Marker for risk assessment of vitreous retinal lymphoma and application thereof

Abstract

The invention relates to the technical field of biomedicine, in particular to a marker for risk assessment of vitreous retinal lymphoma and application of the marker. The marker comprises mutation sites of vitreoretinal lymphoma related genes, and the vitreoretinal lymphoma related genes comprise BTG1, BTG2, DMD, HLA-A, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, VHL, ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, according to the application, the markers are screened out, a corresponding detection object is designed based on the screened markers, and a vitreous retinal lymphoma risk assessment method with high sensitivity and reliability and a corresponding risk assessment product are constructed. According to the method, an accurate risk judgment result can be obtained based on a comprehensive score of VRL related somatic cell gene mutation in a to-be-detected sample.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and in particular to a biomarker for risk assessment of vitreoretinal lymphoma and its application. Background Technology

[0002] Vitreoretinal lymphoma (VRL) is a rare ocular tumor, pathologically classified as large B-cell lymphoma. The clinical phenotype of VRL is extremely similar to uveitis, with both presenting with nonspecific signs such as vitreous cell infiltration, vitreous opacity, retinal edema or infiltration, and retinal pigment epithelial lesions, making differential diagnosis quite challenging.

[0003] In recent years, numerous studies have been conducted to differentiate VRL from uveitis. For example, intraocular fluid interleukin testing combined with the intraocular lymphoma diagnostic interleukin score (ISOLD) can aid in the differential diagnosis of VRL to some extent. However, interleukin testing alone cannot be used for definitive diagnosis, nor can it serve as a basis for initiating chemotherapy.

[0004] Currently, the gold standard for diagnosing malignant tumors is still cytopathological examination. However, the detection rate of cytopathology in VRL is relatively low, reported in different literature as only about 40-80%. The main reasons for its limited detection rate include: (1) the small volume of the eyeball, which limits the amount of intraocular fluid sample that can be collected, and the total number of tumor cells contained therein is small; (2) the mechanical force during vitrectomy to obtain samples can easily damage tumor cells; (3) large B cells are inherently fragile and spontaneously die shortly after being removed from the body; (4) in cases of misdiagnosed uveitis, the use of glucocorticoids can cause tumor cell necrosis, further reducing the number of cells that can be used for pathological diagnosis.

[0005] In summary, there is currently a lack of highly sensitive and reliable risk assessment methods for vitreoretinal lymphoma. Summary of the Invention

[0006] In view of the shortcomings of the prior art described above, and in order to solve the problem that there is still a lack of highly sensitive and reliable risk assessment methods for vitreoretinal lymphoma, the purpose of this application is to provide a novel biomarker for risk assessment of vitreoretinal lymphoma and its application, in order to solve the problems in the prior art.

[0007] To achieve the above and other related objectives, this application first provides a biomarker for risk assessment of vitreoretinal lymphoma, the biomarker including mutation sites of vitreoretinal lymphoma-related genes, wherein the vitreoretinal lymphoma-related genes are selected from one or more of BTG1, BTG2, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, and VHL.

[0008] Preferably, the vitreoretinal lymphoma-related genes include BTG1, BTG2, DMD, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, VHL, ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, and TP53.

[0009] This application further provides the use of the above-mentioned biomarkers and / or their detection substances in the preparation of vitreoretinal lymphoma detection products.

[0010] In addition, this application provides a vitreoretinal lymphoma detection product, the detection product comprising a substance for detecting the above-mentioned markers.

[0011] This application also provides a vitreoretinal lymphoma risk assessment product, selected from any of the following:

[0012] 1) A vitreoretinal lymphoma risk assessment device, comprising the following modules: The data acquisition module is used to provide the detection results of the target biomarker of the sample to be tested, and to confirm the mutated gene of the sample to be tested based on the detection results, wherein the target biomarker is the aforementioned biomarker; The risk assessment module is used to assess the risk of vitreoretinal lymphoma in the individual corresponding to the test sample based on the VRL somatic gene mutation comprehensive score.

[0013] 2) A computer-readable storage medium storing computer instructions that, when executed by a processor, implement a method for assessing the risk of vitreoretinal lymphoma, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results, wherein the target biomarker is the biomarker mentioned above; Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0014] 3) An electronic terminal, comprising: a processor, a memory, a network interface, and a user interface; the memory for storing a computer program, and the processor for executing the computer program stored in the memory, to cause the terminal to perform a vitreoretinal lymphoma risk assessment method, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results. The target biomarker is the biomarker mentioned above. Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0015] 4) A computer program product, comprising a computer program that, when executed by a processor, implements a vitreoretinal lymphoma risk assessment method, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results, wherein the target biomarker is the biomarker mentioned above; Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0016] Compared with the prior art, the beneficial effects of this application are as follows:

[0017] 1. This invention screens biomarkers for risk assessment of vitreoretinal lymphoma (VRL), designs corresponding detection methods based on the screened biomarkers, and constructs a highly sensitive and reliable VRL risk assessment method and corresponding risk assessment products. This method, based on a comprehensive score of VRL-related somatic gene mutations in the test sample, can obtain accurate risk assessment results.

[0018] 2. This invention also screens out viruses associated with uveitis and tumorigenesis, and simultaneously develops detection agents for these viruses, enabling "one-stop" virus detection while detecting gene markers, providing important auxiliary evidence for the differential diagnosis of VRL and infectious uveitis.

[0019] 3. The detection sample of the present invention is cell-free DNA (cfDNA), which avoids consuming the limited cell entities used for cytopathological examination. Attached Figure Description

[0020] Figure 1 This is the technology roadmap for this application.

[0021] Figure 2 This is a flowchart illustrating the interpretation of the results of the vitreoretinal lymphoma risk assessment based on the testing products in this application.

[0022] Figure 3 This is the ROC curve diagram in Embodiment 4 of this application. Detailed Implementation

[0023] To make the inventive objectives, technical solutions, and beneficial effects of this application clearer, the following description, in conjunction with embodiments, further illustrates this application. It should be understood that the embodiments described are for illustrative purposes only and are not intended to limit the scope of the application. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, and those skilled in the art can easily understand other advantages and effects of this application from the content disclosed in this description.

[0024] The first aspect of this application provides a biomarker for risk assessment of vitreoretinal lymphoma, the biomarker including mutation sites of vitreoretinal lymphoma-related genes, the vitreoretinal lymphoma-related genes being selected from one or more of BTG1, BTG2, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, and VHL.

[0025] In some specific embodiments, the vitreoretinal lymphoma-related genes also include one or more of the following: ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, TP53, and DMD.

[0026] In some specific embodiments, the vitreoretinal lymphoma-related genes include BTG1, BTG2, DMD, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, VHL, ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, and TP53.

[0027] In some specific embodiments, the mutation site is a site where a nonsynonymous mutation occurs relative to the reference transcript of the vitreoretinal lymphoma-related gene.

[0028] In this application, the term "reference transcript" refers to a gene coding sequence from an authoritative genome database (such as RefSeq) used as a standard for alignment.

[0029] In this application, the term "non-synonymous mutation" refers to a nucleotide variation that can directly cause changes in the amino acid sequence of a protein.

[0030] In some specific embodiments, the accession number of the reference transcript for BTG1 is NM_001731; the accession number of the reference transcript for BTG2 is NM_006763; the accession number of the reference transcript for DMD is NM_004023; the accession number of the reference transcript for HLA-A is NM_002116; the accession number of the reference transcript for LATS1 is NM_001350339; the accession number of the reference transcript for LRP1B is NM_018557; the accession number of the reference transcript for PTPN11 is NM_080601; the accession number of the reference transcript for PTPRD is NM_001171025; and the accession number of the reference transcript for STAT is NM_001731. The accession number for the reference transcript of 5A is NM_003152; the accession number for the reference transcript of TGFBR2 is NM_001024847; the accession number for the reference transcript of VHL is NM_001354723; the accession number for the reference transcript of ARID1A is NM_139135; the accession number for the reference transcript of ARID1B is NM_001371656; the accession number for the reference transcript of ATM is NM_00005; the accession number for the reference transcript of BCL2 is NM_000633; the accession number for the reference transcript of BCL6 is NM_001706; and the accession number for the reference transcript of BCOR is NM_0 01123383; the accession number of the reference transcript for BRAF is NM_001378474; the accession number of the reference transcript for CARD11 is NM_032415; the accession number of the reference transcript for CD79A is NM_001783; the accession number of the reference transcript for CD79B is NM_001039933; the accession number of the reference transcript for CDKN2A is NM_000077; the accession number of the reference transcript for CDKN2B is NM_078487; the accession number of the reference transcript for EP300 is NM_001429; the accession number of the reference transcript for ETV6 is NM_001987; the accession number of EZH The accession number for the reference transcript of JAK1 is NM_001203247; the accession number for the reference transcript of GNA13 is NM_006572; the accession number for the reference transcript of IRF4 is NM_002460; the accession number for the reference transcript of JAK1 is NM_002227; the accession number for the reference transcript of JAK2 is NR_169763; the accession number for the reference transcript of JAK3 is NM_000215; the accession number for the reference transcript of KMT2D is NM_003482; the accession number for the reference transcript of MYC is NM_001354870; and the accession number for the reference transcript of MYD88 is NM_00246.The accession number for the reference transcript of NOTCH1 is NM_017617; the accession number for the reference transcript of NOTCH2 is NM_024408; the accession number for the reference transcript of PIK3CA is NM_006218; the accession number for the reference transcript of PIM1 is NM_002648; the accession number for the reference transcript of PRDM1 is NM_001198; and the accession number for the reference transcript of RHOA is... The accession number for the reference transcript of SOCS1 is NM_001313943; the accession number for the reference transcript of STAT3 is NM_003745; the accession number for the reference transcript of STAT5B is NM_139276; the accession number for the reference transcript of STAT5B is NM_012448; the accession number for the reference transcript of TET2 is NM_001127208; and the accession number for the reference transcript of TP53 is NM_000546.

[0031] In this application, the term "accession number" refers to a unique identifier assigned to a specific biological sequence by an authoritative bioinformatics database (such as RefSeq), through which the corresponding sequence record can be explicitly queried and referenced.

[0032] The second aspect of this application provides the use of the markers and / or their detectors described in the first aspect in the preparation of vitreoretinal lymphoma detection products.

[0033] In some specific embodiments, the detectable substance includes any one or more of the following: 1) A probe for detecting BTG1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 31; 2) A probe for detecting BTG2, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 32; 3) A probe for detecting HLA-A, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 82; 4) A probe for detecting LATS1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 113; 5) A probe for detecting LRP1B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 114~130; 6) A probe for detecting PTPN11, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 157~161; 7) A probe for detecting PTPRD, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 162-167; 8) A probe for detecting STAT5A, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 183-187; 9) A probe for detecting TGFBR2, the nucleotide sequence of which contains the sequence shown in any or more of SEQ ID No. 196; 10) A probe for detecting VHL, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 202-204.

[0034] Preferably, the detectable substance further includes any one or more of the following: 11) A probe for detecting DMD, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 44-59; 12) A probe for detecting ARID1A, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 1 to 5; 13) A probe for detecting ARID1B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 6-8; 14) A probe for detecting ATM, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 9-21; 15) A probe for detecting BCL2, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 22; 16) A probe for detecting BCL6, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 23; 17) A probe for detecting BCOR, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 24; 18) A probe for detecting BRAF, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 25-30; 19) A probe for detecting CARD11, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 33-37; 20) A probe for detecting CD79A, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 38; 21) A probe for detecting CD79B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 39-40; 22) A probe for detecting CDKN2A, the nucleotide sequence of which includes the sequence shown in SEQ ID No. 41; 23) A probe for detecting CDKN2B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 42-43; 24) A probe for detecting EP300, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 60-68; 25) A probe for detecting ETV6, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 69-70; 26) A probe for detecting EZH2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 71-78; 27) A probe for detecting GNA13, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 79-81; 28) A probe for detecting IRF4, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 83-85; 29) A probe for detecting JAK1, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 86-90; 30) A probe for detecting JAK2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 91-96; 31) A probe for detecting JAK3, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 97~100; 32) A probe for detecting KMT2D, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 101-112; 33) A probe for detecting MYC, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 131-132; 34) A probe for detecting MYD88, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 133; 35) A probe for detecting NOTCH1, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 134-140; 36) A probe for detecting NOTCH2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 141-148; 37) A probe for detecting PIK3CA, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 149-154; 38) A probe for detecting PIM1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 155; 39) A probe for detecting PRDM1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 156; 40) A probe for detecting RHOA, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 168-171; 41) A probe for detecting SOCS1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 172; 42) A probe for detecting STAT3, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 173-182; 43) A probe for detecting STAT5B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 188~192; 44) A probe for detecting TET2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 193-195; 45) A probe for detecting TP53, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 197~201.

[0035] In some specific embodiments, the detection substance includes: 1) A probe for detecting BTG1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 31; 2) A probe for detecting BTG2, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 32; 3) A probe for detecting HLA-A, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 82; 4) A probe for detecting LATS1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 113; 5) A probe for detecting LRP1B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 114~130; 6) A probe for detecting PTPN11, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 157~161; 7) A probe for detecting PTPRD, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 162-167; 8) A probe for detecting STAT5A, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 183-187; 9) A probe for detecting TGFBR2, the nucleotide sequence of which contains the sequence shown in any or more of SEQ ID No. 196; 10) A probe for detecting VHL, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 202-204; 11) A probe for detecting DMD, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 44-59; 12) A probe for detecting ARID1A, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 1 to 5; 13) A probe for detecting ARID1B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 6-8; 14) A probe for detecting ATM, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 9-21; 15) A probe for detecting BCL2, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 22; 16) A probe for detecting BCL6, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 23; 17) A probe for detecting BCOR, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 24; 18) A probe for detecting BRAF, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 25-30; 19) A probe for detecting CARD11, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 33-37; 20) A probe for detecting CD79A, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 38; 21) A probe for detecting CD79B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 39-40; 22) A probe for detecting CDKN2A, the nucleotide sequence of which includes the sequence shown in SEQ ID No. 41; 23) A probe for detecting CDKN2B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 42-43; 24) A probe for detecting EP300, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 60-68; 25) A probe for detecting ETV6, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 69-70; 26) A probe for detecting EZH2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 71-78; 27) A probe for detecting GNA13, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 79-81; 28) A probe for detecting IRF4, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 83-85; 29) A probe for detecting JAK1, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 86-90; 30) A probe for detecting JAK2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 91-96; 31) A probe for detecting JAK3, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 97~100; 32) A probe for detecting KMT2D, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 101-112; 33) A probe for detecting MYC, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 131-132; 34) A probe for detecting MYD88, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 133; 35) A probe for detecting NOTCH1, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 134-140; 36) A probe for detecting NOTCH2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 141-148; 37) A probe for detecting PIK3CA, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 149-154; 38) A probe for detecting PIM1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 155; 39) A probe for detecting PRDM1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 156; 40) A probe for detecting RHOA, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 168-171; 41) A probe for detecting SOCS1, the nucleotide sequence of which contains the sequence shown in SEQ ID No. 172; 42) A probe for detecting STAT3, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 173-182; 43) A probe for detecting STAT5B, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 188~192; 44) A probe for detecting TET2, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 193-195; 45) A probe for detecting TP53, the nucleotide sequence of which contains the sequence shown in any one or more of SEQ ID No. 197~201.

[0036] A third aspect of this application provides a vitreoretinal lymphoma detection product, the detection product comprising a substance for detecting the markers described in the first aspect.

[0037] In some specific embodiments, the substance used to detect the marker is the same as the detector used in the second aspect.

[0038] In some specific embodiments, the detection product also includes a substance for detecting viruses associated with the etiology of vitreoretinal lymphoma.

[0039] In this application, the detection of viruses associated with the etiology of vitreoretinal lymphoma can further aid in etiological identification.

[0040] Preferably, the vitreoretinal lymphoma etiology-associated virus includes one or more of the following: herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, human herpesvirus type 4, human cytomegalovirus, human herpesvirus type 6, human herpesvirus type 8, and circovirus.

[0041] More preferably, the viruses associated with vitreoretinal lymphoma include herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, human herpesvirus type 4, human cytomegalovirus, human herpesvirus type 6, human herpesvirus type 8, and circovirus.

[0042] In some specific embodiments, the testing product further includes any one or more of the following: 1) A probe for detecting herpes simplex virus type 1, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 205-206; 2) A probe for detecting herpes simplex virus type 2, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 207~208; 3) A probe for detecting varicella-zoster virus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 209~210; 4) A probe for detecting human herpesvirus type 4, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 211-212; 5) A probe for detecting human cytomegalovirus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 213-214; 6) A probe for detecting human herpesvirus type 6, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 215-216; 7) A probe for detecting human herpesvirus type 8, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 217-218; 8) A probe for detecting circovirus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 219-220.

[0043] More preferably, the testing product includes: 1) A probe for detecting herpes simplex virus type 1, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 205-206; 2) A probe for detecting herpes simplex virus type 2, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 207~208; 3) A probe for detecting varicella-zoster virus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 209~210; 4) A probe for detecting human herpesvirus type 4, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 211-212; 5) A probe for detecting human cytomegalovirus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 213-214; 6) A probe for detecting human herpesvirus type 6, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 215-216; 7) A probe for detecting human herpesvirus type 8, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 217-218; 8) A probe for detecting circovirus, the nucleotide sequence of which contains the sequence shown in any one or both of SEQ ID No. 219-220.

[0044] The fourth aspect of this application provides a vitreoretinal lymphoma risk assessment product, selected from any of the following:

[0045] 1) A vitreoretinal lymphoma risk assessment device, comprising the following modules: The data acquisition module is used to provide the detection results of the target biomarker of the sample to be tested, and to confirm the mutated gene of the sample to be tested based on the detection results, wherein the target biomarker is the biomarker described in the first aspect; The risk assessment module is used to assess the risk of vitreoretinal lymphoma in the individual corresponding to the test sample based on the VRL somatic gene mutation comprehensive score of the test sample.

[0046] 2) A computer-readable storage medium storing computer instructions that, when executed by a processor, implement a method for assessing the risk of vitreoretinal lymphoma, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results, wherein the target biomarker is the biomarker described in the first aspect; Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0047] 3) An electronic terminal, comprising: a processor, a memory, a network interface, and a user interface; the memory for storing a computer program, and the processor for executing the computer program stored in the memory to cause the terminal to perform a vitreoretinal lymphoma risk assessment method, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results, wherein the target biomarker is the biomarker described in the first aspect; Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0048] 4) A computer program product, characterized in that it includes a computer program, which, when executed by a processor, implements a vitreoretinal lymphoma risk assessment method, the method comprising: The detection results of the target biomarker data of the sample to be tested are obtained, and the mutated gene of the sample to be tested is confirmed based on the detection results, wherein the target biomarker is the biomarker described in the first aspect; Based on the VRL somatic gene mutation comprehensive score of the test sample, the risk of vitreoretinal lymphoma is assessed for the individual corresponding to the test sample.

[0049] In some specific embodiments, the VRL somatic gene mutation comprehensive score is the sum of the comprehensive importance scores (Stotal) corresponding to each mutated gene.

[0050] Preferably, the total value of the BTG1 mutation is 0.224, the total value of the BTG2 mutation is 0.3495, the total value of the DMD mutation is 0, the total value of the HLA-A mutation is 0.225, the total value of the LATS1 mutation is 0.0375, the total value of the LRP1B mutation is 0.0375, the total value of the PTPN11 mutation is 0.15, the total value of the PTPRD mutation is 0.075, the total value of the STAT5A mutation is 0.3495, the total value of the TGFBR2 mutation is 0.313, and the total value of the VHL mutation is 0. 1875, the total for the ARID1A mutation is 0.0375, the total for the ARID1B mutation is 0.0375, the total for the ATM mutation is 0.225, the total for the BCL2 mutation is 0.15, the total for the BCL6 mutation is 0.075, the total for the BCOR mutation is 0.0375, the total for the BRAF mutation is 0.1875, the total for the CARD11 mutation is 0.1255, the total for the CD79A mutation is 0.0375, the total for the CD79B mutation is 0.313, and the total for the CDKN2A mutation is... The total is 0.4555. The total for the CDKN2B mutation is 0.1875, the total for the EP300 mutation is 0.269, the total for the ETV6 mutation is 0.1255, the total for the EZH2 mutation is 0.0375, the total for the GNA13 mutation is 0.0375, the total for the IRF4 mutation is 0.2135, the total for the JAK1 mutation is 0.2625, the total for the JAK2 mutation is 0.4125, the total for the JAK3 mutation is 0.1125, and the total for the KMT2D mutation is 0.1695. The total for the MYC mutation was 0.15, the total for the MYD88 mutation was 0.7, the total for the NOTCH1 mutation was 0.1125, the total for the NOTCH2 mutation was 0.075, the total for the PIK3CA mutation was 0.0375, the total for the PIM1 mutation was 0.0375, the total for the PRDM1 mutation was 0.225, the total for the RHOA mutation was 0.0375, the total for the SOCS1 mutation was 0.1875, the total for the STAT3 mutation was 0.224, and the total for the STAT5B mutation was 0.The total value for the TET2 mutation was 0.0375, and the total value for the TP53 mutation was 0.4555.

[0051] In some specific embodiments, the mutated gene is determined by detection using the detection product described in the third aspect.

[0052] In some specific implementations, the higher the VRL somatic gene mutation comprehensive score, the higher the risk of the individual corresponding to the test sample having vitreoretinal lymphoma.

[0053] Preferably, the VRL somatic gene mutation comprehensive score of the test sample is compared with the cutoff value. When the VRL somatic gene mutation comprehensive score of the test sample is greater than or equal to the cutoff value, it is determined to be positive for vitreoretinal lymphoma; when the VRL somatic gene mutation comprehensive score of the test sample is less than the cutoff value, it is determined to be negative for vitreoretinal lymphoma.

[0054] More preferably, the cutoff value is 0.69025.

[0055] In some specific embodiments, the sample to be tested is cfDNA in intraocular fluid.

[0056] Preferably, the intraocular fluid is aqueous humor and / or vitreous humor.

[0057] The present application will be further illustrated by the following examples, but these examples do not limit the scope of the present application.

[0058] Example 1: Screening of genes and viruses related to vitreoretinal lymphoma and probe design

[0059] During the gene screening phase, based on the sequencing data of 69 lymphoma cases that had been obtained at that time, and compared with the database of normal individuals, a total of 526 mutated genes specific to lymphoma patients were selected. Subsequently, the genes were sorted according to the frequency of mutation occurrence, and at the same time, the presence of evidence of lymphocytic diseases in existing databases (TCGA, COSMIC, OncoKB), literature, and guidelines for these genes was checked. Finally, based on a comprehensive evaluation of the frequency of occurrence and the number of records in literature / databases, the top 45 genes with the highest matching degree were selected.

[0060] They are BTG1, BTG2, DMD, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, VHL, ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, and TP53.

[0061] This application further screens for viruses associated with the occurrence of vitreoretinal lymphoma based on databases, literature guidelines, and local databases.

[0062] These are herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, human herpesvirus type 4, human cytomegalovirus, human herpesvirus type 6, human herpesvirus type 8, and circovirus.

[0063] Subsequently, this application designed probes targeting specific regions of 45 genes associated with vitreoretinal lymphoma (see Table 1) and 8 viruses associated with tumorigenesis (see Table 2).

[0064] Example 2: Establishment of a risk assessment method for vitreoretinal lymphoma

[0065] To address the challenge that existing technologies often fail to utilize single-gene somatic mutations as tumor markers and assessment indicators, this application first extracts relevant signaling pathways from 45 selected genes. The pathway information is sourced from publicly available databases such as KEGG, Reactome, and Pathway Commons, or obtained manually through literature review. These pathways encompass core signaling pathways closely related to tumors, such as PI3K-AKT, MAPK, WNT, TP53, and Notch.

[0066] Subsequently, the importance of 45 genes was scored based on the following factors: 1) relevance to tumor characteristics such as immune infiltration, drug resistance, and metabolism (Stumor); 2) node degree of the signaling pathway (Spath); and 3) clinical relevance (such as survival rate, recurrence rate, or detection rate) (Sclin). Due to the inconsistent scoring units for each dimension, this invention employs min-max normalization, placing all score values ​​between 0 and 1. Each scoring factor for all candidate genes is independently normalized, as shown in the following formula:

[0067] Si′=

Si-min(Si)

max(Si)-min(Si)

[0068] The final total importance score (Stotal) is calculated using a weighted summation method:

[0069] Stotal=Wtumor×Stumor′+Wpath×Spath′+Wclin×Sclin′

[0070] Where Wi represents the weight of each scoring dimension. The weights are set manually based on the general consensus on the importance of each factor in the literature. In this invention, Wclin is preset to 0.4, Wtumor to 0.3, and Wpath to 0.3. Optionally, the optimal weights of each factor can also be determined based on unsupervised learning methods (such as principal component analysis) and unsupervised learning methods (random forest or support vector machine).

[0071] The importance scores of the 45 genes selected in this embodiment are shown in Table 3.

[0072] After extracting cell-free extracellular DNA (cfDNA) from intraocular fluid (aqueous humor or vitreous humor) samples, probes designed in Example 1 were used to detect nonsynonymous mutations relative to their respective reference transcripts in 45 genes associated with vitreoretinal lymphoma. A coefficient k was set to characterize the mutation status: if a gene has a nonsynonymous mutation, k is set to 1; if not, k is set to 0.

[0073] In this embodiment, the accession number of the reference transcript for BTG1 is NM_001731; the accession number of the reference transcript for BTG2 is NM_006763; the accession number of the reference transcript for DMD is NM_004023; the accession number of the reference transcript for HLA-A is NM_002116; the accession number of the reference transcript for LATS1 is NM_001350339; the accession number of the reference transcript for LRP1B is NM_018557; the accession number of the reference transcript for PTPN11 is NM_080601; the accession number of the reference transcript for PTPRD is NM_001171025; the accession number of the reference transcript for STAT5A is NM_003152; TG The accession number for the reference transcript of FBR2 is NM_001024847; the accession number for the reference transcript of VHL is NM_001354723; the accession number for the reference transcript of ARID1A is NM_139135; the accession number for the reference transcript of ARID1B is NM_001371656; the accession number for the reference transcript of ATM is NM_00005; the accession number for the reference transcript of BCL2 is NM_000633; the accession number for the reference transcript of BCL6 is NM_001706; the accession number for the reference transcript of BCOR is NM_001123383; the accession number for the reference transcript of BRAF is NM_001378474; CARD11 The accession number for the reference transcript of CD79A is NM_032415; the accession number for the reference transcript of CD79B is NM_001783; the accession number for the reference transcript of CD79B is NM_001039933; the accession number for the reference transcript of CDKN2A is NM_000077; the accession number for the reference transcript of CDKN2B is NM_078487; the accession number for the reference transcript of EP300 is NM_001429; the accession number for the reference transcript of ETV6 is NM_001987; the accession number for the reference transcript of EZH2 is NM_001203247; the accession number for the reference transcript of GNA13 is NM_006572; and the accession number for the reference transcript of IRF4 is NM_001039933; the accession number for the reference transcript of IRF4 is NM_001039933; the accession number for the reference transcript of ETV6 is NM_001987; the accession number for the reference transcript of EZH2 is NM_001203247; the accession number for the reference transcript of GNA13 is NM_006572; and the accession number for the reference transcript of IRF4 is NM_001039933; the accession number for the reference transcript of E ... The accession number for the reference transcript of JAK1 is NM_002460; the accession number for the reference transcript of JAK2 is NM_002227; the accession number for the reference transcript of JAK3 is NR_169763; the accession number for the reference transcript of JAK3 is NM_000215; the accession number for the reference transcript of KMT2D is NM_003482; the accession number for the reference transcript of MYC is NM_001354870; the accession number for the reference transcript of MYD88 is NM_00246; the accession number for the reference transcript of NOTCH1 is NM_017617; the accession number for the reference transcript of NOTCH2 is NM_024408; and the accession number for the reference transcript of PIK3CA is NM_006218.The accession number for the reference transcript of PIM1 is NM_002648; the accession number for the reference transcript of PRDM1 is NM_001198; the accession number for the reference transcript of RHOA is NM_001313943; the accession number for the reference transcript of SOCS1 is NM_003745; the accession number for the reference transcript of STAT3 is NM_139276; the accession number for the reference transcript of STAT5B is NM_012448; the accession number for the reference transcript of TET2 is NM_001127208; and the accession number for the reference transcript of TP53 is NM_000546.

[0074] Subsequently, the VRL somatic gene mutation comprehensive score (S) was calculated, which is the sum of the comprehensive importance scores (Stotal) corresponding to all mutated genes among the 45 genes.

[0075] S=k(ARID1A)×Stotal(ARID1A)+ k(ARID1B)×Stotal(ARID1B)+ k(ATM)×Stotal(ATM)+ k(BCL2)×Stotal(BCL2)+k(BCOR)×Stotal (BCOR)+k(BRAF)×Stotal(BRAF)+ k(BTG1)×Stotal(BTG1)+k(ARID1A)×Stotal(ARID1A)+k(BTG2)×Stotal(BTG2)+k(CARD11)×Stota l(CARD11)+k(CD79A)×Stotal(CD79A)+k(CD78B)×Stotal(CD79B)+k(CDKN2A)×Stotal(CDKN2A)+…+ k(VHL)×Stotal(VHL).

[0076] Example 3: Performance Verification of VRL Testing Products

[0077] cfDNA was extracted from intraocular fluid (aqueous humor and vitreous humor) samples using the QIAamp Circulating Nucleic Acid Kit (QIAGEN), and then a library was constructed. Subsequently, liquid-phase hybridization capture was performed using the probes designed in Example 1 to enrich the target gene regions. The enriched library was quantified using the Illumina Novaseq sequencing platform, and the data were further analyzed for bioinformatics. Raw sequencing data were quality assessed using FastQC, and adapters and low-quality fragments were filtered using Trimmomatic / fastp.

[0078] The virus identification process is as follows: the human genome (hg19) and the viral reference genome are merged into a single index library, the alignment tool Kraken / Braken is used to retain the reads that can be mapped, the virus-specific reads are extracted, and the virus species are identified.

[0079] The tumor-related gene mutation detection workflow is as follows: Clean reads are aligned to the human reference genome (hg19) using BWA-MEM (v0.7.17) to generate a SAM file. The results are sorted, PCR duplicates are removed (MarkDuplicates), and format conversion (SAM to BAM) is performed using Samtools and Picard. Somatic SNVs (single nucleotide variants) and InDels (insertion / deletion variants) are detected using GATK Mutect2 (v4.2 and above). A combined tumor-normal sample analysis (matched tumor-normal mode) improves mutation detection accuracy. Contamination estimation, panel of normals (PoN) filtering, and Germline Resource filtering are performed during the detection process. Mutation results are annotated using ANNOVAR or Ensembl VEP to obtain the functional site, gene information, and pathogenicity prediction of the mutation. Further databases such as OncoKB, ClinVar, and COSMIC are used to identify clinically relevant mutations, known driver mutations, or drug target variants.

[0080] Statistical analysis of the frequency of different types of mutations (C>T, T>A, etc.).

[0081] To investigate the performance of the detection product of this invention (containing probes targeting 45 related genes and 8 tumor-associated viruses) in detecting mutation sites, frozen samples from 20 retrospective clinical cases were selected and tested using the detection product and protocol of this invention. The results were compared with previous genomic sequencing results for these 20 cases to evaluate the accuracy of the detection product of this invention. The results showed that the detection product of this invention had a sensitivity of 100% (16 / 16) and a specificity of 100% (4 / 4) for detecting tumor-associated mutation sites, consistent with previous tests.

[0082]

[0083] Example 4: Performance evaluation of VRL somatic gene mutation comprehensive score (S)

[0084] This embodiment provides a method for assessing the risk of tumors using the VRL somatic gene mutation scoring system using a receiver operating characteristic curve (ROC curve), which is used to characterize the performance of the scoring system in assisting clinical identification of potential risks.

[0085] A total of 121 retrospective samples were used for the evaluation, including 35 normal controls and 86 clinically diagnosed VRL samples. Each sample underwent hybridization capture sequencing using the detection product of this invention (containing probes targeting 45 related genes and 8 tumor-associated viruses). Sequencing data were filtered using the standard GATK workflow, followed by somatic SNV and indel annotation information. Site annotation was performed using ANNOVAR or Ensembl VEP to obtain mutation information such as pathogenicity prediction.

[0086] Furthermore, the obtained variant sites are used to calculate the somatic variant comprehensive score of the sample based on the VRL gene mutation comprehensive scoring system. Specifically, taking sample 1 as an example, the calculation process of the VRL gene mutation comprehensive scoring system is demonstrated. Sample 1, detected by the detection product of this invention, yielded the following gene variant sites:

[0087]

[0088] The combined importance scores of the genes detected at the above loci are summed to obtain the VRL somatic mutation comprehensive score S for this sample.

[0089] S=S total（LRP1B） + S total（IRF4） + S total（HLA-A） + S total（PRDM1） + S total（MYD88） =0.0375+0.2135+0.225+0.0815+0.7=1.2575.

[0090] The VRL somatic mutation comprehensive score S for each sample was calculated using the method described above and used as a predictive variable. VRL samples were marked as 1 for positive samples and 0 for negative control samples. The true positive rate (TPR) and false positive rate (FPR) of the scoring system were then calculated using the pROC package in R language. ROC curves were plotted and the area under the curve (AUC) was calculated to characterize the system's performance. The ROC curve of this invention is shown below. Figure 3As shown, the results demonstrate that the VRL comprehensive score exhibits good discriminative ability. When the cutoff value is 0.69025, the method has a sensitivity of 91.86%, a specificity of 82.86%, and an AUC value of 0.905, indicating that it has high accuracy in identifying potential VRL risks.

[0091] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit this application. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of this application. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in this invention should still be covered by the claims of this application.

[0092] Table 1. Genes related to primary vitreoretinal lymphoma and their probe design.

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099] Table 2. Viruses and specific sequence probes associated with vitreoretinal lymphoma.

[0100]

[0101] Table 3. Overall Importance Score of 45 Genes

[0102]

Claims

1. A marker for vitreoretinal lymphoma risk assessment, comprising a mutation site of a vitreoretinal lymphoma-associated gene selected from one or more of BTG1, BTG2, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, and VHL.

2. The marker of claim 1, wherein, The vitreoretinal lymphoma-associated gene further comprises one or more of ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, TP53, and DMD; preferably, the vitreoretinal lymphoma-associated gene comprises BTG1, BTG2, DMD, HLA-A, LATS1, LRP1B, PTPN11, PTPRD, STAT5A, TGFBR2, VHL, ARID1A, ARID1B, ATM, BCL2, BCL6, BCOR, BRAF, CARD11, CD79A, CD79B, CDKN2A, CDKN2B, EP300, ETV6, EZH2, GNA13, IRF4, JAK1, JAK2, JAK3, KMT2D, MYC, MYD88, NOTCH1, NOTCH2, PIK3CA, PIM1, PRDM1, RHOA, SOCS1, STAT3, STAT5B, TET2, and TP53.

3. Marker according to claim 1 or 2, characterized in that The mutation site is a site of non-synonymous mutation relative to a reference transcript of the vitreoretinal lymphoma-associated gene.

4. The marker of claim 3, wherein, The reference transcript of BTG1 has accession number NM_001731; the reference transcript of BTG2 has accession number NM_006763; the reference transcript of DMD has accession number NM_004023; the reference transcript of HLA-A has accession number NM_002116; the reference transcript of LATS1 has accession number NM_001350339; the reference transcript of LRP1B has accession number NM_018557; the reference transcript of PTPN11 has accession number NM_080601; the reference transcript of PTPRD has accession number NM_001171025; the reference transcript of STAT5A has accession number NM_003152; the reference transcript of TGFBR2 has accession number NM_001024847; the reference transcript of VHL has accession number NM_001354723; the reference transcript of ARID1A has accession number NM_139135; the reference transcript of ARID1B has accession number NM_001371656; the reference transcript of ATM has accession number NM_00005; the reference transcript of BCL2 has accession number NM_000633; the reference transcript of BCL6 has accession number NM_001706; the reference transcript of BCOR has accession number NM_001123383; the reference transcript of BRAF has accession number NM_001378474; the reference transcript of CARD11 has accession number NM_032415; the reference transcript of CD79A has accession number NM_001783; the reference transcript of CD79B has accession number NM_001039933; the reference transcript of CDKN2A has accession number NM_000077; the reference transcript of CDKN2B has accession number NM_078487; the reference transcript of EP300 has accession number NM_001429; the reference transcript of ETV6 has accession number NM_001987; the reference transcript of EZH2 has accession number NM_001203247; the reference transcript of GNA13 has accession number NM_006572; the reference transcript of IRF4 has accession number NM_002460; the reference transcript of JAK1 has accession number NM_002227; the reference transcript of JAK2 has accession number NR_169763; the reference transcript of JAK3 has accession number NM_000215; the reference transcript of KMT2D has accession number NM_003482; the reference transcript of MYC has accession number NM_001354870; the reference transcript of MYD88 has accession number NM_00246; the reference transcript of NOTCH1 has accession number NM_017617;The reference transcript of NOTCH2 has accession number NM_024408; the reference transcript of PIK3CA has accession number NM_006218; the reference transcript of PIM1 has accession number NM_002648; the reference transcript of PRDM1 has accession number NM_001198; the reference transcript of RHOA has accession number NM_001313943; the reference transcript of SOCS1 has accession number NM_003745; the reference transcript of STAT3 has accession number NM_139276; the reference transcript of STAT5B has accession number NM_012448; the reference transcript of TET2 has accession number NM_001127208; the reference transcript of TP53 has accession number NM_000546. 5.Use of the marker and / or its detection agent of any one of claims 1-4 in the preparation of a vitreoretinal lymphoma detection product.

6. Use according to claim 5, characterized in that, The detection agent comprises any one or more of the following: 1) a probe for detecting BTG1, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 31; 2) a probe for detecting BTG2, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 32; 3) a probe for detecting HLA-A, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 82; 4) a probe for detecting LATS1, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 113; 5) a probe for detecting LRP1B, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 114-130; 6) a probe for detecting PTPN11, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 157-161; 7) a probe for detecting PTPRD, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 162 to 167; 8) a probe for detecting STAT5A, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 183 to 187; 9) a probe for detecting TGFBR2, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID No. 196; 10) a probe for detecting VHL, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 202 to 204; Preferably, the detection agent further comprises any one or more of: 11) a probe for detecting DMD, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 44 to 59; 12) a probe for detecting ARID1A, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 1 to 5; 13) a probe for detecting ARID1B, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 6 to 8; 14) a probe for detecting ATM, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 9 to 21; 15) a probe for detecting BCL2, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 22; 16) a probe for detecting BCL6, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 23; 17) a probe for detecting BCOR, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 24; 18) a probe for detecting BRAF, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 25 to 30; 19) a probe for detecting CARD11, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 33 to 37; 20) a probe for detecting CD79A, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 38; 21) a probe for detecting CD79B, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 39 to 40; 22) a probe for detecting CDKN2A, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 41; 23) a probe for detecting CDKN2B, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 42 to 43; 24) a probe for detecting EP300, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 60 to 68; 25) a probe for detecting ETV6, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 69 to 70; 26) a probe for detecting EZH2, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 71 to 78; 27) a probe for detecting GNA13, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 79 to 81; 28) a probe for detecting IRF4, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 83 to 85; 29) a probe for detecting JAK1, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 86 to 90; 30) a probe for detecting JAK2, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 91 to 96; 31) a probe for detecting JAK3, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 97 to 100; 32) a probe for detecting KMT2D, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 101 to 112; 33) a probe for detecting MYC, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 131 to 132; 34) a probe for detecting MYD88, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 133; 35) a probe for detecting NOTCH1, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 134 to 140; 36) a probe for detecting NOTCH2, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 141 to 148; 37) a probe for detecting PIK3CA, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 149 to 154; 38) a probe for detecting PIM1, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 155; 39) a probe for detecting PRDM1, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 156; 40) a probe for detecting RHOA, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 168 to 171; 41) a probe for detecting SOCS1, the nucleotide sequence of which comprises a sequence represented by SEQ ID No. 172; 42) a probe for detecting STAT3, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 173 to 182; 43) a probe for detecting STAT5B, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 188 to 192; 44) a probe for detecting TET2, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 193 to 195; 45) a probe for detecting TP53, the nucleotide sequence of which comprises a sequence represented by any one or more of SEQ ID Nos. 197 to 201.

7. A vitreoretinal lymphoma detection product, the detection product comprising a substance for detecting a marker according to any one of claims 1 to 4.

8. The test product of claim 7, wherein, The substance for detecting the marker comprises any one or more of: 1) a probe for detecting BTG1, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 31; 2) a probe for detecting BTG2, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 32; 3) a probe for detecting HLA-A, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 82; 4) a probe for detecting LATS1, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 113; 5) a probe for detecting LRP1B, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 114-130; 6) a probe for detecting PTPN11, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 157-161; 7) a probe for detecting PTPRD, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 162-167; 8) a probe for detecting STAT5A, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 183-187; 9) a probe for detecting TGFBR2, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID No. 196; 10) a probe for detecting VHL, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 202-204; Preferably, the substance for detecting the marker further comprises any one or more of the following: 11) a probe for detecting DMD, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 44-59; 12) a probe for detecting ARID1A, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 1-5; 13) a probe for detecting ARID1B, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 6-8; 14) a probe for detecting ATM, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 9-21; 15) a probe for detecting BCL2, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 22; 16) a probe for detecting BCL6, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 23; 17) a probe for detecting BCOR, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 24; 18) a probe for detecting BRAF, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 25-30; 19) a probe for detecting CARD11, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 33-37; 20) a probe for detecting CD79A, the nucleotide sequence of which comprises the sequence shown in SEQ ID No. 38; 21) a probe for detecting CD79B, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 39-40; 22) a probe for detecting CDKN2A, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 41; 23) a probe for detecting CDKN2B, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 42-43; 24) a probe for detecting EP300, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 60-68; 25) a probe for detecting ETV6, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 69-70; 26) a probe for detecting EZH2, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 71-78; 27) a probe for detecting GNA13, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 79-81; 28) a probe for detecting IRF4, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 83-85; 29) a probe for detecting JAK1, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 86-90; 30) a probe for detecting JAK2, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 91-96; 31) a probe for detecting JAK3, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 97-100; 32) a probe for detecting KMT2D, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 101-112; 33) a probe for detecting MYC, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 131-132; 34) a probe for detecting MYD88, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 133; 35) a probe for detecting NOTCH1, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 134-140; 36) a probe for detecting NOTCH2, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 141-148; 37) a probe for detecting PIK3CA, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 149-154; 38) a probe for detecting PIM1, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 155; 39) a probe for detecting PRDM1, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 156; 40) a probe for detecting RHOA, the nucleotide sequence of which comprises the sequence set forth in any one or more of SEQ ID Nos. 168-171; 41) a probe for detecting SOCS1, the nucleotide sequence of which comprises the sequence set forth in SEQ ID No. 172; 42) a probe for detecting STAT3, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 173-182; 43) a probe for detecting STAT5B, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 188-192; 44) a probe for detecting TET2, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 193-195; 45) a probe for detecting TP53, the nucleotide sequence of which comprises the sequence shown in any one or more of SEQ ID Nos. 197-201.

9. The test product of claim 7, wherein, The detection product also includes a substance for detecting a virus related to the etiology of vitreoretinal lymphoma; Preferably, the virus related to the etiology of vitreoretinal lymphoma includes one or more of herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, human herpes virus type 4, human cytomegalovirus, human herpes virus type 6, human herpes virus type 8, and circovirus.

10. The test product of claim 7, wherein, The detection product also includes any one or more of the following: 1) a probe for detecting herpes simplex virus type 1, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 205-206; 2) a probe for detecting herpes simplex virus type 2, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 207-208; 3) a probe for detecting varicella-zoster virus, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 209-210; 4) a probe for detecting human herpes virus type 4, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 211-212; 5) a probe for detecting human cytomegalovirus, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 213-214; 6) a probe for detecting human herpes virus type 6, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 215-216; 7) a probe for detecting human herpes virus type 8, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 217-218; 8) a probe for detecting circovirus, the nucleotide sequence of which comprises the sequence shown in any one or both of SEQ ID Nos. 219-220.

11. A vitreoretinal lymphoma risk assessment product selected from any one of the following: 1) a vitreoretinal lymphoma risk assessment device comprising the following modules: a data acquisition module for providing detection results of target markers of a sample to be tested, and confirming the mutation gene of the sample to be tested based on the detection results, the target markers being the markers according to any one of claims 1-4; a risk assessment module for assessing the risk of an individual corresponding to the sample to be tested of suffering from vitreoretinal lymphoma based on the comprehensive score of VRL somatic gene mutations of the sample to be tested; 2) A computer readable storage medium, the storage medium stores computer instructions, when the computer instructions are executed by a processor, a method for assessing the risk of vitreoretinal lymphoma is implemented, the method comprises: obtaining the detection result of the target marker data of the sample to be tested, and confirming the mutant gene of the sample to be tested based on the detection result, the target marker is the marker of any one of claims 1-4; based on the VRL somatic gene mutation comprehensive score of the sample to be tested, the risk of vitreoretinal lymphoma of the individual corresponding to the sample to be tested is evaluated; 3) An electronic terminal, comprising: a processor, a memory, a network interface and a user interface; the memory is used to store computer programs, and the processor is used to execute the computer programs stored in the memory, so that the terminal executes a vitreoretinal lymphoma risk assessment method, the method comprises: obtaining the detection result of the target marker data of the sample to be tested, and confirming the mutant gene of the sample to be tested based on the detection result, the target marker is the marker of any one of claims 1-4; based on the VRL somatic gene mutation comprehensive score of the sample to be tested, the risk of vitreoretinal lymphoma of the individual corresponding to the sample to be tested is evaluated; 4) A computer program product, comprising a computer program, the computer program is executed by a processor to implement a vitreoretinal lymphoma risk assessment method, the method comprises: obtaining the detection result of the target marker data of the sample to be tested, and confirming the mutant gene of the sample to be tested based on the detection result, the target marker is the marker of any one of claims 1-4; based on the VRL somatic gene mutation comprehensive score of the sample to be tested, the risk of vitreoretinal lymphoma of the individual corresponding to the sample to be tested is evaluated.

12. The risk assessment product of claim 11, wherein, The VRL somatic gene mutation comprehensive score is the sum of the comprehensive importance score Stotal corresponding to each mutant gene.

13. The risk assessment product of claim 12, wherein, The Stotal for the BTG1 mutation is 0.224, the Stotal for the BTG2 mutation is 0.3495, the Stotal for the DMD mutation is 0, the Stotal for the HLA-A mutation is 0.225, the Stotal for the LATS1 mutation is 0.0375, the Stotal for the LRP1B mutation is 0.0375, the Stotal for the PTPN11 mutation is 0.15, the Stotal for the PTPRD mutation is 0.075, the Stotal for the STAT5A mutation is 0.3495, the Stotal for the TGFBR2 mutation is 0.313, the Stotal for the VHL mutation is 0.1875, the Stotal for the ARID1A mutation is 0.0375, the Stotal for the ARID1B mutation is 0.0375, the Stotal for the ATM mutation is 0.225, the Stotal for the BCL2 mutation is 0.15, the Stotal for the BCL6 mutation is 0.075, the Stotal for the BCOR mutation is 0.0375, the Stotal for the BRAF mutation is 0.1875, the Stotal for the CARD11 mutation is 0.1255, the Stotal for the CD79A mutation is 0.0375, the Stotal for the CD79B mutation is 0.313, the Stotal for the CDKN2A mutation is 0.4555, the Stotal for the CDKN2B mutation is 0.1875, the Stotal for the EP300 mutation is 0.269, the Stotal for the ETV6 mutation is 0.1255, the Stotal for the EZH2 mutation is 0.0375, the Stotal for the GNA13 mutation is 0.0375, the Stotal for the IRF4 mutation is 0.2135, the Stotal for the JAK1 mutation is 0.2625, the Stotal for the JAK2 mutation is 0.4125, the Stotal for the JAK3 mutation is 0.1125, the Stotal for the KMT2D mutation is 0.1695, the Stotal for the MYC mutation is 0.15, the Stotal for the MYD88 mutation is 0.7, the Stotal for the NOTCH1 mutation is 0.1125, the Stotal for the NOTCH2 mutation is 0.075, the Stotal for the PIK3CA mutation is 0.0375, the Stotal for the PIM1 mutation is 0.0375, the Stotal for the PRDM1 mutation is 0.225, the Stotal for the RHOA mutation is 0.0375, the Stotal for the SOCS1 mutation is 0.1875, the Stotal for the STAT3 mutation is 0.224, the Stotal for the STAT5B mutation is 0.1255, the Stotai of the TET2 mutation is 0.0375, and the Stotai of the TP53 mutation is 0.4555; And / or, the mutant gene is detected and determined by the detection product of any one of claims 7-10.

14. The risk assessment product of claim 11, wherein, The higher the VRL somatic gene mutation comprehensive score of the sample to be tested, the higher the risk of vitreoretinal lymphoma of the individual corresponding to the sample to be tested; preferably, the VRL somatic gene mutation comprehensive score of the sample to be tested is compared with the cutoff value, when the VRL somatic gene mutation comprehensive score of the sample to be tested is greater than or equal to the cutoff value, it is determined that the vitreoretinal lymphoma is positive; when the VRL somatic gene mutation comprehensive score of the sample to be tested is less than the cutoff value, it is determined that the vitreoretinal lymphoma is negative; more preferably, the cutoff value is 0.69025.

15. The risk assessment product of claim 11, wherein, The sample to be tested is cfDNA in intraocular fluid; preferably, the intraocular fluid is aqueous humor and / or vitreous humor.

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