Application of substance specifically combined with NSE protein or coding gene thereof in preparation of ARN diagnosis product or prognosis product

Abstract

The invention relates to the field of medicine, in particular to application of a substance specifically combined with NSE protein or a coding gene thereof to preparation of an ARN diagnosis product or a prognosis product. By detecting the NSE protein or the encoding gene thereof, optic neuropathy caused by acute retinal necrosis can be informed, and doctors can recognize ARN diseases earlier, so that proper medical intervention can be implemented quickly. Therefore, the risk of retinal abscission and visual loss can be remarkably reduced, clinical doctors can be assisted in accurately evaluating the severity of the illness state and predicting the possibility of visual recovery, and individuation and precision of treatment can be promoted. In addition, by avoiding unnecessary medicine use and medical resource waste, economic benefits are brought, and both the patient and the society are benefited.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to the use of a substance that specifically binds to NSE protein or its encoding gene in the preparation of ARN diagnostic products or prognostic products. Background Art

[0002] Acute Retinal Necrosis (ARN) is a rare but potentially devastating intraocular inflammatory disease that can affect people of all ages. ARN is mainly caused by viral infections, including varicella-zoster virus (VZV), herpes simplex virus (HSV), and cytomegalovirus (CMV). The pathological mechanism of ARN involves direct viral invasion of retinal neurons, triggering a local inflammatory response. The virus spreads to the retina through the trigeminal ganglion or through the blood, causing retinitis. The inflammatory response leads to retinal vasculitis, which in turn causes retinal ischemia and tissue necrosis. The clinical manifestations of ARN include vitritis, occlusive retinal arterial vasculitis, and confluent necrotizing retinitis. Patients may experience symptoms such as decreased vision, eye pain, light sensitivity, and floaters. Fundus examination shows white or yellowish-white necrotic lesions on the retina, which are usually located in the mid-periphery of the retina and can expand rapidly. Early diagnosis of ARN faces major challenges. The lack of specificity in early symptoms and the possibility of similarities with other retinal diseases, especially uveitis, make diagnosis difficult. Early literature reports show that the misdiagnosis rate of ARN can be as high as over 70%. Treatment of ARN usually includes antiviral drugs, corticosteroids, and in some cases laser therapy or surgery. However, the timeliness and effectiveness of treatment are crucial to improving prognosis. Currently, problems in treatment include insufficient timeliness of treatment, lack of targeted antiviral treatment, and insufficient control of disease progression. If ARN is not diagnosed and treated promptly and correctly, patients may have a high incidence of retinal detachment, leading to severe vision loss or even blindness. Therefore, early correct diagnosis and timely treatment, and striving to control the progression of the disease before retinal detachment occurs, are of great significance for improving the prognosis of ARN patients.

[0003] In summary, the current diagnosis of acute retinopathy is mainly determined by clinical ophthalmologists based on the patient's clinical manifestations. However, due to the difficulty of observation caused by vitreous opacity and the subjectivity of physical examination, the early correct diagnosis rate of the disease is low, and misdiagnosis and missed diagnosis are prone to occur. Clinically, objective and accurate biomarkers are needed to help with the early correct diagnosis of ARN. Given the severity of ARN and the urgency of treatment, it is necessary to develop new diagnostic tools.

[0004] Enolase is widely present in organisms and is a key enzyme in the glycolysis pathway. It has three subunits with different immune properties: α, β, and γ. The γγ dimer only exists in neurons and neuroendocrine cells and is called neuron-specific enolase (NSE). The content in normal body fluids is very low. Nerve damage caused by viral infection, ischemia and hypoxia can cause NSE to leak from damaged neurons. Its level can initially reflect the severity of nerve damage. The higher the NSE content, the more severe the nerve damage and the worse the prognosis. In addition, NSE has a fast detection speed and can obtain results within half an hour. It has high clinical application value, but there is no public information on the correlation between NSE and ARN in aqueous humor. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a substance that specifically binds to NSE protein or its encoding gene for use in preparing ARN diagnostic products or prognostic products, so as to solve the problems in the prior art.

[0006] To achieve the above-mentioned object and other related objects, the present invention provides the use of a substance that specifically binds to NSE protein or its encoding gene in the preparation of a retinal necrosis diagnostic product or a prognostic product.

[0007] Preferably, the retinal necrosis is acute retinal necrosis (ARN).

[0008] The acute retinal necrosis diagnostic product is a product for distinguishing patients with acute retinal necrosis from healthy people or patients with other diseases such as uveitis.

[0009] The acute retinal necrosis diagnostic product may also be a product for determining the severity of acute retinal necrosis.

[0010] As described above, the use of the substance that specifically binds to NSE protein or its encoding gene in the preparation of ARN diagnostic products or prognostic products of the present invention has the following beneficial effects: it can objectively and accurately observe the damage of optic neuropathy caused by acute retinal necrosis, and by detecting the level of NSE protein in aqueous humor, according to the judgment of clinical diagnostic value, it can help clinical early diagnosis of ARN, timely adopt the correct treatment method, effectively reduce the risk of retinal detachment and blindness, help clinical accurate assessment of disease severity and visual prognosis, achieve precision treatment, reduce drug abuse, and have high economic benefits for patients and society. The application potential of using aqueous humor NSE protein level as a biomarker for evaluating optic nerve damage is not limited to this, and may also cover several other ophthalmic diseases, which may also cause optic nerve damage, such as glaucoma, optic neuritis and infectious endophthalmitis. This discovery provides a new direction for the early diagnosis and treatment of these diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Shown is a graph showing the difference in NSE protein levels in the aqueous humor of ARN and UV patients of the present invention.

[0012] Figure 2 Shown is a ROC curve based on the expression level of NSE protein in aqueous humor samples of ARN patients and control patients.

[0013] Figure 3 Results are shown as the correlation between the level of NSE in aqueous humor and the percentage of retinal necrosis area observed clinically.

[0014] Figure 4 Shown are the expression trend of NSE level in aqueous humor during treatment (left figure) and the trend of necrotic area percentage (right figure).

[0015] Figure 5 Shown are the results of correlation analysis between NSE levels and retinal detachment survival rate. DETAILED DESCRIPTION

[0016] The detection of neuron-specific enolase (NSE) is currently reported to be mainly used in the detection of related diseases in blood and cerebrospinal fluid. There are very few reports on the detection of NSE in human aqueous humor. There are few studies on the diagnosis of ARN by detecting the expression of secretory proteins in the aqueous humor of patients. There is currently no report on the establishment of a connection between NSE and ARN diagnosis. The present invention collects the aqueous humor of patients with ARN in the disease group, and the control group is a uveitis with early symptoms similar to ARN, easy to misdiagnose, and requiring special differential diagnosis, and detects the expression of NSE in the sample to be tested. If the expression of NSE is higher than the diagnostic value of 7.18ng / mL, it can be diagnosed as ARN, and the diagnostic value obtained by the comparative analysis of the disease group control disease has a very high clinical application value. In addition, the correlation analysis of the level of NSE in the aqueous humor and the percentage of retinal necrosis area has obtained the correlation coefficient and linear formula of the two, which can help the clinical understanding of the severity of the disease more objectively.

[0017] Based on the above content, the present invention provides the use of a substance that specifically binds to NSE protein or its encoding gene in the preparation of a retinal necrosis diagnosis product or a prognosis product.

[0018] In certain embodiments of the present invention, the retinal necrosis is acute retinal necrosis (ARN).

[0019] NSE protein is a dimeric enzyme encoded by the NSE gene, which consists of two identical subunits with an α-β-α sandwich structure, which contributes to the stability and catalytic activity of the enzyme. NSE protein catalyzes the conversion of 2-phosphoglycerate (2-PG) into phosphoenolpyruvate (PEP) during glycolysis.

[0020] In certain embodiments of the present invention, the NSE protein is a wild-type NSE protein or a mutant NSE protein.

[0021] The NSE protein is a full-length sequence or a protein fragment, that is, the acute retinal necrosis diagnostic product or prognostic product includes a substance that can specifically bind to the full-length sequence or protein fragment of the NSE protein.

[0022] The NSE protein or its encoding gene is derived from an animal, preferably a mammal; the mammal is preferably a rodent, an artiodactyl, a perissodactyl, a lagomorph, a primate, etc. The mammal includes, for example, humans, non-human primates (such as monkeys), mice, pigs, cows, goats, rabbits, rats, guinea pigs, hamsters, monkeys or other non-human mammals; non-mammals include, for example, non-mammalian vertebrates, such as birds or fish, and non-mammalian invertebrates.

[0023] In a preferred embodiment, the NSE protein or its encoding gene is derived from human. The human NSE gene is located in the human chromosome 1p36.2 region, and the NSE protein encoded by the gene consists of 495 amino acid residues.

[0024] The substance that specifically binds to the NSE protein or its encoding gene is not specifically limited, as long as it can specifically bind to the NSE protein or its encoding gene, for example, it can be an antibody, an antibody functional fragment, a conjugated antibody, a nucleic acid molecule, a lipid, a small chemical molecule, a polypeptide, a protein, a lentivirus, an adeno-associated virus, a nanoparticle, a liposome, an extracellular vesicle or a cell.

[0025] The acute retinal necrosis diagnostic product is a product for distinguishing patients with acute retinal necrosis from healthy people or patients with other diseases such as uveitis.

[0026] The acute retinal necrosis diagnostic product can also be a product for judging the severity of acute retinal necrosis. When the acute retinal necrosis diagnostic product detects the higher the expression of NSE protein or its encoding gene, the more severe the acute retinal necrosis is. Clinically, the severity of acute retinal necrosis can be measured by the percentage of retinal necrosis area.

[0027] The acute retinal necrosis prognosis product refers to a product used to predict the progression of a disease, such as a product that predicts the probability of retinal detachment (RD) leading to vision loss.

[0028] In the diagnostic product or prognostic product, the active ingredient that performs the function may be only a substance that specifically binds to the NSE protein or its encoding gene, and may also contain other substances. That is, the substance used to detect the NSE protein or its encoding gene is the only active ingredient or one of the active ingredients of the product. The product can be used in combination with other products or alone.

[0029] The product can be a single-component substance or a multi-component substance.

[0030] The product can be a commercial reagent or a kit or a homemade reagent. The commercial kit is, for example, an electrochemiluminescence kit. The components of the electrochemiluminescence kit include, for example, one or more of an antibody, an antibody functional fragment, and a conjugated antibody that can specifically bind to NSE, and also include a system buffer, a measuring cell cleaning solution, a pre-cleaning solution, a needle washing solution, a reaction cup, and a sample tip.

[0031] The form of the product is not particularly limited and can be in various forms such as solid, liquid, gel, semi-fluid, aerosol, etc.

[0032] The sample type mainly targeted by the product is aqueous humor.

[0033] The present invention does not specifically limit the etiology, clinical manifestations, severity, and presence or absence of complications of the acute retinal necrosis. As long as it is acute retinal necrosis, the diagnostic product of the present invention can be used for diagnosis.

[0034] the term

[0035] In the present invention, "marker" refers to a biomolecule or biomolecule fragment, whose change and / or detection can be associated with a specific physical condition or state. The terms "marker (marker)", "molecular marker (marker)" or "biomarker (marker)" are used interchangeably throughout the disclosure of the invention.

[0036] In the present invention, the "patient", "subject" or "individual" may refer to an organism, and in some aspects, the subject may be a human. The subject providing the sample may include a population at potential risk of the disease or a population diagnosed with the disease. The disease in the present invention refers to ARN or uveitis.

[0037] In the present invention, the term "specimen" and "sample" are used interchangeably, and include substances obtained from individual or separated tissues, cells or body fluids and suitable for marker detection.

[0038] In the present invention, the "prognosis" refers to the prediction of the possible consequences of trauma or disease (such as ARN), and the prognostic indicators include short-term efficacy indicators and long-term efficacy indicators, including but not limited to HR, ORR, DCR, OS, PFS, DFS, and RFS. Among them, HR refers to the hazard ratio (HR), which is used to compare the risk of an event (such as disease recurrence, death, etc.) occurring in two groups (such as the observation group and the control group) within a specific time. The calculation method of HR is usually based on a survival analysis model, such as the Cox proportional hazard model. In practical applications, statistical software (such as R, SAS, SPSS, etc.) is usually used to construct a Cox model and calculate HR. If HR>1, it means that the risk of the observation group is higher than that of the control group. If HR<1, it means that the risk of the observation group is lower than that of the control group. If HR=1, it means that the risks of the two groups are the same. "OS" refers to "Overall Survival", which refers to the total survival time of all patients in the study; "PFS" refers to "Progression Free Survival", which usually refers to the time when the patient's disease does not progress or become resistant after treatment; "DFS" refers to "Disease Free Survival", which usually refers to the time from randomization to disease recurrence or death (due to any reason); "RFS" refers to "Recurrence Free Survival", which usually refers to the patient's recurrence from the date of initial treatment to the date of recurrence or the last follow-up date. Generally, it is expressed clinically as median OS, median PFS, median RFS, and median DFS, that is, the survival time / progression-free survival / recurrence-free survival / disease-free survival time achieved by 50% of patients.

[0039] In the present invention, the "ROC curve" refers to a receiver operating characteristic curve. In certain specific embodiments of the present invention, the ROC curve refers to a ROC curve between a true positive rate and a false positive rate.

[0040] In the present invention, the "expression level" may refer to the concentration or amount of the gene / protein of the marker of the present invention in a sample.

[0041] In the present invention, the terms "high expression", "high expression level" and the like are interchangeable and should be used to mean at least 5%, 10% or 20%, preferably at least 30% or 50%, more preferably at least 80% or 100% or more significant increase compared to a "control" or "threshold value". For example, Student's T-test can be repeated for genes with at least one expression intensity exceeding the threshold value to determine significance.

[0042] In the present invention, the terms "low expression", "low expression level" and the like are interchangeable and should be used to mean a decrease of at least 5%, 10% or 20%, preferably at least 30% or 50%, more preferably at least 80% or 100% or more significantly compared to a "control" or "threshold value". For example, Student's T-test can be repeated for genes with at least one expression intensity below the threshold to determine significance.

[0043] In the present invention, the setting of the "control" or "threshold" of gene or protein expression is easily set by those skilled in the art based on the main idea of ​​the present invention. Selecting an appropriate "control" or "threshold" is a routine part of experimental design. For example, the expression level of the corresponding gene / protein can be first analyzed with statistical significance based on samples of subjects (patients) whose prognosis / therapy efficacy is clear, and the expression value obtained is used as the "control" or "threshold".

[0044] In the present invention, the "kit" may refer to a system of materials or reagents used to implement the methods disclosed in the present invention.

[0045] The sequence information of the NSE gene can be found in NCBI Gene: 2026, and the present invention may also cover its sequence variants in organisms.

[0046] The markers disclosed in the present invention can be used as markers (markers) to evaluate the development of ARN and the efficacy of therapy, so as to understand the disease state of the individual, evaluate or predict the prognosis of the disease risk, and formulate treatment / medication plans.

[0047] As a method, the method of using the marker to diagnose ARN or evaluate the prognosis of ARN includes: (1) detecting the expression level of NSE protein or gene in the subject sample; (2) based on the expression level obtained in (1): when high expression, it indicates that the patient has ARN, and the higher the expression level, the more serious the disease, or the poorer the prognosis and the shorter the survival period; conversely, ARN does not exist or the prognosis is good.

[0048] In the present invention, when the risk value is higher than a preset threshold, it indicates that ARN exists or the risk of disease prognosis is high; when the risk value is lower than the preset threshold, it indicates that ARN does not exist or the risk of disease prognosis is low.

[0049] The expression level of the markers of the present invention can be determined according to established standard procedures well known in the art (reference). The determination can be performed at the DNA, RNA or protein level, for example, by detecting cDNA levels after RNA reverse transcription using real-time fluorescence quantitative PCR technology; for example, by detecting protein levels using electrochemiluminescence or ELISA.

[0050] As an optional method, depending on the protein of the marker, detection can be achieved using an antibody that specifically binds to the protein.

[0051] As an optional method, primers that specifically amplify the markers can be designed according to their sequences for detection. Polymerase chain reaction (PCR) technology is a technology well known to those skilled in the art, and its basic principle is a method for enzymatically synthesizing specific DNA fragments in vitro. The method of the present invention can be performed using conventional PCR technology.

[0052] The method of amplifying a specific gene fragment by PCR is a well-known technique in the art and is not particularly limited in the present invention. The amplified product can be labeled by a method of amplifying with a primer having a labeling group at the 5' end, or by a method of incorporating a mononucleotide with a labeling group during the amplification process, or by a method of adding a detection probe that specifically binds to the amplified product during hybridization. The labeling group includes, but is not limited to, digoxigenin molecule (DIG), biotin molecule (Bio), fluorescein and its derivative biological molecules (FITC, etc.), other fluorescent molecules (such as Cy3, Cy5, etc.), alkaline phosphatase (AP), horseradish peroxidase (HRP), etc.

[0053] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0054] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0055] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0056] The present invention collected aqueous humor samples from 40 ARN patients and 43 uveitis (UV) patients, and used electrochemiluminescence to detect the NSE expression level in the aqueous humor samples. The detection instrument was Roche cobas e602. The difference in the expression of NSE in the aqueous humor of ARN patients and uveitis patients was statistically analyzed. In addition, the present invention also used GraphPad Prism 9.5.1 to draw a scatter plot of the NSE detection concentration and the Kaplan-Meier survival curve of two differential diagnosis diseases, and used MedCalc 19.6.4 to draw a single ROC curve to analyze the specificity and sensitivity of NSE protein for ARN diagnosis. The details are as follows:

[0057] Example 1

[0058] 1. Experimental Materials

[0059] The neuron-specific enolase (NSE) electrochemiluminescence kit used in this example was purchased from Roche Diagnostics GmbH. The reagent components are as follows:

[0060] 1. Streptavidin-coated microparticles, 1 bottle, 6.5 ml. Contains magnetic microparticles coated with streptavidin, particle concentration 0.72 mg / ml, containing preservatives.

[0061] 2. Biotinylated anti-NSE antibody, 1 bottle, 10 ml, containing biotinylated anti-NSE monoclonal antibody 18E5 (mouse source) 1.0 mg / l, phosphate buffer 50 mmol / l, Ph 7.2; contains preservative.

[0062] 3. Ru(bpy)32+ labeled anti-NSE antibody, 1 bottle, 10 ml. Contains 1.0 mg / l of anti-NSE monoclonal antibody 84B10 (mouse source) labeled with ruthenium complex, concentration 2 mg / l, phosphate buffer 50 mmol / l, pH 7.2, containing preservatives.

[0063] Other required materials:

[0064] 1 System Buffer (ProCell)

[0065] 2. Cleaning fluid for measuring cell (CleanCell)

[0066] 3Preclean

[0067] 4. Probewash

[0068] 5 reaction cups (CUP)

[0069] 6. Tip (TIP)

[0070] 2. Experimental steps

[0071] 1. Collect samples

[0072] Aqueous humor was collected from 40 patients with ARN, and aqueous humor was collected from 43 patients with uveitis (control group). Among them, the aqueous humor of 40 patients with ARN was derived from patients diagnosed by clinical diagnostic criteria; the aqueous humor of 44 patients with uveitis was derived from control patients with no fundus necrotic lesions. The collected aqueous humor was placed in a 1.5 mL EP tube, centrifuged at 4000 × g for 10 minutes, and the supernatant was aspirated and stored in a -80 ° C refrigerator. Before use, the aqueous humor was taken out and thawed in a 4 ° C refrigerator and aliquoted to avoid repeated freezing and thawing.

[0073] 2. Electrochemiluminescence method to detect the content of NSE in aqueous humor

[0074] In the first stage of the reaction, the sample is incubated with biotinylated NSE monoclonal antibody and specific NSE monoclonal antibody labeled with ruthenium complex to form a sandwich complex. In the second step, streptavidin-coated microparticles are added to allow the above-formed complex to bind to the microparticles through the reaction between biotin and streptavidin. The reaction mixture is sucked into the measuring cell, and the microparticles are adsorbed to the electrode through a magnet. The unbound substances are washed away by the Procell, and chemiluminescence is generated after the electrode is pressurized, which is measured by a photomultiplier tube. The instrument automatically calculates the test results through the calibration curve of the detector.

[0075] 3. t-test analysis

[0076] Draw the standard curve: In the Excel worksheet, use the standard concentration as the horizontal axis and the corresponding OD value as the vertical axis to draw the standard linear regression curve, and calculate the NSE concentration value of each sample according to the curve equation. The t-test method was used to analyze whether the NSE protein levels in the ARN group and the control group (UV group) were significantly different. The test results are as follows Figure 1 The results showed that compared with UV patients, the level of NSE protein in the aqueous humor of ARN patients was significantly increased, indicating that the expression level of NSE is closely related to ARN and can be used as a biomarker for the diagnosis of ARN.

[0077] Example 2: ROC curve analysis of accuracy, sensitivity and specificity

[0078] GraphPad Prism 9.5.1 was used to draw ROC curves to analyze the value of NSE protein in the diagnosis of ARN. Based on the expression level of NSE protein in aqueous humor samples of 40 ARN patients and 43 control patients, ROC curves were drawn to evaluate the ability of NSE to distinguish ARN from UV patients.

[0079] The ROC curve for distinguishing ARN and UV patients based on NSE protein diagnosis was used, with the NSE concentration value with the largest Youden index as the cutoff value (i.e., diagnostic value), and the corresponding AUC and 95% confidence interval, sensitivity, and specificity were calculated at the same time. The sensitivity and specificity calculation formulas are as follows:

[0080] TPR=TP / (TP+FN)

[0081] Where TPR is the sensitivity,

[0082] TP is the True Positives, which is the number of samples correctly predicted as positive.

[0083] FN is False Negatives, which is the number of positive samples that are incorrectly predicted as negative.

[0084] FPR=FP / (FP+TN)

[0085] Among them, FPR is specificity,

[0086] FP stands for False Positives, which is the number of negative samples that are incorrectly predicted as positive.

[0087] TN is the True Negatives, which is the number of samples correctly predicted as negative.

[0088] Test results such as Figure 2 When NSE is used for ARN diagnosis, the diagnostic value is 7.18 ng / mL, that is, if the expression level of NSE is higher than the diagnostic value of 7.18 ng / mL, it can be diagnosed as ARN. The AUC value of the ROC curve is 0.975, the sensitivity is 92.5%, the specificity is 95.45%, and the Youden index is 0.880. The results show that the expression level of NSE in human aqueous humor has very high specificity, accuracy and sensitivity in the diagnosis of ARN.

[0089] Example 3. Correlation analysis between markers and percentage of retinal necrosis area

[0090] Patients with confirmed ARN underwent ultra-wide-angle fundus imaging (UWF) examinations using the 200Tx laser scanning ophthalmoscope retinal imaging system from the British Optos company. The collected images were evaluated by two experienced physicians and a consensus was reached on the percentage of active retinal necrosis area. GraphPad Prism 9.5.1 was used to draw a linear regression analysis of the aqueous humor NSE concentration and the percentage of active retinal necrosis area, as well as the dynamic changes of the two results after treatment. The results are shown in Figure 2. Figure 3 , 4 As shown. Figure 3 As shown in Figure 2, the level of NSE in aqueous humor is correlated with the percentage of retinal necrosis area observed clinically, with a correlation coefficient r value of 0.6603, indicating that the expression level of NSE can reflect the severity of the disease. Figure 4 As shown, the level of NSE in aqueous humor showed a significant downward trend in the three detections during the treatment, which was consistent with the downward trend of the percentage of necrosis area.

[0091] Example 4 Prognostic association between NSE and ARN

[0092] To analyze the relationship between aqueous humor NSE concentration and retinal detachment (retinal detachment) survival rate, the aqueous humor NSE concentration of all ARN patients was ranked, and the median value of 400pg / mL was taken as the cutoff value. The patients were divided into two groups: >400pg / mL and <400pg / mL. The patients were followed up for a maximum of one year, and the follow-up was terminated when retinal detachment occurred. GraphPad Prism 9.5.1 was used to draw the Kaplan-Meier curve, and the results showed that there was a significant correlation between aqueous humor NSE levels and the risk of retinal detachment ( Figure 5 ; P = 0.002). Patients with NSE concentrations > 400 pg / mL had a significantly increased risk of developing retinal detachment, with a hazard ratio (HR) of 5.389 (95% confidence interval, 1.853-15.6). This suggests that aqueous humor NSE can be used as a prognostic marker for patients with ARN.

[0093] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and the variations of the methods in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of the present invention.

Claims

1. Use of a substance that specifically binds to NSE protein or its encoding gene in the preparation of a retinal necrosis diagnostic product or a prognostic product.

2. The use according to claim 1, characterized in that The retinal necrosis diagnostic product or prognostic product is an acute retinal necrosis diagnostic product or prognostic product.

3. The use according to claim 1, characterized in that The NSE protein is a full-length sequence or a protein fragment; and / or, the NSE protein is a wild-type NSE protein or a mutant NSE protein.

4. The use according to claim 1, characterized in that The NSE protein or its encoding gene is derived from animals, preferably from mammals; more preferably from humans.

5. The use according to claim 1, characterized in that: The substance that specifically binds to NSE protein or its encoding gene is selected from antibodies, functional fragments of antibodies, nucleic acid molecules, lipids, small chemical molecules, polypeptides, liposomes or cells.

6. The use according to claim 2, characterized in that: The acute retinal necrosis diagnostic product is a product for distinguishing patients with acute retinal necrosis from healthy people or patients with other eye diseases.

7. The use according to claim 6, characterized in that The acute retinal necrosis diagnostic product is a product for distinguishing patients with acute retinal necrosis from patients with uveitis.

8. The use according to claim 2, characterized in that: The acute retinal necrosis diagnostic product is a product used to determine the severity of acute retinal necrosis.

9. The use according to claim 1, characterized in that The retinal necrosis diagnostic product or prognostic product is an electrochemiluminescence kit.

10. The use according to claim 1, characterized in that The sample type of the retinal necrosis diagnostic product or prognostic product is aqueous humor.

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