CN114137214B - Immunodetection kit for predicting occurrence of mental symptoms after stress and application thereof - Google Patents
Immunodetection kit for predicting occurrence of mental symptoms after stress and application thereof Download PDFInfo
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- CN114137214B CN114137214B CN202111475342.5A CN202111475342A CN114137214B CN 114137214 B CN114137214 B CN 114137214B CN 202111475342 A CN202111475342 A CN 202111475342A CN 114137214 B CN114137214 B CN 114137214B
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- G01N2333/914—Hydrolases (3)
- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
- G01N2333/918—Carboxylic ester hydrolases (3.1.1)
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/30—Psychoses; Psychiatry
Abstract
The invention relates to a biomarker for predicting psychological symptoms such as anxiety depression and the like after stress and a detection kit thereof. The present invention provides a group of biomarkers which can predict the occurrence of mental symptoms such as anxiety depression after stress in the blood of a potential patient and are selected from one or more of butyrylcholinesterase, catalase, dopamine-beta-hydroxylase, insulin-like growth factor binding protein 2, neuropil 1 or amino acid sequences with 80% homology with the same. The invention also provides a detection kit and a detection method for detecting the proteins in a blood sample; and uses of the antigens and kits described above; and provides a reference concentration threshold for the risk of developing high levels of post-stress anxiety depression for these proteins. The immune detection kit prepared by the kit can rapidly, simply and accurately detect the concentration of the biomarker in blood, and is suitable for large-scale popularization and application.
Description
Technical Field
The present invention relates to the detection and identification of biomarkers produced in subjects suffering from mental symptoms such as anxiety depression after stress and provides a metric for predicting the risk of differentiating high occurrence in subjects potentially suffering from anxiety depression. The kit and the method are used for providing a reference concentration threshold value of high occurrence risk of the markers, so that reliable detection and identification of the biomarkers for assisting diagnosis and prevention of mental symptoms such as anxiety depression after stress are possible.
Background
In recent years, the incidence rate of stress events such as sudden public health events, various major accidents, natural and artificial disasters and the like is gradually increased, more than 70% of people worldwide can experience traumatic events at least once in life, and event passers-by and rescue workers and other rescuers often have mental and psychological symptoms such as anxiety, depression and the like after the stress events, and partial individuals even develop serious mental diseases. Among them, post-traumatic stress disorder (PTSD) is one of the more serious mental diseases. Post-traumatic stress disorder refers to a mental disorder in which an individual experiences, witnesses, or encounters one or more physical deaths involving itself or another person, or is threatened by death, or is severely injured, or is delayed in appearance and persistence after physical integrity is compromised. Clinically manifested as wound memory invasion, wound-related stimulus avoidance, cognitive negative changes, enhanced arousal, behavioural overstress, and the like. The life-long prevalence of PTSD is approximately 1.3% -12.2% and the annual prevalence is approximately 0.2% -3.8% depending on cultural background and economic development level in different countries. Whereas mental diseases after stress, including PTSD, have become a disease of great concern in today's society, especially during the current time of COVID-19 pandemic. The disease has longer disease course and is easy to relapse, the patient often has actions such as self-disabled, suicide, drug abuse and the like, the life quality of the patient is seriously influenced, and serious psychological, physiological and economic burdens are caused on individuals, families and society.
PTSD is more serious and of greatest concern in post-stress psychiatric disorders, and its clinical diagnostic criteria are currently widely accepted by the American society of psychology DSM-5 and world health organization ICD-11. In DSM-5, the diagnostic criteria for PTSD share 8 large entries, divided into 4 symptom groups and one subtype, where the 4 symptom groups are wound memory invasion, avoidance of wound-related stimuli, negative changes in cognition and emotion, increased arousal, and behavioral overstress. Whereas in ICD-11, diagnosis of PTSD is reduced to 3 symptom groups, 6 classes of symptoms, including constant re-experiencing traumatic events, traumatic stimulation avoidance, and high vigilance. Although the diagnostic criteria for PTSD vary somewhat, the therapeutic modalities mainly include psychological interventions, drug therapies and some other innovative therapies. Psychological interventions are first-line treatment methods recommended as most guidelines, mainly including prolonged exposure therapy, cognitive processing therapy, cognitive behavioral therapy, ocular desensitization, and reprocessing therapy, among others. The drug therapy is generally used as a two-line therapy, including selective 5-hydroxytryptamine reuptake inhibitor, serotonin and norepinephrine reuptake inhibitor, monoamine oxidase, sympatholytic agent, anticonvulsant, benzodiazepines, etc.
Although PTSD is highly prevalent in people experiencing one or more severe wounds, not all individuals experiencing a wound become PTSD patients, with actual PTSD lifelong prevalence far below theoretical speculation suggesting significant individual variability in the incidence of PTSD. PTSD morbidity studies based on different ethnic groups found that the incidence of white ethnic groups was significantly lower than other ethnic groups, while studies based on twins also found that the incidence of syngeneic twins was significantly higher than that of heteroootwins after experiencing traumatic events, suggesting that genetic factors significantly affected the incidence of PTSD. The diagnosis of PTSD is mainly based on the medical history, mental state judgment, symptom duration, clinical mental scale detection and self-description of patients, and the detection of the PTSD is lack of objectively and effectively biomarkers. Considering that most traumatic events are unavoidable, it becomes particularly important to predict the risk factors for PTSD after the occurrence of a wound. However, PTSD has few objective, effective and specific biological markers to be screened, and is still lacking as an objective index for early warning diagnosis and therapeutic efficacy evaluation of PTSD. Detection of biomarkers in blood is considered to be the most potential and convenient way to aid in PTSD diagnosis due to its advantages of convenience, minimal trauma, etc. Studies have reported that decreased cortisol levels in the blood after trauma may be associated with the development of PTSD, and that imbalance of some neurotransmitters in the blood, adrenocorticotropic hormone (ACTH), glucocorticoid (GC), epinephrine, and catecholamine levels, etc. are also considered to be markers associated with PTSD. In addition, other biomarkers that may be associated with PTSD include cholecystokinin (CCK), nitric Oxide Synthase (NOS), neuropeptide Y (NPY), and p11 protein, among others. But the specificity and sensitivity of these markers have not been well verified. Meanwhile, specific markers of mental diseases after stress other than PTSD are more recently reported, so that specific biomarkers for mental symptoms such as anxiety depression and the like after stress are urgently needed to be discovered and developed.
Therefore, in order to effectively predict the occurrence of mental symptoms such as anxiety and depression after stress, it is necessary to identify novel biomarkers and develop a detection kit capable of specifically detecting the biomarkers.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a group of biomarkers capable of predicting occurrence of mental symptoms such as anxiety depression and the like after stress, and a detection kit and a detection method for detecting the biomarkers in a blood sample, wherein the detection kit based on the group of biomarkers can be used for detecting the concentration of the group of biomarkers in blood of a potential stress patient with high sensitivity, high specificity and high stability, and the concentration threshold value provided by the invention prompts high occurrence risk of the mental symptoms such as anxiety depression and the like, so that basis can be brought to doctors for accurately predicting occurrence of mental diseases such as anxiety, depression, PTSD and the like of the stress patient, further treatment can be timely adopted to prevent occurrence of the diseases, pain and burden of the patient can be timely lightened, and the immune detection kit prepared by the kit can be used for rapidly, simply and accurately detecting the concentration of the biomarkers in the blood and is suitable for large-scale popularization and application.
In order to achieve the above object, in a first aspect of the present invention, there is provided an immunoassay kit for predicting occurrence of mental symptoms such as anxiety depression after stress, the immunoassay kit comprising a reagent for quantitatively detecting expression level of a biomarker selected from the group consisting of: butyrylcholinesterase (BCHE), catalase (CAT), dopamine-Beta-Hydroxylase (DBH), insulin-like growth factor binding protein 2 (Insulin Like Growth Factor Binding Protein, IGFBP 2), neuropilin1 (NRP 1), an amino acid sequence having at least 80% homology to the amino acid sequence of BCHE, an amino acid sequence having at least 80% homology to the amino acid sequence of CAT, an amino acid sequence having at least 80% homology to the amino acid sequence of DBH, an amino acid sequence having at least 80% homology to the amino acid sequence of IGFBP2, and an amino acid sequence having at least 80% homology to the amino acid sequence of NRP1.
Preferably, the psychological symptoms such as post-stress anxiety depression include post-stress anxiety, post-stress depression, post-traumatic stress disorder, post-stroke anxiety, post-stroke depression, post-brain trauma anxiety, post-brain trauma depression and other post-stress related mental diseases.
In a second aspect of the invention, there is provided the use of a biomarker for the manufacture of a reagent for detecting the occurrence of a psychological symptom following a predicted stress, said reagent being a reagent for quantitatively detecting the expression level of said biomarker, said biomarker being selected from the group consisting of: one or more of butyrylcholinesterase, catalase, dopamine-beta-hydroxylase, insulin-like growth factor binding protein 2, neuropil 1, an amino acid sequence having at least 80% homology with the amino acid sequence of butyrylcholinesterase, an amino acid sequence having at least 80% homology with the amino acid sequence of catalase, an amino acid sequence having at least 80% homology with the amino acid sequence of dopamine-beta-hydroxylase, an amino acid sequence having at least 80% homology with the amino acid sequence of insulin-like growth factor binding protein 2, and an amino acid sequence having at least 80% homology with the amino acid sequence of neuropil 1.
In a third aspect of the invention, there is provided a method of detecting the level of the biomarker protein described above in whole blood, central nervous system tissue, serum, plasma, cerebrospinal fluid, saliva, sweat, tears, urine, oral samples, or a combination thereof, in an immunoassay kit. Preferably, the body fluid is blood. The method for detecting the above-mentioned proteins can be carried out by a conventional Enzyme-linked immunosorbent assay (Enzyme-linked immunosorbent assay, ELISA), preferably using a sandwich ELISA method.
Preferably, the immunoassay kit further comprises a carrier on which an antigen for detecting the biomarker protein is coated.
Preferably, the immunoassay kit further comprises a primary antibody, wherein the primary antibody is used for detecting the biomarker protein.
Preferably, the immunoassay kit further comprises a labeled secondary antibody of the biomarker protein primary antibody. The label used may be horseradish peroxidase, alkaline phosphatase, fluorescent molecule FITC (or other fluorescent label), chemiluminescent detection. The detection method can be a color development method, a fluorescence method, chemiluminescence and electrochemiluminescence method for qualitative and quantitative analysis.
More preferably, the labeled primary antibody against the biomarker protein is a horseradish peroxidase labeled secondary antibody against the primary antibody against the biomarker protein.
In a fourth aspect of the invention, there is provided the use of an immunoassay kit for the biomarker protein described above for detecting the biomarker protein in human blood, the high risk concentration threshold for the protein being determined by comparing the measured level of the biomarker protein in a subject (e.g. a patient having undergone a traumatic event) with the measured level of the biomarker protein in a quantitative mass spectrometry technique.
The invention has the beneficial effects that: the biological markers for predicting the occurrence of mental symptoms such as anxiety depression after stress are one or more of Butyrylcholinesterase (BCHE), catalase (CAT), dopamine-Beta-Hydroxylase (DOPAmine Beta-Hydroxylase, DBH), insulin-like growth factor binding protein 2 (Insulin Like Growth Factor Binding Protein, IGFBP 2), nerve cellulose 1 (Neuropilin 1, NRP 1), an amino acid sequence with at least 80% homology with an amino acid sequence of CAT, an amino acid sequence with at least 80% homology with an amino acid sequence of DBH, an amino acid sequence with at least 80% homology with an amino acid sequence of IGFBP2, and an amino acid sequence with at least 80% homology with an amino acid sequence of NRP1, and can be used for accurately detecting the biological markers in high specificity and high sensitivity in blood after a stress event, and can be used for detecting the biological markers in time, so that the biological markers can be used for detecting the stress, can be accurately detected in time, and can be used for detecting the stress, and the patients can be prevented from suffering from the diseases and the diseases, and the diseases can be easily detected in time, and the patients.
Drawings
FIG. 1 is a graph comparing protein levels of BCHE in serum from post-traumatic anxiety depression symptom-free patients and post-traumatic anxiety depression symptom-free patients.
FIG. 2 is a graph comparing CAT protein levels in serum of patients without anxiety-depression symptoms after trauma and patients with anxiety-depression symptoms after trauma.
FIG. 3 is a graph comparing protein levels of DBH in serum of a patient without symptoms of anxiety depression after a trauma, a patient with symptoms of anxiety depression after a trauma.
FIG. 4 is a graph comparing protein levels of IGFBP2 in serum of post-traumatic anxiety depression symptom-free patients and post-traumatic anxiety depression symptom-free patients.
FIG. 5 is a graph comparing protein levels of NRP1 in serum of a patient without anxiety-depressive symptoms following a trauma, a patient with anxiety-depressive symptoms following a trauma.
Detailed Description
In order to make the technical contents of the present invention more clearly understood, the following examples are specifically described.
Protein biomarkers
Biomarkers for predicting the occurrence of mental symptoms such as anxiety and depression after stress are Butyrylcholinesterase (BCHE), catalase (CAT), dopamine-Beta-Hydroxylase (DBH), insulin-like growth factor binding protein 2 (Insulin Like Growth Factor Binding Protein, igfbp 2), and neuropil 1 (neuropil 1, nrp 1).
BCHE is an esterase with broad substrate specificity that contributes to the inactivation of the neurotransmitter acetylcholine. BCHE is capable of degrading neurotoxic organophosphates. As shown in fig. 1, the present invention identified that BCHE protein was significantly reduced in whole blood, plasma, serum levels (p < 0.05) in patients experiencing post-traumatic anxiety depression symptoms as compared to levels in patients experiencing post-traumatic anxiety depression symptoms.
CAT is present in almost all organisms that breathe aerobically, and is used to protect cells from the toxic effects of hydrogen peroxide. CAT is capable of promoting cell growth, including T cells, B cells, myeloid leukemia cells, melanoma cells, mast cell tumor cells, and normal and transformed fibroblasts. As shown in fig. 2, the present invention identifies that CAT protein levels in whole blood, plasma, serum of patients experiencing anxiety depression symptoms after trauma are significantly reduced (p < 0.05) compared to levels of patients experiencing anxiety depression symptoms after trauma.
DBH protein is expressed in the nerve secretory vesicles and chromaphilic granules of the adrenal medulla, catalyzing the conversion of dopamine to norepinephrine, a hormone, which is also the main neurotransmitter of the sympathetic nervous system. DBH exists in soluble and membrane-bound forms, depending on the absence or presence of signal peptide, respectively. The gene mutation results in a deficiency of dopamine-beta-hydroxy acids in human patients characterized by defects in autonomic and cardiovascular function, including hypotension and ptosis. The present invention identifies that DBH protein is significantly reduced (p < 0.05) in whole blood, plasma, serum of patients experiencing post-traumatic anxiety depression symptoms as compared to the levels of patients experiencing post-traumatic anxiety depression symptoms.
IGFBP2 inhibits IGF-mediated growth and development rates. IGFBP2 extends the half-life of IGF and has been shown to inhibit or stimulate the growth promoting effect of IGF on cell culture. IGFBP2 alters IGF interaction with its cell surface receptor. The present invention identifies that IGFBP2 protein is significantly reduced (p < 0.05) in whole blood, plasma, serum of patients with symptoms of anxiety depression after a wound compared to the levels of patients without symptoms of anxiety depression after a wound.
NRP1 is a cell surface receptor involved in development of the cardiovascular system, angiogenesis, formation of the neural circuit, and organogenesis outside the nervous system. NRP1 mediates the chemical rejection activity of signalin. NRP1 recognizes the motif R/KXXR/K on its ligand according to the C-terminal rule, leading to cellular internalization and vascular leakage. It binds to the PLGF-2 subtype of signalin 3A, PGF, the VEGF165 subtype of VEGFA and VEGFB. Co-expression of NRP1 with KDR results in increased binding and chemotaxis of VEGF165 with KDR. NRP1 regulates VEGF-induced angiogenesis. NRP1 binds to VEGFA to initiate signaling pathways required for motor neuron axonal guidance and cell body migration, including facial motor neuron migration from diamond 4 to the caudal side of diamond 6 (by similarity) during embryonic development. NRP1 regulates mitochondrial iron transport through interactions with ABCB 8/MITOSUR. NRP1 binds VEGF-165 and may inhibit its binding to cells. NRP1 can induce apoptosis by sequestering VEGF-165. NRP1 can also bind individual members of the signalin family. NRP1 expression has a negative impact on vascular number and integrity. NRP1 acts as a host factor for human coronavirus SARS-CoV-2 infection. NRP1 recognizes and binds to the motif RRAR on SARS-CoV-2 spinous process protein S1 with C-terminal regularity to enhance infection with SARS-CoV-2. The present invention identifies that NRP1 protein is significantly reduced (p < 0.05) in whole blood, plasma, serum of patients with anxiety depression symptoms after trauma compared to the levels of patients without anxiety depression symptoms after trauma.
Kit for detecting a substance in a sample
In the invention, biomarkers (BCHE, CAT, DBH, IGFBP2, NRP 1) for the occurrence of mental symptoms such as anxiety depression and the like after stress in 5 kinds of serum are respectively detected by five enzyme-linked immunoassay kits. The assay was performed by the priority sandwich ELISA method.
1. Establishment of sandwich ELISA method
1) And measuring a plurality of costar strips coated with the first biomarker for resisting mental and psychological symptoms such as anxiety depression after stress and the like according to the number of detection samples.
2) Sample dilutions were added at 100 ul/well.
3) Standard samples, blank and diluted serum samples, 50 ul/well, 37 degrees for 2 hours, shaker 200rpm were added.
4) PBST wash, 400 ul/well, 5 beats, pat dry.
5) Another horseradish peroxidase-labeled primary biomarker antibody, 200 ul/well, was added and incubated at 37℃for 2 hours with shaking table 200rpm.
6) PBST wash, 400 ul/well, 5 beats, pat dry.
7) TMB color development TMB solution A and solution B were mixed in equal volumes, 100 ul/well, 37℃for 20min.
8) Stop solution, 50 ul/well stop, read with a microplate reader at 450 nm.
2. The kit comprises the following components:
the kit comprises the following parts:
1) The costar ELISA plates coated with the biomarker primary antibodies for detecting the occurrence of mental symptoms such as anxiety depression after stress
2) PTSD biomarker standard samples were each one tube (1 ml)
3) One bottle of sample diluent (100 ml)
4) Horseradish peroxidase-labeled anti-biomarker antibodies were each one tube (20 ml)
5) 10 x PBST bottle (100 ml)
6) TMB color development solution A bottle (60 ml), B bottle (60 ml)
7) Stop solution bottle (60 ml)
3. Identification of biomarkers for mental symptoms such as anxiety depression after stress
1) Patient: 11 outpatients experiencing brain trauma were selected and evaluated by 2 psychiatrists based on diagnostic criteria of DSM-5 for their appearance of anxiety-depressive symptoms 6 months after brain trauma, 8 of which were not showing anxiety-depressive symptoms and were not diagnosed as PTSD, and 3 of which were showing anxiety-depressive symptoms and were diagnosed as PTSD. The demographics of 11 patients are shown in Table 1:
table one: demographic profile of 11 outpatient brain trauma patients
| Numbering of brain trauma patients | Sex (sex) | Age of | Diagnosis of |
| PT-071 | Female woman | 25 | Anxiety-free depression, non-PTSD |
| PT-072 | Female woman | 34 | Anxiety-free depression, non-PTSD |
| PT-073 | Female woman | 30 | Anxiety-free depression, non-PTSD |
| PT-076 | Female woman | 24 | Anxiety-free depression, non-PTSD |
| PT-078 | Female woman | 44 | Anxiety-free depression, non-PTSD |
| PT-079 | Male men | 55 | Anxiety-free depression, non-PTSD |
| PT-083 | Male men | 29 | Anxiety-free depression, non-PTSD |
| PT-087 | Female woman | 50 | Anxiety-free depression, non-PTSD |
| PT-077 | Female woman | 49 | Anxiety depression, PTSD |
| PT-082 | Female woman | 25 | Anxiety depression, PTSD |
| PT-095 | Female woman | 29 | Anxiety depression, PTSD |
2) Collection of blood samples: blood samples were collected by venipuncture in a vacuum tube containing heparin for anticoagulation, and 10ml of blood was collected per patient. After centrifugation at 3000rpm for 5 minutes, the blood sample was separated into serum and plasma. All whole blood, serum, plasma were stored in-80 ℃ refrigerator after sub-packaging for subsequent determination.
3) Identifying protein content in serum samples: the serum samples were assayed for protein content using a 4D-Label-free succinylation quantitative proteomics method. The non-standard (Label-free) proteome quantitative technology is a novel protein quantitative technology independent of isotope labeling, and the technology analyzes the proteolysis peptide fragments through liquid chromatography-mass spectrometry without using expensive stable isotope labels as internal labels, only analyzes mass spectrum data generated when large-scale identification of proteins is needed, and can relatively quantify the corresponding proteins by comparing signal intensities of the corresponding peptide fragments in different samples. And taking out the serum sample from the temperature of minus 80 ℃, weighing a proper amount of the sample into a mortar precooled by liquid nitrogen, and adding liquid nitrogen to sufficiently grind the sample into powder. After adding lysis buffer to the powder, each sample was subjected to enzymatic hydrolysis with an equal amount of protein. The peptide fragment obtained by enzymolysis is separated by a NanoElute ultra-high performance liquid system, injected into a Capilliry ion source for ionization and then analyzed by a timsTOF Pro mass spectrum. And (3) carrying out database search by adopting analysis software based on the raw file obtained by mass spectrum detection, carrying out quality control on peptide fragments and protein levels based on search results, carrying out protein annotation, and then carrying out protein quantitative analysis.
4) And (3) results: comparing the protein content of patients with anxiety-depressive symptoms and without anxiety-depressive symptoms who experienced brain trauma, a set of proteins differentially expressed in patients with anxiety-depressive symptoms was identified, as follows:
and (II) table: proteins specific for patients with anxiety depression symptoms
Figure 1 shows a significant reduction in protein levels of BCHE in serum of patients with anxiety-depressive symptoms (p < 0.05) compared to patients without anxiety-depressive symptoms. Figure 2 shows that the protein level of CAT in serum was significantly reduced in patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms (p < 0.05). Figure 3 shows a significant reduction in protein levels of DBH in serum of patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms (p < 0.05). Figure 4 shows that IGFBP2 protein levels were significantly reduced in serum from patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms (p < 0.05). Figure 5 shows a significant reduction in protein levels of NRP1 in serum of patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms (p < 0.05).
5) Summarizing: 5 proteins were found to significantly distinguish between patients with symptoms of anxiety depression and patients without symptoms of anxiety depression after stress. Comprising (1) butyrylcholinesterase BCHE (SwissProt P06276), which was reduced by about 44% in serum samples from patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms, based on the results of DIA mass spectrometry. (2) Catalase CAT (SwissProt P04040), according to the results of DIA mass spectrum, decreased CAT by about 47% in serum samples of patients with anxiety-depressive symptoms compared with patients without anxiety-depressive symptoms. (3) Dopamine-beta-hydroxylase DBH (SwissProt P09172) showed about 52% reduction of DBH in serum samples of patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms, based on the results of DIA mass spectrometry. (4) Insulin-like growth factor binding protein 2IGFBP2 (SwissProt P18065) IGFBP2 was reduced by about 75% in serum samples from patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms, based on the results of DIA mass spectrometry. (5) Neuropil 1NRP1 (SwissProt O14786), NRP1 was reduced by about 23% in serum samples of patients with anxiety-depressive symptoms compared to patients without anxiety-depressive symptoms according to the results of DIA mass spectrometry.
4. Application of biomarker immunodetection kit for mental symptoms such as anxiety depression after stress
1) And (2) subject: use of a kit for recruiting 16 healthy subjects and 15 outpatients who had undergone a traumatic event, the demographics of these subjects are as shown in table three:
table three: demographic profile of subject
2) Collection of blood samples: blood samples were collected by venipuncture in a vacuum tube containing heparin for anticoagulation, and 10ml of blood was collected per patient. After centrifugation at 3000rpm for 5 minutes, the blood sample was separated into serum and plasma. All whole blood, serum, plasma were stored in-80 ℃ refrigerator after sub-packaging for subsequent determination.
3) Protein content of biomarker for detecting mental symptoms such as anxiety depression after stress in serum sample: the serum sample adopts the kit of the invention, and the protein content of butyrylcholinesterase, catalase, dopamine-beta-hydroxylase, insulin-like growth factor binding protein 2 and nerve cellulose 1 is respectively detected by ELISA method.
4) And (3) results: the results of serum testing of 31 subjects are shown in table four:
table four: ELISA detection result of biomarker of mental symptoms such as anxiety depression and the like of subject after stress
5) And (3) data analysis: from the results of table four, the biomarker protein content mean, standard deviation and 95% ci interval for mental symptoms such as anxiety depression in healthy control and post-stress subjects experiencing traumatic events were calculated, respectively, as shown in table five:
table five: marker detection result analysis data sheet for psychological symptoms such as anxiety depression and the like of subject after stress
6) Summarizing: the expression levels of markers of mental symptoms such as anxiety depression after stress in serum of healthy control subjects and subjects who have experienced traumatic events are respectively: BCHE:2748±158.4ng/ml (healthy control) and 3064±237.8ng/ml (wound group); CAT:59.69 + -4.083 pg/ml (healthy control) and 67.24 + -3.718 pg/ml (trauma group); DBH: 24.15.+ -. 5.577ng/ml (healthy control) and 25.29.+ -. 7.149ng/ml (wound group); IGFBP2: 142.2.+ -. 19.85ng/ml (healthy control) and 276.6.+ -. 34.95ng/ml (wound group); NRP1: 265.2.+ -. 11.31ng/ml (healthy control) and 239.2.+ -. 17.66ng/ml (wound group).
5. Determination of biomarker high risk threshold for mental symptoms such as anxiety depression after stress
1) The high risk cutoff threshold of the marker is calculated according to the reduction rate of the expression amount of BCHE, CAT, DBH, IGFBP and NRP1 of the patient with anxiety depression symptoms after stress compared with the patient without anxiety depression symptoms, which is determined by quantitative proteomics, by combining the expression amount of the protein in the serum of the subject with wound event, which is determined by the kit, and the calculation formula is as follows:
the Cutoff calculation formula is: cutoff = mean expression level of trauma group determined by ELISA x (1-rate of decrease in expression of patients with anxiety-depression symptoms determined by quantitative mass spectrometry)
The biomarker high risk threshold for mental symptoms such as anxiety depression after each stress is calculated according to the formula and the average expression level of the trauma group in the table IV, and is respectively smaller than the following expression level:
BCHE：3064×(1-44％)＝1716ng/ml；
CAT：67.24×(1-47％)＝35.64pg/ml；
DBH：25.29×(1-52％)＝12.14ng/ml；
IGFBP2：276.6×(1-75％)＝69.15ng/ml；
NRP1：239.2×(1-23％)＝184.2ng/ml。
2) Judging the incidence risk of the detected stress patient through the cutoff value, wherein the incidence risk of the detected stress patient is considered to be high risk when the incidence risk is lower than the cutoff value, and the incidence risk of mental diseases such as anxiety, depression, PTSD and the like is considered to be high as long as one protein is detected to be high risk.
According to the invention, through a nonstandard (Label-free) proteome quantification technology, the protein content in blood samples of patients suffering from anxiety depression and anxiety depression after brain trauma is compared, and a group of biomarkers capable of effectively predicting occurrence risk of mental symptoms such as anxiety depression after stress are obtained through screening: BCHE, CAT, DBH, IGFBP2 and NRP1. According to the biomarker of mental symptoms such as anxiety depression after stress, which is obtained by identification, an immune kit for detecting the expression level in a blood sample is prepared, and a detection method is established. The blood sandwich enzyme-labeled immunoassay (sandwich ELISA) of the invention is based on the immunochemical principle, and provides an antigen-antibody detection method with high sensitivity, high specificity and high stability. The biomarker detection kit for the psychological symptoms such as anxiety depression and the like after stress with high sensitivity, high specificity and high stability is obtained by repeatedly screening and verifying the prepared kit by using the serum of healthy control patients and patients subjected to traumatic events. And the biomarker concentration threshold value for predicting the high occurrence risk of mental diseases such as anxiety, depression, PTSD and the like in the target population is given by combining analysis mass spectrum quantitative data and ELISA results. The blood ELISA detection kit is quicker and simpler to operate; the accuracy and the precision of the detection result are greatly improved; low cost, easy popularization and suitability for the market. The ELISA detection method can be used as a large-scale screening tool for early screening of mental diseases after stress through optimization of various parameters and experimental procedures.
In summary, the invention provides a group of biomarkers capable of predicting occurrence of mental symptoms such as anxiety depression after stress, and the like, and a detection kit and a detection method for detecting the biomarkers in a blood sample, and potential patients can detect the concentration of the protein in blood with high specificity, high sensitivity and high specificity after experiencing a stress event by using the kit, and the high risk threshold range provided by the invention can bring basis for accurately predicting occurrence of mental diseases such as anxiety, depression, PTSD and the like for a doctor, and further timely take treatment to prevent the occurrence of the diseases, and timely relieve pain and burden of the patient.
In this specification, the invention has been described with reference to specific embodiments thereof. It will be apparent, however, that various modifications and changes may be made without departing from the spirit and scope of the invention. The description is thus to be regarded as illustrative instead of limiting.
Claims (2)
1. Use of a reagent for detecting a biomarker comprising insulin-like growth factor binding protein 2 in a kit for diagnosing the occurrence of symptoms of anxiety-depression in a subject following stress in a traumatic event.
2. The use according to claim 1, wherein the biomarker further comprises one or more of butyrylcholinesterase, catalase, dopamine-beta-hydroxylase, and neuropil 1.
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Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6025368A (en) * | 1997-02-25 | 2000-02-15 | Celtrix Pharmaceuticals, Inc. | Method for treating the symptoms of chronic stress-related disorders using IGF |
| CN1541107A (en) * | 2000-09-22 | 2004-10-27 | ��ʿ����ѧ | Growth factor complex |
| WO2007106685A2 (en) * | 2006-03-10 | 2007-09-20 | Novartis Ag | Targets for depression and bipolar disorders |
| CN101903777A (en) * | 2007-12-19 | 2010-12-01 | 赛诺瓦神经学科技有限公司 | Be used to diagnose and monitor the method and the biomarker of mental illness |
| CN103492590A (en) * | 2011-02-22 | 2014-01-01 | 卡里斯生命科学卢森堡控股有限责任公司 | Circulating biomarkers |
| WO2015082927A1 (en) * | 2013-12-05 | 2015-06-11 | Cambridge Enterprise Limited | Novel biomarker panel for major depressive disease |
| CN106062563A (en) * | 2014-01-28 | 2016-10-26 | 普雷德姆泰克有限公司 | Biomarker and methods for early diagnosis of alzheimer's disease |
| EP3232198A1 (en) * | 2014-12-12 | 2017-10-18 | Seoul National University R&DB Foundation | Biomarker for diagnosis of hepatoma and use thereof |
| CN107921094A (en) * | 2015-06-04 | 2018-04-17 | 圣拉斐尔医院有限公司 | IGFBP3 and application thereof |
| JP2018132526A (en) * | 2017-02-16 | 2018-08-23 | 国立大学法人千葉大学 | Markers, method for inspection, and inspection kit for major depressive disorder and bipolar disorder, and method for screening therapeutic medicines for major depressive disorder and bipolar disorder |
| CN108441551A (en) * | 2017-04-11 | 2018-08-24 | 刘小翠 | A kind of biomarker and its application process of phrenoblabia |
| CN112957453A (en) * | 2021-03-04 | 2021-06-15 | 上海市精神卫生中心（上海市心理咨询培训中心） | Application of insulin-like growth factor binding protein 2 in preparation of medicine for treating mental disorder caused by brain trauma |
Family Cites Families (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003234136A1 (en) * | 2002-04-18 | 2003-11-03 | The General Hospital Corporation | Drg11-responsive (dragon) gene family |
| US20050084880A1 (en) * | 2003-07-11 | 2005-04-21 | Ronald Duman | Systems and methods for diagnosing & treating psychological and behavioral conditions |
| JP5540000B2 (en) * | 2008-10-15 | 2014-07-02 | リッジ ダイアグノスティックス，インコーポレイテッド | Human biomarker hypermapping of depressive disorder |
| US20110294693A1 (en) * | 2008-11-17 | 2011-12-01 | The George Washington University | Compositions and Methods for Identifying Autism Spectrum Disorders |
| RU2011127198A (en) * | 2008-12-04 | 2013-01-10 | Эбботт Лэборетриз | IMMUNOGLOBULINS WITH DOUBLE VARIABLE DOMAINS AND THEIR APPLICATION |
| UY32808A (en) * | 2009-07-29 | 2011-02-28 | Abbott Lab | IMMUNOGLOBULINS AS A DUAL VARIABLE DOMAIN AND USES OF THE SAME |
| GB201021502D0 (en) * | 2010-12-20 | 2011-02-02 | Cambridge Entpr Ltd | Biomarkers |
| EP2656081B1 (en) * | 2010-12-20 | 2017-09-06 | Cambridge Enterprise Ltd. | Method and biomarkers for differentially diagnosing psychotic disorders |
| AU2012308305B2 (en) * | 2011-09-14 | 2017-11-23 | Banyan Biomarkers, Inc. | Processes and kits to detect and monitor for diagnostic biomarkers for post traumatic stress disorder (PTSD) and to differentiate between suicidal and non-suicidal form of the disorder |
| US9733261B2 (en) * | 2012-04-24 | 2017-08-15 | Astute Medical, Inc. | Methods and compositions for diagnosis and prognosis of stroke or other cerebral injury |
| US20150301058A1 (en) * | 2012-11-26 | 2015-10-22 | Caris Science, Inc. | Biomarker compositions and methods |
| BR112016004153A2 (en) * | 2013-08-28 | 2019-09-17 | Caris Life Sciences Switzerland Holdings Gmbh | "method for characterizing a disease or disorder, kit, composition, method for generating an input library, oligonucleotide, plurality of oligonucleotides and method for identifying an aptamer |
| GB201404189D0 (en) * | 2014-03-10 | 2014-04-23 | Cambridge Entpr Ltd | Novel biomarkers |
| US20170356903A1 (en) * | 2014-11-21 | 2017-12-14 | Caris Science, Inc. | Oligonucleotide probes and uses thereof |
| US20170258843A1 (en) * | 2016-03-14 | 2017-09-14 | AngioStem, Inc. | Stem cell mediated neuroregeneration and neuroprotection |
| US20180161377A1 (en) * | 2016-12-09 | 2018-06-14 | Neoneuron Llc | Method of treating neurological disorders with stem cell therapy |
| US20200397828A1 (en) * | 2019-04-29 | 2020-12-24 | The Broad Institute, Inc. | Atlas of choroid plexus cell types and therapeutic and diagnostic uses thereof |
| US20210009639A1 (en) * | 2019-07-12 | 2021-01-14 | Northwestern University | Insulin like growth factor binding protein bioactive peptide fragments |
-
2021
- 2021-12-06 CN CN202311409525.6A patent/CN117451995A/en active Pending
- 2021-12-06 CN CN202111475342.5A patent/CN114137214B/en active Active
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6025368A (en) * | 1997-02-25 | 2000-02-15 | Celtrix Pharmaceuticals, Inc. | Method for treating the symptoms of chronic stress-related disorders using IGF |
| CN1541107A (en) * | 2000-09-22 | 2004-10-27 | ��ʿ����ѧ | Growth factor complex |
| WO2007106685A2 (en) * | 2006-03-10 | 2007-09-20 | Novartis Ag | Targets for depression and bipolar disorders |
| CN101903777A (en) * | 2007-12-19 | 2010-12-01 | 赛诺瓦神经学科技有限公司 | Be used to diagnose and monitor the method and the biomarker of mental illness |
| CN103492590A (en) * | 2011-02-22 | 2014-01-01 | 卡里斯生命科学卢森堡控股有限责任公司 | Circulating biomarkers |
| WO2015082927A1 (en) * | 2013-12-05 | 2015-06-11 | Cambridge Enterprise Limited | Novel biomarker panel for major depressive disease |
| CN106062563A (en) * | 2014-01-28 | 2016-10-26 | 普雷德姆泰克有限公司 | Biomarker and methods for early diagnosis of alzheimer's disease |
| EP3232198A1 (en) * | 2014-12-12 | 2017-10-18 | Seoul National University R&DB Foundation | Biomarker for diagnosis of hepatoma and use thereof |
| CN107921094A (en) * | 2015-06-04 | 2018-04-17 | 圣拉斐尔医院有限公司 | IGFBP3 and application thereof |
| JP2018132526A (en) * | 2017-02-16 | 2018-08-23 | 国立大学法人千葉大学 | Markers, method for inspection, and inspection kit for major depressive disorder and bipolar disorder, and method for screening therapeutic medicines for major depressive disorder and bipolar disorder |
| CN108441551A (en) * | 2017-04-11 | 2018-08-24 | 刘小翠 | A kind of biomarker and its application process of phrenoblabia |
| CN112957453A (en) * | 2021-03-04 | 2021-06-15 | 上海市精神卫生中心（上海市心理咨询培训中心） | Application of insulin-like growth factor binding protein 2 in preparation of medicine for treating mental disorder caused by brain trauma |
| CN112972652A (en) * | 2021-03-04 | 2021-06-18 | 上海市精神卫生中心（上海市心理咨询培训中心） | Application of insulin-like growth factor binding protein 2 in preparation of medicine for resisting mental disorder caused by early stress |
| CN113144173A (en) * | 2021-03-04 | 2021-07-23 | 上海市精神卫生中心（上海市心理咨询培训中心） | Application of insulin-like growth factor binding protein 2 in preparation of medicine for treating mental disorder caused by stress in adult period |
Non-Patent Citations (6)
| Title |
|---|
| Burgdorf J, et al..IGFBP2 Produces Rapid-Acting and Long-Lasting Effects in Rat Models of Posttraumatic Stress Disorder via a Novel Mechanism Associated with Structural Plasticity.The International Journal of Neuropsychopharmacology.2017,全文. * |
| D O Minchenko et al..Expression of insulin-like growth factor binding protein genes and its hypoxic regulation in U87 glioma cells depends on ERN1 mediated signaling pathway of endoplasmic reticulum stress..Endocr Regul..2015,第49卷全文. * |
| Jyoti,Yadav et al..Sialic acid associated with oxidative stress and total antioxidant capacity (TAC) expression level as a predictive indicator in moderate to severe Alzheimer's disease..Exp Gerontol..2020,第141卷全文. * |
| 刘世钰 等.基于组学技术的抑郁症相关生物标志物研究进展.药学实践杂志.2018,(第03期),全文. * |
| 杨慧芳，张长征.创伤后应激障碍与认知.中央编译出版社,2021,108-110. * |
| 樊键 等.精神创伤后应激障碍的神经递质水平相关性.中国健康心理学杂志.2013,(第08期),全文. * |
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