Model, device and equipment for assisting in screening Alzheimer disease and storage medium

Abstract

The invention relates to a model, a device and equipment for assisting in screening Alzheimer's disease, and a storage medium. Input variables of the model comprise a P-tau181 concentration value and an A beta 42 concentration value of an organism, and output variables of the model comprise a ratio of the P-tau181 concentration value to the A beta 42 concentration value. On the basis of a flow fluorescence detection technology, the relevance between the ratio of the P-tau181 concentration value to the Abeta42 concentration value and the neurodegenerative diseases is deeply analyzed, a reference interval is designed, an efficient screening tool is further developed, and high-specificity, high-sensitivity and high-accuracy auxiliary screening of the neurodegenerative diseases such as the Alzheimer's disease is achieved.

Description

Technical Field

[0001] The present invention belongs to the field of biological detection technology and relates to a model, a device, an equipment and a storage medium for assisting in the screening of neurodegenerative diseases. Background Art

[0002] Alzheimer's disease (AD) is one of the most common neurodegenerative diseases and the most common cause of cognitive impairment. The pathogenesis has not yet been fully elucidated. According to the "Guidelines for the Diagnosis and Treatment of Alzheimer's Disease in China", the current screening and evaluation of Alzheimer's disease mainly includes clinical evaluation, imaging examination and laboratory examination. Clinical evaluation is a routine examination with general performance and requires relatively professional and systematic training and long-term growth to enable a clinician to have systematic and standardized cognitive evaluation capabilities. Imaging evaluation, MR or CT are mostly non-specific, and there may be no obvious atrophy and signs of involvement in the early stage. AD has an insidious onset and gradually progresses from the preclinical stage with intact cognition to the prodromal stage of mild cognitive impairment (MCI) and then to the dementia stage. The entire course of the disease can be up to 20 to 30 years.

[0003] At present, the gold standard for Alzheimer's disease detection is Aβ-PET and Tau-PET brain imaging or cerebrospinal fluid testing. Cerebrospinal fluid testing requires puncture, patient compliance is poor, PET is expensive, and only a few cognitive centers have the ability to detect, making it difficult to popularize. In the entire diagnostic logic, Aβ is considered to be the earliest biological marker and the core of AD diagnosis. There is currently a lack of specific and cost-effective biological indicators representing Aβ-PET, and the detection of Aβ42 in cerebrospinal fluid cannot be used as a diagnostic biological marker for clinical diagnosis. Although the International Alzheimer's Disease Conference proposed in the latest diagnosis and staging that P-tau217 is considered a diagnostic biological marker because of its good correlation with Aβ-PET, the cost of detecting P-tau217 is lower than that of combined detection, but it still only has good diagnostic performance on ultra-sensitive platforms such as the SIMOA platform, so it is difficult to apply to community and routine physical examination scenarios.

[0004] In summary, there is an urgent need for biomarkers and diagnostic tools that are easily accessible, economical, sensitive, specific, and simple to use to improve the screening level of community doctors and effectively screen preclinical and AD patients. Summary of the invention

[0005] In view of the deficiencies of the prior art and actual needs, the present invention provides a model, device, equipment and storage medium for assisting in the screening of Alzheimer's disease, develops efficient screening tools, and achieves high specificity, sensitivity and accuracy to assist in the screening of neurodegenerative diseases such as Alzheimer's disease.

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

[0007] In a first aspect, the present invention provides a model for assisting in the screening of Alzheimer's disease, wherein the input variables of the model include the body's P-tau181 concentration value and the Aβ42 concentration value, and the output variables of the model include the ratio of the P-tau181 concentration value to the Aβ42 concentration value (P-tau181 / Aβ42); the criterion for determining a positive Alzheimer's disease is: the ratio of the P-tau181 concentration value to the Aβ42 concentration value is greater than 0.109.

[0008] The present invention detects plasma P-tau181 (phosphorylated Tau181 protein) and Aβ42 (β-amyloid protein 42) of multiple volunteers based on flow cytometry technology, deeply analyzes the correlation between the concentrations and their ratio and neurodegenerative diseases, further establishes a reference interval with the ratio of the concentration value of P-tau181 to the concentration value of Aβ42 as an indicator, and constructs a model that can effectively identify Alzheimer's disease patients (including AD-induced MCI patients), with specificity and sensitivity both reaching more than 90%, thereby achieving effective screening of preclinical and AD patients.

[0009] In a second aspect, the present invention provides use of the model for assisting in screening for Alzheimer's disease described in the first aspect in preparing a product for assisting in diagnosing neurodegenerative diseases.

[0010] It can be understood that the product may be a diagnostic device or a kit, etc., and the kit may include reagents for detecting the concentration value of P-tau181 and the concentration value of Aβ42 in the body.

[0011] In a third aspect, the present invention provides a device for assisting in the diagnosis of neurodegenerative diseases, the device comprising a data acquisition unit, a data processing unit and a determination unit;

[0012] The data acquisition unit is used to perform the following steps:

[0013] Detecting the concentration values ​​of P-tau181 and Aβ42 in the sample to be tested;

[0014] The data processing unit is used to perform the following steps:

[0015] Inputting the P-tau181 concentration value and the Aβ42 concentration value obtained by the data acquisition unit into the model for assisting in screening for Alzheimer's disease described in the first aspect, and outputting the ratio of the P-tau181 concentration value to the Aβ42 concentration value;

[0016] The determination unit is used to perform the following steps:

[0017] The determination is made based on the ratio of the P-tau181 concentration value to the Aβ42 concentration value obtained by the data processing unit and the result is output.

[0018] Based on the constructed model for assisting in the screening of Alzheimer's disease, the present invention further designs a device for assisting in the diagnosis of neurodegenerative diseases, which can screen and differentiate Alzheimer's disease from many other neurodegenerative diseases (such as frontotemporal dementia, vascular dementia and Parkinson's disease, etc.).

[0019] Preferably, the sample to be tested includes plasma.

[0020] Preferably, the detection method comprises flow cytometry.

[0021] The present invention is developed based on the detection results of the flow fluorescence platform, which significantly improves the detection sensitivity and specificity, and has a low detection bottleneck, low instrument and equipment requirements, low reagent cost, is easier to promote, has a short detection cycle, and is more suitable for auxiliary community screening.

[0022] Preferably, when the ratio of the P-tau181 concentration value to the Aβ42 concentration value is greater than 0.109, the result is determined and output as positive for Alzheimer's disease.

[0023] Preferably, when the ratio of the P-tau181 concentration value to the Aβ42 concentration value is ≤0.109, the result is determined and output as negative for Alzheimer's disease, and combined with the medical history, it is confirmed whether it is other degenerative diseases.

[0024] In the present invention, when the ratio of the P-tau181 concentration value to the Aβ42 concentration value is ≤0.109, Alzheimer's disease can be basically ruled out. In this case, if it is to be investigated whether it is other degenerative diseases, a comprehensive analysis such as PET analysis, cerebrospinal fluid analysis, medical history, family history or neurological scale is required.

[0025] Preferably, the other degenerative disease comprises at least one of frontotemporal dementia, vascular dementia or Parkinson's disease.

[0026] In a fourth aspect, the present invention provides an electronic device comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor implements the function of the device for assisting in diagnosing neurodegenerative diseases as described in the third aspect when executing the computer program.

[0027] In a fifth aspect, the present invention provides a computer-readable storage medium storing a computer program, which, when executed by a processor, implements the functions of the device for assisting in diagnosing neurodegenerative diseases as described in the third aspect.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention uses flow fluorescence detection technology to find that P-tau181 / Aβ42 has significant differences in healthy people, AD patients and patients with other degenerative diseases. Based on P-tau181 / Aβ42, the auxiliary diagnosis model is further developed, and the diagnostic accuracy of AD patients can reach 96%, the sensitivity can reach 95%, and the specificity can reach 97%. Based on the flow platform, the detection bottleneck is low, the instrument and equipment requirements are low, the reagent cost is low, it is easier to promote, and the detection cycle is short, which is more suitable for auxiliary community screening. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is the scatter plot of P-tau181 / Aβ42 in AD+MCI samples and negative samples.

[0031] Figure 2 This is the ROC curve of P-tau181 / Aβ42 in diagnosing AD+MCI, SIVD and FTD. DETAILED DESCRIPTION

[0032] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following examples are only simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0033] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0034] Example 1

[0035] 123 normal samples, 60 AD+MCI samples, 34 subcortical ischemic cerebrovascular disease (SIVD) samples and 34 frontotemporal dementia (FTD) samples were collected from multicenters across the country. The concentrations of P-tau181 and Aβ42 were detected based on the flow cytometry technology platform. The specific process includes:

[0036] Based on flow cytometry technology, the capture microsphere mixture in the detection kit contains capture microspheres with different fluorescence intensities; the surfaces of the capture microspheres are coated with Aβ42, Aβ40, and p-Tau181 specific antibodies respectively; the capture microspheres specifically bind to Aβ42, Aβ40, and p-Tau181 in the sample to be tested, and then combine with the biotin-labeled detection antibody, and the capture microspheres, the sample to be tested, and the detection antibody form a double antibody sandwich complex (capture microspheres + sample to be tested + detection antibody), and then react with the added phycoerythrin-labeled streptavidin (SA-PE), and the Aβ42, Aβ40, and p-Tau181 content of the sample to be tested is obtained by analyzing the fluorescence intensity of the complex.

[0037] The concentration ratio of P-tau181 and Aβ42 (P-tau181 / Aβ42) was analyzed. Figure 1 As shown, a reference interval of P-tau181 / Aβ42≤0.109 was further established, 95% of normal people fall within this range, and people with AD can be screened out if their values ​​exceed this range.

[0038] A model for auxiliary screening of Alzheimer's disease was established based on the reference interval P-tau181 / Aβ42≤0.109, and the diagnostic efficacy of the model was further verified. Its accuracy in diagnosing AD can reach 96%, sensitivity can reach 95%, and specificity is 97%.

[0039] Based on the constructed model for assisting in the screening of Alzheimer's disease, a device for assisting in the diagnosis of neurodegenerative diseases can be further designed to screen and differentiate Alzheimer's disease from many other neurodegenerative diseases (such as frontotemporal dementia, vascular dementia, and Parkinson's disease, etc.). The device for assisting in the diagnosis of neurodegenerative diseases includes a data acquisition unit, a data processing unit, and a determination unit; the data acquisition unit is used to execute the following: detecting the P-tau181 concentration value and the Aβ42 concentration value in the sample to be tested; the data processing unit is used to execute the following: inputting the P-tau181 concentration value and the Aβ42 concentration value obtained by the data acquisition unit into the model for assisting in the screening of Alzheimer's disease, and outputting the ratio of the P-tau181 concentration value to the Aβ42 concentration value; the determination unit is used to execute the following: :According to the ratio of the P-tau181 concentration value and the Aβ42 concentration value obtained by the data processing unit, a judgment is made and a result is outputted. When P-tau181 / Aβ42>0.109, the judgment and output result is positive for Alzheimer's disease; when P-tau181 / Aβ42≤0.109, the judgment and output result is negative for Alzheimer's disease. Combined with the medical history and other information, confirm whether it is other degenerative diseases, such as using PET or cerebrospinal fluid analysis for subsequent confirmatory evaluation, and combined with the medical history, family history, neurological scale and other comprehensive analysis.

[0040] ROC curve analysis results ( Figure 2 ) showed that the diagnostic accuracy of P-tau181 / Aβ42 for AD+MCI and FTD can reach 86%, the sensitivity can reach 90%, the specificity can reach 80%, and the AUC can reach 92%. The diagnostic accuracy of P-tau181 / Aβ42 for AD+MCI and SIVD can reach 88%, the sensitivity can reach 100%, the specificity can reach 68%, and the AUC can reach 91%.

[0041] Example 2

[0042] This example verifies the model for assisting in screening for Alzheimer's disease.

[0043] The comparison and analysis of the model determination results of the present invention for assisting in screening for Alzheimer's disease and the gold standard results are as follows:

[0044] A head-to-head comparison was performed on clinical samples sent from multiple cognitive impairment centers. Fifteen plasma samples and five cerebrospinal fluid samples were tested at the same time. The test results showed that four of the five cerebrospinal fluid samples were positive, and the prediction results of the model of the present invention were consistent with the characterization results of the cerebrospinal fluid Aβ42 / Aβ40 ratio of the patients (sample 1: P-tau181 / Aβ42 ratio was 0.264 (reference interval: ≤0.109), and the cerebrospinal fluid Aβ42 / Aβ40 ratio was 0.02; sample 2: P-tau181 / Aβ42 ratio was 0.124 (reference interval: ≤0.109), and the cerebrospinal fluid Aβ42 / Aβ40 ratio was 0.02 ; Sample 3: P-tau181 / Aβ42 ratio was 0.017 (reference interval: ≤0.109), and the cerebrospinal fluid Aβ42 / Aβ40 ratio was 0.09; Sample 4: P-tau181 / Aβ42 ratio was 0.456 (reference interval: ≤0.109), and the cerebrospinal fluid Aβ42 / Aβ40 ratio was 0.01; Sample 5: P-tau181 / Aβ42 ratio was 0.116 (reference interval: ≤0.109), and the cerebrospinal fluid Aβ42 / Aβ40 ratio was 0.03;), all showed positive consistency results, sample 3 was a negative sample, and the prediction result of the model of the present invention also showed negative. The model of the present invention predicted that 10 of the 18 plasma samples tested were negative for P-tau181 / Aβ42 ratio and 8 were positive samples. This result is consistent with the P-tau217 test results of the simoa platform recognized in the current international consensus on Alzheimer's disease (the judgment standard is P-tau217 ≤ 0.47 pg / mL is negative) (Table 1).

[0045] Table 1

[0046]

[0047]

[0048] Example 3

[0049] This embodiment provides an electronic device, which includes a memory, a processor, and a computer program stored in the memory and executable on the processor. When the processor executes the computer program, the function of the device for assisting in diagnosing neurodegenerative diseases as described in Example 1 is implemented.

[0050] Those skilled in the art will appreciate that the apparatus of the present invention may be implemented in various forms of hardware, software, firmware, dedicated processors, or a combination thereof.

[0051] Example 4

[0052] This embodiment provides a computer-readable storage medium, which stores a computer program. The computer program has a program code, and when the program code is executed in a corresponding processor, controller, computing device or terminal, it realizes the function of the device for assisting in diagnosing neurodegenerative diseases as described in Example 1.

[0053] In summary, the present invention found that P-tau181 / Aβ42 was significantly different between healthy people, AD patients and patients with other degenerative diseases based on flow fluorescence detection technology, and further developed an efficient screening tool based on P-tau181 / Aβ42 to achieve high specificity, sensitivity and accuracy to assist in screening neurodegenerative diseases such as Alzheimer's disease. Moreover, based on the flow platform, the detection bottleneck is low, the instrument and equipment requirements are low, the reagent cost is low, it is easier to promote, the detection cycle is short, and it is more suitable for assisting community screening.

[0054] The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention shall fall within the protection scope and disclosure scope of the present invention.

Claims

1. A model for assisting in screening of Alzheimer's disease, characterized in that: The input variables of the model include the body's P-tau181 concentration value and Aβ42 concentration value, and the output variables of the model include the ratio of the P-tau181 concentration value to the Aβ42 concentration value; The criterion for determining a positive diagnosis of Alzheimer's disease is: the ratio of the concentration value of P-tau181 to the concentration value of Aβ42 is greater than 0.

109.

2. Use of the model for assisting in the screening of Alzheimer's disease according to claim 1 in the preparation of products for assisting in the diagnosis of neurodegenerative diseases.

3. A device for assisting in the diagnosis of neurodegenerative diseases, characterized in that The device comprises a data acquisition unit, a data processing unit and a determination unit; The data acquisition unit is used to perform the following steps: Detecting the concentration values ​​of P-tau181 and Aβ42 in the sample to be tested; The data processing unit is used to perform the following steps: Inputting the P-tau181 concentration value and the Aβ42 concentration value obtained by the data acquisition unit into the model for assisting in screening for Alzheimer's disease according to claim 1, and outputting the ratio of the P-tau181 concentration value to the Aβ42 concentration value; The determination unit is used to perform the following steps: The determination is made based on the ratio of the P-tau181 concentration value to the Aβ42 concentration value obtained by the data processing unit and the result is output.

4. The device for assisting in the diagnosis of neurodegenerative diseases according to claim 3, characterized in that: The sample to be tested includes plasma.

5. The device for assisting in the diagnosis of neurodegenerative diseases according to claim 3 or 4, characterized in that: The detection method includes flow cytometry.

6. The device for assisting diagnosis of neurodegenerative diseases according to any one of claims 3 to 5, characterized in that: When the ratio of the P-tau181 concentration value to the Aβ42 concentration value is greater than 0.109, the result is determined and output as positive for Alzheimer's disease.

7. The device for assisting in the diagnosis of neurodegenerative diseases according to any one of claims 3 to 6, characterized in that: When the ratio of the P-tau181 concentration value to the Aβ42 concentration value is ≤0.109, the result is determined and output as negative for Alzheimer's disease, and combined with the medical history, it is determined whether it is other degenerative diseases.

8. The device for assisting diagnosis of neurodegenerative diseases according to claim 7, characterized in that: The other degenerative disease includes at least one of frontotemporal dementia, vascular dementia or Parkinson's disease.

9. An electronic device comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein: When the processor executes the computer program, the function of the device for assisting in diagnosing a neurodegenerative disease as claimed in any one of claims 3 to 8 is realized.

10. A computer-readable storage medium storing a computer program, characterized in that: When the computer program is executed by a processor, the function of the device for assisting in diagnosing a neurodegenerative disease as claimed in any one of claims 3 to 8 is realized.

Images