CN106521012A - Detection reagent kit for detecting MDS (myelodysplastic syndrome)-related gene group - Google Patents
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Abstract
The invention discloses a groups of sequencing reagent kits for screening MDS (myelodysplastic syndrome)-related gene mutation and subsequence medical interpretation databases. The MDS-related gene group includes 16 genes of U2AF1, TP53 and the like. The sequencing reagent kits comprise multiple PCR (polymerase chain reaction) primers for amplifying all exons of the 16 MDS-related genes. The sequencing reagent kits and the medical interpretation databases have the advantages that the reagent kits for screening the MDS gene mutation are high in detection efficiency and detection rate and wide in mutant gene coverage; given medical interpretation reports are accurate and authoritative, and the like.
Description
Technical field
The present invention relates to technical field of molecular biology, further to the detectable of examination MDS associated gene mutations
Box and its medical science unscrambling data storehouse, and in particular to a kind of detection kit of detection MDS related gene groups.
Background technology
MDS (myelodysplastic syndrome) is one group and originates from the different of hemopoietic medullary system committed stem cell or pluripotent stem cell
The Clonal illness of matter, be mainly characterized by ineffective hematopoiesis and it is high-risk develop into acute myeloid leukemia, clinical manifestation is that hemopoietic is thin
Born of the same parents occur different degrees of ANOMALOUS VARIATIONS on quality and quantity.Its concrete clinical manifestation is anemia, can be with infection or bleeding, part
Patient can be asymptomatic.Some patientss can have liver, and spleen, lymph node silght enlargement, small number of patients can have breastbone tenderness, rib or extremity
Arthralgia.Hemogram can be in pancytopenia, or any one is and two is cytopenia.Nineteen eighty-two is advised by FAB cooperative groups
Name of disease is established, and is five types by MDS point：Refractory anemia；Refractory anemia increases with ring-type sideroblast；Refractory anemia
Increase with germinal cell, the refractory anemia with excess of blasts in transformation；Leukemia chronic myelo-monocytic.
About 10/,100,000～12/,100,000 population of MDS sickness rate, involves middle-aged and elderly people more, and the case of more than 50 years old accounts for 50%～
70%, the ratio of men and women is 2：1.MDS30%～60% is converted into leukemia.Its cause of death in addition to leukemia, it is most due to
Infection, especially bleeding, intracranial hemorrhage.
The cause of disease of MDS is still not clear, thus it is speculated that be to cause gene mutation, chromosome due to factors such as biology, chemistry, or physics
Exception makes the cell clonal hypertrophy of certain canceration.Generally acknowledged already, mutagenic agent such as virus, some drugses (such as chemotherapeutic), radiation
(radiotherapy), industrial reaction agent (such as benzene, polyethylene) and environmental pollution etc. can carcinogenesis, mutagenic agent can cause chromosome
Reset or gene rearrangement, it is also possible to only cause the change of gene expression to cause MDS.
MDS, as acute leukemia, is that the malignant clone by derived from the hematopoietic stem cell of an exception grows up
" Clonal disease ".Mainly involve myeloid cell, make bone marrow grain, red and huge three be the invalid DH of cell, its apoptotic cell
Quantity substantially increases.
The method of detection MDS relates generally to cellular morphology, flow cyctometry, cytogeneticss, molecular biology at present.Its
The experimental technique of middle molecular biology mainly has nest-type PRC, generation sequencing etc..PCR method intuitively specifically can not be dashed forward
Become sequence, and the method for generation sequencing is affected by sequencing throughput, it is impossible to disposably detect whole exons of multiple genes
Region.And secondary sequencing disposably can accurately measure the exon sequence of multiple genes as a kind of high-throughout detection method
Row, are possibly realized precisely treatment.
The content of the invention
It is contemplated that for the technological deficiency of prior art, there is provided a kind of detectable of detection MDS related gene groups
Box, is more limited to the diagnostic method for solving MDS in prior art, is lacked from the angle of molecular biology and is diagnosed the foundation of MDS.
The invention solves the problems that another technical problem be prior art MDS diagnostic method accuracy it is relatively low.
To realize above technical purpose, the present invention is employed the following technical solutions：
A kind of detection kit of detection MDS related gene groups, the test kit include following 16 genes：U2AF1,
CEBPA, SF3B1, TP53, SETBP1, NRAS, NPM1, RUNX1, TET2, KRAS, IDH1, DNMT3A, ASXL1, JAK2,
PTPN11, SRSF2
Above-mentioned 16 genes, are made up of 85 hot spot mutations, and its mutational site is the conventional mutational site in this area, preferably
Include indel mutational sites (insertion or the sequence that causes of disappearance change insertion or deletion, indel), nothing
Adopted mutational site, splice region mutational site or missense mutation site, 16 gene extron subregions more preferably of the present invention
Hot spot mutation it is as shown in table 1.
1 hot spot mutation list of table
The hot spot mutation of 16 gene extron subregions of the present invention can be used to make MDS auxiliary diagnosis, ability
Domain MDS conventional diagnostic method has morphology, cytogeneticss and flow cyctometry, and the method for the invention is preferably from molecule
Biology angle makes diagnosis to MDS.
The multiple PCR primer of the present invention one group of detection of offer MDS related genes exon region of the present invention, it is described to draw
The amplification scope of thing includes the exon region of 16 and MDS related genes, and it is that 250bp is left to expand the individual chip length for obtaining
It is right.The multiple PCR primer provided using the present invention, with reference to secondary sequencing technologies, can detect mutant gene group of the present invention
In whole exon regions.
Wherein described secondary sequencing reagent is reagent commonly used in the art, gained sequence is entered as long as disclosure satisfy that
The requirement of the secondary sequencing of row.The preparation method of reagent thereof is this area customary preparation methods, preferably commercially available.
Detection kit of the present invention more preferably also includes that the genomic DNA that will be extracted in detection object is made and is available for surveying
The reagent in the library that sequence is used, this kind of reagent is reagent commonly used in the art, as long as disclosure satisfy that multiplex PCR to genome
The requirement of DNA mass.The preparation method of reagent thereof is this area customary preparation methods, preferably commercially available.
Detection kit of the present invention preferably also includes：DNTP solution, archaeal dna polymerase, for isolated or purified core
Acid is Backman magnetic beads.
Tissue of the sample of the present invention for detection object, as long as the gene of detection object can be extracted from detection sample
Group DNA.The detection sample is preferably one or more in bone marrow, blood, solid tumor sample, it is therefore preferable to bone marrow
Sample.
A kind of using method of test kit of the present invention, which comprises the following steps：
(1) bone marrow of detection object is extracted using detection kit of the present invention, genomic DNA is extracted；
(2) exon region of 16 MDS related genes described in invention in genomic DNA is entered with multiple PCR primer
Row amplification；
(3) fragment of amplification gained in step (2) is made the library for being available for Ion Torrent microarray datasets to be sequenced；
(4) gained DNA library in step (3) is sequenced, obtains lower machine data；
(5) to (4) in lower machine data carry out bioinformatic analysis, then understood using medical science unscrambling data storehouse
Make diagnosis.
The concrete operation method of each step refers to kit specification above.
Reagent and raw material used by the present invention is commercially available.
The present invention positive effect be：Can be used for examination MDS associated gene mutations using what the present invention was provided
Gene group, with reference to high throughput sequencing technologies, it is possible to achieve auxiliary diagnosis are carried out to MDS outside morphology and flow cyctometry,
Assessment is made especially for disease prognosis to make anticipation to the action effect for related target medicine there is special advantage.This
The detection kit of the gene group for examination MDS associated gene mutations of bright offer has detection efficiency height, recall rate height etc.
Advantage.
Specific embodiment
Hereinafter the specific embodiment to the present invention is described in detail.In order to avoid excessive unnecessary details,
Will not be described in detail to belonging to known structure or function in following examples.In addition to being defined, institute in following examples
Technology and scientific terminology are with the identical meanings being commonly understood by with those skilled in the art of the invention.
Material and method：
In hematopathy hospital of the Chinese Academy of Medical Sciences and consonance magnificent medical diagnosiss center in October, 2015 in July, 2016
In the patient of diagnosis, according to following standard, we enter Line Continuity and enter group (194).
Inclusion criteria is：Jing morphocytologyes and flow cyctometry are diagnosed as the trouble of myelodysplastic syndrome (MDS)
Person.
The diagnostic result of 194 patients is as shown in table 2：
2 patient diagnosis the results list of table
Collection 3mL Bone Marrow of Patients extraction genomic DNAs are to be measured, and the detection is through the human relations of hematopathy hospital of the Chinese Academy of Medical Sciences
The approval of reason committee.
Sample process：In sample, genome utilizes Tiangeng DNA extraction kit (article No.：DP318-03) it is stripped, has
Operation instructions of the body operational approach referring to test kit.
Genomic DNA is expanded with multiple PCR primer, use of the Examination on experimental operation referring to Life companies test kit
Description.
The structure in library：The DNA fragmentation obtained to amplification builds storehouse examination using the Ion Torrent platforms that Life companies produce
Agent box carries out the structure of sequencing library, operation instructions of the Examination on experimental operation referring to Life companies test kit.
High-flux sequence：The sequencing library for building is carried out into height Jing after Water-In-Oil process on Ion Torrent platforms
Flux is sequenced, Water-In-Oil processing method and high-flux sequence method refer to high-flux sequence instrument Ion Torrent and which is matched somebody with somebody
The operation instructions of complete equipment One Touch.
Bioinformatic analysis：First, using Ion Report softwares (v4.6, Thermo Fisher, Carlsbad,
CA, USA) low-quality sequencing fragment is filtered, and qualified sequencing fragment is compared into mankind's reference gene group hg19
(http://hgdownload.cse.ucsc.edu/downloads.html), using Torrent Variant Caller
(v4.6.0.7) subprogram detection mutational site, software parameter use the setting of acquiescence.This software has been optimized for
Process the distinctive type of error of Ion Torrent microarray datasets.The finally mutation to finding out includes that SNP and Indel is used
ANNOVAR(http://annovar.openbioinformatics.org/en/latest/) software annotated, in annotation
Appearance includes being mutated the position in genome, the gene of association, gene extron numbering, nucleotide level variation, corresponding egg
White level makes a variation, and the mutation is in dbSNP (http://www.ncbi.nlm.nih.gov/snp/), 1000Genomes data bases
(http://www.1000genomes.org/) and cancer mutation database COSMIC (http://
Cancer.sanger.ac.uk/cosmic the annotation in), PolyPhen (http://genetics.bwh.harvard.edu/
) and SIFT (http pph2/://sift.jcvi.org/) function prediction result etc..Further caused a disease using the screening of principle once
Mutational site：(1) genomic locations being located according to mutation and type, filter out and the prominent of impact are not produced on protein product sequence
Become；(2) using 1000Genomes data, each ratio of mutation in crowd is annotated, if ratio is less than or equal to 1% not
It is considered pleomorphism site；(3) cancer databases COSMIC are retrieved, whether one mutation of inquiry is on the books in COSMIC, its
Occur in hematological system tumor；(4) predict whether the mutation affects egg using protein function forecasting software PolyPhen-2
Contour painting energy；(5) if after (2) (3) (4), a mutation meets " non-polymorphism ", " crossing described in hematological system tumor "
At least two in " impact protein function " this three, will be judged to may be related to disease.Finally, if a though mutation
So it is unsatisfactory for (5) but is recorded as detecting in tumor in COSMIC, then is read the mutation for interrogatory.Finally
If one mutation be expressly recited in the literature not with disease height correlation, be directly judged to hot spot mutation.
Statistical analysiss：The ratio that each gene carries pathogenic mutation is counted, mutation rate highest gene is filtered out, is used
Software is Excel.
Medical science is understood：Molecular pathology is made to the genic mutation type that bioinformatic analysis draw with medical science unscrambling data storehouse
Diagnosis.Medical science unscrambling data storehouse used is as follows：
(1) U2AF1 gene mutation is mainly seen in MDS, AML.U2AF1 mutation can cause its protein level to rise, and change
Its cut mode.The MDS patient of U2AF1 mutation easily progresses to secondary acute marrow series leukemia, but its overall survival is not had
Have an impact.(PMID：22158538)
(2) CEBPA is the important transcription factor for safeguarding the differentiation of granulocyte hemopoietic system.CEBPA gene mutation is mainly seen in
AML patient, especially M1, M2 and M4, are currently known the AML patient prognosis bona of CEBPA dialleles mutation.
(3) SF3B1 gene mutation is mainly seen in CLL, MDS and MPN patients, the prognosis malas in CLL patient.In MDS
In, it is that refractory anemia increases (RARS) with ringed sideroblasts that SF3B1 is mutated modal hypotype, and overall survival is carried
It is high.(PMID：21995386)
(4) TP53 gene mutation is mainly seen in the diseases in the blood system such as MDS, AML, CLL and solid tumor.TP53 genes are dashed forward
The MDS survivals of change are reduced and most prognosis malas.Acting on the related medicine of TP53 and its path has Cisplatin,
Fluorouracil, Docetaxel, Paclitaxel, Aspirin.(PMID：21714648)
(5) SETBP1 gene coded proteins participate in the regulation and control of DNA replication dna.SETBP1 somatic mutatioies are turned with marrow series leukemia
Change related, the prognosis malas in MDS and CMML.(PMID：23832012)
(6) NRAS gene mutation is mainly seen in the diseases in the blood system such as AML, CML, ALL, MPN and solid tumor.RAS eggs
White NRAS is primarily involved in RAS signal path.In AML, if patient carries NRAS, DNMT3A and NPM1 mutation simultaneously, then carry
Show prognosis bona.RAS mutation are still not enough to become the independent prognostic factor in AML.But nearest research prompting, for carrying
The AML patient of RAS mutation, it is probably beneficial after alleviation to carry out after treatment using the cytosine arabinoside of high dose.(PMID：
26376137)
(7) NPM1 mutation are mainly seen in AML, especially the AML of normal karyotype.NPM1 mutation mostly are the frameshit of Exon 12
Mutation, is mutated with A types most commonly seen.NPM1 is individually mutated prompting patient prognosis bona；Dash forward when other prognosis malas genes are merged
During change (such as FLT3, DNMT3A), patient's prognosis malas；Document report is mutated not for the elder patients more than 65 years old, NPM1
Reresent prognosis bona.NPM1 mutation can be additionally used in the monitoring of minimal residual disease.The patient of NPM1 mutation is using heavy dose of soft
Erythromycin for treating may benefit.(PMID：26755712；22417203；26376137；25713434)
(8) RUNX1 mutation are mainly seen in medullary system tumor patient, are especially more common in AML patient.RUNX1 mutation MDS and
AML patient's prognostic level and overall survival can all be reduced.(PMID：19808697；22689681；21714648)
(9) TET2 gene mutation is mainly seen in AML patient, also exists, at present in CML, MDS, MPN and Lymphoma
TET2 gene mutation is the preferable index of prognosis in CMML, and the prognosis malas in MDS/MPN, Secondary cases AML.
(10) KRAS is found in the diseases in the blood system such as MDS, AML, MPN, MM and solid tumor.RAS albumen KRAS
It is primarily involved in RAS signal path.KRAS will not inducing cell morphology and the change of transfer ability, but can strengthen ERK,
The phosphorylation of PDK1 and AKT.RAS mutation are still not enough to become the independent prognostic factor in AML.But nearest research prompting,
For the AML patient for carrying RAS mutation, after treatment is carried out using the cytosine arabinoside of high dose after alleviation possibly beneficial
's.
(11) IDH1 gene mutation is mainly seen in AML, MDS and MPN patients.IDH1 gene codes Isocitrate dehydrogenase 1,
Participate in intracellular oxidative damage defense mechanism.For AML patient, merge possible prognosis when NPM1 is mutated preferable.IDH1 inhibitor AG-
120 have carried out Phase I clinical trial (NCT02632708) in AML patient.(PMID：22915641；27245312；
26660517；22417203)
(12) DNMT3A gene mutation is mainly seen in AML, especially the AML of normal karyotype, is also found in other blood diseases
Disease, or normal old people (occurring in normal person, general load is relatively low, and independent mutation).DNMT3A is used as a table
The gene of genetic correlation is seen, the Preleukemia stage is typically occurred in, its mutational load is general higher.DNMT3A gene Primary mutations
Form is point mutation, is common in the aminoacid of R882 positions more, and modal mutant form is R882H.DNMT3A gene mutation is suffered from
The most prognosis malas of person.DNMT3A mutation patients may be benefited using the daunorubicin of high dose, and DNA methylation transferring enzyme is pressed down
Preparation decitabine has sound response.(PMID：25426837；26755712；22417203；19776405)
(13) ASXL1 gene mutation is had been reported that in MPN patient in AML, CML, MDS, with AML overall patient's survival rates
Reduce correlation (PMID:22417203).
(14) being currently known JAK2exon12/14 abrupt climatic changes contributes to MPN diagnosis, and V617F mutation are found in 90-95%
PV patient and 50-60% ET and PMF patients.JAK2 albumen is the non-receptor related to cytokine receptor signal path
Type tyrosine kinase, participates in the growth of cell, development, differentiation.Ruxolitinib is JAK2 inhibitor, can improve MPN patient
The symptoms such as splenomegaly.(PMID：24740812)
(15) PTPN11 gene mutation is mainly seen in CML, AML, ALL and MPN patient.PTPN11 is protein tyrosine phosphatase
Enzyme, plays an important role in cell growth, differentiation, mitotic cycle and oncogenic transformation.Dodecane-
Trimethylamine can target PTPN11 (in grinding).
(16) SRSF2 mutation are mainly seen in the diseases such as MDS, t-AML and CML, the MDS overall patients life of SRSF2 mutation
Rate reduction is deposited, the risk to AML conversions is improved.(PMID：22389253)
Using test kit testing result of the present invention：
Detected by 194 patients, the positive rate of 16 genes is as shown in table 3 below：
3 16 gene masculine rate lists of table
| Mutant gene | Mutation case load | Mutation rate | Mutant gene | Mutation case load | Mutation rate |
| U2AF1 | 44 | 22.68% | TET2 | 8 | 4.12% |
| CEBPA | 23 | 11.86% | KRAS | 7 | 3.61% |
| SF3B1 | 22 | 11.34% | IDH1 | 6 | 3.09% |
| TP53 | 14 | 7.22% | DNMT3A | 6 | 3.09% |
| SETBP1 | 10 | 5.15% | ASXL1 | 6 | 3.09% |
| NRAS | 9 | 4.64% | JAK2 | 6 | 3.09% |
| NPM1 | 9 | 4.64% | PTPN11 | 4 | 2.06% |
| RUNX1 | 8 | 4.12% | SRSF2 | 4 | 2.06% |
Show Jing after statistics：(being summarized according to above table) as can be seen here, using the method for secondary sequencing to bone marrow
Hypertrophy exception syndrome carries out auxiliary diagnosis, with diagnosis accurately, obtain the characteristics of containing much information.
Above embodiments of the invention are described in detail, but the content have been only presently preferred embodiments of the present invention,
Not to limit the present invention.All any modification, equivalent and improvement made in the application range of the present invention etc., all should
It is included within protection scope of the present invention.
Claims (7)
1. a kind of detection kit of detection MDS related gene groups, it is characterised in that the test kit includes following 16 genes：
U2AF1, CEBPA, SF3B1, TP53, SETBP1, NRAS, NPM1, RUNX1, TET2, KRAS, IDH1, DNMT3A, ASXL1,
JAK2, PTPN11, SRSF2.
2. a kind of detection kit of detection MDS related gene groups according to claim 1, it is characterised in that the detection
Test kit also includes the primer of 16 genes above, and the amplification scope of the primer includes whole exons of 16 genes of the above
Region.
3. the detection kit of a kind of detection MDS related gene groups according to claim 2, it is characterised in that described outer aobvious
Subregion includes 85 hot spot mutations, 85 hot spot mutations and its corresponding relation such as following table institute with 16 genes
Show：
4. a kind of detection kit of detection MDS related gene groups according to claim 3, it is characterised in that the reagent
Box is included for expanding the multiple PCR primer of the mutant gene group exon region, and for the reagent of secondary sequencing.
5. a kind of detection kit of detection MDS related gene groups according to claim 3, it is characterised in that the reagent
Box include by extract in detection object genomic DNA make be available for be sequenced library reagent.
6. a kind of detection kit of detection MDS related gene groups according to claim 3, it is characterised in that the reagent
Box includes dNTP solution, archaeal dna polymerase, for the reagent of isolated or purified nucleic acid, positive control or negative control.
7. a kind of detection kit of detection MDS related gene groups according to claim 3, it is characterised in that the reagent
Box includes the data base of gene group examining report, and the data base contains the medical explanation to the hot spot mutation.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107916290A (en) * | 2017-12-29 | 2018-04-17 | 天津协和华美医学诊断技术有限公司 | A kind of detection kit for detecting hereditary thrombocytopenia related gene group |
| CN111304331A (en) * | 2020-03-09 | 2020-06-19 | 南京实践医学检验有限公司 | MDS detection kit based on multiple PCR (polymerase chain reaction) targeted high-throughput sequencing and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290137A (en) * | 2013-06-26 | 2013-09-11 | 北京迈基诺基因科技有限责任公司 | Screening method of tumor susceptibility gene |
| CN103993096A (en) * | 2014-06-09 | 2014-08-20 | 中国医学科学院血液病医院（血液学研究所） | Kit for diagnosing congenital bone marrow failure diseases |
| CN104862407A (en) * | 2015-06-02 | 2015-08-26 | 上海艾迪康医学检验所有限公司 | Primer and method for detecting EZH2 genes |
| CN105506107A (en) * | 2015-12-30 | 2016-04-20 | 杭州艾迪康医学检验中心有限公司 | Primers and method for detecting polymorphic hotspot mutation condition of DOCK2 gene |
| CN106065414A (en) * | 2016-06-15 | 2016-11-02 | 浙江大学 | Noninvasive cancer of pancreas polygenes detection method and kit based on blood plasma cfDNA detection technique |
-
2016
- 2016-12-29 CN CN201611241375.2A patent/CN106521012A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290137A (en) * | 2013-06-26 | 2013-09-11 | 北京迈基诺基因科技有限责任公司 | Screening method of tumor susceptibility gene |
| CN103993096A (en) * | 2014-06-09 | 2014-08-20 | 中国医学科学院血液病医院（血液学研究所） | Kit for diagnosing congenital bone marrow failure diseases |
| CN104862407A (en) * | 2015-06-02 | 2015-08-26 | 上海艾迪康医学检验所有限公司 | Primer and method for detecting EZH2 genes |
| CN105506107A (en) * | 2015-12-30 | 2016-04-20 | 杭州艾迪康医学检验中心有限公司 | Primers and method for detecting polymorphic hotspot mutation condition of DOCK2 gene |
| CN106065414A (en) * | 2016-06-15 | 2016-11-02 | 浙江大学 | Noninvasive cancer of pancreas polygenes detection method and kit based on blood plasma cfDNA detection technique |
Non-Patent Citations (4)
| Title |
|---|
| 宋陆茜等: "二代测序技术在骨髓增生异常综合征诊治中的临床应用", 《诊断学理论与实践》 * |
| 罗锦霞等: "基因组检测综合分析在骨髓增生异常综合征诊治中的应用", 《中国实验血液学杂志》 * |
| 许婧: "SFB1、SRSF2及U2AF1基因突变在骨髓增生异常综合征中的表达及临床意义", 《中国优秀硕士学位论文全文数据库》 * |
| 阴常欣: "高通量测序技术在急性髓系白血病临床检测中的方法学建立及其初步应用", 《中国博士学位论文全文数据库》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107916290A (en) * | 2017-12-29 | 2018-04-17 | 天津协和华美医学诊断技术有限公司 | A kind of detection kit for detecting hereditary thrombocytopenia related gene group |
| CN111304331A (en) * | 2020-03-09 | 2020-06-19 | 南京实践医学检验有限公司 | MDS detection kit based on multiple PCR (polymerase chain reaction) targeted high-throughput sequencing and preparation method thereof |
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