CN114410774A - Risk assessment method applied to Alzheimer's disease - Google Patents
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Abstract
The invention discloses a disease risk assessment method applied to Alzheimer's disease, belonging to the technical field of preventive medicine and comprising the following steps: s1, acquiring medical history of the evaluation object; s2, performing mild cognitive impairment assessment on the assessment subjects; s3, screening the evaluation object for further cognitive disorder evaluation; s4, carrying out routine laboratory examination on the evaluation object; s5, carrying out APOE gene detection on the evaluation object; s6, analyzing and detecting the cerebrospinal fluid biomarker of the evaluation object; s7, completing the evaluation analysis of the risk of Alzheimer' S disease. According to the method for evaluating the risk of the Alzheimer's disease, a patient is firstly subjected to medical history collection, two-layer screening of mild cognitive impairment and cognitive impairment is carried out, risk pre-evaluation is carried out based on evaluation content, multiple detections in a laboratory are finally carried out, and the risk of the Alzheimer's disease is finally evaluated and judged according to multiple parameters.
Description
Technical Field
The invention belongs to the technical field of preventive medicine, and particularly relates to a disease risk assessment method applied to Alzheimer's disease.
Background
Alzheimer's disease, the chinese name "senile dementia", is a neurodegenerative disease caused by the death of nerve cells in the brain, and is mostly occurred in people over 65 years old. According to the statistics of the United states, the rate of Alzheimer's disease in 65-69 years old is about 4%, and in the elderly over 85 years old, the rate is as high as 36%. Meanwhile, AD ranks 9 th among causes of death in elderly over 65 years old. There are about 400 million patients with alzheimer's disease in the whole united states. And in China, the number of patients reaches 900 ten thousand. The total number of AD patients in the whole world is up to 2100-3500 ten thousand. It is expected that by 2030, the number of Alzheimer's disease patients worldwide may increase to 7000 million. With age, the risk of Alzheimer's disease increases.
The occurrence of alzheimer's disease is a progressive process. Symptoms at the initial stage of onset may be overlooked because they are mistakenly considered to be amnesia due to age. With the gradual development of the disease, the learning and memory ability, cognitive judgment ability and basic ability of many daily activities of the patient are gradually degraded, and meanwhile, symptoms such as temperament change and behavior difficulty can appear. In the advanced stages of the disease, alzheimer's disease can lead to the loss of intelligence and ultimately death of the patient. The average survival time per patient after diagnosis was approximately 8 years. In the united states, the cost of care for patients alone, in addition to the cost of medication, is as high as $ 1000 billion. The various costs for each AD patient are on average $ 174,000. Such high costs are an extremely heavy burden to both individuals and society. Because of the progressive nature of alzheimer's disease, the patient himself and his family members often are unaware that they have exhibited symptoms of the disease in their early stages of onset and are often mistaken for the normal phenomena that occur as a result of aging. When a patient has already shown obvious abnormal symptoms and is in medical service, the method usually comprises behavioral tests, clinical tests and the like to determine whether the cranial nerves of the patient have abnormalities. Early screening is very important because Alzheimer's disease has already progressed to a certain stage and often the best treatment opportunity has been missed.
Disclosure of Invention
In view of the deficiencies of the prior art, the present invention provides a method for assessing risk of alzheimer's disease, which solves the above problems in the background art.
In order to achieve the purpose, the invention provides the following technical scheme: a method for assessing the risk of Alzheimer's disease comprises the following steps:
s1, acquiring medical history of the evaluation object, wherein the medical history acquisition comprises current medical history, past medical history and family history;
s2, performing mild cognitive impairment assessment on the assessment objects by adopting a Montreal cognitive assessment scale, wherein the assessment contents comprise attention, executive functions, memory, language functions, visual structure functions, abstract thinking, calculation and orientation force;
s3, screening the evaluation object for further cognitive disorder evaluation;
s4, carrying out routine laboratory examination on the evaluation object;
s5, carrying out APOE gene detection on the evaluation object by adopting a real-time fluorescent quantitative PCR method;
s6, analyzing and detecting the cerebrospinal fluid biomarker of the evaluation object;
s7, and finishing the evaluation analysis of the risk of Alzheimer' S disease based on the detection results of the steps S5 and S6.
Further optimizing the technical scheme, in the step S1, the current medical history includes onset time, onset form and clinical symptoms, wherein the clinical symptoms include impaired symptoms of different cognitive domains including memory, executive and attention, language and visual space; past history includes other diseases that may lead to alzheimer's disease, including central nervous system disease and history of hypertension, hyperglycemia, and hyperlipidemia; family history includes a family history of dementia, including alzheimer's disease.
Further optimizing the technical scheme, in the S4, routine laboratory tests comprise blood routine, biochemical ten items, blood coagulation four items, folic acid, VB12, Jiagong items, HIV and syphilis screening and other diseases which can cause cognitive decline caused by Alzheimer' S disease.
In the S5, the APOE coding gene is located on chromosome 19, and 6 different APOE genotypes including 3 homozygous types and 3 heterozygous types exist, which are determined by single nucleotide polymorphism sites rs429358 and rs 7412.
Further optimizing the technical scheme, in S5, the real-time fluorescent quantitative PCR method includes the following specific operation procedures:
s501, extracting DNA of an evaluation object, wherein the DNA is derived from extracellular free DNA in human body fluid;
s502, carrying out PCR reaction on the APOE gene fragment by taking the extracted free DNA as a template, and identifying the base types of two SNP loci rs429358 and rs7412 of the APOE gene;
and S503, carrying out PCR reaction by using a probe with FAM and TET fluorescent luminescent groups, and detecting a fluorescent result after the PCR reaction is started.
Further optimizing the technical scheme, the Alzheimer disease risk is high when the fluorescence detection result is as follows:
1) both the FAM at the rs429358 site and the TET fluorescent luminescent group have reaction in detection, and both the FAM at the rs7412 site and the TET fluorescent luminescent group have reaction in detection;
2) FAM at the rs429358 site and TET fluorescent luminescent groups are both reacted, and only the TET fluorescent luminescent groups at the rs7412 site are reacted;
3) the rs429358 site only has TET fluorescent luminous group detection reaction, and the rs7412 site only has TET fluorescent luminous group detection reaction.
In the S6, the AD cerebrospinal fluid biomarkers include total tau protein T-tau, phosphorylated tau protein P-tau, and beta-amyloid precursor protein APP cleavage fragment a β 42 consisting of 42 amino acids.
Further optimizing the technical scheme, in the step S3, the screening method for further cognitive impairment assessment includes a simple mental state scale and a long-valley dementia scale.
Further optimizing the technical scheme, the evaluation content of the simple mental state scale comprises orientation ability, immediate recall, attention, calculation ability, delayed recall, language function and visual space sense.
Further optimizing the technical scheme, the evaluation content of the ChangYuchuan dementia scale comprises orientation force, memory function, common knowledge, calculation and object inscription naming memory.
Compared with the prior art, the invention provides a disease risk assessment method applied to Alzheimer's disease, which has the following beneficial effects:
according to the method for evaluating the risk of the Alzheimer's disease, the patient is firstly subjected to medical history collection, the mild cognitive disorder and the cognitive disorder are screened in two layers, the risk is pre-evaluated based on evaluation content, finally, a plurality of tests in a laboratory are carried out, and the risk of the Alzheimer's disease is finally evaluated and judged according to a plurality of parameters, so that the evaluation accuracy of the method is greatly improved.
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FIG. 1 is a schematic flow chart of a risk assessment method for Alzheimer's disease according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
referring to fig. 1, a method for assessing risk of alzheimer's disease comprises the following steps:
and S1, acquiring medical history of the evaluation object, wherein the medical history acquisition comprises the current medical history, the past medical history and the family history.
Wherein the current medical history comprises onset time, onset form and clinical symptoms, wherein the clinical symptoms comprise impaired symptoms of different cognitive domains including memory, executive and attention, language and visual space; past history includes other diseases that may lead to alzheimer's disease, including central nervous system disease and history of hypertension, hyperglycemia, and hyperlipidemia; family history includes a family history of dementia, including alzheimer's disease.
And S2, performing mild cognitive impairment assessment on the assessment subjects by adopting a Montreal cognitive assessment scale, wherein the assessment contents comprise attention, executive functions, memory, language functions, visual structure functions, abstract thinking, calculation and orientation.
The screening method for further cognitive disorder assessment comprises a simple mental state scale and a long-valley dementia scale. The evaluation content of the simple mental state scale comprises orientation ability, immediate recall, attention, calculation ability, delayed recall, language function and visual space feeling. The evaluation content of the ChangYuchuan dementia scale comprises orientation force, memory function, common knowledge, calculation and object inscription naming memory. In the simple mental state scale, the normal score is more than or equal to 27, the mild dementia score is 21-26, the severe dementia score is 10-20, the severe dementia score is less than 10, the score is related to education level, and a clear corresponding score relation exists.
And S3, screening the evaluation object for further cognitive disorder evaluation.
And S4, performing routine laboratory examination on the evaluation object.
Wherein, routine laboratory examination includes blood routine, biochemical ten items, blood coagulation four items, folic acid, VB12, first-degree full item, HIV and syphilis screening, and diseases which can cause cognitive decline caused by Alzheimer's disease
And S5, carrying out APOE gene detection on the evaluation object by adopting a real-time fluorescence quantitative PCR method.
Wherein, the APOE coding gene is positioned on chromosome 19, and 6 different APOE genotypes exist, including 3 homozygous types and 3 heterozygous types, determined by single nucleotide polymorphism sites rs429358 and rs 7412.
The real-time fluorescent quantitative PCR method comprises the following specific operation procedures:
s501, extracting DNA of an evaluation object, wherein the DNA is derived from extracellular free DNA in human body fluid;
s502, carrying out PCR reaction on the APOE gene fragment by taking the extracted free DNA as a template, and identifying the base types of two SNP loci rs429358 and rs7412 of the APOE gene;
and S503, carrying out PCR reaction by using a probe with FAM and TET fluorescent luminescent groups, and detecting a fluorescent result after the PCR reaction is started.
Further, the primer sequence and the probe sequence for rs429358 site amplification in the PCR reaction are as follows:
rs429358F：5’-ACGGCTGTCCAAGGAGCTGC-3’
rs429358R：5’-AGGCGCACCCGCAGCTCCTC-3’
the above is the primer sequence.
prob-1T：5’-FAM-AGGACGTGTGCGGCCGCCTG-MGB Eclipse-3’
prob-1C：5’-TET-AGGACGTGCGCGGCCGCCTG-MGB Eclipse-3’
The probe sequences are provided, and the probes are provided with FAM or TET fluorescent luminescent groups and MGB Eclipse fluorescent quenching groups, so that a PCR system can not emit fluorescence when the PCR reaction is not carried out; when the PCR reaction is initiated, the PCR product and the fluorescent probe bind to initiate fluorescence and have a sufficiently strong light intensity to be detected and recorded.
Further, the primer sequence and the probe sequence for rs7412 site amplification in the PCR reaction are:
rs7412F：5’-TGCGGGTGCGCCTCGCCTCCC-3’
rs7412R：5’-AGGCGCTCGCGGATGGCGCTG-3’
the above is the primer sequence.
prob-2T：5’-FAM-CAGAAGTGCCTGGCAGTGTA-MGB Eclipse-3’
prob-2C：5’-TET-CAGAAGCGCCTGGCAGTGTA-MGB Eclipse-3’
The above is a probe sequence.
Further, the risk of Alzheimer's disease is high when the fluorescence detection result is as follows:
1) both the FAM at the rs429358 site and the TET fluorescent luminescent group have reaction in detection, and both the FAM at the rs7412 site and the TET fluorescent luminescent group have reaction in detection;
2) FAM at the rs429358 site and TET fluorescent luminescent groups are both reacted, and only the TET fluorescent luminescent groups at the rs7412 site are reacted;
3) the rs429358 site only has TET fluorescent luminous group detection reaction, and the rs7412 site only has TET fluorescent luminous group detection reaction.
And S6, analyzing and detecting the cerebrospinal fluid biomarker of the evaluation subject.
Wherein the AD cerebrospinal fluid biomarkers comprise total tau protein T-tau, phosphorylated tau protein P-tau and beta-amyloid precursor protein APP decomposition fragment Abeta 42 consisting of 42 amino acids. Total tau protein T-tau and phosphorylated tau protein P-tau in cerebrospinal fluid of patients with Alzheimer's disease are both obviously increased, and the content of beta-amyloid precursor protein APP decomposition fragment Abeta 42 is obviously reduced.
S7, and finishing the evaluation analysis of the risk of Alzheimer' S disease based on the detection results of the steps S5 and S6.
The invention has the beneficial effects that:
according to the method for evaluating the risk of the Alzheimer's disease, the patient is firstly subjected to medical history collection, the mild cognitive disorder and the cognitive disorder are screened in two layers, the risk is pre-evaluated based on evaluation content, finally, a plurality of tests in a laboratory are carried out, and the risk of the Alzheimer's disease is finally evaluated and judged according to a plurality of parameters, so that the evaluation accuracy of the method is greatly improved.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A method for assessing the risk of Alzheimer's disease comprises the following steps:
s1, acquiring medical history of the evaluation object, wherein the medical history acquisition comprises current medical history, past medical history and family history;
s2, performing mild cognitive impairment assessment on the assessment objects by adopting a Montreal cognitive assessment scale, wherein the assessment contents comprise attention, executive functions, memory, language functions, visual structure functions, abstract thinking, calculation and orientation force;
s3, screening the evaluation object for further cognitive disorder evaluation;
s4, carrying out routine laboratory examination on the evaluation object;
s5, carrying out APOE gene detection on the evaluation object by adopting a real-time fluorescent quantitative PCR method;
s6, analyzing and detecting the cerebrospinal fluid biomarker of the evaluation object;
s7, and finishing the evaluation analysis of the risk of Alzheimer' S disease based on the detection results of the steps S5 and S6.
2. The method for assessing risk of developing alzheimer' S disease as claimed in claim 1, wherein in S1, the medical history includes onset time, onset form and clinical symptoms, wherein the clinical symptoms include impaired symptoms of different cognitive domains including memory, executive and attention, language and visual space; past history includes other diseases that may lead to alzheimer's disease, including central nervous system disease and history of hypertension, hyperglycemia, and hyperlipidemia; family history includes a family history of dementia, including alzheimer's disease.
3. The method as claimed in claim 1, wherein in step S4, the disease causing cognitive decline due to Alzheimer' S disease includes routine laboratory tests including blood routine, biochemical tests, coagulation tests, folic acid, VB12, Jiagong tests, HIV tests and syphilis tests.
4. The method of claim 1, wherein the APOE-encoding gene is located on chromosome 19 in S5 and 6 different APOE genotypes including 3 homozygous and 3 heterozygous forms are present, depending on the single nucleotide polymorphism sites rs429358 and rs 7412.
5. The method for assessing risk of developing alzheimer' S disease as claimed in claim 1, wherein in S5, the real-time fluorescence quantitative PCR method comprises the following specific procedures:
s501, extracting DNA of an evaluation object, wherein the DNA is derived from extracellular free DNA in human body fluid;
s502, carrying out PCR reaction on the APOE gene fragment by taking the extracted free DNA as a template, and identifying the base types of two SNP loci rs429358 and rs7412 of the APOE gene;
and S503, carrying out PCR reaction by using a probe with FAM and TET fluorescent luminescent groups, and detecting a fluorescent result after the PCR reaction is started.
6. The method for assessing the risk of developing alzheimer's disease as claimed in claim 5, wherein the risk of developing alzheimer's disease is high when the fluorescence detection result is the following result:
1) both the FAM at the rs429358 site and the TET fluorescent luminescent group have reaction in detection, and both the FAM at the rs7412 site and the TET fluorescent luminescent group have reaction in detection;
2) FAM at the rs429358 site and TET fluorescent luminescent groups are both reacted, and only the TET fluorescent luminescent groups at the rs7412 site are reacted;
3) the rs429358 site only has TET fluorescent luminous group detection reaction, and the rs7412 site only has TET fluorescent luminous group detection reaction.
7. The method of claim 1, wherein the biomarkers in cerebrospinal fluid of AD comprise total tau protein T-tau, phosphorylated tau protein P-tau, and beta-amyloid precursor protein APP degradation fragment A β 42 consisting of 42 amino acids in S6.
8. The method as claimed in claim 1, wherein the screening method for cognitive impairment assessment in S3 includes methods of the gratuitous mental state scale and the long-valley dementia scale.
9. The method as claimed in claim 8, wherein the evaluation content of the mental state scale includes orientation ability, immediate memory, attention, calculation ability, delayed memory, language function and visual space sense.
10. The method as claimed in claim 8, wherein the evaluation content of the Long Guchuan dementia Scale comprises orientation, memory, general knowledge, calculation, object name and memory.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012053255A1 (en) * | 2010-10-20 | 2012-04-26 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Biomarker for alzheimer's disease or mild cognitive impairment |
| CN108004315A (en) * | 2017-12-22 | 2018-05-08 | 佰世凯（杭州）生物科技有限公司 | Appraisal procedure for Alzheimer's disease risk |
| CN111524602A (en) * | 2020-04-28 | 2020-08-11 | 西安玖诚玖谊实业有限公司 | Old person's memory and cognitive function aassessment screening early warning system |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012053255A1 (en) * | 2010-10-20 | 2012-04-26 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Biomarker for alzheimer's disease or mild cognitive impairment |
| CN108004315A (en) * | 2017-12-22 | 2018-05-08 | 佰世凯（杭州）生物科技有限公司 | Appraisal procedure for Alzheimer's disease risk |
| CN111524602A (en) * | 2020-04-28 | 2020-08-11 | 西安玖诚玖谊实业有限公司 | Old person's memory and cognitive function aassessment screening early warning system |
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