Marker, kit, method, system and application for ALS

Abstract

The invention provides a marker, a kit, a method, a system and application for ALS, and the marker comprises N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide and N-(hexadecanoyl)-1-de-sample sphingomyelin. The invention solves the problems that the pathogenicity of different variation sites of children cannot be accurately confirmed through SPT mutation factors, the conformity with clinical symptoms cannot be evaluated and the early recognition of disease progression cannot be realized in the prior art.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical detection, and particularly relates to a biomarker, kit, method, system and application for ALS. Background Art

[0002] Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease that selectively affects upper motor neurons such as the motor cortex and pyramidal tract, as well as lower motor neurons such as brainstem motor nuclei and anterior horn of the spinal cord. The clinical manifestations mainly include muscle weakness, muscle atrophy, bulbar palsy and pyramidal tract signs, etc. The peak incidence age of amyotrophic lateral sclerosis is 55 - 75 years old, and the median survival period is about 27.5 months. Although ALS is rare in children, there are continuous case reports, and its symptoms and disease course are similar to those of adult ALS, including muscle weakness, atrophy and loss of motor function.

[0003] The pathological mechanism of amyotrophic lateral sclerosis (ALS) is complex, mainly involving multiple factors such as abnormal protein aggregation, abnormal DNA / RNA function, changes in the cytoskeleton and axonal dynamics, etc. There is currently no single mechanism to explain its pathogenesis. Pediatric ALS is often associated with gene mutations, such as mutations in SPTLC1 and SPTLC2, etc. These clear monogenic etiologies can provide unique insights into the pathophysiology of the disease and guidance for the design of treatment strategies. The diagnosis of pediatric ALS depends on clinical evaluation, neuroimaging examination and genetic testing. Due to the possible overlap of symptoms with other neurological diseases, the diagnosis is often delayed. With the continuous progress of gene detection technology, mutation screening for specific pathogenic genes can achieve accurate diagnosis, further revealing the pathological mechanism of the disease, which not only helps in the diagnosis, but also provides an important basis for formulating individualized treatment plans.

[0004] However, there is still an urgent need to propose a scientific biomarker detection scheme, using the biomarker as a potential biomarker for pediatric ALS, aiming to confirm the pathogenicity of different variant sites, evaluate the consistency with clinical symptoms, and achieve early identification of disease progression. This detection scheme will provide important support for the early diagnosis, pathological mechanism analysis and individualized treatment of pediatric ALS. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides a biomarker, kit, method, system and application for ALS, so as to solve the problems that the existing technology cannot accurately confirm the pathogenicity of different variant sites in children through SPT mutation factors, cannot evaluate the consistency with clinical symptoms, and cannot achieve early identification of disease progression.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a biomarker and its combination for diagnosing or prognosticating childhood amyotrophic lateral sclerosis, and the biomarker is selected from any one or a combination of: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine.

[0008] In a second aspect, a kit for diagnosing childhood amyotrophic lateral sclerosis is further provided, and the components of the kit include:

[0009] the biomarker or its combination as described in the first aspect; and

[0010] an isotope label.

[0011] As a further improvement of the present invention, the isotope label is a mixed liquid composed of at least two (preferably 5) biomarkers among the biomarkers.

[0012] As a further improvement of the present invention, the isotope label is a mixed liquid composed of N-oleoyl sphingomyelin and C18 ceramide.

[0013] As a further improvement of the present invention, the components of the kit include:

[0014] a buffer;

[0015] a quality control product; and,

[0016] a positive sample and a negative sample.

[0017] In a third aspect, a method for detecting a biomarker for childhood amyotrophic lateral sclerosis is further provided, including:

[0018] obtaining a sample to be tested;

[0019] injecting the sample to be tested into a liquid chromatograph to separate each component in the sample to be tested through the liquid chromatograph;

[0020] injecting the separated sample to be tested into a mass spectrometer, and ionizing the separated components entering therein through the mass spectrometer and performing mass analysis on the ionized components to determine the target component as a biomarker based on the result of the mass analysis of the ionized components;

[0021] wherein the target component includes or is selected from: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine.

[0022] Fourth aspect, there is also provided a prognosis method for childhood amyotrophic lateral sclerosis, including:

[0023] Detecting the content of the biomarker described in the first aspect contained in the body fluid of ALS patients;

[0024] Predicting the condition of ALS patients based on the detected content of the biomarker components.

[0025] Fifth aspect, there is also provided a biomarker detection system for predicting or prognosticating childhood amyotrophic lateral sclerosis, including:

[0026] An acquisition unit for acquiring a sample to be tested;

[0027] A liquid chromatograph for receiving the sample to be tested to separate each component in the sample to be tested;

[0028] A mass spectrometer for receiving the separated sample to be tested, ionizing the separated components entering therein, and performing mass analysis on the ionized components, so as to determine the target component as a biomarker based on the result of the mass analysis of the ionized components;

[0029] Wherein, the target components include: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine.

[0030] Sixth aspect, there is also provided a use of a biomarker in the preparation of a product for evaluating the prognosis risk of childhood amyotrophic lateral sclerosis, wherein the biomarker includes or is selected from N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine.

[0031] Seventh aspect, there is also provided a computer storage medium, characterized in that it includes: a computer program is stored on the computer-readable storage medium, and when the computer program is executed by a processor, the steps of the method described in the third aspect are implemented.

[0032] Eighth aspect, there is provided a method for constructing a prognosis risk assessment model for childhood amyotrophic lateral sclerosis, wherein the prognosis risk assessment model uses the biomarker described in the third aspect as a parameter and is constructed through a regression equation.

[0033] Ninth aspect, there is also provided a use of the biomarker described in the first aspect or a combination thereof in the preparation of a kit for predicting or prognosticating childhood amyotrophic lateral sclerosis.

[0034] In a specific embodiment of the present invention, the components in the kit are shown in Table 1.

[0035] Table 1 Details of the Kit Components

[0036] Component Serial Number Name Specification Quantity 1 Standard (Mixed liquid of 6 sphingolipids) 1 mL 5 2 Isotope internal standard (Mixed solution of 2 sphingolipids) 1 mL 1 3 Buffer 1 500 mL 1 4 Buffer 2 500 mL 1 5 Quality control product 1 mL 1 6 Positive sample 1 mL 1 7 Negative sample 1 mL 1

[0037] In a specific embodiment of the present invention, the details of the information of 6 candidate markers are shown in Table 2.

[0038] Table 2: Details of the Information of 6 Candidate Markers

[0039]

[0040] In a specific embodiment of the present invention, the details of the information of 6 standards are shown in Table 3.

[0041] Table 3: Details of the Information of 6 Standards

[0042]

[0043] In a specific embodiment of the present invention, the isotope internal standard information is shown in Table 4.

[0044] Table 4 Isotope Internal Standard Information Table

[0045]

[0046] The present invention has the following beneficial effects compared with the prior art:

[0047] The present invention uses a marker combination composed of five preferred markers as potential biomarkers for childhood amyotrophic lateral sclerosis, to confirm the pathogenicity of different mutation sites through the changes in the levels of markers in human body fluid samples, and to identify the consistency with clinical symptoms, so as to be able to early identify the progression of the disease.

[0048] Moreover, the kit of the present invention has important value in the diagnosis and differential diagnosis of childhood ALS. It not only provides a non-invasive method to detect biomarkers in the early stage of the disease to help accurately diagnose the disease early, and can evaluate the disease progression and treatment effect by monitoring the changes in the marker levels, but also can provide data support for clinical research to further understand the pathological mechanism of amyotrophic lateral sclerosis, making the kit a powerful tool in the diagnosis and management of childhood amyotrophic lateral sclerosis. Description of the Drawings

[0049] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0050] Figure 1 Graph of the content of candidate biomarker components in a test sample of human plasma.

[0051] Figure 2 Schematic flowchart of a prognosis method for childhood amyotrophic lateral sclerosis.

[0052] Figure 3 ROC curve graph after logistic regression for predicting the risk of ALS using biomarkers in human plasma samples.

[0053] Figure 4 Schematic structural block diagram of a biomarker detection system for childhood amyotrophic lateral sclerosis according to an embodiment of the present invention.

[0054] Figure 5 Topological structure diagram of a computer-readable storage medium disclosed by the present invention. Detailed implementation manners

[0055] To make the above objects, features, and advantages of the present invention more obvious and understandable, the following will describe the detailed implementation manners of the present invention in conjunction with the accompanying drawings. Many specific details are set forth in the following description in order to fully understand the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below. The technical features in each embodiment of the present invention can be combined correspondingly without conflict.

[0056] Example 1:

[0057] This embodiment provides a method for screening and validating biomarkers for childhood amyotrophic lateral sclerosis. Based on the UPLC-MS detection platform, metabolomics analysis of plasma is an important means to discover biomarkers. The samples in this embodiment are from the included children with ALS and their fathers and mothers, and blood samples of the ALS patients and their fathers and mothers are collected. Those clinically diagnosed with childhood ALS are used as the case group, and those without any ALS symptoms are used as healthy controls. Other diseases are excluded.

[0058] Detect the sphingolipidome in the metabolome of the blood samples.

[0059] Reagents and instruments The reagents and detection instruments used for metabolomics detection are shown in Tables 5 and 6.

[0060] Table 5: Main reagents for sphingolipid detection

[0061] Reagent Name Model Manufacturer Isopropanol (Chromatographic grade) 4L Fisher Methanol (Chromatographic grade) 4L Fisher Water 4L Fisher Formic acid 50 mL Shanghai Aladdin Biochemical Technology Co., Ltd. Ammonium formate 100g Shanghai Aladdin Biochemical Technology Co., Ltd.

[0062] Table 6: Main instruments for sphingolipid detection

[0063]

[0064] Sample pretreatment method

[0065] Take 100 μL of whole blood sample, add it to a 1.5 mL centrifuge tube, add 400 μL of methanol solution, seal and mix well, and perform ultrasonic oscillation for 10 min. Then put the ultrasonicated centrifuge tube into a refrigerated centrifuge, set it at 12000 r / min and centrifuge at 4 °C for 10 min. Take 200 μL of the supernatant and transfer it to a glass injection vial with an inner insert for measurement.

[0066] Chromatographic parameters

[0067] Mobile phase:

[0068] A: Water + 0.1% formic acid + 10 mM ammonium formate

[0069] B: Isopropanol: methanol (9:1) + 0.1% formic acid + 10 mM ammonium formate Chromatographic system: UPLC (ACQUITY UPLC I-Class, Waters)

[0070] Chromatographic column: BEH Amide, 1.7 μm, 2.1X100 mm (Waters)

[0071] Column temperature: 35 °C

[0072] Sample chamber temperature: 10 °C

[0073] Table 7: Chromatographic elution gradient:

[0074] Time (min) Flow rate (mL / min) A(％) B(％) Curve Initial 0.25 5 95 Initial 0.50 0.25 5 95 6 3.00 0.25 30 70 6 4.00 0.25 50 50 6 4.10 0.25 5 95 6 5.00 0.25 5 95 6

[0075] Mass spectrometry parameters Mass spectrometer: Triple quadrupole tandem mass spectrometer (ACQUITY TQ-S, Waters) Ionization mode: ESI+ (positive ion mode)

[0076] Capillary voltage (kV): 2.0

[0077] Desolvation gas temperature (°C): 500

[0078] Source temperature (°C): 150

[0079] Desolvation gas flow rate (L / h): 800

[0080] Cone gas flow rate (L / h): 150

[0081] Acquisition mode: Multiple reaction monitoring (MRM)

[0082] Table 8: MRM and its parameters

[0083]

[0084] Statistical analysis

[0085] Statistical analyses were all completed using specialized statistical analysis software (R language, version R-4.4.1). The statistical significance level was set at p < 0.05, and all statistical tests were two-sided tests. A bar chart is a graphical statistical report with the length of a rectangle as a variable, showing the data distribution by a series of vertical bars with unequal heights, and the error bars represent the standard error. The bar charts in the present invention were made using GraphPad Prism 9.0 software. Machine learning and ROC were analyzed and plotted using the plotROC package in the R language.

[0086] Figure 1 It is a chart showing the component content in a test sample of human plasma. Six groups of bar charts are presented in the figure. Each group of bar charts compares the concentrations of different types of compounds in two samples, which are "Control" (control group) and "ALS" (amyotrophic lateral sclerosis patient group) respectively. It can be found that there is no statistical significance in the concentration difference between the two groups in group E (marked as ns, indicating not significant), suggesting that Cer(m18:0 / 16:0) (i.e., N-(hexadecanoyl)-1-deoxysphingosine) may have a relatively small association with ALS. The concentrations in the ALS group in the five groups A - D and F are significantly higher than those in the control group, that is, the concentrations of the other 5 types of compounds in the ALS patient group are significantly higher than those in the control group, indicating that these 5 types of compounds play important roles in the pathological process of ALS.

[0087] It can be seen that through the above embodiments, 5 types of compounds are obtained as markers for childhood amyotrophic lateral sclerosis, which helps to early identify the progression of the disease, evaluate the disease progression and treatment effect. That is, the present embodiment can also provide a marker for childhood amyotrophic lateral sclerosis, which includes: N-stearoylsphingomyelin, N-oleoylsphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-1-deoxysphinganine.

[0088] Thus, the five components detected by the marker detection method of the present embodiment, as potential biomarkers for childhood amyotrophic lateral sclerosis, can confirm the pathogenicity of different mutation sites through the changes in the levels in human body fluids (including blood or cerebrospinal fluid, preferably plasma) samples, and identify the consistency with clinical symptoms, so as to be able to early identify the progression of the disease. Thereby solving the problems in the prior art that the pathogenicity of different mutation sites in children cannot be accurately confirmed by SPT mutation factors, the consistency with clinical symptoms cannot be evaluated, and the early identification of disease progression cannot be achieved.

[0089] Further confirmation of biomarkers: Another 10 pediatric ALS patients were selected and analyzed and evaluated using the ROC curve. Figure 3 The ROC analysis results of the 5-biomarker combination (N-stearoylsphingomyelin, N-oleoylsphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine) are shown, and the results indicate that the AUC value of the biomarker combination is 1.0.

[0090] It should be noted that the samples involved in this example (such as the original data of the test samples) are from the included pediatric ALS patients and their fathers and mothers, and blood samples of the ALS patients and their fathers and mothers were collected. Those clinically diagnosed with pediatric ALS were used as the case group, and those without any ALS symptoms were used as healthy controls, excluding other diseases.

[0091] Example 2:

[0092] This example also provides a kit for pediatric amyotrophic lateral sclerosis. The components of the kit include: the biomarkers described in Example 1 and isotope labels. The isotope label is a mixed liquid composed of at least two of the biomarkers. In one example, the isotope label is a mixed liquid composed of N-oleoylsphingomyelin and C18 ceramide.

[0093] In the above example, the components of the kit include: two buffers; a quality control product; and positive and negative samples. Among them, the buffer includes water and isopropanol containing formic acid and ammonium formate.

[0094] It can be understood that the kit in this example has important value in the diagnosis and differential diagnosis of pediatric ALS. It not only provides a non-invasive method to detect biomarkers in the early stage of the disease to help with early and accurate disease diagnosis, but also can evaluate disease progression and treatment effects by monitoring changes in the levels in the kit, and can also provide data support for clinical research to further understand the pathological mechanism of amyotrophic lateral sclerosis, making the kit a powerful tool in the diagnosis and management of pediatric amyotrophic lateral sclerosis.

[0095] It should be noted that the scheme or principle involved in the kit of this example is the same as that of Example 1. For specific content details, refer to Example 1 and will not be elaborated here.

[0096] Example 3:

[0097] As Figure 2 shown, this example also provides a prognostic method for pediatric amyotrophic lateral sclerosis, which includes:

[0098] Step 202. Detect the component content contained in the body fluid of ALS patients based on the biomarker described in Embodiment 1;

[0099] Step 204. Predict the condition of ALS patients based on the detected component content.

[0100] Thus, the prognosis method of this embodiment helps to accurately diagnose the disease in the early stage of ALS patients, evaluate the disease progression and treatment effect, and helps to further understand the pathological mechanism of amyotrophic lateral sclerosis, facilitating the effective diagnosis and treatment of childhood amyotrophic lateral sclerosis.

[0101] It should be noted that the solution or principle involved in this embodiment is the same as that in Embodiment 1 and / or Embodiment 2. For the specific content details, refer to Embodiment 1 and / or Embodiment 2, and no further elaboration will be provided here.

[0102] Example 4:

[0103] As Figure 4 shown, this embodiment also provides a biomarker detection system 400 for childhood amyotrophic lateral sclerosis, including: an acquisition unit 401 for acquiring a sample to be tested; a liquid chromatograph 402 for receiving the sample to be tested to separate each component in the sample to be tested; a mass spectrometer 403 for receiving the separated sample to be tested, ionizing the separated components entering it, and performing mass analysis on the ionized components to determine the target component as a biomarker based on the results of the mass analysis of the ionized components. Among them, the target components include: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxyl sphinganine. Among them, the acquisition unit 401 includes an ultrasonic oscillator 401a and a refrigerated centrifuge 401b. The ultrasonic oscillator 401a performs ultrasonic oscillation on the mixed and sealed solution of whole blood sample and methanol, and the refrigerated centrifuge 401b separates the solid particles or liquids with different densities in the ultrasonicated mixed liquid, thereby forming a sample to be tested.

[0104] The system 400 of this embodiment also includes a data processing platform 404 for extensive acquisition and analysis of mass spectrometry data, providing a comprehensive environment for processing, analyzing, visualizing, and reporting mass spectrometry data. Thus, through an intuitive user interface and an automated workflow, the efficiency of data acquisition, analysis, and report generation can be improved, supporting basic qualitative and quantitative analysis to complex multi-stage mass spectrometry data processing.

[0105] It should be understood that the five components detected by the biomarker detection system of this embodiment are used as potential biomarkers for childhood amyotrophic lateral sclerosis, so as to confirm the pathogenicity of different mutation sites through the changes in the levels in human body fluid samples, and to identify the consistency with clinical symptoms, so as to be able to identify the progression of the disease at an early stage.

[0106] The kit made of the five components detected by the biomarker detection system of this embodiment has important value in the diagnosis and differential diagnosis of childhood ALS. It not only provides a non-invasive method to detect biomarkers in the early stage of the disease to help accurately diagnose the disease at an early stage, but also can evaluate the disease progression and treatment effect by monitoring the changes in the levels in the kit, and can also provide data support for clinical research to further understand the pathological mechanism of amyotrophic lateral sclerosis, so that the kit becomes a powerful tool in the diagnosis and management of childhood amyotrophic lateral sclerosis.

[0107] It should be noted that the solutions or principles involved in this embodiment are the same as those in Embodiment 1 and / or Embodiment 2. For the specific content details, refer to those described in Embodiment 1 and / or Embodiment 2, and will not be elaborated here.

[0108] Example 5:

[0109] This embodiment also provides an application of a biomarker in the preparation of a product for evaluating the prognosis risk of childhood amyotrophic lateral sclerosis. The biomarker includes N-stearoylsphingomyelin, N-oleoylsphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-deoxysphingosine.

[0110] The products of this embodiment are selected from: reagents, kits, test strips, gene chips, high-throughput sequencing platforms, antibody chips, instrument platforms. The instrument platform includes a measurement module for measuring the content of the protein expressed by the combined genes in the sample to be tested.

[0111] It can be understood that this embodiment uses the biomarker composed of five components as a potential biomarker for childhood amyotrophic lateral sclerosis, so as to confirm the pathogenicity of different mutation sites through the changes in the levels in human body fluid samples, and to identify the consistency with clinical symptoms, so as to be able to identify the progression of the disease at an early stage.

[0112] The kit of this embodiment has important value in the diagnosis and differential diagnosis of childhood ALS. It not only provides a non-invasive method to detect biomarkers in the early stage of the disease to help accurately diagnose the disease at an early stage, but also can evaluate the disease progression and treatment effect by monitoring the changes in the levels in the kit, and can also provide data support for clinical research to further understand the pathological mechanism of amyotrophic lateral sclerosis, so that the kit becomes a powerful tool in the diagnosis and management of childhood amyotrophic lateral sclerosis.

[0113] It should be noted that the solution or principle involved in this embodiment is the same as that of Embodiment 1 and / or Embodiment 2. For the specific content details, please refer to Embodiment 1 and / or Embodiment 2, and thus will not be elaborated here.

[0114] Example 6:

[0115] This embodiment also provides a terminal device, which may include a processor, a memory, and a computer program stored on the memory and executable on the processor. When the computer program is executed by the processor, it implements each process of the above Figure 1 marker detection method embodiment shown, and can achieve the same technical effects. To avoid repetition, it will not be elaborated here.

[0116] Example 7:

[0117] Combined with Figure 5 shown, this embodiment also discloses a specific implementation of a computer-readable storage medium 500. The computer-readable storage medium 500 may be configured in whole or in part in a physically formed computer, server, cluster server, or data center.

[0118] In this embodiment, computer program instructions 501 are stored in the computer-readable storage medium 500. When the computer program instructions 501 are read and run by a processor 502, the steps in the marker detection method disclosed in Embodiment 1 are executed.

[0119] Optionally, the computer-readable storage medium 500 may be configured as a server, and the server runs on a physical device for building a private cloud, hybrid cloud, or public cloud. At the same time, the computer-readable storage medium 500 may also be configured as a random access memory (RAM), read-only memory (ROM), programmable read-only memory (PROM), erasable programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), etc.

[0120] The computer-readable storage medium 500 is used to store a program. After receiving an execution instruction, the processor 502 executes the marker detection method disclosed in Embodiment 1.

[0121] Meanwhile, the processor 502 disclosed in this embodiment may be an integrated circuit chip with signal processing capabilities. The processor 502 may be a general-purpose processor, including a central processing unit (CPU), a network processor (NP), etc.; it may also be a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components. It can implement or execute the various methods, steps, and logic block diagrams disclosed in the embodiments of the present invention. The general-purpose processor may be a microprocessor or the general-purpose processor may also be any conventional processor.

[0122] For the technical solutions of the same parts in the computer-readable storage medium 500 disclosed in this embodiment as in Embodiment 1 and / or Embodiment 2, please refer to what is described in Embodiment 1 and / or Embodiment 2, and details will not be repeated here.

[0123] The above-described embodiments are only a preferred solution of the present invention, but they are not intended to limit the present invention. Those of ordinary skill in the relevant technical fields can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, all technical solutions obtained by means of equivalent replacement or equivalent transformation fall within the protection scope of the present invention.

[0124] The serial numbers of the above-described embodiments of the present invention are only for description and do not represent the advantages or disadvantages of the embodiments.

[0125] Those of ordinary skill in the art can understand that all or part of the steps for implementing the above embodiments can be completed by hardware, or can be completed by a program instructing relevant hardware. The program can be stored in a computer-readable storage medium. The above-mentioned storage medium can be a read-only memory, a magnetic disk, an optical disc, or the like.

[0126] The above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A marker for diagnosis or prognosis of amyotrophic lateral sclerosis in children and a combination thereof, characterized in that: The marker is selected from any one or a combination of N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide and N-(hexadecanoyl)-l-ceramide.

2. A kit for diagnosing amyotrophic lateral sclerosis in children, characterized in that: The components of the kit include: The marker according to claim 1 or a combination thereof; and Isotope markers.

3. The kit according to claim 2, characterized in that The isotope label is a mixed liquid consisting of at least two (preferably five) of the markers.

4. The kit according to claim 3, characterized in that The isotope label is a mixed liquid consisting of N-oleoyl sphingomyelin and C18 ceramide.

5. The kit according to claim 2, characterized in that The components of the kit include: Buffer; Quality control products; and, Positive samples and negative samples.

6. A method for detecting markers for amyotrophic lateral sclerosis in children, characterized in that: include: Obtaining samples to be tested; Injecting the sample to be tested into a liquid chromatograph to separate the components in the sample to be tested by the liquid chromatograph; injecting the separated sample to be tested into a mass spectrometer, and ionizing the separated components entering therein by the mass spectrometer and performing mass analysis on the ionized components, so as to determine the target component as a marker based on the result of the mass analysis on the ionized components; The target components include: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide and N-(hexadecanoyl)-l-desphingomyelin.

7. Use of the marker or the combination thereof according to claim 1 in the preparation of a kit for predicting or prognosing amyotrophic lateral sclerosis in children.

8. A marker detection system for predicting or prognosing amyotrophic lateral sclerosis in children, comprising: An acquisition unit acquires a sample to be tested; A liquid chromatograph, used for receiving the sample to be tested to separate the components in the sample to be tested; A mass spectrometer, used for receiving the separated sample to be tested, ionizing the separated components entering therein, and performing mass analysis on the ionized components, so as to determine the target component as a marker based on the result of the mass analysis on the ionized components; The target components include: N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide and N-(hexadecanoyl)-l-desphingomyelin.

9. Use of a marker in the preparation of a product for assessing the prognosis risk of amyotrophic lateral sclerosis in children, characterized in that: The markers include N-stearoyl sphingomyelin, N-oleoyl sphingomyelin, C18 ceramide, N-(oleoyl)-ceramide, and N-(hexadecanoyl)-l-ceramide.

10. A computer storage medium, characterized in that: include: The computer readable storage medium stores a computer program, and when the computer program is executed by a processor, the steps of the method according to claim 6 are implemented.

11. A method for constructing a prognostic risk assessment model for amyotrophic lateral sclerosis in children, characterized in that: The prognostic risk assessment model uses the markers as described in claim 1 or a combination thereof as parameters and is constructed through a regression equation.

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