Application of cerebrospinal fluid CD38 protein and its specific antibody in the preparation of diagnostic products for neurosyphilis or nerve damage

Abstract

The present invention relates to the field of biomedical technology, and specifically to the use of cerebrospinal fluid CD38 protein and its specific antibody in the preparation of neurosyphilis or nerve damage diagnosis products. By measuring the concentration of CD38 protein in cerebrospinal fluid, the concentration is used as an indicator for early diagnosis of neurosyphilis, symptomatic neurosyphilis, asymptomatic neurosyphilis, typing detection of neurosyphilis and latent syphilis, and evaluation of the degree of nerve damage. The indicator can be used alone or as a main reference indicator in combination with other indicators to complete the diagnostic screening, typing and nerve damage evaluation of syphilis. Compared with the existing technology, this marker has the advantages of good sensitivity and high specificity, which is conducive to the diagnosis and management of neurosyphilis.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the application of cerebrospinal fluid CD38 protein and its specific antibody in the preparation of neurosyphilis or nerve damage diagnosis products. Background Art

[0002] Syphilis is a systemic infectious disease caused by the invasion of Treponema pallidum into the human body. Treponema pallidum spreads to the central nervous system within a few days after infection and may cause asymptomatic meningitis and severe symptomatic neurosyphilis.

[0003] Neurosyphilis is a group of clinical syndromes caused by Treponema pallidum invading the nervous system and causing damage to the meninges, brain, blood vessels or spinal cord. It can occur at all stages of the course of syphilis, not just in tertiary syphilis. The clinical manifestations of neurosyphilis vary, ranging from no clinical symptoms to complex clinical manifestations with extensive involvement of the nervous system. Clinically, it is usually divided into 5 types: asymptomatic, meningeal syphilis, meningeal vascular syphilis, tabes dorsalis and paralytic dementia (brain parenchymal syphilis). Although latent syphilis has no clinical symptoms, its harmfulness should be taken seriously. It can become recurrent syphilis and may progress to visceral and neurosyphilis. Patients with asymptomatic neurosyphilis have no obvious symptoms or signs, but there are abnormal changes in cerebrospinal fluid. If the abnormal cerebrospinal fluid persists for more than 5 years, the probability of developing symptomatic neurosyphilis is as high as 87%. In the study of primary and secondary syphilis, it was found that 20% of patients had abnormal cerebrospinal fluid, that is, neurosyphilis can occur in the early stage of syphilis. Early neurosyphilis includes syphilitic meningitis and vascular syphilis, and late neurosyphilis includes paralytic dementia, tabes dorsalis, etc. The incubation period of paralytic dementia is 10-15 years. Paralytic dementia is mainly manifested by mental and intellectual decline, such as memory loss, abnormal behavior, personality changes, and severe patients have epilepsy, stroke-like onset, secondary hemiplegia and aphasia.

[0004] Early diagnosis of neurosyphilis is very important. Finding sensitive biomarkers is helpful for the diagnosis of neurosyphilis and is of great significance for preventing serious and irreversible sequelae. At present, the diagnosis of neurosyphilis mainly relies on the examination of cerebrospinal fluid, including CSF-RPR, CSF white blood cell count and protein level. CSF-RPR is the same as the rapid plasma reagin ring card test in plasma. A positive result of this indicator can diagnose neurosyphilis. Important reference indicators for neurosyphilis also include cerebrospinal fluid cell count or protein level. When the CSF-RPR result is negative, if there are clinical signs of neurosyphilis, positive plasma tests and abnormal cerebrospinal fluid cell count, neurosyphilis can also be diagnosed. The increased protein content in the cerebrospinal fluid of patients with neurosyphilis is positively correlated with the severity of the disease and negatively correlated with the long-term prognosis. The increase in leukocytosis and the increase in protein content in the cerebrospinal fluid indicate the destruction of the blood-brain barrier and the presence of inflammatory reaction in the skull, which plays an auxiliary role in the diagnosis of neurosyphilis. However, the sensitivity of CSF-RPP was found to be only 30%-70% during the diagnosis process, which is low and insensitive to the diagnosis of early asymptomatic neurosyphilis. Currently, in the clinical diagnosis and treatment process, neurosyphilis is mainly diagnosed and treated in patients with symptoms of neurosyphilis, and there is a lack of sensitive, reliable and effective cerebrospinal fluid biological markers for distinguishing neurosyphilis. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of the technical problem that the existing CSF-RPR diagnostic method has low sensitivity, especially insensitivity to the diagnosis of early asymptomatic neurosyphilis, the present invention provides the use of cerebrospinal fluid CD38 protein and its specific antibody in the preparation of neurosyphilis or nerve damage diagnostic products. The use of this marker can realize early diagnosis, screening, typing detection and nerve damage assessment of neurosyphilis. Compared with the existing technology, it has higher sensitivity and can screen neurosyphilis and symptomatic neurosyphilis.

[0007] (II) Technical solution

[0008] In order to achieve the above object, the main technical solutions adopted by the present invention include:

[0009] In a first aspect, the present invention relates to CD38 as a cerebrospinal fluid marker for early diagnosis, screening and typing of patients with neurosyphilis.

[0010] In a second aspect, the present invention provides the use of CD38 protein and its specific antibodies in the preparation of diagnostic products for neurosyphilis screening, typing detection or assessment of the degree of nerve damage; the diagnostic product is a detection kit, antibody chip, antibody probe or detector with CD38 protein or its specific antibody as the target molecule.

[0011] According to a preferred embodiment of the present invention, the kit uses CD38 protein or its specific antibody as the detection target.

[0012] According to a preferred embodiment of the present invention, the kit is an enzyme-linked immunosorbent assay kit or an immunohistochemistry kit.

[0013] According to a preferred embodiment of the present invention, the kit comprises: a primary antibody that can specifically recognize CD38, a polypeptide dye, magnetic beads, and a biotin-labeled secondary antibody.

[0014] According to a preferred embodiment of the present invention, the reagents include citrate buffer, PBS washing solution, peroxidase blocker, goat serum, streptomycin antibiotic-peroxidase, DAB solution and hematoxylin solution.

[0015] According to a preferred embodiment of the present invention, the kit further comprises reagents or materials for obtaining cerebrospinal fluid from the patient.

[0016] It should be noted that when detecting the expression level of CD38 protein in cerebrospinal fluid, the Olink ultra-high sensitivity protein detection platform can also be used to quantify the expression level of CD38.

[0017] The third aspect is a product used for early diagnosis, screening, typing detection or assessment of the degree of nerve damage in patients with neurosyphilis; the product or device can quantitatively detect the concentration of CD38 in the patient's ex vivo cerebrospinal fluid; the product is a detection kit, antibody chip, antibody probe or detector.

[0018] Preferably, the product comprises antibodies that specifically bind to CD38 and other reagents that can quantitatively detect the concentration of CD38 protein in serum and cerebrospinal fluid. The experimental method used in the examples of this application is Olink neurology panel (95801).

[0019] In a fourth aspect, the present invention also relates to an assessment system for the risk of nerve damage in patients with neurosyphilis, comprising a detection module and a judgment module; wherein the detection module is used to obtain the concentration value of cerebrospinal fluid CD38; the judgment module comprises a readable carrier storing a judgment rule;

[0020] The judgment rule is: when the cerebrospinal fluid CD38 concentration value is less than 5.002NPX value, the risk of neurosyphilis is low, which is a low risk of brain damage;

[0021] When 5.400NPX>cerebrospinal fluid CD38 concentration value ≥5.002NPX value, it is judged that the risk of neurosyphilis is high and the risk of neurological damage is moderate;

[0022] When the cerebrospinal fluid CD38 concentration value is ≥5.400NPX, it is judged that the risk of symptomatic neurosyphilis is high and the risk of nerve damage is high.

[0023] Preferably, the reference value is a CD38 cut-off value obtained from a syphilis patient population. Preferably, the cerebrospinal fluid cut-off value for the diagnosis of non-neurosyphilis and neurosyphilis (including asymptomatic neurosyphilis and symptomatic neurosyphilis) is 5.002NPX. When the cerebrospinal fluid CD38 detection value is greater than or equal to 5.002NPX, the risk of neurosyphilis increases and the degree of nerve damage increases; preferably, the cerebrospinal fluid cut-off value for the diagnosis of asymptomatic neurosyphilis and symptomatic neurosyphilis is 5.400NPX. When the cerebrospinal fluid CD38 detection value is greater than or equal to 5.400NPX, the risk of symptomatic neurosyphilis increases and the degree of nerve damage further increases. These cut-off values ​​are the critical values ​​of CD38 statistical analysis obtained from 88 syphilis patients.

[0024] Syphilis typing includes symptomatic neurosyphilis, asymptomatic neurosyphilis or non-neurosyphilis, etc. In this application, non-neurosyphilis and latent syphilis have the same meaning.

[0025] In a fifth aspect, the present invention also provides a syphilis detection kit, which comprises: a first detection reagent composition for detecting serum or cerebrospinal fluid CD38 protein levels; and a second detection reagent composition for detecting cerebrospinal fluid leukocytes and / or cerebrospinal fluid proteins.

[0026] In the above scheme, when diagnosing neurosyphilis (including early diagnosis, typing and staging, and nerve damage assessment), the cerebrospinal fluid CD38 protein level can be combined with two or three indicators of cerebrospinal fluid leukocytes and cerebrospinal fluid proteins related to nerve damage in patients with neurosyphilis for early diagnosis, typing and staging, and nerve damage assessment of neurosyphilis. Among them, the CD38 level in the cerebrospinal fluid is determined by Olink precision proteomics, i.e., antibody immunoassay combined with quantitative real-time PCR. The number of cerebrospinal fluid leukocytes and the amount of cerebrospinal fluid protein can be detected by any known method.

[0027] (III) Beneficial effects

[0028] The present invention provides a new cerebrospinal fluid CD38 for early diagnosis screening, typing detection and nerve damage assessment of neurosyphilis. By detecting the expression level of CD38 in the patient's cerebrospinal fluid, a quantitative assessment of the degree of nerve damage in patients with neurosyphilis can be achieved; or by detecting the expression level of CD38 in the patient's cerebrospinal fluid, neurosyphilis can be diagnosed, including early diagnosis and typing detection. Compared with the prior art, the sensitivity of this marker for the diagnosis of neurosyphilis reaches 80.00, which is conducive to the rapid and accurate screening of latent syphilis and neurosyphilis and the assessment of the risk of nerve damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The Olink method was used to detect the expression of CD38 protein in the cerebrospinal fluid and serum of three groups of syphilis patients with different types: latent syphilis, asymptomatic neurosyphilis and symptomatic neurosyphilis.

[0030] Figure 2 The results showed that the level of CD38 protein in cerebrospinal fluid was positively correlated with that in serum.

[0031] Figure 3 The results showed that the expression of CD38 in cerebrospinal fluid was significantly positively correlated with cerebrospinal fluid protein and leukocytes.

[0032] Figure 4 The results showed that the expression of CD38 in serum was positively correlated with cerebrospinal fluid protein and leukocytes.

[0033] Figure 5 The AUC of different levels of cerebrospinal fluid and serum CD38 concentrations was used to estimate the probability of a confirmed diagnosis of neurosyphilis when the diagnosis of neurosyphilis (including asymptomatic neurosyphilis and symptomatic neurosyphilis) was used as the clinical outcome.

[0034] Figure 6 When the diagnosis of symptomatic neurosyphilis was used as the clinical outcome, the AUC of different levels of cerebrospinal fluid and serum CD38 concentrations were used to estimate the probability of a confirmed diagnosis of symptomatic neurosyphilis.

[0035] Figure 7 To distinguish asymptomatic neurosyphilis from non-neurosyphilis when asymptomatic neurosyphilis was the clinical outcome, the AUC of the probability of a confirmed diagnosis of asymptomatic neurosyphilis was evaluated at different levels of cerebrospinal fluid and serum CD38 concentrations.

[0036] Figure 8 All syphilis patients were divided into two groups based on the cerebrospinal fluid CD38 cutoff value of 5.002NPX value, and the differences in cerebrospinal fluid protein and leukocytes, the indicators of neurological damage, between the two groups were found.

[0037] Fig. 9 Neurosyphilis patients were divided into two groups based on the cerebrospinal fluid CD38 cutoff value of 5.400NPX value, and the differences in cerebrospinal fluid protein and leukocytes, the nerve damage indicators, between the two groups of patients. DETAILED DESCRIPTION

[0038] In order to better explain the present invention, the present invention is now described in detail.

[0039] Early diagnosis of neurosyphilis is very important. Finding sensitive biomarkers is helpful for the diagnosis of neurosyphilis and is of great significance for preventing serious and irreversible sequelae. Therefore, the inventors screened multiple indicators related to neurological function in the serum and cerebrospinal fluid of patients with neurosyphilis and obtained CD38, which has significant diagnostic value in cerebrospinal fluid.

[0040] After research, it was found that the levels of CD38 protein in the cerebrospinal fluid of patients with different types of syphilis were significantly different. The level of CD38 protein in the cerebrospinal fluid was correlated with the type of neurosyphilis and the occurrence of nerve damage. The difference between the levels of CD38 protein in the cerebrospinal fluid of patients with latent syphilis (or non-neurosyphilis) and those with asymptomatic neurosyphilis and symptomatic neurosyphilis was statistically significant. The levels of CD38 protein in the cerebrospinal fluid of patients with both types of neurosyphilis increased significantly, including patients with asymptomatic neurosyphilis, indicating that the level of CD38 protein changed during the stage of neurosyphilis. Therefore, this biomarker can effectively distinguish neurosyphilis from latent syphilis.

[0041] In order to clarify the relationship between serum and cerebrospinal fluid CD38 and neurosyphilis, this example studied 88 syphilis patients (15 cases of latent syphilis, 30 cases of asymptomatic neurosyphilis, and 43 cases of paralytic dementia). The research process used the receiver operating characteristic curve (ROC) to evaluate the diagnostic potential of CD38 for neurosyphilis. The results were as follows: the area under the curve (AUC) of cerebrospinal fluid CD38 was 0.8128, P = 0.0001, which was considered statistically significant, with a statistical critical value of 5.002NPX, a sensitivity of 80.00, and a specificity of 63.01; while the AUC of serum CD38 was 0.5365, P = 0.6571, which was considered statistically insignificant. Accordingly, the occurrence of neurosyphilis can be better evaluated according to the level of CD38 in the cerebrospinal fluid of syphilis patients, and the sensitivity is very high.

[0042] Furthermore, in order to clarify the relationship between serum and cerebrospinal fluid CD38 and symptomatic neurosyphilis, the study used ROC to evaluate the diagnostic potential of cerebrospinal fluid CD38 for symptomatic neurosyphilis in 73 patients with neurosyphilis (30 cases of asymptomatic neurosyphilis and 43 cases of paralytic dementia). The results were as follows: the AUC of cerebrospinal fluid CD38 was 0.6419, P = 0.0402, which was considered statistically significant, with a statistical critical value of 5.400NPX, a sensitivity of 66.67, and a specificity of 55.81; the AUC of serum CD38 was 0.6434, P = 0.0381, which was considered statistically significant, with a statistical critical value of 3.833NPX, a sensitivity of 70.00, and a specificity of 60.47. The above results show that the levels of cerebrospinal fluid and serum CD38 in patients with neurosyphilis can be used to diagnose the occurrence of symptomatic neurosyphilis.

[0043] In addition, screening for asymptomatic neurosyphilis is a difficult point in clinical diagnosis and treatment. It is difficult to diagnose without clinical symptoms, so all syphilis patients need to be screened. In order to clarify the relationship between serum and cerebrospinal fluid CD38 and asymptomatic neurosyphilis, 45 patients with latent syphilis and asymptomatic neurosyphilis (15 cases of latent syphilis and 30 cases of asymptomatic neurosyphilis) were studied in the following examples, and ROC was used to evaluate the diagnostic potential of CD38 for the occurrence of asymptomatic neurosyphilis. The results of the study were: the AUC of cerebrospinal fluid CD38 was 0.7511, P = 0.0065, which was considered statistically significant, the statistical critical value was 4.723NPX, the sensitivity was 73.33, and the specificity was 66.67; the AUC of serum CD38 was 0.5422, P = 0.6473, which was considered not statistically significant. The above results indicate that the occurrence of asymptomatic neurosyphilis can be better evaluated based on the level of CD38 in the cerebrospinal fluid of syphilis patients, while serum CD38 cannot be used for diagnosis for the time being and requires verification with more samples.

[0044] In summary, all patients were divided into two groups according to the patient's cerebrospinal fluid CD38 Cut-Off value (5.002NPX). Patients with cerebrospinal fluid damage-related indicators such as cerebrospinal fluid protein and leukocytes increased significantly above this value. It was further found that when the CD38 cerebrospinal fluid was lower than 5.400NPX, the risk of symptomatic neurosyphilis in neurosyphilis patients was lower, and the risk of brain damage was lower; while above this value, the risk of symptomatic neurosyphilis was higher, and the risk of brain damage increased. Therefore, the specific cerebrospinal fluid Cut-off value for the diagnosis of neurosyphilis is 5.002NPX, the cerebrospinal fluid Cut-off value for symptomatic neurosyphilis is 5.400NPX, and the Cut-off value for the diagnosis of non-neurosyphilis or latent syphilis is 4.723NPX. When the cerebrospinal fluid CD38 is lower than this value, it indicates a lower risk of neurosyphilis.

[0045] Based on the above research results, the present invention provides the application of CD38 as a cerebrospinal fluid marker in the early diagnosis of neurosyphilis and the assessment of nerve damage. Specifically, the level of CD38 protein in the cerebrospinal fluid of syphilis patients is used as a reliable and effective primary reference indicator or auxiliary reference indicator for the early diagnosis, typing and nerve damage assessment of syphilis. In medical practice, by detecting the expression level of CD38 in the cerebrospinal fluid and comparing it with the statistical critical value, it is possible to achieve high-sensitivity detection and typing of syphilis, especially neurosyphilis, and accurately quantify the patient's nerve damage, supervision and treatment.

[0046] In order to better understand the above technical solution, exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to enable a clearer and more thorough understanding of the present invention and to fully convey the scope of the present invention to those skilled in the art.

[0047] The experimental methods in the following experimental examples are conventional methods unless otherwise specified. The raw materials and reagents used in the following examples are commercially available products unless otherwise specified. Related technical contents include:

[0048] 1. Research subjects

[0049] With informed consent and meeting the inclusion criteria, 88 cases were selected for inclusion, including 15 cases of latent syphilis, 30 cases of asymptomatic neurosyphilis and 43 cases of paralytic dementia, to establish a neurosyphilis cohort of different severity. The diagnostic criteria were implemented according to the WS273-2007 syphilis diagnostic criteria.

[0050] 2. Sample collection

[0051] According to the conditions of the enrolled patients, lumbar puncture and cubital venipuncture were performed to obtain cerebrospinal fluid and blood for routine tests, and cerebrospinal fluid and serum samples were collected. With the informed consent of the patients, 5 mL of cerebrospinal fluid was collected by lumbar puncture, and 10 mL of blood samples were collected by vein. After the cerebrospinal fluid was collected by lumbar puncture, it was packaged in cryopreservation tubes and coded and labeled; after the blood was collected by venipuncture, it was placed in a 4°C centrifuge and centrifuged for 10 minutes at 2000 rpm to quickly separate the upper serum and place it in a cryopreservation tube.

[0052] 3. Detection of CD38 concentration in serum and cerebrospinal fluid using the Olink method

[0053] Olink neurology panel (95801) was used to detect 92 proteins related to neurological function, including CD38 protein. Olink precision proteomics is a precision proteomics research tool that is very suitable for large-scale screening, with good sensitivity, rapidity, high-throughput analysis, and specificity at the multiple channel level. Therefore, this example uses Olink to detect markers in the serum and cerebrospinal fluid of patients with different types of neurosyphilis, and it is found that CD38 protein has significant differences in cerebrospinal fluid. The detection of cerebrospinal fluid proteins and cerebrospinal fluid leukocytes was carried out according to the method described in WS 273-2007.

[0054] Statistics and analysis of experimental results:

[0055] (1) CD38 protein significantly increases in cerebrospinal fluid and serum of patients with neurosyphilis

[0056] Olink precision proteomics was used to detect the differences in the expression of nerve-related proteins in serum and cerebrospinal fluid, and CD38 protein with significant differences in cerebrospinal fluid was screened. Figure 1 As shown in Table 1, the level of CD38 in the cerebrospinal fluid of patients with symptomatic neurosyphilis (symptomatic) and asymptomatic neurosyphilis (asymptomatic) was significantly higher than that of patients with latent syphilis. Therefore, the level of CD38 in the cerebrospinal fluid can be used to categorize syphilis and indicate its severity ( Figure 1 ).Depend on Figure 1 As can be seen from Figure (a), the CD38 concentration in the cerebrospinal fluid of patients with symptomatic neurosyphilis>the CD38 concentration in the cerebrospinal fluid of patients with asymptomatic neurosyphilis>the CD38 concentration in the cerebrospinal fluid of patients with latent syphilis; Figure 1 (b) shows that unlike the situation in cerebrospinal fluid, there is no significant difference in the CD38 concentration in the serum of patients with neurosyphilis (symptomatic and asymptomatic) and that in the serum of patients with latent syphilis. Only the serum CD38 concentration between asymptomatic and symptomatic neurosyphilis patients is different.

[0057] Table 1: Statistical data of the difference in CD38 expression in cerebrospinal fluid and serum among three different types of syphilis

[0058] Table 1 shows the statistical data of the difference in CD38 expression in cerebrospinal fluid and serum in three different types of syphilis. Among them, cerebrospinal fluid, latent syphilis & asymptomatic represent the statistical values ​​of the difference in CD38 concentration between latent syphilis and asymptomatic neurosyphilis in cerebrospinal fluid. The mean difference reflects the average difference between each marker value and the arithmetic mean. The difference P value can show whether there is a significant difference between the two groups, and the difference multiple can show the multiple of this difference. It can be seen from the statistics in Table 1 that there is a significant difference in the cerebrospinal fluid CD38 protein level between patients with neurosyphilis and patients with latent syphilis, and the difference is statistically significant.

[0059] (2) There is no significant positive correlation between cerebrospinal fluid CD38 protein and serum CD38 protein

[0060] Figure 2 The results showed that the CD38 protein in serum was positively correlated with that in cerebrospinal fluid, but the correlation was not significant (R=0.1579, p=0.1418). Therefore, the CD38 protein in serum cannot accurately indicate the increase of CD38 protein in cerebrospinal fluid.

[0061] (3) CSF CD38 protein is significantly positively correlated with brain injury indicators

[0062] Combination Figure 3From (a) and (b), we can see that the expression level of CD38 protein in cerebrospinal fluid is significantly positively correlated with the protein in the cerebrospinal fluid (R=0.4704, p<0.0001) and the number of leukocytes (R=0.4322, p<0.0001). Therefore, CD38 protein in cerebrospinal fluid can indicate the abnormality of protein and leukocytes in cerebrospinal fluid. In practice, cerebrospinal fluid cell count and / or protein level can be used as a reference indicator, together with the CD38 protein level in cerebrospinal fluid, as a means of early diagnosis and screening of neurosyphilis, typing detection and quantitative assessment of the degree of nerve damage.

[0063] (4) There is no correlation between serum CD38 and brain injury indicators

[0064] Combination Figure 4 From (a) and (b), we can see that the expression level of CD38 protein in serum has no significant correlation with the protein in the patient's cerebrospinal fluid (R=0.2639, p=0.0130) and the number of white blood cells (R=-0.02137, p=0.8433). Therefore, CD38 protein in serum is different from CD38 protein in cerebrospinal fluid, and CD38 protein in serum cannot indicate abnormalities in protein and white blood cells in cerebrospinal fluid.

[0065] (6) Diagnostic role of cerebrospinal fluid CD38 and serum CD38 in neurosyphilis detection

[0066] In order to further clarify the relationship between cerebrospinal fluid CD38 and serum CD38 and neurosyphilis, this example studied 88 patients with syphilis. First, according to the level of CD38 in the patient's cerebrospinal fluid, the 88 patients were divided into two groups using Cut-Off (critical value = 5.002NPX). Then, the receiver operating characteristic curve (ROC) was used to evaluate the diagnostic potential of CD38 for neurosyphilis. The results showed that the area under the curve (AUC) of cerebrospinal fluid CD38 (0.8128 [95%CI 0.7082-0.9174]), P = 0.0001, was considered statistically significant, with a sensitivity of 80.00 and a specificity of 63.01 (see Figure 5 (a)). Then, according to the level of CD38 in the patients' cerebrospinal fluid, the 88 patients were divided into two groups using Cut-Off (critical value = 3.779NPX). The results showed that the area under the curve (AUC) of serum CD38 (0.5365 [95% CI 0.3739-0.6992]), P = 0.6571, was considered not statistically significant (see Figure 5 (b)). The above results show that the diagnosis of neurosyphilis can be well achieved based on the level of CD38 in the cerebrospinal fluid of syphilis patients with high sensitivity.

[0067] In addition to the above-mentioned role in detecting neurosyphilis, the level of cerebrospinal fluid CD38 can also be used to diagnose symptomatic neurosyphilis in neurosyphilis. In order to further clarify the relationship between serum CD38 and cerebrospinal fluid CD38 and symptomatic neurosyphilis, this example studied 73 patients with neurosyphilis. According to the patient's cerebrospinal fluid CD38 Cut-Off value (critical value = 5.400NPX) and serum CD38 Cut-Off value (critical value = 3.833NPX), these 73 patients were divided into two groups with high CD38 concentration and low CD38 concentration. ROC was used to evaluate the diagnostic potential of CD38 for symptomatic neurosyphilis in patients with neurosyphilis (including symptomatic and asymptomatic neurosyphilis patients). The results showed that the area under the curve (AUC) of cerebrospinal fluid CD38 (0.6419 [95% CI 0.5102-0.7736]), P = 0.0402, was considered statistically significant (see Figure 6 (a)); the area under the curve (AUC) of serum CD38 (0.6434 [95% CI 0.5174-0.7695]), P = 0.0381, was also considered statistically significant (see Figure 6 The above results indicate that the occurrence of symptomatic neurosyphilis can be well diagnosed based on the level of CD38 in the serum and cerebrospinal fluid of patients with neurosyphilis.

[0068] In order to further clarify the relationship between serum CD38 and cerebrospinal fluid CD38 and asymptomatic neurosyphilis, this example studied 45 syphilis patients (15 latent syphilis patients and 30 asymptomatic neurosyphilis). According to the cut-off value of cerebrospinal fluid CD38 (critical value = 4.723NPX) and serum CD38 cut-off value (critical value = 3.754NPX), the 45 patients were divided into two groups with high CD38 concentration and low CD38 concentration, and ROC was used to evaluate the diagnostic potential of CD38 in the above 45 syphilis patients. Figure 7 As shown in the figure, the experimental results were: the area under the curve of cerebrospinal fluid CD38 AUC (0.7511 [95% CI 0.6050-0.8972]), P = 0.0065, which was considered statistically significant (see Figure 7 (a)); the area under the curve of serum CD38 AUC (0.5422 [95% CI 0.3492-0.7352]), P = 0.6473, no significant significance (see Figure 7 The above results show that the occurrence of asymptomatic neurosyphilis can be diagnosed based on the level of CD38 in the cerebrospinal fluid of patients with neurosyphilis, with a sensitivity of 73.33.

[0069] (7) CSF CD38 levels can indicate the degree of brain damage

[0070] All patients were divided into two groups according to the cut-off value of the patient's cerebrospinal fluid CD38 level (5.002NPX), among which the cerebrospinal fluid protein of patients with cerebrospinal fluid CD38 higher than 5.002NPX (see Figure 8 (a)) and a significant increase in white blood cells (see Figure 8 Therefore, patients with CSF CD38 values ​​below 5.002NPX have a lower risk of neurosyphilis and a lower risk of brain damage, while those with CSF CD38 values ​​above this value have a higher risk of neurosyphilis and an increased risk of brain damage.

[0071] It was further found that when CSF CD38 ≥ 5.400NPX, CSF protein increased (see Fig. 9 (a)), but the increase in white blood cells is not significant (see Fig. 9 (b) At this time, the patient has a higher risk of developing symptomatic neurosyphilis and a further increased risk of brain damage. When CSF CD38 is lower than 5.400NPX, the risk of neurosyphilis patients developing symptomatic neurosyphilis is relatively low, and the risk of brain damage is small.

[0072] In summary, the concentration of CD38 protein in cerebrospinal fluid can be used as a biological indicator for early diagnosis of neurosyphilis, typing of symptomatic neurosyphilis, asymptomatic neurosyphilis, neurosyphilis and latent syphilis, and evaluation of the degree of nerve damage. It can be used alone or as a main reference indicator in combination with other indicators to complete the diagnostic screening, typing and nerve damage evaluation of syphilis.

[0073] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the embodiments, it should be understood by those skilled in the art that any modification or equivalent replacement of the technical solutions of the present invention does not depart from the spirit and scope of the technical solutions of the present invention and should be included in the scope of the claims of the present invention.

Claims

1. The use of CD38 protein and its specific antibody in the preparation of diagnostic products for screening, typing or evaluating the degree of nerve damage in patients with neurosyphilis; characterized in that: The diagnostic product is a detection kit, antibody chip, antibody probe or detector with CD38 protein or its specific antibody as the target molecule; the typing is symptomatic neurosyphilis, asymptomatic neurosyphilis and latent syphilis; the detection sample is cerebrospinal fluid.

2. The use according to claim 1, characterized in that: The kit uses CD38 protein or its specific antibody as a detection target.

3. The use according to claim 1, characterized in that: The kit is an enzyme-linked immunosorbent assay kit or an immunohistochemistry kit.

4. The use according to claim 1, characterized in that: The kit comprises: a primary antibody capable of specifically recognizing CD38, a polypeptide dye, magnetic beads and a biotin-labeled secondary antibody.

5. The use according to claim 4, characterized in that: The reagents include citrate buffer, PBS washing solution, peroxidase blocking agent, goat serum, streptomycin antibiotic-peroxidase, DAB solution and hematoxylin solution.

6. The use according to claim 1, characterized in that: The kit also includes reagents or materials for obtaining cerebrospinal fluid from the neurosyphilis patient.

7. A system for assessing the risk of nerve damage in patients with neurosyphilis, characterized in that: It includes a detection module and a judgment module; wherein the detection module is used to obtain the concentration value of the cerebrospinal fluid CD38 protein; the judgment module includes a readable carrier storing the judgment rules; The judgment rule is: when the cerebrospinal fluid CD38 concentration value is less than 5.002NPX value, the risk of neurosyphilis is low, which is a low risk of brain damage; When 5.400NPX>cerebrospinal fluid CD38 concentration value ≥5.002NPX value, it is judged that the risk of neurosyphilis is high and the risk of neurological damage is moderate; When the cerebrospinal fluid CD38 concentration value is ≥5.400NPX, it is judged that the risk of symptomatic neurosyphilis is high and the risk of nerve damage is high.

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