathed 120 min, then 95 DEG C of heating 15 min.
3. hybridization, washing and result detect
95 DEG C of sex change 10 min of fluorescently-labeled PCR product, be placed in immediately on ice, for hybridization, hybridization 20 μ l systems comprise: fluorescein-labeled PCR product 15 μ l, 20 × SSPE, 1.2 μ l, 1% Triton 0.2 μ l, 10 × Denhandts, 0.9 μ l, methane amide 0.5 μ l, ddH
2o 2.2 μ l.Reaction conditions is that 48 DEG C of temperature are bathed 120 min, then in succession use 1 × lavation buffer solution I(5 × SSC, 0.1% SDS), 1 × lavation buffer solution II(2 × SSC, 0.1% SDS) and 1 × lavation buffer solution III(1 × SSC) at 42 DEG C of each washing 10 min, finally use ddH
2o washs 0.5 min.
Chip after washing, after drying, scans (also can with other laser scanner) with GenePix 4000B confocal laser scanner.Chip after scanning hybridization obtains results of hybridization, then obtains data file with GenePix Pro processing image, then data file is analyzed and just can be obtained person under inspection's predicting the outcome to antitumor drug curative effect and toxic side effect.
Embodiment 4: curative effect is estimated
Sample: person under inspection (male sex, 41 years old)
Detection operating process according to embodiment 1-3 detects, and wherein, the electrophoresis detection that the electrophoresis detection after this person under inspection's goal gene multiplex PCR amplification the results are shown in Figure after 1, PCR product fragmentation the results are shown in Figure 2, and detection chip results of hybridization is shown in Fig. 3.Gene chip check result is imported to the susceptibility assessment of analysis software acquisition person under inspection common cancer.This person under inspection is higher to the susceptibility of lung cancer and liver cancer, the smoking and drinking of need keeping under strict control.
Sequence table
Bo Aobangke bio tech ltd, medicine city, <110> Taizhou
<120> common cancer tumor susceptibility gene detection chip
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Claims (8)
1. common cancer tumor susceptibility gene detection chip, comprises step:
1) extract person under inspection's sample DNA;
2) sample DNA is carried out to pcr amplification, purifying, the fragmentation and fluorescein-labelled of heritable variation, described heritable variation comprises: rs505922, rs2294008, rs2279744, rs738409, rs2274223, rs667282, rs6983267 and rs2231142;
3) fluorescein-labeled PCR product carries out gene chip hybridization, the SNP site of testing goal gene;
4) the gene type result obtaining according to step 3), analyzes curative effect and the toxic side effect of person under inspection to antitumor drug.
2. common cancer tumor susceptibility gene detection chip as claimed in claim 1, it is characterized in that: described step 2) in the primer of rs505922 be the primer of sequence shown in SEQ ID NO:1～2, the primer of rs2294008 is the primer of sequence shown in SEQ ID NO:3～4, the primer of rs2279744 is the primer of sequence shown in SEQ ID NO:5～6, the primer of rs738409 is the primer of sequence shown in SEQ ID NO:7～8, the primer of rs2274223 is the primer of sequence shown in SEQ ID NO:9～10, the primer of rs667282 is the primer of sequence shown in SEQ ID NO:11～12, the primer of rs6983267 is the primer of sequence shown in SEQ ID NO:13～14, the primer of rs2231142 is the primer of sequence shown in SEQ ID NO:15～16.
3. common cancer tumor susceptibility gene detection chip as claimed in claim 1, is characterized in that: described step 2) in fragmentation be by the PCR product of purifying after measured after concentration, carry out fragmentation with DNase I;
Described fluorescein-labelled be that fragmentation PCR product utilization deoxynucleotidyl transferase is carried out fluorescein-labelled at 3 ' end.
4. common cancer tumor susceptibility gene detection chip as claimed in claim 1, it is characterized in that: the preparation of described step 3) gene chip is that the probe designing and synthesizing is in advance downloaded to by contact point sample or ink jet type point of sample on the solid phase carrier sheet base of slide or silicon chip material, wherein, probe is the DNA probe of sequence shown in SEQ ID NO:17～SEQ ID NO:32.
5. the detection chip as described in claim 1 or 4, is characterized in that, described probe is DNA, RNA, DNA-RNA mosaic, PNA or derivatives thereof.
6. primer as claimed in claim 2, is characterized in that, nucleotide chain or its complementary strand that described primer contains sequence shown in SEQ ID NO:1～SEQ ID NO:16.
7. common cancer tumor susceptibility gene detection chip as claimed in claim 1, is characterized in that: the gene type result root in described step 4) imports analysis software the susceptibility of person under inspection's common cancer is assessed.
8. the common cancer tumor susceptibility gene detection chip as described in claim 1 or 7, is characterized in that: described common cancer comprises mammary cancer, lung cancer, cancer of the stomach, prostate cancer, carcinoma of the pancreas, bladder cancer, liver cancer, leukemia, the esophageal carcinoma and colorectal cancer.
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Cited By (7)
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| 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 |
| CN104711368A (en) * | 2015-04-13 | 2015-06-17 | 玉峰惠仁生物医药科技（北京）有限公司 | Noninvasive polygenic hereditary colorectal cancer detection kit based on next-generation sequencing technology |
| CN106434979A (en) * | 2016-11-24 | 2017-02-22 | 深圳市核子基因科技有限公司 | Kit for detecting gastric cancer susceptibility and SNP marker thereof |
| CN106434980A (en) * | 2016-11-24 | 2017-02-22 | 深圳市核子基因科技有限公司 | Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit |
| CN107012232A (en) * | 2017-04-26 | 2017-08-04 | 成都中创清科医学检验所有限公司 | Primer and detection method for detecting the related SNP site of gastric cancer susceptibility |
| CN108977548A (en) * | 2018-08-24 | 2018-12-11 | 北京青航基因科技有限公司 | SNP site combination and the application in the product of preparation detection tumor susceptibility gene |
| CN109996891A (en) * | 2017-01-05 | 2019-07-09 | 南京帝基生物科技有限公司 | Method for carrying out the early detection of colon cancer and/or colon cancer precursor and for monitoring colon cancer recurrence |
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2012
- 2012-12-25 CN CN201210570393.0A patent/CN103898197A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN104593499B (en) * | 2015-01-20 | 2017-12-05 | 汕头大学医学院 | A kind of detection kit of the efficient molecule parting of cancer of the esophagus susceptibility loci |
| CN104711368A (en) * | 2015-04-13 | 2015-06-17 | 玉峰惠仁生物医药科技（北京）有限公司 | Noninvasive polygenic hereditary colorectal cancer detection kit based on next-generation sequencing technology |
| CN106434979A (en) * | 2016-11-24 | 2017-02-22 | 深圳市核子基因科技有限公司 | Kit for detecting gastric cancer susceptibility and SNP marker thereof |
| CN106434980A (en) * | 2016-11-24 | 2017-02-22 | 深圳市核子基因科技有限公司 | Kit for detecting esophagus cancer susceptivity and SNP (single nucleotide polymorphism) marker of kit |
| CN109996891A (en) * | 2017-01-05 | 2019-07-09 | 南京帝基生物科技有限公司 | Method for carrying out the early detection of colon cancer and/or colon cancer precursor and for monitoring colon cancer recurrence |
| CN109996891B (en) * | 2017-01-05 | 2020-10-16 | 南京帝准生物科技有限公司 | Methods for performing early detection of colon cancer and/or colon cancer precursor cells and for monitoring colon cancer recurrence |
| CN107012232A (en) * | 2017-04-26 | 2017-08-04 | 成都中创清科医学检验所有限公司 | Primer and detection method for detecting the related SNP site of gastric cancer susceptibility |
| CN108977548A (en) * | 2018-08-24 | 2018-12-11 | 北京青航基因科技有限公司 | SNP site combination and the application in the product of preparation detection tumor susceptibility gene |
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