[go: up one dir, main page]

CN112852966A - Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof - Google Patents

Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof Download PDF

Info

Publication number
CN112852966A
CN112852966A CN202110307861.4A CN202110307861A CN112852966A CN 112852966 A CN112852966 A CN 112852966A CN 202110307861 A CN202110307861 A CN 202110307861A CN 112852966 A CN112852966 A CN 112852966A
Authority
CN
China
Prior art keywords
mapk
pancreatic cancer
gene
detection
calcb
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202110307861.4A
Other languages
Chinese (zh)
Inventor
虞先濬
施思
魏妙艳
梁晨
徐近
王巍
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fudan University Shanghai Cancer Center
Original Assignee
Fudan University Shanghai Cancer Center
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fudan University Shanghai Cancer Center filed Critical Fudan University Shanghai Cancer Center
Priority to CN202110307861.4A priority Critical patent/CN112852966A/en
Publication of CN112852966A publication Critical patent/CN112852966A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention belongs to the technical field of pancreatic cancer polygene detection, and discloses a pancreatic cancer detection panel based on a second-generation sequencing technology, which has events such as gene mutation, copy number variation and the like with definite clinical relevance with pancreatic cancer, makes a polygene screening list into a probe, and detects high-risk genes in a targeted manner through DNA sequencing, so that gene mutation with guiding significance for diagnosis and treatment can be detected more efficiently, mutant genes can be detected in a targeted manner, and the cost of gene detection of patients can be reduced.

Description

Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof
Technical Field
The invention belongs to the technical field of pancreatic cancer polygene detection, and relates to a pancreatic cancer detection panel based on a second-generation sequencing technology, a kit and application thereof.
Background
Pancreatic cancer is a malignant tumor of the digestive tract with high malignancy and difficulty in diagnosis and treatment, and about 90% of pancreatic cancer is ductal adenocarcinoma originating from the epithelium of the glandular duct, and the morbidity and mortality of pancreatic cancer have increased remarkably in recent years. Survival rate < 1% for 5 years is one of the worst-prognosis malignancies. The early diagnosis rate of pancreatic cancer is low, the operative mortality rate is high, and the cure rate is low.
The application of gene sequencing to determine the targeted treatment scheme of a patient is increasingly common in pancreatic cancer, but the existing sequencing of pancreatic cancer is not focused on specific genes, namely whole exome sequencing, the specific genes in the existing sequencing are few in number and low in coverage rate, and large-area consideration is difficult to achieve; the latter gene is abundant, mostly plays no guiding role in pancreatic cancer treatment, and is relatively costly. Therefore, there is an urgent need for a method for detecting pancreatic cancer that combines genes related to pancreatic cancer therapy with genes that are not limited to a small number of targeted therapeutic genes.
Disclosure of Invention
In order to realize accurate detection of pancreatic cancer and provide more effective information for diagnosis, the invention selects gene variation data of multiple platforms such as TCGA database, MSKCC-IMPACT database and the like, enriches important exon regions and partial intron regions of 685 genes based on a second-generation sequencing technology by using a biotin probe hybridization method, and performs high-depth sequencing, thereby finding out events such as gene mutation, copy number variation and the like with definite clinical relevance to pancreatic cancer, and making a multiple gene screening list into a probe.
In a first aspect, the present invention provides a pancreatic cancer detection panel based on a next-generation sequencing technology, said panel comprising a gene for detecting a mutation gene, a copy number variation gene and a gene for occurrence of a rearrangement event that are relevant for pancreatic cancer prognosis.
In certain embodiments, the genetic mutations include the following genes: ABCB1, ABCC 1, ABL1, ACVR 11, ACVR 21, ADGRA 1, AFF1, AGO 1, AHNAK, AKT1, ALOX12 1, AMER1, ANKRD1, APC, APCDD1, AR, ARAF, ARFRP1, ARHGEF1, ARID 11, ARID 51, ARMC 1, ARRDC 1, ASH 11, ASXL1, ATF1, ATR 1, ATM, ATP1A1, ATP2B 1, ATRCR 36AURK, AURKB 1, AXIN1, AXB 2 CSF, BAC 1, CABCB 1, CABCC 1, CABCCDC 1, CABCCDK 1, CALCBCCDK 1, 363672, CALCBCCDK 1, CALCBCCDK 363672, 36363636363672, CALCBCCDK 3636363672, CALCBCCDK 1, 3636363672, 363672, 363636363672, CALCBCCDK 36363636363672, CALCBCCDK 1, 363636363672, 1, 36363636363636363636363636363636363672, CALCBCCDK 1, CALCBCCDK 363672, CALCBCCDK 1, CALCBCCDK 363672, 1, 363636363636363636363636363636363636363636363636363636363636363636363672, CALCBCCDK 1, CALCBCCDK 1, 1, CTLA, CTNNA, CTNNB, CUL4, CUX, CXCR, CYLD, CYP17A, CYP19A, CYP2C, CYP2D, CYSLTR, DAXX, DCUN1D, DDR, DEPDC, DICER, DIS, DMC, DNJB, DNMT3, DOT1, DPYD, DROSHA, DUSP, DYNC2H, E2F, EED, EGFL, EGFR, EIF1, EIF4A, EIF4, ELF, ELK, ELOC, EME, EMSY, EP300, EPAS, EPCAM, EPHA, EPHB, ERBB, ERCC, ERCG, ERF, ERG, ERV, FAS, ERV, ERFV, CUV, CU, FGFR, FGAT, EPHA, FGAT, GPS2, GREM1, GRM 1, GSK3 1, GSTM1, GSTP1, GSTT1, H3F3 1, HDAC1, HGF, HIST1H 11, HIST1H 21, HIST1H3 MDMA 1, HIST1H3 1, HIST2H3 MDMA 1, HIST2H 1, KM 36K 1, KM 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K, MAPK 36K 1, MAPK 36K, MDCT, KM K36K KM, KM 36K 1, KM 1, MAPK 36K 1, KM 1, MAPK 36K 1, MAPK 36K 1, MAPK3 MDCT, KM 1, MAPK 36K 1, KM 1, MAPK 36K KM, MSH6, MSI1, MSI2, MST1, MST 11, MTAP, MTHFR, MTOR, MUS 1, MUTYH, MYB, MYC, MYCL, MYCN, MYD 1, MYD RADD 1, NBN, NCOA 1, NCOR1, NEB, NEGR1, NF1, NFE2L 1, NFKBIA, NKX 1-1, NOTCH1, OTCH1, NQO1, NR3C1, NR4A 1, NRAS, NRG1, NSD1, NT5C 1, NTHL1, NTRK 36RK 1, PTP 1, PRP 36P 1, PARP1, PSP 36K 1, PSP 1, PS, RAD51, RAD54, RAF, RANBP, RARA, RASSF, RB, RBM, RECQL, REL, RET, RFWD, RHEB, RHOA, RICTOR, RIT, RNF, ROBO1ROBO, ROS, RPA, RPL, RPS6KA, RPS6KB, RPTOR, RRAS, RRAM, RSPO, RTEL, RUNX1T, RXRRYBP, SCN7, SDC, SDHA, SDHAF, SDHB, SDHC, SDHD, SEMA3, SESN, SETD, SESF, SF3A, SGK, SH2B, SH2D1, SHOC, SHQ, SLC34A, SLCO1B, SPET, SLTP 1, SLTP, SLSP, SLTP, SMTP, SMAD, SMSC, SMTP, SMAD, SMSC, SMTP, SMSC, SMAD, SMTP, SMSC, SMTP, SMAD, SMSC, SMTP, SMAD, SMTP, SMSC, SMTP, SMSC, SMTP, SMSC, SMTP, SMRT, SMSC, SMTP, SMSC, TRIM33, TRRAP, TSC1, TSC2, TSHR, TYMS, TYRO3, U2AF1, U2AF2, UGT1A1, UMPS, UPF1, USP6, USP8, VEGFA, VEGFB, VEGFC, VHL, VTCN1, WHSC1, WHSC1L1, WISP3, WRN, WT1, WWTR1, XIAP, XPC, XPO1, XRCC1, XRCC2, XRCC3XRCC4, XRCC6, YAP1, YES1, YY1, ZBTTB 2, ZFH HX3, ZNF217, ZNF292, ZNF703, ZNRF3, ZNRF 2.
In certain embodiments, the gene copy number variation comprises the following genes: CDK4, CDK6, CYP2D6, ERBB2, MDM2, MDM4, MET, MYC.
In certain embodiments, the gene in which the rearrangement event occurs is: ALK, BCL2, BCR, BRAF, BRCA1, BRCA2, CD274, CD74, CD86, EGFR, ERBB4, ETV4, ETV5, ETV6, EWSR1, EZR, FGFR1, FGFR2, FGFR3, KIT, KMT2A, MET, MSH2, MYB, MYC, NOTCH2, NRG1, NTRK1, NTRK2, NTRK3, NUTM1, PDGFRA, RAF1, RARA, RET, ROS1, RSPO2, SDC4, SLC34A2, TMPRSS 2.
In another aspect, the invention provides a detection kit for detecting pancreatic cancer based on second-generation sequencing, the kit comprises a detection probe and a detection reagent, and the detection probe is a probe for detecting gene mutation or copy number variation of panel according to the first aspect of the invention.
In certain embodiments, the probe is an RNA probe.
In certain embodiments, the gene mutation detection comprises point mutations, deletions, insertions, fusions, and the like.
On the other hand, the invention also provides application of the detection panel in preparation of a pancreatic cancer detection device.
In another aspect, the present invention also provides a pancreatic cancer detection apparatus, including: the sequencing module is used for extracting DNA of a sample to be tested and carrying out high-throughput sequencing to obtain a sequencing result; the comparison module is used for processing the high-throughput sequencing result and comparing the data with the panel to obtain mutation information; and the analysis module is used for analyzing the obtained mutation information to obtain a medication scheme. A
In another aspect, the present invention also provides a method for diagnosing and detecting pancreatic cancer, comprising the steps of,
step S1: obtaining a detection sample DNA (the sample DNA comprises DNA tissue sample DNA and cfDNA of a blood sample), extracting the DNA of a focus tissue and a paracarcinoma or blood tissue of a patient, and optionally extracting paraffin-embedded tissues.
Step S2: constructing a sample DNA library, randomly fragmenting DNA into small fragments of several hundred bases or less, and then preparing the library by adopting a KAPA library construction kit. The method mainly comprises the following steps: DNA breaking, filling in the tail end, adding ' A ' at the 3 ' end, connecting by a joint, purifying, amplifying a library and purifying. After library preparation was complete, it was stored at-20 ℃.
Step S3: constructing a detection probe for detecting the panel, hybridizing with a DNA library, capturing and sequencing; whole genome or whole exome sequencing can also be performed directly.
Step S4: and (4) carrying out biological information analysis on the sequencing result to obtain a sample mutation result.
Wherein the sequencing result is a fastq file, and the sequencing result is analyzed by using an algorithm for acquiring the gene mutation issued by Broad to obtain the gene mutation result and annotate the gene mutation result. The main steps include the quality control of fastq files, genome association, analysis of somatic mutation (somatic mutation) and germ cell mutation (germline mutation), and annotation. The software used respectively had: the fastqc and fastx _ toolkit are used for quality control; bwa, and gatk and mutect2 obtain somatic mutation; obtaining embryo cell mutation by a HaplotpypeCaller method; ANNOVAR is annotated.
Step S5: comparing the sample mutation result with the detection panel to obtain the sample gene mutation result, and combining clinical information to obtain the final diagnosis.
Compared with the prior art, the invention has the beneficial effects that:
1) the detection panel of the invention not only considers the genes related to pancreatic cancer treatment, but also is not limited to a few targeted treatment genes, and brings the genes related to the pancreatic cancer development mechanism into the gene list of targeted sequencing, so that the practicability is much higher than that of full exome sequencing, and the panel can provide useful gene information no matter the guide of clinical medication or the subsequent search of new targets of pancreatic cancer.
2) The gene set provided by the invention is used for preparing a detection probe of the multi-gene panel, so that the accurate sequencing is realized, and the abnormal condition of the gene listed by the invention can be used as a reference for clinical diagnosis, so that doctors can be helped to deeply know the cancer development condition of patients, the patients can know the condition of self gene variation, and the design of a treatment scheme is also helped.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments will be briefly described below.
FIG. 1 is a diagram showing the results of quality evaluation of DNA sample extraction and sequencing in examples.
FIG. 2 shows the results of detection of gene mutation in leukocyte of blood cells in examples.
FIG. 3 shows the results of detection of multiple gene mutations (physiological mutation) in tissues.
FIG. 4 shows the results of the detection of multiple gene copy number variation in tissues.
FIG. 5 shows the results of the detection of tissue gene rearrangement.
FIG. 6 is a reference diagram of gene mutation versus drug application information.
Detailed Description
The invention will be better understood from the following examples. However, it is easily understood by those skilled in the art that the description of the embodiment is only for illustrating and explaining the present invention and is not for limiting the present invention described in detail in the claims. Unless otherwise specified, reagents, methods and equipment used in the present invention are conventional methods, and test materials used therein are available from commercial companies, unless otherwise specified.
Examples
Step one, extracting DNA from cancer tissues and Blood samples of a patient, and using kits which are respectively TIANGEN TGuide Cells/Tissue Genomic DNAkit and TIANGEN TGuide Large Volume Blood Genomic DNAkit (1-3 ml).
Step two, preparing a DNA sample library. The DNA was randomly disrupted using a bioruptortucd-300 non-contact fully automatic sonicator, which is not required for ctDNA. DNA400ng was taken, diluted to 50. mu.l with nucleose-free water, transferred to a 0.5mL Eppendorf Lobind Tube, mixed well, centrifuged briefly and placed on ice for use. Placing a sample: and symmetrically placing centrifuge tubes (if single tubes exist, adding water into the empty tubes for balancing), screwing the rotating head, placing the centrifuge tubes on an ice box, and precooling for 1-2 min. And (3) carrying out ultrasonic disruption of 150-200 bp. The previous ultrasonication step was repeated for a total of 9 cycles, ending about 90 min. Then, the KAPA library construction kit is adopted for library preparation.
Step three, preparing an RNA probe. The oligo pool of the target gene was obtained from the company, and diluted to 1 ng/. mu.l with 1 XTE (pH 8.0). The oligo sequence was amplified with the Herculase II Fusion DNA Polymerase kit. Then Ambion SP6 megascript kit is used for RNA transcription, and the preparation of the probe is completed. The RNA probe was stored at-80 ℃ and a portion of the RNA probe library was diluted to 100 ng/. mu.l. The probe prepared by the method can be applied to 200-300 samples, and the cost of sequencing is greatly reduced.
Step four, hybridization capture and sample delivery sequencing.
1. Mixing 95 μ l Block and DNA library with total amount of not less than 500ng and volume of about 5 μ l, centrifuging, marking as "DNA-Block", placing on PCR instrument, covering hot cover, and keeping at 95 deg.C for 5 min; hold at 65 ℃.
2. And matching a 'bait' Mix reaction system in the PCR reaction tube, marking as 'bait', placing the 'bait' tube on the PCR instrument for incubation when the temperature of the PCR instrument is reduced to 65 ℃ for 2.5min, and covering a thermal cover.
TABLE 1 PCR reaction System
Figure BDA0002988592730000091
3. Placing the PCR tube 'bait' into a PCR instrument for 2.5min, sucking 13 mul Hyb Buffer from the 'Hyb Buffer' and transferring the Hyb Buffer to a 'DNA-block' sample, sucking 6 mul from a 'bait' hole and transferring the Hyb Buffer to the DNA-block sample, sucking and beating the mixture for 10 times by light, fully mixing the mixture uniformly, avoiding generating a large amount of bubbles, pasting a membrane, covering a hot cover of the PCR instrument, and incubating the mixture overnight for 24h at 65 ℃. The capture beads are then ready. According to the requirement of 3 multiplied by 165 mu l of high-stringencybuffer for each sample, subpackaging the high-stringencybuffer in a 96-well plate, opening a drying instrument, adjusting the temperature of the drying instrument to 65 ℃, and placing the drying instrument in a drying instrument for preheating when the temperature of the drying instrument is stabilized at 65 ℃. Then, capture of the target region DNA library is performed, and enrichment of the DNA library is completed (Post-PCR reaction). The library was then mixed and sample-fed sequenced.
Step five, data analysis
The original file for analysis is fastq format data. Bwa is used for matching, and a haplotypeCaller method is used for obtaining germline mutation; somatic mutation analysis of tissue samples, using muttec 2, was then annotated using ANNOVAR.
And step six, after obtaining the annotation results of all the tissue samples, arranging the annotation results as shown in the attached drawing.
In which, fig. 1 shows the quality control of tissue samples, which mainly includes quality evaluation of DNA sample extraction and quality evaluation of sequencing, which are the most basic parts of the whole experiment, and the failure of data quality will affect the accuracy and reliability of subsequent results.
FIG. 2 shows the result of detecting the gene mutation of leukocyte in blood cells, which is a specific case of the gene mutation of the mutated leukocyte in blood cells (germline mutation) related to the gene list listed in the present invention in the bioinformatics analysis result described before after the sequencing of the DNA sample of the patient.
FIG. 3 shows the result of detection of multiple gene mutations (genetic mutation) in a tumor tissue.
FIG. 4 shows the result of detecting the variation of the copy number of multiple genes in tissue, and the method is the same as that in FIG. two and FIG. three.
FIG. 5 shows the results of detection of tissue gene rearrangement
FIG. 6 is a reference of gene mutation to drug application information, which can refer to the past drug application information of the corresponding mutation according to germline mutation, and the genetic mutation and the copy number variation of the key gene, so as to conveniently make a treatment plan for a patient. In conclusion, through the gene detection panel of the invention, the whole genome or whole exon sequencing is carried out on the sample, and the mutation conditions of all genes are obtained after analysis, so that the genes in the gene panel listed in the invention are found for further analysis by combining clinic. According to the gene provided by the invention, a multi-gene panel probe can be manufactured, accurate sequencing is carried out, the result is obtained, and then information such as clinic and the like of a patient sample is combined, so that a powerful reference can be provided for clinical diagnosis of the patient, the cancer development condition of the patient can be further understood, and a treatment scheme can be conveniently and accurately formulated.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are given by way of illustration of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (8)

1. Pancreatic cancer detection panel based on a next-generation sequencing technology, characterized in that the detection panel comprises a mutant gene, a copy number variation gene and a gene with rearrangement events for detecting pancreatic cancer prognosis related.
2. The method of detecting panel according to claim 1, wherein said genetic mutations comprise the following genes: ABCB1, ABCC 1, ABL1, ACVR 11, ACVR 21, ADGRA 1, AFF1, AGO 1, AHNAK, AKT1, ALOX12BAMER1, ANKRD1, APC, APCDD1, AR, ARAF, ARFRP1, AREF 12ARID 11, ARID 51, ARMC 1, ARRDC 1, ASH1 BPA 1, ASXL1ASXL 1, ATF1, ATM, ATP1A1, ATP2B 1, ATR 36NNNN 1, AURK 1, AX3672, AX 36AX 1, AX B36L 1, AXB 36X B2, BAC B1, CABCB 1, CABCC 1, CABCKN 1, CALCB 36OCB 1, CALCB 1, CALCB 3636363672, 363636363672, CALCB 1, CALCB 363636363636363636363636363672, 1, CALCB 1, CALCB 1, CALCB 1, CA, CUL, CUL4, CUX, CXCR, CYLD, CYP17A, CYP19A, CYP2C, CYP2D, CYSLTR, DAXX, DCUN1D, DDR, DEPDC, DICER, DIS, DMC, DNAJB, DNMT3, DOT1, DPYD, DROSHA, DUSP, DYNC2H, E2F, EED, EGFL, EGFR, EIF1, EIF4A, EIF4, ELF, ELK, ELOC, EME, EMSY, EP300, EPAS, EPCAM, EPHA, EPHB, ERBB, ERCC, ERF, ERRRFI, ETETV, FASV, EXA, GFZFH, FGFR, GSK3B, GSTM1, GSTP1, GSTT1, H3F3 1, HDAC1, HGF, HIST1H 11, HIST1H 21, HIST1H3 1, MEST 2H3 1, HIST3H 1, HLA-1-1, HNF 11, HOHOL 1, HRAS 1, KM 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 363672, MAPK 36K 1, MAPK 36K 36363672, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MAPK 36K 1, MTAP, MTHFR, MTOR, MUS81, MUTYH, MYB, MYC, MYCL, MYCN, MYD88, MYOD1, NBN, NCOA3, NCOR 3, NEB, NEGR 3, NF 3, NFE2L 3, NFKBIA, NKX3-1, NOTCH3, OTCH3, NPM 3, NQO RADD 3, NR3C 3, NRAS, NRG 3, NSD 3, NT5C 3, NTHL 3, PRORK 3, NTP 3, PTP 3, PRK 3, PRP 3, PSP, RANBP2, RARA, RASA 2, RASSF 2, RB 2, RBBP 2, RBM 2, RECQL 2, REL, RET, RFWD2, RHEB, RHOA, RICTOR, RIT 2, RNF 2, ROBO1ROBO2, ROS 2, RPA 2, RPL2, RPS6KA 2, RPS6KB2, RPTOR, RRAGCRRAS, RRAS2, RRPO 2, RSPO2, RTEL 2, RUNX 2, NX1T 2, RXRAYBP, SCN7 2, SDC 2, SDHA, SDHAF2, SDHB, SDHC, SDHD, SEMA3 2, SESN 36SN 2, SESSN 2, SESTTSTP 2, SETSTC 2, SUTSCP 2, TSTSTSTSTC 2, TSTSTSTSTSTSTSTSTC 2, TSTSTSTSTSTSTSTSTSTSTSS 2, TSTSTSTSTSTSTSTSTSS 2, TSTSTSTSS 2, TSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSS 2, TSTSTSTSTSTSTSTSTSTSTSSSF 2, TSTSTSTSTSTSTSTSTSSSF 2, TSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSSST 2, TSSST 2, TSTSTSTSTSTSTSTSTSTSTSTSSST 2, TSSST 2, TSTSTSSST 2, TSSST 2, TSTSTSTSTSTSTSTSTSSST 2, TSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSSST 2, TSSST 2, TSTSTSTSTSTSTSS 2, TSS 2, TSTSTSTSTSTSTSTSTSTSS 2, TSTSS 2, TSTSTSTSS 2, TSTSTSTSTSTSTSS 2, TSTSTSTSTSS 2, TSTSTSSST 2, TSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSS 2, TSS 2, TSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSTSS 2, TSS 2, TSTSTSTSTSTSTSTSS 2, TSS 2, TSTSTSTSTSSSP 2, TSS 2, TSTSTSS 2, TSS 2, TSTSTSTSTS, U2AF2, UGT1A1, UMPS, UPF1, USP6, USP8, VEGFA, VEGFB, VEGFC, VHL, VTCN1, WHSC1, WHSC1L1, WISP3, WRN, WT1, WWTR1, XIAP, XPC, XPO1, XRCC1, XRCC2, XRCC3XRCC4, XRCC6, YAP1, YES1, YY1, ZBTB2, ZFH 3, ZNF217, ZNF292, ZNF703, ZNRF3, ZRSR 2.
3. The test panel of claim 1, wherein the gene copy number variation comprises the following genes: CDK4, CDK6, CYP2D6, ERBB2, MDM2, MDM4, MET, MYC.
4. The detection panel of claim 1, wherein the genes for which a rearrangement event occurs are: ALK, BCL2, BCR, BRAF, BRCA1, BRCA2, CD274, CD74, CD86, EGFR, ERBB4, ETV4, ETV5, ETV6, EWSR1, EZR, FGFR1, FGFR2, FGFR3, KIT, KMT2A, MET, MSH2, MYB, MYC, NOTCH2, NRG1, NTRK1, NTRK2, NTRK3, NUTM1, PDGFRA, RAF1, RARA, RET, ROS1, RSPO2, SDC4, SLC34A2, TMPRSS 2.
5. A detection kit for detecting pancreatic cancer based on second-generation sequencing, which comprises a detection probe and a detection reagent, wherein the detection probe is directed to any one of the probes for detecting gene mutation or copy number variation of panel according to claims 1-4.
6. The kit of any of claims 1-4 for detecting panel or claim 5, wherein said genetic mutation detection comprises point mutations, deletions, insertions, fusions and the like.
7. Use of a panel according to any one of claims 1 to 4 for the preparation of a pancreatic cancer detection device.
8. A pancreatic cancer detection device, comprising:
the sequencing module is used for extracting DNA of a sample to be tested and carrying out high-throughput sequencing to obtain a sequencing result;
a comparison module for processing the result of the high-throughput sequencing and comparing the data with the panel of any one of claims 1-4 to obtain mutation information;
and the analysis module is used for analyzing the obtained mutation information to obtain a medication scheme.
CN202110307861.4A 2021-03-23 2021-03-23 Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof Pending CN112852966A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110307861.4A CN112852966A (en) 2021-03-23 2021-03-23 Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110307861.4A CN112852966A (en) 2021-03-23 2021-03-23 Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof

Publications (1)

Publication Number Publication Date
CN112852966A true CN112852966A (en) 2021-05-28

Family

ID=75992319

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110307861.4A Pending CN112852966A (en) 2021-03-23 2021-03-23 Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof

Country Status (1)

Country Link
CN (1) CN112852966A (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113388679A (en) * 2021-08-04 2021-09-14 杭州浙大迪迅生物基因工程有限公司 Primer probe set and kit for RT-PCR detection of human leukotriene receptor CysLTR2mRNA
CN113416785A (en) * 2021-07-27 2021-09-21 武汉良培医学检验实验室有限公司 Application of gene marker combination in preparation of probe combination and/or kit and/or system for detecting solid tumors of children
CN113652484A (en) * 2021-08-03 2021-11-16 苏州京脉生物科技有限公司 Application of sequencing panel, kit and preparation method of sequencing library
CN114045286A (en) * 2021-11-22 2022-02-15 上海交通大学医学院附属瑞金医院 Gene set for molecular typing of pancreatic cancer and its application
CN114796524A (en) * 2022-03-31 2022-07-29 武汉大学中南医院 Application of BMS1P4 gene in preparation of medicines for inhibiting bladder cancer cells
CN115449555A (en) * 2022-10-26 2022-12-09 山东大学 Application of ADGRA2 as a biomarker for breast cancer chemotherapy efficacy and prognosis evaluation
CN115927592A (en) * 2022-11-07 2023-04-07 中南大学湘雅医院 Application of Csde1 gene as target in preparation of medicine for treating non-alcoholic fatty liver disease

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5852759B1 (en) * 2015-04-01 2016-02-03 株式会社キュービクス Detection of pancreatic cancer by gene expression analysis
CN107922973A (en) * 2015-07-07 2018-04-17 远见基因组系统公司 Method and system for the modification detection based on sequencing
CN110904235A (en) * 2019-12-20 2020-03-24 深圳市新合生物医疗科技有限公司 Gene panel for detecting tumor targeted drug related gene mutation, method, application and kit
WO2020226333A1 (en) * 2019-05-03 2020-11-12 주식회사 디시젠 Method of predicting cancer prognosis and composition for same
CN111996257A (en) * 2020-09-07 2020-11-27 复旦大学附属肿瘤医院 Gastric cancer detection panel based on next-generation sequencing technology and application thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5852759B1 (en) * 2015-04-01 2016-02-03 株式会社キュービクス Detection of pancreatic cancer by gene expression analysis
CN107922973A (en) * 2015-07-07 2018-04-17 远见基因组系统公司 Method and system for the modification detection based on sequencing
WO2020226333A1 (en) * 2019-05-03 2020-11-12 주식회사 디시젠 Method of predicting cancer prognosis and composition for same
CN110904235A (en) * 2019-12-20 2020-03-24 深圳市新合生物医疗科技有限公司 Gene panel for detecting tumor targeted drug related gene mutation, method, application and kit
CN111996257A (en) * 2020-09-07 2020-11-27 复旦大学附属肿瘤医院 Gastric cancer detection panel based on next-generation sequencing technology and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
杨吉勇等: "胰腺癌生物学诊断治疗进展", 《现代中西医结合杂志》 *
高杰等: "差异表达miRNA在胰腺癌预后判断中的价值", 《癌变？畸变？突变》 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113416785A (en) * 2021-07-27 2021-09-21 武汉良培医学检验实验室有限公司 Application of gene marker combination in preparation of probe combination and/or kit and/or system for detecting solid tumors of children
CN113652484A (en) * 2021-08-03 2021-11-16 苏州京脉生物科技有限公司 Application of sequencing panel, kit and preparation method of sequencing library
CN113388679A (en) * 2021-08-04 2021-09-14 杭州浙大迪迅生物基因工程有限公司 Primer probe set and kit for RT-PCR detection of human leukotriene receptor CysLTR2mRNA
CN114045286A (en) * 2021-11-22 2022-02-15 上海交通大学医学院附属瑞金医院 Gene set for molecular typing of pancreatic cancer and its application
CN114796524A (en) * 2022-03-31 2022-07-29 武汉大学中南医院 Application of BMS1P4 gene in preparation of medicines for inhibiting bladder cancer cells
CN114796524B (en) * 2022-03-31 2023-04-25 武汉大学中南医院 Application of BMS1P4 gene in preparation of medicines for inhibiting bladder cancer cells
CN115449555A (en) * 2022-10-26 2022-12-09 山东大学 Application of ADGRA2 as a biomarker for breast cancer chemotherapy efficacy and prognosis evaluation
CN115449555B (en) * 2022-10-26 2023-10-13 山东大学 Application of ADGRA2 as a biomarker for breast cancer chemotherapy efficacy and prognosis evaluation
CN115927592A (en) * 2022-11-07 2023-04-07 中南大学湘雅医院 Application of Csde1 gene as target in preparation of medicine for treating non-alcoholic fatty liver disease

Similar Documents

Publication Publication Date Title
CN112852966A (en) Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof
CN109427412B (en) Sequence combination for detecting tumor mutation load and design method thereof
US11001837B2 (en) Low-frequency mutations enrichment sequencing method for free target DNA in plasma
Liu et al. The contribution of hereditary cancer-related germline mutations to lung cancer susceptibility
US11211144B2 (en) Methods and systems for refining copy number variation in a liquid biopsy assay
WO2024138956A1 (en) Minimal residual disease detection method and apparatus, device, and storage medium
Kuo et al. Validation and implementation of a modular targeted capture assay for the detection of clinically significant molecular oncology alterations
US20240191282A1 (en) Tissue-specific methylation marker
CN108753967A (en) A kind of gene set and its panel detection design methods for liver cancer detection
CN111996257A (en) Gastric cancer detection panel based on next-generation sequencing technology and application thereof
CN112626213A (en) Liver cancer detection panel based on next-generation sequencing technology, kit and application thereof
US20220392579A1 (en) Classifier models to predict tissue of origin from targeted tumor dna sequencing
CN114250303A (en) A breast cancer multi-gene detection panel, kit and application thereof
CN113832231A (en) Multi-gene panel for gynecological tumor detection, kit and application thereof
CN110806479A (en) Detection panel of breast cancer related kinase mutation and application thereof
US20240279745A1 (en) Systems and methods for multi-analyte detection of cancer
CN111334505A (en) Standard substance for pan-tumor gene detection and preparation method and application thereof
CN111424087A (en) Detection Panel for pan-cancer species detection or targeted drug application based on next-generation sequencing, kit and application
D′ Antonio et al. Identifying DNase I hypersensitive sites as driver distal regulatory elements in breast cancer
CN117043358A (en) Detecting the presence or absence of multiple types of cancer
CN117418003A (en) Marker, probe and application thereof
WO2016049929A1 (en) Method for constructing sequencing library and application thereof
AU2020268861B2 (en) Chromosome conformation markers of prostate cancer and lymphoma
Saldivar et al. Analytic validation of NeXT Dx™, a comprehensive genomic profiling assay
CN111172288A (en) Colorectal cancer polygene screening probe and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination