Congenital agomphosis gene detection panel and application thereof

Abstract

The invention discloses a congenital agomphosis gene detection panel and application thereof, and belongs to the technical field of gene detection. The congenital odontia gene detection panel comprises probes for detecting or hybridizing 104 congenital odontia related genes shown in a capture table 1, a sample DNA and the panel are subjected to solution / chip hybridization, sequencing is carried out after DNA enrichment of a target area is completed, harmful sites or genes related to congenital odontia are screened / predicted according to a determination result, and the congenital odontia gene detection panel is used for detecting or hybridizing the 104 congenital odontia related genes. The expansibility, speed and resolution required for evaluating the target gene can be met, multiple genes in multiple samples can be evaluated at the same time, time is saved, and cost is reduced. Besides, the panel sequencing is high in coverage rate, strong in pertinence, high in detection sensitivity, higher in effectiveness, easier to manage and capable of more accurately, comprehensively and effectively detecting the related genes of the congenital odontoprisis and mutation conditions of the related genes, and accurate diagnosis and risk assessment of the congenital odontoprisis are achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of gene detection, and in particular relates to a congenital tooth agenesis detection panel and an application thereof. Background Art

[0002] Tooth agenesis (TA) is a common developmental disease caused by genetic and environmental factors that result in impaired tooth germ development and a subsequent reduction in the number of teeth. Excluding the third molars, the prevalence of simple tooth agenesis ranges from 2.6% to 11.3%.

[0003] At present, the diagnosis of congenital tooth agenesis is mainly based on clinical diagnosis, that is, diagnosis is made according to the number of missing teeth. The advantage of this method is that it is simple and clear, but the disadvantage is that it is not conducive to the accurate diagnosis and treatment of the disease, and may cause misdiagnosis or delayed diagnosis. The reason is that congenital tooth agenesis is not only manifested as an abnormal number of teeth. Many patients with congenital tooth agenesis are accompanied by structural or functional abnormalities of other teeth or oral cavity, and even many systemic diseases. It is difficult to diagnose these systemic diseases simply by the number of missing teeth, so it is easy to cause missed diagnosis. In this case, it is often necessary to use genetic testing to achieve accurate diagnosis. The disadvantage of lack of molecular diagnosis is that the risk of recurrence of congenital tooth agenesis and congenital tooth agenesis-related systemic diseases cannot be judged, and intervention treatment cannot be taken in the early stage of malformation embryo formation; for individuals who have been born, it is impossible to prevent their diseases and symptoms in the early stage to prevent further aggravation of the disease; in addition, many genetic diseases related to congenital tooth agenesis have complex phenotypes, and it is difficult to diagnose simply through 1 to 2 clinical phenotypes. It is necessary to use molecular diagnosis to form a final diagnosis, so as to carry out appropriate treatment methods.

[0004] At present, the main molecular diagnostic technologies include first-generation sequencing (Sanger sequencing) technology and high-throughput sequencing technology (Next Generation Sequence, NGS, also known as second-generation sequencing). Sanger sequencing technology is based on the principle of dideoxy chain termination method, combining DNA polymerase synthesis fragments, isotope-labeled nucleotides and gel electrophoresis to identify nucleotides, and is the gold standard for sequencing. However, the disadvantages of low throughput and high cost limit its further large-scale application. Commonly used second-generation sequencing technologies mainly include: whole genome sequencing, whole exome sequencing, etc. NGS is the most widely used sequencing technology at present, which can sequence hundreds of thousands to millions of DNA molecules at a time. Its core idea is to sequence while synthesizing, and determine the sequence of DNA by capturing the markers of the newly synthesized ends. Compared with traditional first-generation sequencing technology, it has advantages such as high output and high resolution, but its disadvantage is that the amount of data is large, and the enrichment, screening and interpretation of single disease-causing genes require a certain genetic and clinical background.

[0005] Since congenital tooth agenesis is a disease with obvious genetic heterogeneity and complex phenotype, there are more than 100 pathogenic genes. It is difficult to determine which gene is involved simply by PCR and Sanger sequencing of a gene fragment. At present, second-generation sequencing has been used as a technical means for genetic research on congenital tooth agenesis and related diseases at home and abroad, but it has not yet been widely used for clinical molecular diagnosis. The reason is that for patients with congenital tooth agenesis encountered in clinical practice, if the existing sequencing methods are directly adopted, there are still disadvantages such as high price, high data analysis cost, incomplete gene coverage, and low analysis accuracy. In addition, it is difficult for ordinary clinicians to screen out which genes are related to congenital tooth agenesis from a large amount of second-generation sequencing data in a short period of time, and which genes not only cause congenital tooth agenesis, but are more likely to cause systemic diseases associated with congenital tooth agenesis. Therefore, it is necessary to clarify which genes are directly related to congenital tooth agenesis based on a large amount of clinical and genetic data, and enrich these genes in a detection product so that ordinary clinicians can make clinical judgments based on their genetic test results in a short period of time, but there is no product specifically used for genetic diagnosis of congenital tooth agenesis. Summary of the invention

[0006] In order to overcome the shortcomings of the above-mentioned prior art, the purpose of the present invention is to provide a congenital tooth agenesis gene detection panel and its application, so as to solve the technical problems of the existing detection methods such as high cost, incomplete gene coverage and low analysis accuracy.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention discloses a congenital tooth agenesis detection panel, wherein the panel is composed of hybridization probes for detecting congenital tooth agenesis-related genes, wherein the genes are ADAMTS2, ANOS1, ANTXR1, ATP6V1B2, AXIN2, BCL11B, BCOR, BMP2, BMP4, CACNA1S, CCDC120, CDH1, CEP152, CFAP418, CHD7, CLCN7, CLDN16, COL 17A1, COL1A1, COL1A2, COL3A1, CREB3L1, CREBBP, CTNNB1, CTNND1, DKK1, DSP, EDA, EDAR, EDARADD, EVC, EVC 2. EYA1, FAM20A, FAM83H, FGF3, FGFR1, FGFR2, GJA1, GLI2, GLI3, GNAS, GREM2, GRHL2, GUSB, HNRNPK, HSPA9, IFIH1, IFITM5, IKBKG, IRF6, JAG1, KCNJ2, KDF1, KIF4A, KMT2C, KMT2D, KREMEN1, KRT17, LAMB3, LEMD2, LMNA , LRP5, LRP6, LTBP3, MESD, MMP20, MSX1, NECTIN1, NKX2-1, NSD1, OFD1, OPN3, PAX9, PIGA, PIK3C2A, PITX2, P OC1A, POLR3A, POLR3B, POLR3GL, POLR3K, PORCN, POSTN, PPP1R15B, PRKACA, PRKACB, PTH1R, RUNX2, SATB1, S H3BP2, SLC10A7, SLC13A5, SLC4A4, SMOC2, TBX3, TCOF1, TP63, TRIO, TSPEAR, UBR1, WDR35, WNT10A and WNT10B.

[0009] The second aspect of the present invention discloses a method for preparing a congenital tooth agenesis detection panel, wherein the exon region of a gene related to congenital tooth agenesis is used as a target region to design a panel probe to obtain a congenital tooth agenesis detection panel.

[0010] Preferably, the congenital tooth agenesis gene detection panel is fixed on a silicon wafer or a glass slide of a gene chip, or a liquid phase reaction pool is used.

[0011] The third aspect of the present invention discloses the use of the above-mentioned congenital tooth agenesis detection panel in the detection of congenital tooth agenesis.

[0012] Preferably, the sample DNA is solution / chip hybridized with the congenital tooth agenesis detection panel, and after the target region DNA is enriched, high-throughput sequencing technology is used for sequence determination, and the harmful sites or genes related to congenital tooth agenesis are screened / predicted based on the determination results.

[0013] More preferably, the solution / chip hybridization product is detected using an Agilent liquid phase capture system.

[0014] Preferably, the target region DNA is the DNA of the exon region of the target gene.

[0015] Preferably, DNBSEQ is used TM High-throughput sequencing technology.

[0016] A fourth aspect of the present invention discloses a gene chip or kit for detecting congenital tooth agenesis, comprising the above-mentioned congenital tooth agenesis gene detection panel.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention provides a congenital tooth agene detection panel. Compared with whole genome sequencing and whole exome sequencing, the congenital tooth agene detection panel reduces the number of detected genes from more than 20,000 whole genomes to 104 genes related to congenital tooth agenesis, and the detection area is located from the whole gene to the exon region accounting for about 1.5% of the gene. By capturing the specific regions of 104 specific pathogenic genes, the detection analysis accuracy is high, the sequencing depth is large, and the sequencing depth can ensure more than 1000×, which meets the scalability, speed and resolution required for evaluating target targeted genes, and multiple genes in multiple samples can be evaluated at the same time, thereby saving time and reducing costs. The sequencing cost is greatly reduced: compared with the clinical 3,000-4,000 yuan / case whole exome sequencing, which requires screening in tens of thousands of genes, the congenital tooth agenesis detection panel can directly detect 104 congenital tooth agenesis-related pathogenic genes at one time, and the panel detection cost for each sample does not exceed 1,000 yuan, and the time to determine the known congenital tooth agenesis pathogenic genes is greatly shortened. It can be seen that the panel inspection is low in cost, efficient, and widely applicable, and is more suitable for rapid clinical diagnosis. In addition, panel targeted gene sequencing has higher coverage, stronger specificity, higher detection sensitivity, higher effectiveness, and is easier to manage. Therefore, it can more accurately, comprehensively, and effectively detect and easily analyze congenital tooth agenesis-related genes and their mutations, thereby achieving accurate diagnosis and risk assessment of congenital tooth agenesis, filling the gap in the field of clinical rapid molecular diagnosis of congenital tooth agenesis.

[0019] The application of the above-mentioned congenital agenesis gene detection panel provided by the present invention in the detection of congenital agenesis is to carry out solution / chip hybridization between sample DNA and the congenital agenesis gene detection panel, complete DNA enrichment in the target area and then use high-throughput sequencing technology to perform sequence determination, which can be used to detect various gene structures of congenital agenesis, help to explain the molecular mechanism of congenital agenesis and related diseases, and provide a more comprehensive framework and foundation for conducting research on the functionality, genetic family verification, molecular mechanism of pathogenesis, etc. of congenital agenesis and related diseases; and it can be applied to the concomitant diagnosis of congenital agenesis and related diseases, and the obtained precise targeted sequencing results can accurately evaluate the genetic risk, recurrence risk, disease risk, etc. of congenital agenesis and related diseases. DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used in the text are the common meanings understood by those skilled in the art for the present invention. In the event of a conflict, the definition in this specification shall prevail.

[0021] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0022] Herein, all features such as values, quantities, contents and concentrations defined in the form of numerical ranges or percentage ranges are for simplicity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to have included and specifically disclosed all possible secondary ranges and individual values ​​within the range (including integers and fractions).

[0023] In this document, unless otherwise specified, “includes,” “including,” “contains,” “has,” or similar terms cover the meanings of “consisting of” and “mainly consisting of,” for example, “A includes a” covers the meanings of “A includes a and other” and “A only includes a.”

[0024] In this document, in order to make the description concise, not all possible combinations of various technical features in various embodiments or examples are described. Therefore, as long as there is no contradiction in the combination of these technical features, the various technical features in various embodiments or examples can be combined arbitrarily, and all possible combinations should be considered to be within the scope of this specification.

[0025] In this article, gene detection panel is a term used after the development of high-throughput gene detection and gene sequencing. It refers to designing probes corresponding to several genes on the same capture chip to capture target DNA for subsequent gene sequencing. In the detection, not only one site or one gene is detected, but multiple genes and multiple sites are detected at the same time. These sites and genes need to be selected and combined according to a standard, which requires a detection panel.

[0026] As used herein, the terms "gene detection panel", "gene panel" or "panel" have the same meaning and can be used interchangeably, particularly referring to a collection of DNA probes or a combination of hybridization probes used as a means of detecting multiple genes.

[0027] Herein, the gene panel is composed of hybridization probes for detecting congenital tooth agenesis information and related genes, and its common solid phase carrier can be a silicon wafer or glass slide of a gene chip. Of course, it can also be a liquid phase probe reaction pool, in which various reactants and probes can be reacted, and no specific limitation is made here.

[0028] In this article, the congenital tooth agenesis gene detection panel includes specific hybridization probes for detecting or hybridizing and capturing congenital tooth agenesis-related genes; among them, congenital tooth agenesis-related genes include 104 genes, covering the genes currently reported to be related to non-syndromic congenital tooth agenesis and syndromic congenital tooth agenesis.

[0029] Herein, the selection of capture system includes, but is not limited to, using Agilent liquid phase capture system.

[0030] In this article, the selection of high-throughput sequencing technology includes but is not limited to using DNBSEQ TM Technology is carried out.

[0031] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0032] The following examples use conventional instruments and equipment in the art. The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The various raw materials used in the following examples, unless otherwise specified, are conventional commercial products, and their specifications are conventional specifications in the art.

[0033] It should be noted that the genetic testing using the congenital tooth agenesis detection panel provided by the present invention can be performed using a conventional whole exon capture method. For example, in one embodiment of the present invention, the whole exon capture method of BGI is used.

[0034] The congenital tooth agenesis detection panel provided by the present invention uses the Agilent liquid phase capture system to efficiently capture and enrich the DNA in the exon region of the target gene, and then uses DNBSEQ TM Technology for high-throughput sequencing. The specific implementation steps are as follows:

[0035] 1. Preparation of the congenital tooth agenesis detection panel

[0036] 1. Screening of genes related to congenital tooth agenesis

[0037] The existing congenital tooth agenesis-related genes were collected and sorted out. The collection methods included collecting related genes obtained by sequencing samples of congenital tooth agenesis patients, collecting related genes disclosed in databases, and collecting related genes disclosed in literature. A total of 104 gene detection target genes were screened and obtained (the information is shown in Table 1) for constructing the congenital tooth agenesis gene detection panel in this case.

[0038] Table 1: Information about the 104 target genes

[0039]

[0040]

[0041]

[0042] 2. Detection target area selection

[0043] Based on the 104 genes obtained in step 1, 1858 exons of the 104 genes (including 10 bases extended at the 3' and 5' ends of each exon) were used as target regions, with a total size of 349.906 kbp.

[0044] 3. Detection panel probe design

[0045] Using probe design software, 16,215 probes with a total size of 486.707 kbp were designed to form a genetic testing panel for congenital tooth agenesis, which was then synthesized by BGI.

[0046] Probe evaluation: Align the probe design region with all target regions and calculate the coverage of all target regions by each probe. The calculation formula is: Coverage = Number of reads on the alignment / Number of reads for target sequencing. Probes with a coverage of ≥90% of all target regions are selected for panel synthesis.

[0047] The designed panel probes were evaluated according to the evaluation criteria, and the evaluation results (as shown in Table 2) showed that the coverage of the above 104 genes reached 99.45957%, the coverage uniformity was >99%, and the sequencing depth was >1000×.

[0048] Table 2: Summary of probe coverage of exon regions of 104 target genes

[0049]

[0050]

[0051]

[0052]

[0053] 2. Application of congenital tooth agenesis test panel

[0054] The congenital tooth agenesis gene detection panel prepared by the above method (synthesized by BGI) was used to test the blood samples of congenital tooth agenesis patients, as follows:

[0055] 1. Genomic DNA Extraction

[0056] In this case, QIAamp DNA Blood Mini (Qiagen, Valencia, USA) blood genomic DNA extraction kit was used to extract DNA from blood samples. The specific steps are as follows:

[0057] (1) Sample processing: 200 μL whole blood DNA extraction: Pipette 200 μL blood into the correspondingly labeled 1.5 mL EP tube and proceed directly to the next step.

[0058] (2) Add 20 μL of Proteinase K to the above solution and mix well.

[0059] (3) Add 200 μL of Buffer GL and vortex until thoroughly mixed.

[0060] (4) Incubate at 56°C for 10 min, inverting several times to mix.

[0061] (5) Add 200 μL of anhydrous ethanol, invert several times to mix, and centrifuge briefly.

[0062] (6) Add all the solution obtained in step (5) to the adsorption column (Spin Columns DM) in the collection tube, centrifuge at 12000 rpm for 1 minute, pour out the waste liquid in the collection tube, and put the adsorption column back into the collection tube.

[0063] (7) Add 500 μL of Buffer GW1 to the adsorption column, centrifuge at 12,000 rpm for 1 minute, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube.

[0064] (8) Add 500 μL of Buffer GW2 to the adsorption column, centrifuge at 12,000 rpm for 1 minute, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube. Repeat this step once.

[0065] (9) Centrifuge at 12,000 rpm for 2 minutes, discard the waste liquid in the collection tube, and place the adsorption column at room temperature for several minutes to completely dry.

[0066] (10) Place the adsorption column in a new centrifuge tube, add 50-200 μL Buffer GE or sterile water, leave at room temperature for 2-5 minutes, centrifuge at 12,000 rpm for 1 minute, collect the DNA solution, and store at -20°C for later use.

[0067] 2. DNA Library Preparation

[0068] (1) Qualified DNA samples were randomly sheared by an ultrasonic high-performance sample processing system (Covaris), and fragments were selected using the Agencourt AMPure XP-Medium kit so that the sample bands were concentrated around 150-250 bp. The amount of purified DNA samples was detected using Qubit.

[0069] (2) Repair the double-stranded ends of the DNA and add an "A" base to the 3' end. Prepare an adapter ligation reaction system to connect the adapter index to the two ends of the DNA.

[0070] (3) The linker-ligated library is subjected to linear amplification (LM-PCR) to prepare a hybridization library.

[0071] (4) The library with specific index is hybridized with hundreds of thousands of biotin-labeled probes in liquid phase. The biotin-labeled DNA fragments are adsorbed by magnetic beads, and then the magnetic beads are eluted and the RNA probes are degraded to finally obtain the DNA fragments in the exon region.

[0072] (5) Prepare the PCR reaction system and set the reaction program to perform PCR linear amplification on the products after hybridization elution, and then recover the products. PCR products were detected using Agilent 2100 Bioanalyzer.

[0073] (6) Take an appropriate amount of PCR product and denature it into a single strand, prepare a circularization reaction system, mix it thoroughly, and react it at an appropriate temperature for a certain period of time to obtain a single-stranded circular product. After digesting the linear DNA molecules that have not been circularized, the final library is obtained.

[0074] (7) Use Agilent 2100 Bioanalyzer (Agilent DNA 1000 Reagents) to detect the fragment size and concentration of the library.

[0075] 3. Sequencing

[0076] Using DNBSEQ TM High-throughput sequencing is performed using the technology. The single-stranded circular DNA molecules are replicated by rolling circles and linearly amplified by 2 to 3 orders of magnitude to form a DNA nanoball (DNBs) containing multiple copies. The obtained DNBs are added to the mesh holes on the chip using high-density DNA nanochip technology and sequenced using the double-end sequencing method (MDA-PE) of combined probe anchor polymerization technology (cPAS) multiple displacement amplification.

[0077] 4. Analysis of Sequencing Results

[0078] The sequencing data is aligned to the reference genome, SNP detection and InDel analysis are performed, and then the variant set is filtered out through database annotation. Information analysis starts with the raw data of the sequencing, as follows:

[0079] (1) Filter the raw data containing sequencing adapter sequences, bases with very low sequencing quality, and unmeasured bases (represented by N) to obtain clean data.

[0080] (2) Use the alignment software (Burrows-Wheeler Aligner, BWA) to align the clean data of each sample to the human reference genome (GRCh37 / GRCh38) to obtain the initial alignment result file in BAM format. Then, perform genome-level variation detection according to the optimal variation detection analysis software HaplotypeCaller software recommended by the GATK official website. Based on the alignment results, use the Picard tool to remove duplicate reads, and use GATK to perform local realignment and base quality recalibration.

[0081] (3) Use GATK+SAMtools to perform single sample variation detection and perform statistics and annotation on the results.

[0082] (4) Variant screening and prediction of disease relevance: The results of variant detection are analyzed through three analysis strategies based on variant harmfulness, sample conditions, and gene functional phenotype to screen out harmful sites or genes associated with the disease.

[0083] 5. Principles of variant screening

[0084] (1) Screening according to ACMG genetic variation classification standards and guidelines. According to the American College of Medical Genetics and Genomics (ACMG) genetic variation classification standards and guidelines, the variants are classified into pathogenic, likely pathogenic, uncertain significance, likely benign, and benign according to different conditions. There are 28 evidence categories in the ACMG variation classification system, and the harmfulness of the variant site is classified according to the combination of the 28 evidences.

[0085] (2) Selection based on gene frequency: SNPs and InDels with a frequency ≤ 0.01 or not present in the 1000 Genomes, ESP6500, and gnomAD population databases were retained.

[0086] (3) Filter based on the importance of mutations. The importance of mutations (Putative_impact: a simple estimate of the assumed impact / harmfulness) is divided into LOW, MODIFIER, MODERATE, and HIGH according to the mutation type. Generally, mutations with HIGH or MODERATE importance are focused on.

[0087] (4) Selection based on harmfulness prediction software.

[0088] SIFT4G_Pred prediction value and judgment: The smaller the number, the more significant the impact. D stands for Damaging, and T stands for Tolerated. A standardized probability less than 0.05 is predicted to be harmful ("D"), and greater than or equal to 0.05 is predicted to be tolerable ("T").

[0089] Polyphen2_HDIV_Pred predicted value and judgment: The larger the number, the more significant the impact. D means possibly damaging, P means possibly damaging, and B means benign.

[0090] Polyphen2_HVAR_Pred predicted value and judgment: The larger the number, the more significant the impact. D means possibly damaging, P means possibly damaging, and B means benign.

[0091] Screening was performed based on the condition that the prediction results of the above three harmfulness prediction software were predicted as harmful (or #N / A) by more than two methods.

[0092] (5) The 104 genes in the congenital tooth agenesis panel were selected based on their classification. On the one hand, the syndromic and non-syndromic types were distinguished according to the patient's phenotype to screen for pathogenic genes; on the other hand, patients with non-syndromic congenital tooth agenesis were further screened based on gene priority.

[0093] (6) Focus on known mutations in known genes, gene mutations in signal pathways related to congenital tooth agenesis, etc.

[0094] (7) Query databases such as NCBI, OMIM, and PUBMED and literature reports.

[0095] 6. Sanger sequencing verification

[0096] Primers were synthesized according to the DNA fragments to be sequenced, amplified by polymerase chain reaction (PCR), and sequenced by Sanger sequencing using ABI3730xl sequencer (Applied Biosystems, USA). The sequencing results were compared with the reference sequence using "Sequencher" software.

[0097] The above-mentioned congenital agenesis panel was used for sequencing of 8 congenital agenesis patients, and the above-mentioned variation screening principle was used for variation screening. The results of pathogenic variation analysis of the 8 patients are shown in Table 3. As can be seen from Table 3, all 8 patients were found to carry the pathogenic gene of congenital agenesis, and some of them carried multiple pathogenic genes. These results reflect that this panel can quickly, accurately and comprehensively screen the relevant pathogenic genes in congenital agenesis patients, and is an excellent molecular diagnostic product for identifying congenital agenesis.

[0098] Table 3: Panel sequencing results of 8 patients with congenital tooth agenesis

[0099]

[0100] Note: Type I congenital edentulism refers to the number of congenitally missing teeth ≤ 5; Type II congenital edentulism refers to the number of congenitally missing teeth ≥ 6; Type III congenital edentulism refers to the loss of all deciduous and permanent teeth

[0101] The above examples have verified the gene panel detection scheme of the present invention, and the results show that the congenital tooth agenesis gene detection panel provided by the present invention can directly detect congenital tooth agenesis, and the detection results are consistent with the actual diagnosis.

[0102] It is understandable that, according to different detection or research design requirements, the preparation method of the present application can also be used to obtain more types of gene panels or gene panels for special purposes, and it is not even limited to gene panels for congenital tooth agenesis detection, and no specific limitation is made here.

[0103] The above contents are only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. A congenital tooth agenesis detection panel, characterized in that: The panel is composed of hybridization probes for detecting congenital tooth agenesis-related genes, and the genes are ADAMTS2, ANOS1, ANTXR1, ATP6V1B2, AXIN2, BCL11B, BCOR, BMP2, BMP4, CACNA1S, CCDC120, CDH1, CEP152, CFAP418, CHD7, CLCN7, CLDN16, COL17A1, COL1A1, COL1A2, COL 3A1, CREB3L1, CREBBP, CTNNB1, CTNND1, DKK1, DSP, EDA, EDAR, EDARADD, EVC, EVC2, EYA1, FAM20A, FAM8 3H, FGF3, FGFR1, FGFR2, GJA1, GLI2, GLI3, GNAS, GREM2, GRHL2, GUSB, HNRNPK, HSPA9, IFIH1, IFITM5, IK BKG, IRF6, JAG1, KCNJ2, KDF1, KIF4A, KMT2C, KMT2D, KREMEN1, KRT17, LAMB3, LEMD2, LMNA, LRP5, LRP6, LTBP3, MESD, MMP20, MSX1, NECTIN1, NKX2-1, NSD1, OFD1, OPN3, PAX9, PIGA, PIK3C2A, PITX2, POC1A, PO LR3A, POLR3B, POLR3GL, POLR3K, PORCN, POSTN, PPP1R15B, PRKACA, PRKACB, PTH1R, RUNX2, SATB1, SH3B P2, SLC10A7, SLC13A5, SLC4A4, SMOC2, TBX3, TCOF1, TP63, TRIO, TSPEAR, UBR1, WDR35, WNT10A and WNT10B.

2. The method for preparing a congenital tooth agenesis detection panel according to claim 1, characterized in that: The panel probes were designed with the exon regions of congenital tooth agenesis-related genes as the target regions to obtain a congenital tooth agenesis gene detection panel.

3. The method for preparing a congenital tooth agenesis detection panel according to claim 2, characterized in that: The congenital tooth agenesis gene detection panel is fixed on a silicon wafer or a glass slide of a gene chip, or a liquid phase reaction pool is used.

4. The congenital tooth agenesis detection panel according to claim 1, and the congenital tooth agenesis detection panel prepared by the preparation method according to claim 2 or 3, and their use in detecting congenital tooth agenesis.

5. The use according to claim 4, characterized in that: The sample DNA is hybridized with the congenital tooth agenesis gene detection panel in solution / chip. After the target area DNA is enriched, high-throughput sequencing technology is used for sequence determination. Based on the determination results, harmful sites or genes related to congenital tooth agenesis are screened / predicted.

6. The use according to claim 5, characterized in that: The solution / chip hybridization products were detected using the Agilent liquid capture system.

7. The use according to claim 5, characterized in that: The target region DNA is the DNA in the exon region of the target gene.

8. The use according to claim 5, characterized in that: Using DNBSEQ TM High-throughput sequencing technology.

9. A gene chip or kit for detecting congenital tooth agenesis, characterized in that: Including the congenital tooth agenesis detection panel as described in claim 1.

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