CN110320368A - Diagnostic kit based on TREM2 and its application on diagnosis of Parkinson disease product - Google Patents

Abstract

The invention discloses a diagnostic kit and its application in the preparation of Parkinson's disease clinical diagnostic products. The diagnostic kit includes: a specific antibody and one or more detection reagents; the specific antibody has the ability to specifically bind to the TREM2 protein, and the detection reagent is used to cooperate with the specific antibody to detect a sample TREM2 gene expression level. This diagnostic kit uses TREM2 protein as a biomarker for early diagnosis of PD. Compared with the traditional diagnosis method of Parkinson's disease, it has the advantages of timeliness, specificity and sensitivity. It can timely know the disease risk in the early stage of the disease, and take corresponding preventive and treatment measures according to the risk level, which is conducive to improving the quality of life. patient outcomes.

Description

TREM2-based diagnostic kit and its application in Parkinson's disease diagnostic products

technical field

The invention relates to the technical field of biological detection, in particular to a diagnostic kit and its application in the preparation of Parkinson's disease clinical diagnostic products.

Background technique

Parkinson's disease (PD) is a degenerative disease of the central nervous system with clinical manifestations including resting tremor, bradykinesia, rigidity and postural gait disturbance. Key features of PD pathology include loss of dopaminergic neurons (DA) in the central nervous system (CNS) nigrostriatal, abnormal activation of microglia, and neuronal α-synuclein (α-synuclein) deposition.

Currently, the gold standard for the diagnosis of Parkinson's disease relies on brain biopsy, while traditional clinical diagnosis mainly relies on comprehensive evaluation of medical history, symptoms, signs and imaging. There are many deficiencies in traditional clinical diagnostic evaluation methods.

First, relying on the traditional clinical diagnosis system of PD, when the patient shows symptoms and signs, most of the DA neurons in the nigrostriatal region have been lost, and the opportunity for early intervention and neuron protection has been missed. Secondly, the traditional clinical diagnosis system has the disadvantages of high misdiagnosis rate and long time for diagnosis.

In order to overcome the shortcomings of traditional diagnostic methods, appropriate biomarkers can be selected to achieve early diagnosis of PD and improve the accuracy and sensitivity of PD diagnosis.

Among them, α-synuclein is a popular molecule for the current candidate PD diagnostic markers. Alpha-synuclein can be abnormally deposited in the gut, digestive glands, and skin of PD patients. However, in the early diagnosis of PD, the accuracy and sensitivity of cerebrospinal fluid α-synuclein as a diagnostic marker are not ideal, and the range of individual differences is large.

In addition, α-synuclein exists in various forms, including monomers, oligomers of different degrees of polymerization, such as dimers, trimers, tetramers, and the like. Too many forms of existence increase the difficulty of clinical application of α-synuclein.

Therefore, it is urgent to provide ideal biomarkers to assist in the early clinical diagnosis of PD.

SUMMARY OF THE INVENTION

In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a diagnostic kit and its application in Parkinson's disease diagnostic products, aiming to solve the problem of poor early diagnosis effect of PD in the prior art.

In order to achieve the above objective, a first aspect of the embodiments of the present invention provides a diagnostic kit. The diagnostic kit includes: a specific antibody and one or more detection reagents; the specific antibody has the ability to specifically bind to the TREM2 protein, and the detection reagent is used to cooperate with the specific antibody to detect the presence of the sample. TREM2 gene expression levels.

Optionally, the specific antibody is selected from monoclonal and/or polyclonal antibodies.

Optionally, the sample is selected from human cerebrospinal fluid and blood.

Optionally, the diagnostic kit is an immunodetection kit; the detection reagent and the specific antibody are used to quantitatively detect the water-soluble TREM2 protein in the sample by an immunodetection method.

Optionally, the TREM2 protein is an expression product of the TREM2 gene, including soluble human TREM2 protein and functional equivalents of human TREM2 protein.

Optionally, the TREM2 protein is selected from one or more of the following proteins:

(1) a protein composed of the amino acid sequence shown in SEQ ID No.1;

(2) A protein derived from the amino acid sequence shown in SEQ ID No. 1 and having the same function as the amino acid sequence shown in SEQ ID No. 1;

(3) A protein composed of an amino acid sequence having a homology of 80% or more with the amino acid sequence shown in SEQ ID No. 1.

Optionally, the protein derived from the amino acid sequence shown in SEQ ID No.1 and having the same function as the amino acid sequence shown in SEQ ID No.1 specifically includes:

A protein derived from the amino acid sequence shown in SEQ ID No. 1 by substitution and/or deletion and/or addition of one or more amino acids.

Optionally, the number of amino acids substituted and/or deleted and/or added is 1-50.

Optionally, a protein composed of an amino acid sequence having a homology of more than 90% to 95% with the amino acid sequence shown in SEQ ID No. 1.

The second aspect of the embodiments of the present invention provides an application of the above-mentioned diagnostic kit in preparing a clinical diagnostic product for Parkinson's disease.

The detection kit provided in the embodiment of the present invention uses TREM2 protein as a biomarker for early diagnosis of PD. Compared with the traditional diagnosis method of Parkinson's disease, it has the advantages of timeliness, specificity and sensitivity. It can timely know the disease risk in the early stage of the disease, and take corresponding preventive and therapeutic measures according to the risk level, which is conducive to improving the quality of life. patient outcomes.

Description of drawings

One or more embodiments are exemplified by the pictures in the corresponding drawings, and these exemplifications do not constitute limitations of the embodiments, and elements with the same reference numerals in the drawings are denoted as similar elements, Unless otherwise stated, the figures in the accompanying drawings do not constitute a scale limitation

1 is a schematic diagram of the correlation between plasma soluble TREM2 protein and synuclein in the control group of Example 1 of the present invention;

Figure 2 is a schematic diagram of the correlation between the cerebrospinal fluid soluble TREM2 protein and synuclein in the control group of Example 1 of the present invention;

3 is a schematic diagram of the correlation between the cerebrospinal fluid soluble TREM2 protein and the level of synuclein in Example 1 of the present invention;

4 is a schematic diagram of the correlation between plasma soluble TREM2 protein and synuclein levels in Example 1 of the present invention;

Fig. 5 is the ROC curve that the level of soluble TREM2 in cerebrospinal fluid in Example 1 of the present invention differentiates PD and normal elderly;

FIG. 6 is a schematic diagram showing the sensitivity and accuracy of the level of soluble TREM2 in the cerebrospinal fluid in Example 1 of the present invention in differentiating PD from normal elderly people.

The amino acid sequence of human TREM2 protein is: MEPLRLLILLFVTELSGAHNTTVFQGVAGQSLQVSCPYDSMKHWGRRKAWCRQLGEKGPCQRVVSTHNLWLLSFLRRWNGSTAITDDTLGGTLTITLRNLQPHDAGLYQCQSLHGSEADTLRKVLVEVLADPLDHRDAGDLWFPGESESFEDAHVEHSISRSLLEGEIPFPPTSILLLLACIFLIKILAASALWAAAWHGQKPGTHPP (NoSELHDDCY)

Detailed ways

In order to make the objectives, technical solutions and effects of the present invention clearer and clearer, the present invention will be further described in detail below with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

Unless otherwise specified, scientific and technical terms used in this specification have the meanings commonly understood by those skilled in the art. Moreover, the protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology related terms and laboratory procedures used herein are the terms and routine procedures widely used in the corresponding fields.

The numerical values and numerical ranges disclosed in the embodiments of the present invention are approximations, not definite values. All values within the limits of error may be included where error or experimental conditions allow. The numerical ranges provided in the present examples are used to represent the relative amounts of the components in the mixture as well as the ranges of temperatures or other parameters recited in other method examples.

In this specification, "diagnosing Parkinson's disease (PD)" or "assessing Parkinson's disease" includes both judging whether a subject has Parkinson's disease, and judging whether a subject is at risk of developing Parkinson's disease . According to the change of the state of the disease, it can include the remission of the disease and the complete cure of the disease.

As described in the Background Art, alpha-synuclein is considered to be the causative agent of PD. α-synuclein has strong self-polymerization ability and can form oligomers and fibers that are more toxic than α-synuclein monomers. Therefore, to determine whether a gene or its expression product has a correlation with Parkinson's disease, it can be indirectly determined by detecting the correlation between the gene or its expression product and α-synuclein.

In order to diagnose or evaluate Parkinson's disease, the applicant creatively provides a diagnostic kit. The diagnostic kit includes: specific antibodies and one or more detection reagents.

Wherein, the specific antibody has the ability to specifically bind to the TREM2 protein, and the detection reagent is used to cooperate with the specific antibody to detect the expression level of the TREM2 gene in the sample.

TREM2 is a microglia-specific receptor, which can help regulate the function of microglia to engulf abnormal proteins in the brain, including abnormal α-synuclein. The applicant has confirmed through experiments that it has a certain correlation with abnormal α-synuclein, can be used as a molecular marker for early diagnosis of PD, and does not have some practical application problems faced by α-synuclein.

In the present specification, the term "antibody" refers to an immunoglobulin molecule generally composed of two identical pairs of polypeptide chains, each pair having one "light" (L) chain and one "heavy" (H) chain. Antibody light chains can be classified as kappa and lambda light chains. Heavy chains can be classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively.

Within the light and heavy chains, the variable and constant regions are linked by a "J" region of about 12 or more amino acids, and the heavy chain also contains a "D" region of about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH).

The heavy chain constant region consists of 3 domains (CH1, CH2 and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain, CL. The constant regions of the antibodies mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.

The VH and VL regions can also be subdivided into regions of high variability called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of 3 CDRs and 4 FRs arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from amino terminus to carboxy terminus. The variable regions (VH and VL) of each heavy/light chain pair, respectively, form the antibody binding site. The assignment of amino acids to regions or domains follows the Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia Definitions in et al. (1989) Nature 342:878-883.

In the present specification, the specific antibody is not limited by any particular method of producing the antibody. Examples are recombinant antibodies, monoclonal antibodies and polyclonal antibodies. The specific antibody can also be an antibody of a different type, eg, an IgG (eg, IgGl, IgG2, IgG3 or IgG4 subtype), IgAl, IgA2, IgD, IgE or IgM antibody.

In this example, the specific binding ability of the specific antibody is derived from the "antigen-binding portion". The antigen-binding portion refers to one or more portions of a full-length antibody that retain the ability to bind the same antigen to which the antibody binds (eg, TREM2) to compete with the intact antibody for specific binding to the antigen.

In particular, antigen-binding portions can be generated by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. In some embodiments, antigen binding moieties include Fab Fab', F(ab')2, Fd, Fv, dAb and complementarity determining region (CDR) fragments, single chain antibodies (eg, scFv), chimeric antibodies, diabodies (diabody) and polypeptides comprising at least a portion of an antibody sufficient to confer specific antigen-binding ability to the polypeptide

It should be noted that the specific antibody or fragment thereof only needs to retain the ability to specifically bind to TREM2 (ie, the antigen-binding portion). Those skilled in the art can select and use any suitable method or technical means to prepare and obtain the specific antibody or its fragment according to the needs of the actual situation.

Based on the inventive idea disclosed in the embodiments of the present invention, those skilled in the art can use the diagnostic kit to specifically and quantitatively detect the expression level of TREM2 protein or TREM2 gene to assist in diagnosing PD and assessing the risk of Parkinson's disease.

In a preferred embodiment, the TREM2 protein in the cerebrospinal fluid and peripheral blood of the subject can be detected at the same time, and according to whether the TREM2 level in the cerebrospinal fluid and the TREM2 level in the peripheral blood are normal, it helps to judge the state of microglia and α-synapses in the CNS The condition of nucleoprotein can assist in diagnosing the risk of PD in the tested person.

Extracellular soluble fragments of TREM2 (Soluble TREM2, sTREM2) can be detected in cerebrospinal fluid and peripheral blood. The level of sTREM2 in the cerebrospinal fluid is closely related to the abnormal protein level in the central nervous system, while the level of sTREM2 in the peripheral blood more reflects the changes of monocytes and macrophages in the peripheral system.

In other words, the samples of the diagnostic kit come from the cerebrospinal fluid and blood of the patient to be tested. The diagnostic kit can simultaneously detect TREM2 protein in cerebrospinal fluid and blood using the same or different methods.

Compared with the method of detecting the amount of TREM2 protein or gene expression level in peripheral blood or cerebrospinal fluid alone, it often cannot correctly reflect the situation of the central nervous system due to the influence of peripheral mononuclear macrophages, and cannot accurately assess intracranial α-synapses. Nucleoproteins and intracranial lesions. A more accurate assessment of intracranial α-synuclein and intracranial lesions can be obtained by simultaneously detecting cerebrospinal fluid and blood samples.

It should be noted that the diagnostic kit provided in the embodiments of the present invention can specifically implement the quantitative detection of TREM2 protein based on any suitable principle and in any suitable manner. Any kit for early diagnosis of PD based on the specific detection of TREM2 protein, or related detection equipment, device or combination of reagents thereof belong to one of the realization forms of the diagnostic kit disclosed in the present invention.

In some embodiments, the diagnostic kit may be an immunoassay kit. The detection reagent and the specific antibody quantitatively detect the water-soluble TREM2 protein in the sample by the method of enzyme-linked immunoassay.

In other words, the diagnostic kit can contain a variety of reagents suitable for ELISA, and cooperate with each other to complete the detection of TREM2. Of course, in the ELISA process, the specific detection reagents used are well known to those skilled in the art, and appropriate reagents can be selected and used according to actual needs.

In some embodiments, the TREM2 protein is the expression product of the TREM2 gene, and contains functional domains related to Parkinson's disease, such as soluble human TREM2 protein and functional equivalents of human TREM2 protein.

The functional equivalents are polypeptides with the same or similar functions, including but not limited to conservative variant proteins of human TREM2 protein, or active fragments thereof, or active derivatives thereof, allelic variants, natural mutants, induced mutants, Proteins encoded by DNAs that can hybridize to human TREM2 DNA under high or low stringency conditions.

Specifically, the TREM2 protein is selected from one or more of the following three proteins:

(1) A protein composed of the amino acid sequence shown in SEQ ID No. 1.

(2) A protein derived from the amino acid sequence shown in SEQ ID No. 1 and having the same function as the amino acid sequence shown in SEQ ID No. 1.

In some embodiments, the derived protein with the same function may be a protein derived from the amino acid sequence shown in SEQ ID No. 1 by substitution and/or deletion and/or addition of one or more amino acids.

Typically, the number of amino acids substituted and/or deleted and/or added is 1-50. Preferably, it can be controlled between 1-30. More preferably, it is selected from 1-20. In the preferred embodiment, it is controlled at 1-10 amino acids.

(3) A protein composed of an amino acid sequence having a homology of 80% or more with the amino acid sequence shown in SEQ ID No. 1.

Homology can also be referred to as sequence identity, indicating the degree of similarity between two amino acid sequences. In a preferred embodiment, it can be an amino acid sequence with more than 90%-95% homology with the amino acid sequence shown in SEQ ID No.1. In other embodiments, it can be specifically selected to be a polypeptide consisting of an amino acid sequence with 96%, 97%, 98%, and 99% homology to the amino acid sequence shown in SEQ ID No. 1.

It should be noted that modification of one or more amino acids in a particular protein does not affect the function of the protein. Those skilled in the art will appreciate that changes in a single amino acid or a small percentage of amino acids or individual additions, deletions, insertions, substitutions to an amino acid sequence are conservative modifications.

Alterations in proteins result in proteins with similar functions, and conservative substitution tables providing amino acids with similar functions are well known in the art.

For example, fusion proteins of TREM2 proteins are proteins modified by the addition of one amino acid or more amino acid residues. In the embodiments of the present invention, there is no restriction on the peptide or protein fused with the TREM2 protein, as long as the obtained fusion protein retains the biological activity of the TREM2 protein.

In other embodiments, the TREM2 protein provided in the embodiments of the present invention further includes non-conservative modifications to the amino acid sequence shown in SEQ ID No. 1, as long as the modified protein can still retain the biological activity of the TREM2 protein. Typically, the number of amino acids mutated in such modified proteins may be 10 or less, such as 6 or less, such as 3 or less.

In the examples of the present invention, the inventive idea of differentially expressing TREM2 gene through total protein in cerebrospinal fluid and peripheral blood is provided, and the correlation between the level of TREM2 protein and Parkinson's disease is revealed. Those skilled in the art can understand that cells expressing TREM2 in brain tissue will fall off into cerebrospinal fluid for circulation, that is, soluble TREM2 protein, so the level of soluble TREM2 in cerebrospinal fluid can reflect the expression of TREM2 gene in brain tissue.

Thus, with the diagnostic kit provided in the embodiments of the present invention, the expression level of soluble TREM2 gene in cerebrospinal fluid and peripheral blood can be detected, and further used to determine whether a subject suffers from Parkinson's disease.

The diagnostic kit provided in the embodiment of the present invention can simultaneously detect the protein level of TREM2 in the cerebrospinal fluid and peripheral blood of the subject, and can be used as a PD clinical diagnostic product for judging whether the subject has Parkinson's disease or whether the subject suffers from Parkinson's disease or not. Whether there is a risk of Parkinson's disease in the test subject, so as to guide the clinician to provide the subject with a preventive plan or a treatment plan.

The diagnostic kit provided by the embodiment of the present invention and its use effect in the early diagnosis of PD will be described in detail below with reference to specific examples. It should be noted that, for the sake of simplicity of presentation, the experimental methods that do not specify specific conditions in the specific examples are usually in accordance with conventional conditions, such as those described in Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989). conditions described, or as recommended by the reagent or equipment manufacturer.

Example 1:

1) Collection of peripheral blood and cerebrospinal fluid samples:

The PD patients in the experimental group were from the First Affiliated Hospital of Guangzhou Medical University, with a total of 50 cases. Aged 57-66 years old, all cases were diagnosed as PD, and the diagnostic criteria were based on the UK Brain Bank clinical diagnostic criteria for Parkinson's disease.

A total of 40 cases in the control group were selected from the routine physical examination population of the First Affiliated Hospital of Guangzhou Medical University. All the subjects were excluded from diseases such as blood lipid metabolism and chronic degenerative diseases of the nervous system, and the age ranged from 55 to 68 years old.

All research subjects have signed the informed consent form for this testing item, and provided peripheral blood and cerebrospinal fluid for gene testing.

2) Construct a detection kit for the determination of soluble TRME2 protein in cerebrospinal fluid and blood:

21) Quantification using two different monoclonal anti-TRME2 antibodies against the extracellular portion of human TRME2 protein (using anti-human TRME2 monoclonal antibody as capture antibody and biotinylated mouse anti-human TRME2 monoclonal antibody as detection antibody) The content of soluble TRME2 was determined.

22) The capture antibody was embedded overnight in PBS buffer at room temperature (RT) at a final concentration of 2.0 μg/ml on a 96-well assay plate.

23) After washing three times with Wash Buffer (0.05% Tween20 in PBS, PH=7.2), the detection plate was blocked with Reagent Diluents (1% BSA in PBS) for 1 h at 25°C.

3) Construct a detection kit for the determination of synuclein:

31) Using two different monoclonal anti-TRME2 antibodies against human synuclein protein (using anti-human synuclein monoclonal antibody as capture antibody and biotinylated mouse anti-human synuclein monoclonal antibody as detection antibody) to quantify the content of synuclein.

32) Embed the capture antibody at a final concentration of 4.0 μg/ml in PBS buffer overnight on a 96-well assay plate at room temperature.

33) After washing three times with Wash Buffer (0.05% Tween20 in PBS, pH=7.2), the detection plate was blocked with Reagent Diluents (1% BSA in PBS) for 1 h at 25°C.

4) Determination of soluble TRME2 in serum and cerebrospinal fluid (soluble human TRME2-FC and human a-synuclein-FC were used as standards in all experiments):

41) Take 250 μl (or 0.25 g) of whole blood or cerebrospinal fluid into an RNase-Free filter column, centrifuge at 13,000 rpm for 2 minutes, and collect the supernatant.

42) Add 100 μl of freshly rewarmed CSF, serum and standard to the coated plate to be tested, incubate at room temperature for 2 hours, wash with Wash Buffer, and add to 100 μl of detection antibody with a final concentration of soluble TRME2 protein (final concentration: 35.0ng /ml) and a-synuclein detection antibody (final concentration: 25.0ng/ml) and incubated at room temperature for 2h.

43) After washing with Wash Buffer, add 100 μl of Streptavidin labeled with horseradish peroxidase and incubate at room temperature for 20 min in the dark.

44) After washing with Wash Buffer, add 100 μl of Substrate Solution (R&D Catalog #DY999) and incubate at room temperature for 20 min in the dark.

45) Color development was stopped by adding 50 μl of 2N sulfuric acid (R&D Catalog #DY994).

46) Measure the OD value of each well at 450 nm, and use the OD value measured at 540 nm to correct.

5) Statistical methods:

Each experiment was repeated three times, and the data were expressed in the form of mean ± standard deviation. SPSS21.0 statistical software was used for statistical analysis. The difference between the two was tested by t test, and it was considered to be statistically significant when P<0.05.

6) Experimental results:

As shown in Figure 1, compared with the normal population, the blood levels of soluble TREM2 and synuclein in the control group showed an increasing trend, and the difference was not statistically significant (P<0.05).

As shown in Figure 2, compared with the normal population, the level of soluble TREM2 synuclein in the cerebrospinal fluid of the control group was significantly increased, and the difference was statistically significant (P<0.05).

As shown in Figure 3, the amount of soluble TREM2 protein in cerebrospinal fluid was positively correlated with the level of synuclein, and the difference was statistically significant (P<0.05).

As shown in Figure 4, plasma soluble TREM2 and synuclein levels were positively correlated, and the difference was not statistically significant (P<0.05).

As shown in Figure 5, the ROC curve (AUC=0.92) of CSF soluble TREM2 levels discriminating PD from normal elderly.

As shown in Figure 6, the sensitivity and accuracy of CSF-soluble TREM2 when used as PD diagnosis are shown.

The above experimental results show that, compared with the normal population, the level of soluble TREM2 in the cerebrospinal fluid of patients with Parkinson's disease is increased, and the level of soluble TREM2 in peripheral blood has no significant relationship, indicating that measuring the level of soluble TREM2 in the cerebrospinal fluid and blood of the subjects can help To determine the risk of a subject suffering from Parkinson's disease. Thus, TREM2 can be used as a molecular marker for evaluating PD disorders.

Embodiments of the present invention also provide a diagnostic kit and applications thereof. The diagnostic kit simultaneously quantitatively detects TREM2 protein in peripheral blood and cerebrospinal fluid, and evaluates the risk of Parkinson's disease in subjects through the expression level of TREM2 gene, so as to improve the clinical diagnosis and treatment of Parkinson's disease.

It can be understood that for those of ordinary skill in the art, equivalent replacements or changes can be made according to the technical solutions of the present invention and the inventive concept, and all these changes or replacements should belong to the protection scope of the appended claims of the present invention.

sequence listing

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Claims (10)

1. a kind of diagnostic kit characterized by comprising specific antibody and one or more kinds of detection reagents；It is described Specific antibody has the ability in conjunction with TREM2 protein-specific, and the detection reagent with the specific antibody for matching It closes, the TREM2 gene expression dose of test sample.

2. diagnostic kit according to claim 1, which is characterized in that the specific antibody is selected from monoclonal antibody And/or polyclonal antibody.

3. diagnostic kit according to claim 1, which is characterized in that the sample is selected from the cerebrospinal fluid and blood of people.

4. diagnostic kit according to claim 1, which is characterized in that the diagnostic kit is immunologic function test reagent Box；The detection reagent and the specific antibody, the water solubility by the method for immune detection, in sample described in quantitative detection TREM2 albumen.

5. diagnostic kit according to claim 1, which is characterized in that the TREM2 albumen is the expression of TREM2 gene Product, the functional equivalent including soluble people TREM2 albumen and people's TREM2 albumen.

6. diagnostic kit according to claim 1, which is characterized in that the TREM2 albumen is in following protein One or more:

(1) protein of the composition of the amino acid sequence as shown in SEQ ID No.1；

(2) derived from the amino acid sequence as shown in SEQ ID No.1, and have and amino acid shown in SEQ ID No.1 The protein of sequence function having the same；

(3) albumen that there is the amino acid sequence of 80% or more homology to constitute with amino acid sequence shown in SEQ ID No.1 Matter.

7. diagnostic kit according to claim 6, which is characterized in that the amino acid as shown in SEQ ID No.1 Derived from sequence, and there is the protein with amino acid sequence function having the same shown in SEQ ID No.1, it is specific to wrap It includes:

Amino acid sequence shown in SEQ ID No.1 by the substitution of one or more amino acid and/or missing and/or is added Add and derivative protein.

8. diagnostic kit according to claim 7, which is characterized in that the ammonia of the substitution and/or deletion and/or addition Base acid quantity is 1-50.

9. diagnostic kit according to claim 6, which is characterized in that with amino acid sequence shown in SEQ ID No.1 The protein that amino acid sequence with 90%-95% or more homology is constituted.

10. a kind of if the described in any item diagnostic kits of claim 1-9 are on preparation Parkinson's disease clinical diagnosis product Using.