KR20160006432A - Biomarker for diagnosing Mild Cognitive Impairment - Google Patents
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Abstract
The present invention relates to an MCI diagnostic kit comprising a biomarker for MCI (mild cognitive impairment) diagnosis and an agent for measuring mRNA of the gene or a protein expression level thereof, and a method for diagnosing MCI using the same. Since the PAI1 biomarker for diagnosis of MCI of the present invention is significantly increased in a person with MCI compared to a normal person, it can be usefully used for a biomarker for MCI diagnosis, an MCI diagnostic kit and an MCI diagnosis method.
Description
The present invention relates to a biomarker for the diagnosis of mild cognitive impairment.
Alzheimer's disease is the most common degenerative brain disease that causes dementia, characterized by gradual onset and gradual progression. Alzheimer's disease patients show generalized atrophy due to loss of neurons. Neuritic plaques and neurofibrillary tangles are characteristic lesions in brain tissue. In the early stages of the disease, these brain pathologies are confined mainly to the hippocampus, which is the main brain region responsible for memory, and the olfactory cortex, but gradually expand to the entire brain. It develops seriously as a result of various clinical symptoms such as disorientation, thinking judgment ability, ability to perform daily life, and mental disorder behavior.
The precise mechanisms and causes of Alzheimer's disease are not known precisely. It is thought that beta-amyloid is over-produced and is harmful to brain cells as it is deposited in the brain. It is thought that hyperphosphorylation, inflammation, oxidative damage of tau protein is also a cause . The nervous system (or the elderly) is involved in the deposition of beta amyloid protein, and the nerve fiber bundle is associated with tau protein hyperphosphorylation.
Aβ 40 (Amyloid β40) and Aβ 42 (Amyloid β42), which are known to accumulate in the nervous system of Alzheimer's disease patients, induce the expression of tPA (tissue plasminogen activator) (Periz G et al., EMBO Rep, 1: 477-478, 2000), which converts a plasminogen to a plasmin, an active serine protease. In normal cases, the activated plasmin is able to remove Aβ, but the plasmin activity is reported to be very low despite the increased tPA expression in the neurons cultured from the brain of the Tg2576 transgenic mice bearing the neuronal half (Tucker et al. J. Neurosci, 20: 3937-3946, 2000). This is understood to mean that the tPA / plasmin cascade is inhibited in Alzheimer's disease patients. PAI1 (Plasmin activator inhibitor 1) is a serpin (serine protease inhibitor) family of genes that binds irreversibly to the above-mentioned tPA and functions to completely inhibit the enzyme activity of tPA (Gils A et al., Curr Med Chem., 11: 2323-2334, 2004). PAI1 has been shown to increase expression at the site of amyloid densities in the brain (Hino H et al., Neurosci Lett, 297: 105-108, 2001) (McGeer PL et al., Brain Res Brain Res Rev, 21: 195-218, 1995). PAI1 expression is increased in aged mice and its activity is also observed in mice with increased A [beta]. In this regard, PAI1 is thought to inhibit tPA and reduce the production of plasmin in Alzheimer's disease patients, thereby inhibiting the efficient elimination of Aβ.
Mild Cognitive Impairment (MCI) refers to the pre-dementia stage in which cognitive function, especially memory, is reduced compared to the same age group, but the ability to perform everyday life is maintained. People with mild cognitive impairment are identified as being at high risk for progression to Alzheimer's disease. Thus, mild cognitive impairment is the stage in which Alzheimer's disease is the earliest discoverable stage. Recently, a new type of Alzheimer's disease drug has been shown to be more effective early in the term than Alzheimer's disease, Early detection has clinical significance. In addition, the prevalence of Alzheimer's disease in men over 65 years is 33% and in women it is 45%. Along with the increase in life expectancy due to population growth and medical advances, It is important to select a group that is likely to progress to Alzheimer's disease early in life and to establish a proper treatment system for it, as well as the life aspect of a healthy old age life, as well as the enormous social and economic costs Which can lead to savings.
To date, the diagnosis of Alzheimer's disease has been mainly based on the history of caregiving through neuropsychologists, neuropsychological evaluation, cognitive test, brain imaging test, etc. However, it is possible to understand the onset and progress of Alzheimer's disease and relatively simple and reliable biomarker The development of the Ideally, biomarkers are easily identifiable from blood samples. Considering that patients with Alzheimer's disease are generally older, lumber punctures for obtaining cerebrospinal fluid are potentially dangerous and are not suitable for routine small to medium-sized hospitals. Blood samples are usually obtained easily and safely at home or in a general hospital and can be inspected, and do not require other expensive equipment or surgical procedures.
Therefore, the present inventors sought to find a simple and efficient biomarker for early selection of a high-risk group capable of progressing to Alzheimer's disease in the pre-stage of Alzheimer's disease, and found that the expression of PAI1 gene was significantly increased in the MCI stage The present invention has been accomplished by confirming that it can be easily confirmed only by a sample.
It is an object of the present invention to provide a MCI (Mild Cognitive Impairment) diagnostic biomarker composed of PAI1 (Plasmin activator inhibitor 1).
Yet another object of the present invention is to provide an MCI diagnostic kit comprising an agent for measuring the mRNA of the PAI1 gene or a protein expression level thereof.
Yet another object of the present invention is to provide a method for detecting a PAI1 gene, comprising: (1) measuring the expression level of a PAI1 gene or the expression level of a protein from a biological sample; And (2) comparing the expression level of the PAI1 gene or the expression level of the protein of step (1) with a normal control sample.
It is another object of the present invention to provide an MCI diagnostic strip comprising the antibody of the PAI1 protein.
In order to achieve the above object, there is provided a diagnostic microbiological marker for MCI (Mild Cognitive Impairment) composed of PAI1 (Plasmin activator inhibitor 1).
The present invention also provides an MCI diagnostic kit comprising an agent for measuring mRNA of the PAI1 gene or a protein expression level thereof.
(1) measuring the expression level or protein expression level of any one or more genes selected from the biomarker from the biological sample; And
(2) comparing the expression level of the gene of step (1) or the expression level of the protein with a normal control sample.
The present invention also relates to a sample pad in which a sample is absorbed;
A conjugate pad that binds to the protein in the sample;
A reaction membrane on which a test line and a control line containing the antibody of the protein are treated;
An absorption pad on which a sample of the remaining amount is absorbed; And
Provides an MCI diagnostic strip comprising a support.
The present invention confirms that the expression of PAI1 is increased in Koreans with MCI and provides a biomarker for the diagnosis of MCI by selecting the high risk group which is likely to develop Alzheimer's disease in advance to diagnose Alzheimer's disease early It can be very useful in providing the basis for treatment.
FIG. 1 is a graph showing the relative expression level of PAI1 expression in blood plasma in a normal group, a group with a mild cognitive impairment, and a group of Alzheimer's patients by Western blot analysis compared with the globulin expression level ( * P < 0.05, # P < 0.05, *** P <0.001);
FIG. 2 is a graph showing the mean and standard deviation of the inter-individual variation of plasma PAI1 expression by ELISA in a normal group, a group with mild cognitive impairment, and a group with Alzheimer's disease ( *** P < 0.001 , ### P <0.001);
FIG. 3 is a graph showing the expression levels of individual individuals by ELISA by examining the inter-individual variation of plasma PAI1 expression in a normal group, a group with mild cognitive impairment, and an Alzheimer's patient group ( * P < 0.05, ### P < 0.001, *** P < 0.001).
Hereinafter, terms used in the present invention will be described.
As used herein, the term "diagnostic biomarker" is a bio-derived substance capable of distinguishing MCI (mild cognitive impairment) patients from normal individuals. MCI A nucleic acid or a protein. The marker of the present invention can be used as a molecular indicator for the onset and progression of MCI, and can be used to pre-screen high-risk groups that are likely to develop Alzheimer's disease in the future.
The term " measuring mRNA or its protein expression level " used in the present invention refers to a method for determining the expression level of a protein and its mRNA, which is an MCI diagnostic biomarker, in a biological sample to diagnose MCI, Measure the amount.
The term " antibody " as used herein also refers to a specific protein molecule directed against an antigenic site. For purposes of the present invention, antibodies that specifically bind to the PAI1 protein may be included.
The term " gene expression level measurement " used in the present invention refers to the step of determining the expression level of a protein as an MCI diagnostic biomarker in a biological sample and measuring the amount of the gene.
The term " ELISA " used in the present invention refers to a method of measuring an amount of an antigen or an antibody using an antigen-antibody reaction using an enzyme as a marker.
The term "
Hereinafter, the present invention will be described in detail.
The present invention provides a MCI (Mild Cognitive Impairment) diagnostic biomarker composed of PAI1 (Plasmin activator inhibitor 1).
The amount of PAI1 may be increased in MCI patients, but is not limited thereto.
The PAI1 may be measured at the mRNA or protein level, but is not limited thereto.
The PAI1 may be used to determine whether or not it progresses to Alzheimer's disease, but the present invention is not limited thereto.
In addition, the present invention provides an MCI diagnostic kit comprising an agent for measuring the mRNA of the PAI1 gene or a protein expression level thereof.
The agent for measuring the mRNA level of the PAI1 gene is a primer, a probe, or an antisense nucleotide that specifically binds to the PAI1 gene. Since the nucleic acid information of the gene is known by GeneBank et al., A person skilled in the art, A primer or a probe capable of specifically amplifying the primer or probe can be designed, but is not limited thereto.
The agent for measuring the PAI1 protein level may be an antibody that specifically binds to the protein, but the present invention is not limited thereto.
The antibody may comprise both a polyclonal antibody of PAI1 and a monoclonal antibody.
Polyclonal antibodies can be prepared by injecting an external host with the PAI1 protein or fragment thereof as an antigen according to conventional methods known to those skilled in the art. Such external hosts include mammals such as mice, rats, sheep, and rabbits, and immunogens are generally administered by intramuscular, intraperitoneal, or subcutaneous injection with adjuvants to increase antigenicity Administered to immunize an external host. Serum can be collected periodically from the immunized external host to collect the serum exhibiting improved activity and specificity for the antigen or separating and purifying the antibody therefrom to produce a polyclonal antibody specific for PAI1.
Monoclonal antibodies can be prepared using an immortalized cell line generation method (Kohler G et al., Nature, 256: 495-497, 1975) by fusion known to those skilled in the art. To briefly describe the above method, a mouse is first immunized with a pure PAI1 protein or a fragment thereof, or a peptide thereof is synthesized and bound to bovine serum albumin and immunized with a mouse. The antibody-producing B lymphocytes isolated from immunized mice are fused with human or mouse myeloma cells to produce immortalized hybridoma cells. Then, the production of monoclonal antibody of hybridoma cells was examined by enzyme-linked immunosorbent assay (ELISA), and positive clones were selected. After the cultivation, the antibody was isolated and purified, or injected into the abdominal cavity of rats A plurality of monoclonal antibodies specific for PAI1 can be prepared by collecting the plurality.
In addition, the antibody used in the detection of the PAI1 protein of the present invention comprises a functional fragment of an antibody molecule as well as a complete form having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least antigen binding ability, and includes Fab, F (ab ') 2, F (ab') 2 and Fv.
Antibodies that can specifically bind to the PAI1 or fragments thereof can be bound to a solid substrate to facilitate subsequent steps such as washing or separation of the complex. Solid substrates include, for example, well plates made of synthetic resin, nitrocellulose, glass substrates, metal substrates, glass fibers, microspheres and micro beads. The synthetic resin includes polyester, polyvinyl, polyvinyl chloride, polystyrene, polypropylene, PVDF, and nylon. In addition, when a sample obtained from an individual is contacted with an antibody capable of specifically binding to PAI1 or a fragment thereof bound to a solid substrate, the sample can be diluted to an appropriate degree before contact with the antibody.
The kit may further comprise a detector that specifically binds to an antibody capable of specifically binding to the PAI1 or fragment thereof. The detector may be a conjugate labeled with a coloring enzyme, a fluorescent substance, a radioactive isotope or a colloid, and preferably binds specifically to an antibody capable of specifically binding to PAI1 or a fragment thereof Which may be a secondary antibody. For example, the chromogenic enzyme may be a peroxidase, an alkaline phosphatase or an acid phosphatase (e.g., horseradish peroxidase); (FITC), Rhodamine-B-isothiocyanate (RITC), fluorescein thiourea (FTH), 7 (fluorescein isothiocyanate) -Acetoxycoumarin-3-yl, fluorescein-5-yl, fluorescein-6-yl, 2'7'-dichlorofluorescein-5-yl, 2 ', 7'-dichlorofluorescein Yl, tetramethylrhodamine-6-yl, 4,4-difluoro-5,7-dimethyl-4- 4-diaza-s-indacene-3-ethyl or 4,4-difluoro-5,7-diphenyl-4-bora-3a, 4a-diaza- -Ethyl and the like can be used, but the present invention is not limited thereto.
The kit can diagnose MCI by measuring the antigen-antibody reaction of the antigen-antibody complex or measuring the amount of the antibody after treating the antibody-antibody complex with the detector. Examples of the antigen-antibody reaction include enzyme immunoassay (ELISA), immunoprecipitation, fluorescence immunoassay, enzyme substrate staining, antigen antibody aggregation and the like. The amount of the detection substance or the presence thereof can be detected by color development, fluorescence, (chemiluminescence), absorbance, reflection or transmission.
The high throughput screening (HTS) system is preferably used as a method for searching for the amount of the detecting substance. In this method, a substrate for inducing color development of a chromogenic enzyme bound to a detecting substance is treated to measure a color reaction Way; A fluorescence method carried out by detecting fluorescence by attaching a fluorescent substance to a detector, or a radiation method carried out by detecting radiation with a radioisotope attached to the detector; It is preferable to use a surface plasmon resonance (SPR) method for measuring the change in plasmon resonance of the surface in real time without a label of the detector, or a surface plasmon resonance imaging (SPRI) method for imaging the SPR system.
For example, it is preferable to select a substrate that induces color development according to a chromogenic enzyme. TMB (3,3 ', 5,5'-tetramethyl bezidine), ABTS [2,2'-azino-bis (3- ethylbenzothiazoline-6-sulfonic acid], OPD (o-phenylenediamine), and the like. At this time, it is more preferable that the coloring agent substrate is provided in a dissolved state in a buffer solution (0.1 M NaAc, pH 5.5). The chromogenic substrate such as TMB is decomposed by the chromogenic enzyme bound to the detector to generate a chromophore, and the presence or absence of the PAI1 protein is detected by visually observing the degree of deposition of the chromophore. In addition, the fluorescence method is a method of spotting a detection body labeled with the fluorescent substance using a fluorescent scanner program to confirm the signal, and the degree of coupling can be confirmed by applying this method. Unlike the fluorescence method, the SPR system does not require that the sample be labeled with a fluorescent material, but it is possible to analyze the binding degree of the antibody in real time, but it has a disadvantage that it is impossible to simultaneously analyze samples. In the case of SPRI, it is possible to perform simultaneous multiple sample analysis by using the fine alignment method, but the detection strength is low.
The kit may further include a washing solution for removing the remaining substances after the antigen-antibody binding reaction and the binding reaction of the detection body. Preferably, the wash liquor comprises a phosphate buffer solution, NaCl and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20 But is not limited thereto. After the antigen-antibody binding reaction, the antibody-antibody conjugate is reacted with the detection antibody, and an appropriate amount of the antibody is added to the solid body and washed 3 to 6 times. The reaction stop solution is preferably, but not limited to, a sulfuric acid solution (H2SO4).
The kit may further comprise a negative control comprising a positive control comprising the PAIl standard antigen and an antiserum of the animal not infected with the antigen.
The MCI diagnostic kit can be used in a variety of ways, including a reverse transcription polymerase chain reaction (RT-PCR) kit, a DNA chip kit, an enzyme-linked immunosorbent assay (ELISA) kit, a sandwich ELISA kit, a protein chip kit, a rapid kit, ) Kit. However, the present invention is not limited to this, and it may be accomplished through any known method known to those skilled in the art if a method of complementary binding to mRNA or protein is used.
In addition,
(1) measuring the expression level of the PAI1 gene or the expression level of the protein from the biological sample; And
(2) comparing the expression level of the PAI1 gene or the expression level of the protein of step (1) with a normal control sample, to provide information necessary for diagnosis of MCI.
The expression level of the PAI1 gene may be, but is not limited to, the mRNA expression level of the PAI1 gene.
The mRNA level may be measured by a reverse transcriptase polymerase, a competitive reverse transcriptase polymerase, a real-time reverse transcriptase polymerase, an RNase protection assay, northern blotting, or a DNA chip, but is not limited thereto.
The measurement of the protein expression level may be performed using an antibody that specifically binds to the protein, but is not limited thereto.
Further, the protein expression level of the present invention can be measured by protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Desorption / Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption / Flock Mass Spectrometry analysis, radioimmunoassay, radial immunodiffusion, Oucheronin immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, complement fixation, two-dimensional electrophoresis, liquid chromatography- But are not limited to, mass spectrometry, LC-MS, liquid chromatography-mass spectrometry / mass spectrometry, Western blot, enzyme linked immunosorbent assay and sandwich ELISA.
The biological sample may be blood, but is not limited thereto.
In addition,
A sample pad on which the biological sample is absorbed;
A conjugate pad that binds to PAI1 in the sample;
A reaction membrane on which a test line and a control line containing each antibody of the protein are treated;
An absorption pad on which a sample of the remaining amount is absorbed; And an MCI diagnostic strip comprising a support.
The biological sample of the present invention may be blood, but is not limited thereto.
The sample pad is a sample, that is, the above-mentioned biological sample is applied by dripping or the like so as to be absorbed into the strip, and the absorbed sample passes through the bonding pad and moves along the reaction membrane by capillary phenomenon.
The reaction membrane of the present invention may be, but is not limited to, nitrocellulose, cellulose, poly ethylene terephthalate (PVDF), polyethersufone (PES), glass fiber or nylon.
In addition, the absorbent pad of the present invention is characterized by being a cellulose, an anionic or a hydrophilic porous polymer. However, the absorbent pad may include a material utilizing a capillary phenomenon which is obvious to a person skilled in the art.
In addition, the support of the present invention is for supporting the elements of the strip and may preferably be plastic.
In a specific example of the present invention, the present inventors used plasma proteins from normal subjects, MCI patient groups, and Alzheimer's disease patient groups to identify biomarkers for the diagnosis of MCI patients, a high-risk group likely to progress to Alzheimer's disease Extraction, and Western blot. As a result, the expression of PAI1 protein was significantly increased. Specifically, it was found that the expression level of the Alzheimer's disease patients was increased compared to that of the MCI patients (Fig. 1).
In addition, the present inventors performed ELISA analysis to examine the expression level of PAI1 protein from the blood of the subjects in each of the above groups. As a result, the expression level of the PAI1 protein in the normal group, the MCI patient group and the Alzheimer's disease group (Fig. 2 and Fig. 3).
Therefore, the PAI1 gene of the present invention shows a significant increase in the expression level of MCI patients than that of normal individuals. Therefore, the PAI1 gene of the present invention can be effectively used as an MCI diagnostic biomarker for MCI diagnostic kits, MCI diagnostic methods, and MCI diagnostic strips.
Hereinafter, the present invention will be described in detail with reference to the following experimental examples.
However, the following experimental examples are intended to illustrate the present invention, and the contents of the present invention are not limited by the following experimental examples.
<Experimental Example 1> Statistical data and apoE genotype analysis of normal subjects, MCI patients and Alzheimer's disease patients
Age, education score, Mini-Mental State Examination (MMSE), Clinical Dementia Rating (CDR), and clinical dementia ratings for the normal, MCI, and Alzheimer patients. Apo E (Apolipoprotein E) genotypes were analyzed.
The questionnaires were collected in the order of the first (6), middle (3), high (3), university (4), and graduate (2) And the number of years. MMSE scores ranged from 0 to 19 for mild cognitive impairment, 20 to 23 for mild cognitive impairment, and 24 to 30 for normal Respectively. ApoE genotyping was performed according to the manufacturer's protocol using the SNaPshot system (Applied Biosystems, Forster City, CA, USA). ApoE is a risk gene that increases the risk of developing Alzheimer's disease. If the gene has E4 (ApoE4) genotype in Korea, it is about 2.7 times when it has one allele gene compared with a person without this genotype, and about 17.4 times when it has two It is reported that the risk of Alzheimer's disease increases. In brief, the PCR was performed by using an extension primer to remove the primers and dNTP that were not used in the synthesis, and the PCR products were purified. The product is again The SNaPshot ready Reaction Mix and SNaPshot primer were mixed with each other to perform a secondary PCR reaction, and the PCR product was sequenced to determine the genotype. CDR score was calculated as 0 for normal, 0.5 for suspected dementia, 1 for mild dementia, and 2 or more for severe dementia.
As a result, the education score and the MMSE score were statistically significant and were lower in the MCI patient group and the Alzheimer disease group, respectively, and the CDR score was increased in the MCI and Alzheimer patients with statistical significance. There was no statistical significance in sex and total protein content. Age was statistically significantly higher in the Alzheimer's group (Table 1). ApoE genotype analysis showed that apo E4 allele ratio was higher in patients with MCI and Alzheimer's disease (Table 2).
All values are expressed as mean ± standard deviation.
* P <0.05, *** P <0.001 compared with normal subjects in MCI patients.
* P <0.05, *** P <0.001 in patients with Alzheimer's disease compared with normal subjects.
In the case of Alzheimer's disease patients, compared with MCI patients, ## P <0.01, ### P <0.001.
EXPERIMENTAL EXAMPLE 2 Analysis of Expression of PAI1 Protein by Western Blot in Normal Subjects, MCI Patients and Alzheimer's Disease
In order to identify the expression biomarkers for MCI diagnosis, the degree of PAI-1 expression was examined by western blotting in the Alzheimer's disease patient group, the MCI patient group and the normal group.
Proteins were extracted from plasma from Alzheimer's disease, MCI and normal groups to measure plasma PAI-1 levels. Proteins were electrophoresed on a 4-12% gradient Nupage gel (Invitrogen, Carlsbad, CA, USA) and transferred to a PVDF membrane (Millipore, Billerica, MA, USA). After blocking for 1 hour at room temperature with TBST (50 mM Tri-HCl, pH 7.6, 150 mM NaCl, 0.1% Tween-20) containing 5% skimmed milk powder, the primary antibody of PAI- Lt; / RTI > overnight. (HRP-conjugated goat anti-mouse IgG antibody: Jackson ImmunoResearch, PA, USA) was reacted with primary antibody bound to the membrane for 30 minutes at room temperature, and then subjected to Pierce ECL Western Blotting Substrate (Thermo Scientific, Waltham, Mass., USA).
As a result, it was confirmed that the expression level of PAI1 protein increased from the normal group to the MCI patient group and the Alzheimer's disease group (Fig. 1).
EXPERIMENTAL EXAMPLE 3 Expression Analysis of PAI1 Protein by ELISA in Normal Subjects, MCI Patients and Alzheimer's Disease
To elucidate the expression biomarkers for MCI diagnosis, the level of PAI-1 expression was examined by ELISA in the Alzheimer's disease patient group, the MCI patient group and the normal group.
Plasma PAI-1 levels were measured by ELISA using a Human Serpin E1 / PAI-1 Duoset (R & D systems, Minneapolis, MN, USA) according to the manufacturer's protocol. Briefly, 100 μl of a plasma sample prepared in a suitable diluent was loaded into a plate well and reacted at room temperature for 2 hours. After washing the plate, the detection antibody was added and reacted at room temperature for 2 hours. After the plate was washed, Streptavidin-HRP was added and reacted at room temperature for 20 minutes and the plate was washed again. The substrate solution was added and reacted at room temperature for 20 minutes. The stop solution was added to stop the reaction, and the absorbance was measured at 450 nm with a microplate reader. The results were expressed as means ± SD. All data were analyzed by one-way ANOVA (post-hoc analysis) and post-hoc test (post-hoc analysis).
As a result, it was confirmed that the expression level of PAI1 protein was statistically significantly increased from the normal group to the MCI patient group and the Alzheimer's disease group (FIGS. 2 and 3). The expression level (mean ± SD) was 0.48 ± 0.22 for the normal group, 0.8 ± 0.39 for the MCI patient group ( *** P <0.001 compared to the normal group) and 1.36 ± 0.51 for the Alzheimer patient group ( *** P <0.001; compared to the normal group, ### P <0.001; compared to the MCI patient group).
Claims (14)
(2) a method for measuring the expression level of PAI1 to provide information necessary for MCI (Mild Cognitive Impairment) diagnosis, comprising the step of comparing the expression level of the PAI1 gene or the protein expression level of the above step (1) with a normal control sample .
A conjugate pad that binds to PAI1 (Plasmin activator inhibitor 1) in the sample;
A reaction membrane on which a test line and a control line containing each antibody of the protein are treated;
An absorption pad on which a sample of the remaining amount is absorbed; And
MCI (Mild Cognitive Impairment) diagnostic strip containing a support.
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2014
- 2014-07-09 KR KR1020140085985A patent/KR20160006432A/en not_active Application Discontinuation
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