Application of reagent for detecting S100A4 in preparation of pancreatitis diagnosis tool

Abstract

The invention discloses application of a reagent for detecting S100A4 in preparation of a pancreatitis diagnosis tool, the S100A4 serving as a biomarker for human pancreatitis diagnosis can be used for detecting the content of the S100A4 through the diagnosis tool for detecting the blood S100A4 protein level, the S100A4 is used for distinguishing and diagnosing acute pancreatitis (AP) and chronic pancreatitis (CP), the accuracy is high, patients can be found in the early stage, and the clinical application prospect is wide. The patient prognosis is improved. As a biomarker, the S100A4 provides more accurate auxiliary capability and better support for clinical medicine judgment of acute pancreatitis and chronic pancreatitis.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical biotechnology, and in particular to application of a reagent for detecting S100A4 in preparing a diagnostic tool for pancreatitis. Background Art

[0002] Pancreatitis is an inflammatory disease caused by abnormal activation of pancreatic digestive enzymes, which digests the pancreatic tissue itself. The condition can be acute or chronic. Acute pancreatitis (AP) usually has an acute onset, and patients will feel severe abdominal pain, which is often located in the left upper abdomen or the entire abdomen, and is often accompanied by symptoms such as nausea, vomiting, and fever. Patients with mild pancreatitis usually have a good prognosis after timely treatment, while patients with severe pancreatitis may face a higher risk of complications and mortality. Chronic pancreatitis (CP) is a multifactorial fibro-inflammatory syndrome. Recurrent pancreatic inflammation leads to extensive fibrotic tissue replacement, resulting in chronic pain, pancreatic exocrine and endocrine insufficiency, decreased quality of life, and shortened life expectancy. The occurrence of pancreatitis is related to a variety of factors, such as biliary disease, alcohol intake, hyperlipidemia, drug reactions, and certain metabolic abnormalities. The key to treating pancreatitis is early diagnosis and timely treatment to alleviate symptoms, prevent complications, and restore the normal function of the pancreas.

[0003] The diagnosis of pancreatitis mainly relies on clinical manifestations, laboratory tests and imaging examinations. The clinical symptoms may be similar to other diseases, such as gastrointestinal diseases and biliary diseases, which makes diagnosis difficult. Among them, AP blood tests usually show elevated levels of amylase and lipase, but 19% to 30% of AP patients still have normal blood amylase, especially in the early stage of the disease, the index is not sensitive, and blood amylase can also be increased due to other intestinal acute abdomen, lung, uterus and lactating breast and other organ lesions, and the specificity is not enough; imaging examinations, such as CT scans or MRI, are helpful to observe the shape, size and surrounding conditions of the pancreas, but the sensitivity of early diagnosis is insufficient; traditional examination methods such as ultrasound examinations may also be interfered with, affecting the accuracy of diagnosis. At the same time, clear CP can only be diagnosed through clinical symptoms and imaging examinations. Indicators such as blood amylase and lipase cannot be used to diagnose CP. The gold standard for diagnosing CP is to obtain tissue samples through pancreatic puncture biopsy or surgery for pathological examination. At present, there is a lack of effective blood markers for diagnosis in clinical practice. Therefore, the diagnosis and treatment of pancreatitis urgently need the discovery and application of new biomarkers.

[0004] Biomarkers refer to certain quantifiable or observable substances or phenomena in organisms, which can be used as indicators in drug treatment, prevention, diagnosis, screening, and monitoring of diseases. With the deepening of biological and medical research, more and more biomarkers have been discovered and applied in clinical practice as a non-invasive detection method.

[0005] At present, S100A4 has been used in the diagnosis of many diseases. Studies have shown that S100A4 plays an important role in the process of tumor metastasis, fibrosis and other diseases. However, the application of S100A4 in the diagnosis of pancreatitis has not been reported. The present invention has discovered for the first time that S100A4 can be used as a specific biomarker for the diagnosis of pancreatitis. Summary of the invention

[0006] The purpose of the present invention is to solve the technical problems existing in the above-mentioned pancreatitis diagnostic tools and disclose the use of a reagent for detecting S100A4 in the preparation of a pancreatitis diagnostic tool.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the use of a reagent for detecting S100A4 in preparing a diagnostic tool for pancreatitis.

[0008] Optionally, the reagent for detecting S100A4 is a reagent used for detecting the content of S100A4 based on enzyme-linked immunosorbent assay, Western Blot or immunofluorescence method.

[0009] Optionally, the diagnostic tool includes a kit, a chip or a test strip for detecting the content of S100A4 in the blood of a subject.

[0010] Optionally, the diagnostic tool comprises a reagent for detecting S100A4, wherein the reagent comprises: a microplate coated with an anti-S100A4 monoclonal antibody, S100A4 capture antibody 10X, S100A4 detection antibody 10X, S100A4 lyophilized recombinant protein, antibody diluent, sample diluent and TMB substrate solution.

[0011] Optionally, the pancreatitis is acute pancreatitis or chronic pancreatitis.

[0012] Optionally, the diagnostic reference value of the diagnostic tool is: for acute pancreatitis, the diagnostic reference value is less than 293.04 ng / ml; for chronic pancreatitis, the diagnostic reference value is greater than 870.64 ng / ml.

[0013] Optionally, the diagnostic tool can also be used to differentiate acute pancreatitis from chronic pancreatitis, with the cutoff value for differentiation being 572.92 ng / ml.

[0014] The present invention also provides a S100A4 detection tool, comprising a microplate coated with an anti-S100A4 monoclonal antibody, a S100A4 capture antibody 10X, a S100A4 detection antibody 10X, a S100A4 freeze-dried recombinant protein, an antibody diluent, a sample diluent and a TMB substrate solution.

[0015] The present invention also provides a system for detecting the S100A4 content in blood to diagnose pancreatitis, the system comprising:

[0016] A data collection module, used for collecting the S100A4 protein expression data in the blood sample of the subject and transmitting it to the data processing module;

[0017] A data processing module, used to draw a standard curve for the received blood S100A4 expression data and convert the concentration value;

[0018] The analysis module is used to perform statistical analysis and ROC curve analysis on the processed data and calculate the area under the curve, sensitivity and specificity;

[0019] The judgment and output module is used to judge whether the data meets the preset judgment conditions; when the blood S100A4 concentration is less than 293.04 ng / ml, it is judged as acute pancreatitis; when the blood S100A4 concentration is greater than 870.64 ng / ml, it is judged as chronic pancreatitis; when used for identification, when the blood S100A4 concentration is greater than 572.92 ng / ml, it is judged as chronic pancreatitis.

[0020] The present invention also provides a computer-aided method for diagnosing pancreatitis by detecting the S100A4 content in blood, comprising the following steps:

[0021] Receive S100A4 expression data in blood, draw a standard curve and convert concentration values;

[0022] Statistical analysis and ROC curve analysis were performed on the processed data to calculate the area under the curve, sensitivity, and specificity;

[0023] Determine whether the data meets the preset determination conditions;

[0024] When the blood S100A4 concentration is less than 293.04 ng / ml, it is diagnosed as acute pancreatitis;

[0025] When the blood S100A4 concentration is greater than 870.64 ng / ml, it is diagnosed as chronic pancreatitis;

[0026] When used for identification, blood S100A4 concentration greater than 572.92 ng / ml is diagnosed as chronic pancreatitis.

[0027] The present invention also provides a computer-readable storage medium storing a computer program. When the computer program is executed by a processor, the functions of the above-mentioned system or the steps of the above-mentioned method can be realized.

[0028] The present invention also provides an electronic device, comprising a memory and a processor, wherein the memory stores a computer program, and the processor is used to execute the computer program to implement the functions of the above-mentioned system, or to implement the steps of the above-mentioned method.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The present invention provides the use of a reagent for detecting S100A4 in the preparation of a pancreatitis diagnostic tool, and a S100A4 detection tool, which can provide a new blood biomarker for pancreatitis diagnosis, and as a non-invasive detection method, can complete the determination of the type of pancreatitis through a simple blood test, and compared with traditional diagnostic methods, it has the advantages of simple operation, rapid detection, and less trauma to patients. At the same time, the computer system and diagnostic method developed by the present invention realize the standardization of the pancreatitis diagnosis process by automatically processing blood S100A4 expression data, statistical analysis and diagnostic determination, and significantly improve the detection efficiency.

[0031] The diagnostic tool of the present invention sets specific diagnostic reference values ​​for different types of pancreatitis. When the blood S100A4 concentration is less than 293.04 ng / ml, it can be accurately determined as acute pancreatitis, with a diagnostic sensitivity of 59.1% and a specificity of 86.1%; when the blood S100A4 concentration is greater than 870.64 ng / ml, it can be accurately determined as chronic pancreatitis, with a diagnostic sensitivity of 55.7% and a specificity of 94.4%. More importantly, the present invention also establishes a critical value of 572.92 ng / ml for distinguishing acute pancreatitis from chronic pancreatitis. Through clear judgment conditions and diagnostic reference values, the accuracy and objectivity of the diagnostic results are guaranteed, providing a reliable auxiliary diagnostic tool for clinical diagnosis and treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1A This is a graph showing a comparison of blood S100A4 levels between a control group and patients with acute pancreatitis in an embodiment of the present invention;

[0033] Figure 1B This is a graph showing the characteristic results of analyzing blood S100A4 as a biomarker for acute pancreatitis through AUC curve in an embodiment of the present invention;

[0034] Figure 1C This is a diagram showing the analysis of the differences between S100A4 in different AP periods in an embodiment of the present invention;

[0035] Figure 2A This is a graph showing a comparison of blood S100A4 levels between a control group and patients with chronic pancreatitis in an embodiment of the present invention;

[0036] Figure 2B This is a graph showing the characteristic results of analyzing blood S100A4 as a pancreatic disease biomarker through AUC curve in an embodiment of the present invention;

[0037] Figure 2C This is a diagram showing the difference between S100A4 in different CP periods according to an embodiment of the present invention;

[0038] Figure 3A This is a comparative display of blood S100A4 levels of patients with acute pancreatitis and chronic pancreatitis in an embodiment of the present invention;

[0039] Figure 3B This is a graph showing the results of analyzing blood S100A4 using the AUC curve to distinguish acute pancreatitis from chronic pancreatitis in an embodiment of the present invention. DETAILED DESCRIPTION

[0040] The application of the reagent for detecting S100A4 of the present invention in preparing a pancreatitis diagnostic tool will be described in more detail below in conjunction with a schematic diagram, wherein a preferred embodiment of the present invention is shown, and it should be understood that those skilled in the art can modify the present invention described herein while still achieving the beneficial effects of the present invention. Therefore, the following description should be understood as being widely known to those skilled in the art and not as a limitation of the present invention.

[0041] For the sake of clarity, not all features of the actual embodiments are described. In the following description, well-known functions and structures are not described in detail because they would clutter the invention with unnecessary detail. It should be recognized that in the development of any actual embodiment, a large number of implementation details must be made to achieve the developer's specific goals, such as changing from one embodiment to another according to the limitations of the relevant system or the relevant business. In addition, it should be recognized that such development work may be complex and time-consuming, but it is just a routine task for those skilled in the art.

[0042] The present invention is described in more detail in the following paragraphs by way of example with reference to the accompanying drawings. The advantages and features of the present invention will become more apparent from the following description. It should be noted that the accompanying drawings are in very simplified form and are not in exact proportions, and are only used to facilitate and clearly assist in explaining the purpose of the embodiments of the present invention.

[0043] S100A4 is a member of the S100 calcium-binding protein family and is encoded by the S100A4 gene. S100A4 is mainly involved in angiogenesis, regulating cell apoptosis, promoting cell movement and invasion, and participating in inflammatory responses, and plays an important role in the occurrence and development of the disease. The present invention found that the level of S100A4 in the blood of AP patients was significantly reduced, and the level in the blood of CP patients was significantly increased; S100A4, as a biomarker for the diagnosis of pancreatitis, not only has the significance of assisting the clinical diagnosis of AP and CP, but also can efficiently distinguish AP and CP.

[0044] The reagent for detecting S100A4 is a reagent used for detecting the content of S100A4 based on enzyme-linked immunosorbent assay, Western Blot or immunofluorescence method.

[0045] The present invention also provides a diagnostic tool, including a test kit, a chip or a test paper, for detecting the content of S100A4 in the blood of a subject.

[0046] The diagnostic tool comprises a reagent for detecting S100A4, wherein the reagent comprises: a microplate coated with an anti-S100A4 monoclonal antibody, S100A4 capture antibody 10X, S100A4 detection antibody 10X, S100A4 lyophilized recombinant protein, an antibody diluent, a sample diluent and a TMB substrate solution.

[0047] Furthermore, the reagents also include a stop solution and a washing buffer.

[0048] The diagnostic reference value of the diagnostic tool is: for acute pancreatitis, the diagnostic reference value is less than 293.04 ng / ml; for chronic pancreatitis, the diagnostic reference value is greater than 870.64 ng / ml.

[0049] The diagnostic tool can also be used to differentiate acute pancreatitis from chronic pancreatitis, with the cutoff value for differentiation being 572.92 ng / ml.

[0050] The present invention also provides an S100A4 detection tool for detecting the S100A4 content in blood, comprising a microplate coated with an anti-S100A4 monoclonal antibody, a S100A4 capture antibody 10X, a S100A4 detection antibody 10X, a S100A4 freeze-dried recombinant protein, an antibody diluent, a sample diluent and a TMB substrate solution.

[0051] Furthermore, the detection tool also includes a stop solution and a washing buffer solution.

[0052] The detection tool includes: a test kit, a test paper or a chip.

[0053] The present invention finds that S100A4 can be used as a biomarker for diagnosing pancreatitis, establishes a specific diagnostic reference value range, and achieves effective differentiation between AP and CP.

[0054] Furthermore, the present invention also provides a computer-aided method for diagnosing pancreatitis by detecting the S100A4 content in the blood, comprising the following steps:

[0055] S1. Receive blood S100A4 expression data, draw a standard curve and convert concentration values;

[0056] S2. Perform statistical analysis and ROC curve analysis on the processed data and calculate the area under the curve, sensitivity and specificity;

[0057] S3. Determine whether the data meets preset determination conditions.

[0058] in:

[0059] S31. When the blood S100A4 concentration is less than 293.04 ng / ml, it is diagnosed as acute pancreatitis;

[0060] S32. When the blood S100A4 concentration is greater than 870.64 ng / ml, it is diagnosed as chronic pancreatitis;

[0061] S33. When used for identification, blood S100A4 concentration greater than 572.92 ng / ml is diagnosed as chronic pancreatitis.

[0062] Furthermore, the present invention provides a system for detecting the content of S100A4 in blood to diagnose pancreatitis. The system mainly includes a data collection module, a data processing module, an analysis module and a judgment output module, and each module is connected through a data bus.

[0063] The data collection module is used to collect the S100A4 protein expression data in the blood sample of the subject. Specifically, the module can receive the expression data output by the detection device through the data interface, or read the stored detection data file through the data import function, and transmit the data to the data processing module. The data input method can include but is not limited to: automatic collection, file import, manual entry, etc.

[0064] After receiving the S100A4 expression data, the data processing module processes the data according to the following steps:

[0065] Calculate the average absorbance value of the blank control (zero) standard;

[0066] The average blank control value was subtracted from all sample absorbance values;

[0067] A standard curve was drawn with the standard protein concentration as the abscissa (x-axis) and the corresponding corrected absorbance value as the ordinate (y-axis);

[0068] The S100A4 concentration of the sample to be tested was converted according to the standard curve.

[0069] It is worth noting that samples whose absorbance values ​​exceed the range of the standard curve need to be appropriately diluted and retested: when the absorbance value is greater than the highest standard value, it should be further diluted; when the absorbance value is lower than the lowest standard value, the dilution factor should be reduced and retested.

[0070] The analysis module uses SPSS software (IBM 25.0) to perform statistical analysis and ROC curve analysis on the processed data.

[0071] Specifically include:

[0072] Descriptive statistics were performed, and mean ± standard deviation (SD) or median with interquartile range (IQR) were calculated;

[0073] One-way ANOVA or Kruskal-Wallis test was performed according to the data distribution;

[0074] ROC curve analysis was performed to calculate the area under the curve, sensitivity, and specificity.

[0075] The judgment output module judges the analysis result according to the preset judgment conditions.

[0076] Specifically:

[0077] When the blood S100A4 concentration is less than 293.04 ng / ml, it is diagnosed as acute pancreatitis;

[0078] When the blood S100A4 concentration is greater than 870.64 ng / ml, it is diagnosed as chronic pancreatitis;

[0079] When it is necessary to distinguish between acute and chronic pancreatitis, 572.92 ng / ml is used as the critical value. If it is greater than this value, it is diagnosed as chronic pancreatitis.

[0080] The judgment output module can generate a standardized diagnosis report, the content of which includes but is not limited to: basic information of the patient, test data, analysis results, diagnosis conclusions, etc. The report output method can include screen display, print output or data export and other forms.

[0081] The system ensures the accuracy of the diagnosis results through standardized data processing and analysis processes; the system uses clear diagnostic reference values ​​to make the judgment of acute and chronic pancreatitis more objective and reliable; through automated data processing and analysis, it significantly improves work efficiency and reduces human errors. It has been verified that the system has a diagnostic sensitivity of 59.1% and a specificity of 86.1% for acute pancreatitis; and a diagnostic sensitivity of 55.7% and a specificity of 94.4% for chronic pancreatitis.

[0082] Furthermore, the present invention verifies the effect of S100A4 as a biomarker through the following experiments: The methods in the examples are conventional methods unless otherwise specified, and the reagents used are conventional commercial reagents or reagents prepared according to conventional methods unless otherwise specified.

[0083] Human S100A4 SimpleStep used in the following examples The composition of the Kit (ab283547, Abcam (Shanghai) Trading Co., Ltd.) is: a microplate coated with anti-S100A4 monoclonal antibody, S100A4 capture antibody 10X, S100A4 detection antibody 10X, S100A4 lyophilized recombinant protein, antibody diluent CPI2, sample diluent 25BS, sample diluent NS, TMB substrate solution, stop solution and washing buffer.

[0084] Sample requirements: Collect plasma samples from healthy controls and pancreatitis patients (including acute pancreatitis and chronic pancreatitis).

[0085] Steps for detecting S100A4:

[0086] a. Reagent preparation: All reagents are equilibrated to room temperature for at least 30 minutes. Standard working solution, antibody mixture, and working wash solution are prepared as required.

[0087] b. Sample processing: Plasma samples were diluted at a ratio of 1:150.

[0088] c. Operation steps:

[0089] (1) Sample addition: 50 μl / well of the diluted sample and standard were added to the wells of the plate in sequence; then 50 μl of the antibody mixture was added to the wells containing the calibrator and sample, mixed well, covered with a sealing film, and incubated on a shaker at room temperature at 400 rpm for 1 hour.

[0090] (2) Washing: Discard the liquid in the wells, add 350 μl of 1X washing solution to each well, soak for at least 10 seconds, aspirate or pour out all the washing solution, repeat 3 times, and after the last wash, thoroughly pat dry the residual liquid in the ELISA wells on absorbent paper.

[0091] (3) Color development: Add 100 μl / well TMB substrate solution to each well, cover with a sealing film, and incubate in the dark on a shaker at 400 rpm for 5 to 20 minutes.

[0092] (4) Stop: Add 100 μL of stop solution to each well and shake on a shaker for 1 minute to mix.

[0093] (5) Detection: Use an ELISA reader to measure the OD value of each well at a wavelength of 450 nm.

[0094] Data Analysis: Calculate the average absorbance value for the blank (zero) standards. Subtract the average blank standard absorbance value from all other absorbance values. Create a standard curve by plotting the average blank minus absorbance value for each standard concentration (y-axis) versus the target protein concentration (x-axis) of the standard protein. Use plotting software to draw the best smooth curve through these points to construct the standard curve. Samples that produce absorbance values ​​greater than the highest standard value should be further diluted and reanalyzed.

[0095] Likewise, samples that yield absorbance values ​​below the lowest standard should be retested at a lower dilution.

[0096] Statistical analysis: Statistical analysis was performed using SPSS software (25.0 from IBM). Continuous variables were expressed as mean ± standard deviation (SD) or median with interquartile range (IQR). One-way analysis of variance or Kruskal-Wallis test was used to compare multiple groups of continuous variables according to the data distribution. Chi-square test was used for analysis of categorical variables. The Spearman correlation coefficient was used to analyze the correlation between S100A4 levels and clinical parameters. Odds ratios (ORs) and their 95% confidence intervals (CIs) were calculated to quantify the strength of the association. The receiver operating characteristic curve (ROC) was used to analyze the diagnostic efficacy of S100A4. The area under the curve (AUC), sensitivity and specificity of the optimal cut-off point were obtained by analysis. p value < 0.05 was considered statistically significant.

[0097] Sample preparation: Plasma was collected from the First Affiliated Hospital of Naval Medical University.

[0098] Testing steps: Follow the above testing steps.

[0099] Result analysis:

[0100] 1) Baseline information:

[0101] A total of 297 participants were included in the experiment provided in this embodiment. The registration form of the participants is shown in Table 1. They were divided into three groups: a healthy control group (n=72), patients with acute pancreatitis (AP) (n=110), and patients with chronic pancreatitis (CP) (n=115).

[0102] The demographic and clinical characteristics of each group are shown in the table below.

[0103] Among them, there were 193 men and 104 women in the study, with a median age of 42 years (12-88 years), and a total of 72 patients with pancreatitis had diabetes.

[0104] Table 1

[0105]

[0106] 2) Plasma testing:

[0107] Compared with healthy controls, the plasma levels of patients with acute pancreatitis were significantly lower (p<0.001) (see Figure 1A ). The AUC of S100A4 as a biomarker for the diagnosis of acute pancreatitis alone was 0.752, with high accuracy (95% CI: 0.683-0.821, cutoff value <293.07ng / ml, sensitivity: 59.1%, specificity: 86.1%) (please refer to Figure 1B ).

[0108] According to the classification criteria of the "Guidelines for the Diagnosis and Treatment of Acute Pancreatitis in China (2021)", 49 cases were classified as mild, 31 cases were moderately severe, and 30 cases were severe. The analysis found that there was no significant difference in S100A4 in different stages of AP (P>0.05) (please refer to Figure 1C ).

[0109] Compared with the healthy control group, the plasma S100A4 level in patients with chronic pancreatitis was significantly increased (p<0.001) (see Figure 2A ). The AUC of S100A4 alone for diagnosing chronic pancreatitis was 0.807, with high accuracy (95% CI: 0.748-0.867, cutoff value>870.64ng / ml, sensitivity: 55.7%, specificity: 94.4%) (please refer to Figure 2B ).

[0110] According to the Guidelines for the Diagnosis and Treatment of Chronic Pancreatitis (Guangzhou, 2018), the disease course and clinical phenotype were staged: 10 cases in stage 0 (subclinical stage), 37 cases in stage 1 (no pancreatic insufficiency), 35 cases in stage 2 (partial pancreatic insufficiency), 20 cases in stage 3 (complete pancreatic insufficiency), and 13 cases in stage 4 (painless end stage). The analysis found that there was no significant difference in S100A4 in different stages of CP (P>0.05) (please refer to Figure 2C ).

[0111] 3) Differentiation and Identification:

[0112] Compared with the acute pancreatitis group, the plasma S100A4 level in patients with chronic pancreatitis was significantly increased (p<0.001) (see Figure 3A ). The AUC of S100A4 for identifying chronic pancreatitis was 0.915, with high accuracy (95% CI: 0.881-0.949, cutoff value>572.92ng / ml, sensitivity: 757%, specificity: 88.2%) (please refer to Figure 3B ).

[0113] An embodiment of the present invention further provides an electronic device, which may be a server, a computer or other device.

[0114] The electronic device may include: a processor, a storage medium and a bus, the storage medium stores machine-readable instructions executable by the processor, when the electronic device is running, the processor and the storage medium communicate through the bus, the processor executes the machine-readable instructions to execute the computer program to implement the functions of the system for detecting the blood S100A4 content to diagnose pancreatitis, or implement the steps of the computer-assisted method for diagnosing pancreatitis by detecting the blood S100A4 content described in the embodiment. The specific implementation method and technical effect are similar and will not be repeated here.

[0115] For ease of explanation, only one processor is described in the above electronic device. However, it should be noted that in some embodiments, the electronic device in the present invention may also include multiple processors, so the steps performed by one processor described in the present invention may also be performed jointly or individually by multiple processors. For example, if the processor of the electronic device executes step A and step B, it should be understood that step A and step B may also be performed jointly by two different processors or individually in one processor. For example, the first processor executes step A, the second processor executes step B, or the first processor and the second processor execute steps A and B together, etc.

[0116] In some embodiments, the processor may include one or more processing cores (e.g., a single-core processor (S) or a multi-core processor (S)). By way of example only, the processor may include a central processing unit (CPU), an application specific integrated circuit (ASIC), an application specific instruction set processor (ASIP), a graphics processing unit (GPU), a physical processing unit (PPU), a digital signal processor (DSP), a field programmable gate array (FPGA), a programmable logic device (PLD), a controller, a microcontroller unit, a reduced instruction set computer (RISC) or a microprocessor, or any combination thereof.

[0117] Accordingly, other embodiments of the present application may also provide a computer-readable storage medium in which computer executable instructions are stored, and when the computer executable instructions are executed by the processor, the various method embodiments of the present application are implemented. Computer-readable storage media include permanent and non-permanent, removable and non-removable media, and information storage can be implemented by any method or technology. Information can be a computer-readable instruction, a data structure, a module of a program, or other data. Examples of computer storage media include, but are not limited to, phase change memory (PRAM), static random access memory (SRAM), dynamic random access memory (DRAM), other types of random access memory (RAM), read-only memory (ROM), electrically erasable programmable read-only memory (EEPROM), flash memory or other memory technology, read-only compact disk read-only memory (CD-ROM), digital versatile disk (DVD) or other optical storage, magnetic cassettes, tape disk storage or other magnetic storage devices or any other non-transmission media that can be used to store information that can be accessed by a computing device.

[0118] In summary, the present invention discloses the use of a reagent for detecting S100A4 in the blood in the preparation of a diagnostic tool for pancreatitis. By detecting the level of S100A4 in the blood, the diagnostic tool can accurately distinguish acute pancreatitis from chronic pancreatitis, wherein the diagnostic sensitivity for acute pancreatitis is 59.1% and the specificity is 86.1%, and the diagnostic sensitivity for chronic pancreatitis is 55.7% and the specificity is 94.4%. As a new type of biomarker diagnostic method, the present invention breaks through the limitations of traditional diagnostic methods, not only can achieve early diagnosis of pancreatitis, but also can effectively distinguish acute from chronic pancreatitis, providing a more reliable diagnostic basis for clinical diagnosis and treatment, and has important clinical application value.

[0119] In addition, the system and method provided by the present invention for detecting the S100A4 content in the blood to diagnose pancreatitis, by automatically processing the blood S100A4 expression data, drawing a standard curve, performing statistical analysis and ROC curve analysis, and finally outputting the diagnosis result according to the preset judgment conditions. The present invention also provides a corresponding computer-aided diagnosis method, including the steps of data reception and processing, statistical analysis and result determination. Both the system and the method use clear diagnostic reference values. The system and the method realize the automation and standardization of the pancreatitis diagnosis process, significantly improve the diagnostic efficiency, ensure the accuracy and objectivity of the diagnostic results through statistical analysis and clear diagnostic reference values, and provide a reliable auxiliary diagnostic tool.

[0120] The above is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any technician in the relevant technical field, without departing from the scope of the technical solution of the present invention, makes any form of equivalent replacement or modification to the technical solution and technical content disclosed in the present invention, which does not depart from the content of the technical solution of the present invention and still falls within the protection scope of the present invention.

Claims

1. Application of a reagent for detecting S100A4 in the preparation of a diagnostic tool for pancreatitis.

2. The use according to claim 1, characterized in that: The reagent for detecting S100A4 is a reagent used for detecting the content of S100A4 based on enzyme-linked immunosorbent assay, Western Blot or immunofluorescence method.

3. The use according to claim 1, characterized in that: The diagnostic tool includes a test kit, a chip or a test paper, which is used to detect the content of S100A4 in the blood of a subject.

4. The use according to claim 1, characterized in that The diagnostic tool comprises a reagent for detecting S100A4, wherein the reagent comprises: a microplate coated with an anti-S100A4 monoclonal antibody, S100A4 capture antibody 10X, S100A4 detection antibody 10X, S100A4 lyophilized recombinant protein, an antibody diluent, a sample diluent and a TMB substrate solution.

5. The use according to any one of claims 1 to 4, characterized in that: The pancreatitis is acute pancreatitis or chronic pancreatitis.

6. The use according to claim 5, characterized in that The diagnostic reference value of the diagnostic tool is: for acute pancreatitis, the diagnostic reference value is less than 293.04 ng / ml; for chronic pancreatitis, the diagnostic reference value is greater than 870.64 ng / ml.

7. The use according to claim 5, characterized in that The diagnostic tool can also be used to differentiate acute pancreatitis from chronic pancreatitis, with the cutoff value for differentiation being 572.92 ng / ml.

8. A S100A4 detection tool, characterized in that: The invention comprises a microplate coated with anti-S100A4 monoclonal antibody, S100A4 capture antibody 10X, S100A4 detection antibody 10X, S100A4 lyophilized recombinant protein, antibody diluent, sample diluent and TMB substrate solution.

9. A system for detecting blood S100A4 content to diagnose pancreatitis, characterized in that: The system comprises: A data collection module, used for collecting S100A4 protein expression data in the blood samples of the subjects and transmitting the data to the data processing module; A data processing module, used to draw a standard curve for the received blood S100A4 expression data and convert the concentration value; The analysis module is used to perform statistical analysis and ROC curve analysis on the processed data and calculate the area under the curve, sensitivity and specificity; The judgment and output module is used to judge whether the data meets the preset judgment conditions; when the blood S100A4 concentration is less than 293.04 ng / ml, it is judged as acute pancreatitis; when the blood S100A4 concentration is greater than 870.64 ng / ml, it is judged as chronic pancreatitis; when used for identification, when the blood S100A4 concentration is greater than 572.92 ng / ml, it is judged as chronic pancreatitis.

10. A computer-aided method for diagnosing pancreatitis by detecting blood S100A4 content, characterized in that: The following steps are involved: Receive blood S100A4 expression data, draw a standard curve and convert concentration values; Statistical analysis and ROC curve analysis were performed on the processed data to calculate the area under the curve, sensitivity, and specificity; Determine whether the data meets the preset determination conditions; When the blood S100A4 concentration is less than 293.04 ng / ml, it is diagnosed as acute pancreatitis; When the blood S100A4 concentration is greater than 870.64 ng / ml, it is diagnosed as chronic pancreatitis; When used for identification, blood S100A4 concentration greater than 572.92 ng / ml is diagnosed as chronic pancreatitis.

11. A computer-readable storage medium storing a computer program, characterized in that: When the computer program is executed by a processor, the computer program can implement the functions of the system according to claim 9 or the steps of the method according to claim 10.

12. An electronic device comprising a memory and a processor, wherein the memory stores a computer program, wherein: The processor is configured to execute the computer program to implement the functions of the system according to claim 9 or the steps of the method according to claim 10.

Images