CN114137225A - Serum polypeptide diagnostic molecular composition for primary depression and application thereof - Google Patents
Serum polypeptide diagnostic molecular composition for primary depression and application thereof Download PDFInfo
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Abstract
The invention discloses a serum polypeptide molecular composition for patients with primary depression and application thereof, wherein the amino acid sequence of ITIH4 in the composition is shown in SEQ.ID.NO.1, and the amino acid sequence of TUBB in the composition is shown in SEQ.ID.NO. 2. ITIH4 is human alpha-trypsin inhibitor heavy chain 4, with an exact molecular weight of 1062.35 daltons; TUBB is human β -tubulin, with an exact molecular weight of 1868.21 daltons. The composition of the ITIH4 and the TUBB shows specific high expression in the serum detection of patients with primary depression, and the composition of the ITIH4 and the TUBB is detected by a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or the expression level of the composition of the ITIH4 and the TUBB is detected by an ELISA method, so that the composition can be used as a detection method for the serum of patients with primary depression.
Description
Technical Field
The invention belongs to the technical field of biomarkers, and particularly relates to a serum polypeptide molecular composition for primary depression, and a detection method and application thereof.
Background
Depression is described by scientists as "cancer of the 21 st century", which is one of the main causes of global disease burden at present, and the worldwide prevalence varies from 3% to 17%, which has great influence on physical and mental health and social life of patients. Like other psychiatric disorders, the etiology of depression is extremely complex, involving psychosocial, genetic, epigenetic, neuroendocrine, and neuroimmune factors. This complexity directly affects our accuracy in diagnosing depression and its subtypes, affects our understanding of their pathophysiology, and the ability to select effective therapeutic strategies. At present, the clinical diagnosis of depression comprises medical history evaluation, mental examination and the like, the commonly used standardized patient self-rating scale and clinical scale can be used for evaluating the severity of depression symptoms of patients, the mental scale comprises PHQ-9, SDS, BDI, QIDS-SR and the like, the diagnosis strategy for early onset of depression is lacked, and the existing diagnosis method lacks sensitivity and specificity for early screening and diagnosis of depression. Therefore, the accurate molecular mechanism of the occurrence and development of the depression is discussed, and the search of a new specific diagnostic marker has important clinical significance for early diagnosis and treatment of the depression and improvement of the life quality of patients.
Before any pathological change occurs in any disease, the intracellular proteins are altered in composition and quantity and are reflected by the pattern of proteins in the serum. Therefore, by comparing the expression of different proteins in the serum of different disease populations, it is possible to screen out disease-related marker molecules. Serum proteomics refers to the study of all proteins expressed in the serum of a selected target population, and on the basis of establishing a normal Protein Expression Map (PEM), differential protein spots of the proteins are searched, and disease-related proteins are identified, so that the structure and function of the proteins are further studied, and a new way is developed for studying major disease pathophysiological mechanisms, specific markers for early diagnosis, drug action targets and the like. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
Serum diagnosis is considered to be the most recent and effective method for early diagnosis of cancer. The method judges the occurrence and development of the tumor by searching tumor markers in blood, particularly protein markers in the blood, so as to realize early diagnosis of the tumor. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a serum polypeptide molecular composition for diagnosing primary depression, a detection method and application thereof, wherein the molecular polypeptide composition comprises (i) human alpha-trypsin inhibitor heavy chain 4(Isoform 2of Inter-alpha-trypsin inhibitor heavywain H4 precorsor, ITIH4) and (ii) human beta-Tubulin (Tubulin beta chain, TUBB), and the composition is a serum marker of the primary depression.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses a serum polypeptide diagnostic molecule composition, which comprises polypeptide ITIH4 and polypeptide TUBB, wherein the amino acid sequence of ITIH4 is shown in SEQ.ID.NO.1, and the amino acid sequence of TUBB is shown in SEQ.ID.NO. 2.
Preferably, the molecular weight of the ITIH4 is 1062.35 daltons and the molecular weight of the TUBB is 1868.21 daltons.
The invention also discloses application of the serum polypeptide diagnostic molecule composition in preparing a serum diagnostic reagent for primary depression, wherein the detection parameter of the ITIH4 in the serum of a primary depression patient is 35.03-88pg/mL, and the detection parameter in the serum of a normal healthy population is 18.705-56.255 pg/mL; the detection parameter of the TUBB in the serum of the patients with the primary depression is 47.475-197.875pg/mL, and the detection parameter in the serum of the normal healthy people is 27.6-93.155 pg/mL.
Preferably, ITIH4 is significantly highly expressed with TUBB in serum samples from patients with primary depression.
Preferably, the serum diagnostic marker for primary depression is a serum polypeptide molecule for ELISA detection of primary depression.
Preferably, the serum polypeptide molecule composition ITIH4 and TUBB are novel targets for ELISA detection markers.
The invention also discloses application of a molecule combined with the serum polypeptide diagnostic molecule composition in preparing a serum diagnostic reagent for primary depression, wherein the serum polypeptide diagnostic molecule composition comprises ITIH4 and TUBB, the amino acid sequence of ITIH4 is shown in SEQ ID No.1, and the amino acid sequence of TUBB is shown in SEQ ID No. 2.
The invention also discloses a diagnostic kit for primary depression, which comprises serum polypeptide molecule compositions ITIH4 and TUBB, wherein the amino acid sequence of ITIH4 is shown in SEQ.ID.NO.1, and the amino acid sequence of TUBB is shown in SEQ.ID.NO.2
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses a serum polypeptide molecular composition for primary depression, the amino acid sequence of which is shown as SEQ.ID.NO.1 and SEQ.ID.NO.2, and the molecular composition comprises (I) ITIH4 and (II) TUBB. ITIH4 is human alpha-trypsin inhibitor heavy chain 4, with an exact molecular weight of 1062.35 daltons; TUBB is human β -tubulin, with an exact molecular weight of 1868.21. ITIH4 and TUBB showed specifically high expression in serum testing of patients with primary depression: expression range of ITIH4 in serum of patients with primary depression is: 35.03-88 pg/mL; the range of expression in serum in normal healthy people is: 18.705-56.255pg/mL, and there was a very significant difference in expression between the two groups (p < 0.001); (vii) TUBB is expressed in the serum of patients with primary depression in the range: 47.475-197.875 pg/mL; the expression range in the serum of normal healthy people is as follows: 27.6-93.155pg/mL, and there was a significant difference in expression between the two groups (p < 0.05).
Given the specific high expression of ITIH4 and TUBB in serum of primary depression, the combination of ITIH4 and TUBB can be used as a serum diagnostic marker for primary depression; the expression level of the composition of the ITIH4 and the TUBB is detected by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or an ELISA method, and the composition of the ITIH4 and the TUBB can be used as a method for detecting patients with primary depression. For the diagnosis of ELISA for detecting serum of primary depression, the composition of ITIH4 and TUBB can be used as a new target point for ELISA detection drugs.
Drawings
FIG. 1 shows the protein polypeptide peaks m/z of the present invention: 1062.35 differential protein polypeptide expression in patients with primary depression (red) and in normal healthy population groups (green);
FIG. 2 shows the protein polypeptide peaks m/z of the present invention: 1868.21 differential protein polypeptide expression in patients with primary depression (red) and in normal healthy population groups (green);
FIG. 3 is a gel chromatographic separation chart of a serum protein polypeptide mixture of a primary depression patient of the invention: wherein, the abscissa in the chromatogram represents the sample outflow time, and the ordinate represents the relative abundance of the polypeptide;
FIG. 4 is a MS/MS mass spectrometric identification profile of ITIH4 of the present invention; wherein, m/z: 1062.35, z is 1;
FIG. 5 is a MS/MS mass spectrometric identification profile of the TUBB of the present invention; wherein, m/z: 1868.21, z is 2;
FIG. 6 shows the expression level of ITIH4 protein in serum of patients with primary depression and normal healthy persons according to the present invention; wherein N is 21;
FIG. 7 is the expression level of TUBB protein in serum of patients with primary depression and normal healthy persons according to the present invention; wherein, N is 17.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in 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.
It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the accompanying drawings:
the specific screening method of the serum diagnosis marker for the primary depression comprises the following steps:
firstly, separating and extracting serum protein polypeptides of a primary depression patient and a normal healthy population by using a liquid protein chip technology, capturing serum protein polypeptide spectrograms of the primary depression patient and the normal healthy population by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology, comparing and analyzing the difference of the serum protein polypeptide spectrograms of the serum protein polypeptides of the primary depression patient and the normal healthy population by using ClinProTools2.1 software, finding out protein polypeptide molecules with significant differential expression among groups, and screening out a primary depression serum marker from a protein polypeptide peak with significant low expression in the serum of the primary depression patient.
The primary depression polypeptide serum diagnosis marker screened is verified as follows:
a protein polypeptide mixture separated from the serum of a primary depression patient is divided into 20-30 components by using HPLC (high performance liquid chromatography), secondary mass spectrum identification is carried out on the components, the identified protein polypeptide is subjected to serum regression analysis by using an enzyme-linked immunosorbent assay, and the result of serum regression verification proves that the protein polypeptide is remarkably high in expression and specificity in the serum of the primary depression patient and can be used as a biomarker for screening the serum of the primary depression patient.
1. Sample collection and processing
Samples were collected from 48 (15 men; 33 women) primary depression patients and 48 normal healthy people (18 men; 30 women) in Yanyang Hospital psychiatric department of Yanan university (9-2021-2020). The sample considers factors such as age, sex, collection time, whether the storage conditions are consistent, whether basic diseases exist and the like. Collecting blood of the collected person with fasting state, collecting 5mL whole blood with vacuum blood collection tube (yellow cap, with isolation gel), and standing at room temperature for 30 min; centrifuging at room temperature for 5min (3000g), subpackaging the upper layer serum into 100 μ L/tube, immediately storing at-80 deg.C, and avoiding repeated freeze thawing.
Reagents and instrumentation:
the serum proteins were extracted using the magnetic bead kit "weak cationic" (MB-WCX) from Bruker, Germany, as well as spectrally pure (HPLC grade) acetonitrile, trifluoroacetic acid (Merck, Germany) and alpha-cyano-4-hydroxycinnamic acid (HCCA) (Sigma, USA).
The instrument used for the experiment included a magnetic bead separator, 600/384AnchorChip target plate, and an AutoFlexIII Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry, MALDI-TOF-MS) (Bruker Daltonics, Germany).
2. Preparation of serum protein samples
A weak cation (MB-WCX) magnetic bead kit is used for capturing serum protein polypeptide, and the specific operation steps are as follows:
completely mixing the magnetic bead suspension for 1min by using a mixer;
adding 10 mu L of MB-WCX binding solution and 10 mu L of MB-WCX magnetic beads into a PCR tube, uniformly mixing, adding 5 mu L of serum, uniformly mixing for at least 5 times, and standing for 5 min;
thirdly, placing the PCR tube into a magnetic column separator, making the magnetic beads adhere to the wall for 1min, and removing the supernatant after the liquid is clear;
adding 100 mu L of MB-WCX flushing fluid, moving the PCR tube on a magnetic column separator back and forth for 10 times, removing supernatant after the magnetic beads are attached to the wall, and repeating the steps three and four times;
fifthly, adding 5 mu L of MB-WCX eluent to wash the adherent magnetic beads, repeatedly blowing and beating for 10 times, allowing the magnetic beads to adhere for 2min, and transferring the supernatant into a clean centrifugal tube;
sixthly, adding 5 mu L of MB-WCX stable solution into a centrifuge tube and mixing uniformly, wherein the extracted protein polypeptide can be used for direct MALDI-TOF-MS detection or frozen in a refrigerator at the temperature of-20 ℃ for mass spectrometry within 24 h.
Mass spectrometry analysis:
mixing 1 μ L of the separated and collected protein sample with 10 μ L of matrix alpha-cyano-4-hydroxycinnamic acid, and spotting 1 μ L of the mixture on an Anchorchip target plate (Bruker, Germany), wherein each sample is spotted with three targets for three times. And after drying at room temperature, putting the target plate into a mass spectrometer for flight time mass spectrometry, correcting the standard substance by adopting FlexControl 2.0 software, and then starting sample detection, wherein each sample generates a mass spectrogram after being subjected to laser targeting for 300 times (5 times of point targeting and 2 times of 30 times of targeting each time), so as to obtain protein polypeptide spectrograms consisting of different mass-to-nuclear ratios (m/z). The protein polypeptide maps of the two groups of serum samples are analyzed by using ClinProTools2.1 software, a genetic algorithm and other biological statistics and bioinformatics methods. Carrying out normalization smoothing treatment on the total ion flow diagram, and eliminating chemical and electro-physical noises; analyzing the difference protein among groups, calculating the difference size, and arranging the difference size from large to small to find out the protein polypeptide peak value (P <0.001) with obvious difference in expression among groups.
After the serum samples of the primary depression patients and the normal healthy population are processed by a magnetic bead separation system and are analyzed by MALDI-TOF-MS, protein polypeptide maps of each sample of the primary depression patients and the normal healthy population are drawn, 136 protein polypeptide peak maps are detected in the molecular weight range of 800 Da-10000 Da, and the three-time repeated stability of each sample is high.
Analyzing the serum protein polypeptide spectrums of the primary depression patients and the normal healthy population captured by mass spectrometry by adopting ClinProTools2.1 software, comparing and analyzing the serum polypeptide spectrums of the primary depression patients and the normal healthy population, detecting a protein polypeptide peak diagram (P <0.001) with extremely obvious difference, and analyzing the protein polypeptide with obviously up-regulated expression in the depression patients, wherein the specific formula is shown in Table 1:
TABLE 1 Mass Spectrometry results of peptide mapping of serum proteins for Primary Depression patients and Normal healthy people
| Molecular weight (mass to charge ratio) | P value | Mean expression of Primary Depression | Average expression of normal healthy population |
| 1868.21↑ | <0.000001 | 3.86±2.27 | 2.53±1.85 |
| 1062.35↑ | <0.000001 | 9.09±15.23 | 2.8±0.60 |
The results of Flex analysis software analysis of the specific high expression 2 protein polypeptides in the serum of the primary depression patients in the table 1 are shown in the figure 1 and the figure 2, and are obtained by m/z: 1062.35 and 1868.21 in comparison of expression in patients with primary depression (red) and in normal healthy people (green), m/z: 1062.35 and 1868.21 are significantly highly expressed in the serum of patients with primary depression, so that the sequence identification is carried out and the peak protein polypeptide is preferably further identified as a marker.
3. Sequence identification of serum potential marker of primary depression
Specifically, a technology combining liquid chromatography separation and mass spectrum is adopted to carry out the treatment on the serum polypeptide marker m/z of the patient with the primary depression: 1062.35 and 1868.21, and performing two-dimensional gel chromatography on the serum protein polypeptide remaining after the mass spectrum loading collected by magnetic bead separation by using a nano-liter flow rate HPLC liquid phase system Easy nLC 1200 of Thermo Scientific company, and performing two-dimensional gel chromatography on the serum protein polypeptide m/z with the expression up-regulated in the serum of a patient suffering from primary depression by combining the method with an Orbitrap Fusion mass spectrometer of Thermo Scientific: 1062.35 and 1868.21 for sequence identification.
The specific operation steps are as follows:
3.1 sample Pre-treatment
mu.L of the sample was mixed with 800. mu.L of 5% acetonitrile 0.5% formic acid solution and added to 1.5ml of a less than 10kDa ultrafiltration tube, centrifuged at 7000 rpm for 40min in a cryocentrifuge set at 20 ℃ and concentrated to 500. mu.L. Desalting, eluting polypeptide with 70% methanol and 0.5% formic acid, collecting and concentrating to 100 μ L, and treating with Zip-tip column for concentration.
3.2 chromatographic separation
Each sample was separated using a nanoliter flow rate HPLC liquid system Easy nLC 1200. Buffer solution: the solution A is 0.1% formic acid aqueous solution, and the solution B is 0.1% formic acid acetonitrile solution. The column was equilibrated with 95% of solution A. The sample was applied to a mass spectrometric pre-column C18trap column (C183 m 0.10X 20mm) by an autosampler and separated by an analytical column C18 column (C181.9m 0.15X 120mm) at a flow rate of 600 nl/min. The relevant liquid phase gradients are shown in table 2 and the results of gel chromatography are shown in figure 3.
TABLE 2 associated liquid phase gradients
3.3 Mass Spectrometry identification
Each sample was separated by capillary HPLC and subjected to mass spectrometry using an Orbitrap Fusion mass spectrometer (Thermo scientific). The main parameter settings are shown in table 3, and the mass spectrometry results are shown in fig. 4 and 5:
table 3 main parameter settings for mass spectrometry
3.4 data analysis
Analyzing the RAW data into RAW files by mass spectrometry, performing library searching identification in a database of unidrop-human by using software sequence and a protocol discover 2.5.0(Thermo Scientific), submitting the RAW files to a sequence server through the protocol discover during library searching, selecting the well-established database, and then searching the database, wherein relevant parameters are shown in table 4:
TABLE 4 relevant parameters for database search
The result filtering parameters were: peptide FDR is less than or equal to 0.01.
The search results are shown in table 5:
TABLE 5 search results of database search
Therefore, the matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) is used for detecting the ITIH4 and TUBB composition or the ELISA method is used for detecting the expression level of the ITIH4 and TUBB composition, and the method can be used for detecting patients with primary depression. The ITIH4 and TUBB compositions are suggested to be proteins specifically associated with primary depression, further validated by ELISA assays.
4. ELISA serum validation analysis of primary depression serum ITIH4 expression
4.1 serum samples
Serum verification analysis by ELISA was performed by collecting 21 cases of serum from patients with primary depression (8 cases in males; 13 cases in females) and 21 cases of serum from normal healthy population (10 cases in males; 11 cases in females). All serum samples were obtained from Yan Yang Hospital, Yanan university, and collected from 2020, 9 months to 2021 months.
4.2 detection method
The expression level of the serum ITIH4 of the primary depression patients and the normal healthy people is detected by adopting an enzyme-linked immunosorbent assay (ELISA), and the kit is purchased from China Hengyuan biology company. The kit is used for determining the level of human alpha-trypsin inhibitor heavy chain 4(ITIH4) in a specimen by using a double-antibody sandwich method. Coating a microporous plate with a purified human alpha-trypsin inhibitor heavy chain 4(ITIH4) antibody to prepare a solid phase antibody, sequentially adding the alpha-trypsin inhibitor heavy chain 4(ITIH4) into the micropores coated with the monoclonal antibody, combining with an alpha-trypsin inhibitor heavy chain 4(ITIH4) antibody marked by HRP to form an antibody-antigen-enzyme labeled antibody compound, and adding a substrate TMB for color development after thorough washing. TMB is converted to blue by the catalysis of HRP enzyme and to the final yellow by the action of acid. The shade of the color was positively correlated with the alpha-trypsin inhibitor heavy chain 4(ITIH4) in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the concentration of human alpha-trypsin inhibitor heavy chain 4(ITIH4) in the sample was calculated from the standard curve. The specific experimental steps refer to the kit specification, and the positive judgment standard is defined according to the kit specification.
4.3 statistical methods
The independent sample T test was performed using SPSS20 software.
4.4 analysis of results
The results of enzyme linked immunosorbent assay analysis show that the expression level of ITIH4 in different detection groups is primary depression patients > normal healthy people, and the specific results are shown in Table 6 and FIG. 6, and have very significant difference between the two groups:
TABLE 6 expression levels of ITIH4 protein in sera of different groups
The ELISA detection is carried out on the ITIH4 in the serum of the primary depression patient and the normal healthy people, and the result shows that the expression of the ITIH4 has specificity: the range of expression in serum of patients with primary depression is: 35.03-88 pg/mL; the expression range in the serum of normal healthy people is as follows: 18.705-56.255pg/mL, and there was a very significant difference in expression between the two groups (p < 0.001). This indicates that: the ITIH4 is a protein closely related to the occurrence of patients with primary depression, and can be used as a primary detection index of primary depression lesion.
Therefore, the ITIH4 expression of the serum sample to be detected can be preliminarily judged to be a patient with primary depression (35.03-88pg/mL) or a normal healthy human (18.705-56.255pg/mL) through an ELISA experiment.
5. ELISA serum validation analysis of Primary Depression serum TUBB expression
5.1 serum samples
Serum verification analysis by ELISA was performed by collecting 17 cases of serum from patients with primary depression (9 cases in males; 8 cases in females) and 17 cases of serum from normal healthy population (10 cases in males; 7 cases in females). All serum samples were obtained from Yan Yang Hospital, Yanan university, and collected from 2020, 9 months to 2021 months.
5.2 detection method
The expression level of serum TUBB of patients with primary depression and normal healthy people is detected by adopting an enzyme-linked immunosorbent assay (ELISA), and the kit is purchased from China Hengyuan biological company. The kit is used for determining the level of human beta-Tubulin (TUBB) in a specimen by using a double-antibody sandwich method. Coating a microporous plate with a purified human beta-Tubulin (TUBB) antibody to prepare a solid-phase antibody, sequentially adding the beta-Tubulin (TUBB) into the micropores coated with the monoclonal antibody, combining with an HRP-labeled beta-Tubulin (TUBB) antibody to form an antibody-antigen-enzyme-labeled antibody complex, and adding a substrate TMB for developing after thorough washing. TMB is converted to blue by the catalysis of HRP enzyme and to the final yellow by the action of acid. The shade of the color is positively correlated with β -Tubulin (TUBB) in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the concentration of human β -Tubulin (TUBB) in the sample was calculated from the standard curve. The specific experimental steps refer to the kit specification, and the positive judgment standard is defined according to the kit specification.
5.3 statistical methods
The independent sample T test was performed using SPSS20 software.
5.4 analysis of results
The results of the enzyme-linked immunosorbent assay analysis show that the expression level of TUBB in different detection groups is greater than that of the normal healthy population of the patients with primary depression, and the two groups have significant differences, and the specific results are shown in table 7 and fig. 7:
TABLE 7 expression levels of TUBB protein in sera of different groups
ELISA detection is carried out on TUBB in serum of patients with primary depression and normal healthy people, and the result shows that the expression of TUBB has specificity: the range of expression in serum of patients with primary depression is: 47.475-197.875 pg/mL; the expression range in the serum of normal healthy people is as follows: 27.6-93.155pg/mL, and there was a significant difference in expression between the two groups (p < 0.05). This indicates that: TUBB is a protein closely related to the occurrence of patients with primary depression and can be used as a primary detection index for primary depression lesions.
Therefore, the TUBB expression of the serum sample to be tested can be preliminarily judged to be a patient with primary depression (47.475-197.875pg/mL) or a normal healthy human (27.6-93.155pg/mL) through an ELISA experiment.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Sequence listing
<110> Seawa medical diagnostic services Co., Ltd in Xian
<120> serum polypeptide diagnostic molecular composition for primary depression and application thereof
<141> 2021-11-25
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 12
<212> PRT
<213> Artificial Sequence
<400> 1
Gly Ser Glu Met Val Val Ala Gly Lys Leu Gln Asp
1 5 10
<210> 2
<211> 19
<212> PRT
<213> Artificial Sequence
<400> 2
Lys Met Ala Val Thr Phe Ile Gly Asn Ser Thr Ala Ile Gln Glu Leu
1 5 10 15
Phe Lys Arg
Claims (8)
1. A serum polypeptide diagnostic molecule composition, which comprises polypeptide ITIH4 and polypeptide TUBB, wherein the amino acid sequence of ITIH4 is shown in SEQ ID No.1, and the amino acid sequence of TUBB is shown in SEQ ID No. 2.
2. The serum polypeptide diagnostic molecule composition of claim 1, wherein the molecular weight of the ITIH4 is 1062.35 daltons and the molecular weight of the TUBB is 1868.21 daltons.
3. The use of the serum polypeptide diagnostic molecule composition of claim 1 in the preparation of a serum diagnostic reagent for primary depression, wherein the detection parameter of the ITIH4 in the serum of a primary depression patient is 35.03-88pg/mL, and the detection parameter in the serum of a normal healthy population is 18.705-56.255 pg/mL; the detection parameter of the TUBB in the serum of the patients with the primary depression is 47.475-197.875pg/mL, and the detection parameter in the serum of the normal healthy people is 27.6-93.155 pg/mL.
4. The use of claim 3, wherein ITIH4 and TUBB are significantly highly expressed in serum samples from patients with primary depression.
5. The use of claim 3, wherein the serum diagnostic marker for primary depression is a serum polypeptide molecule for ELISA detection of primary depression.
6. The use of claim 3, wherein the serum polypeptide molecule composition ITIH4 and TUBB are novel targets for ELISA detection markers.
7. The application of molecules combined with the serum polypeptide diagnostic molecule composition in preparing the serum diagnostic reagent for primary depression is characterized in that the serum polypeptide diagnostic molecule composition comprises ITIH4 and TUBB, the amino acid sequence of ITIH4 is shown in SEQ ID No.1, and the amino acid sequence of the TUBB is shown in SEQ ID No. 2.
8. A diagnostic kit for primary depression, comprising the serum polypeptide diagnostic molecule composition according to claim 1.
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| 中国科协学会学术部: "网络药理学-中药现代化的新思路与新方法", 30 November 2014, 中国科学技术出版社, pages: 12 - 14 * |
| 王继贤 等: "临床生化检验", vol. 2, 31 July 1996, 湖南科学技术出版社, pages: 260 - 261 * |
| 程书钧: "癌前病变和癌前癌症", 31 January 2017, 河南科学技术出版社, pages: 39 - 40 * |
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