CN113151443A - Cytokine combined analysis as schizophrenia marker and application thereof - Google Patents

Cytokine combined analysis as schizophrenia marker and application thereof Download PDF

Info

Publication number
CN113151443A
CN113151443A CN202110409624.9A CN202110409624A CN113151443A CN 113151443 A CN113151443 A CN 113151443A CN 202110409624 A CN202110409624 A CN 202110409624A CN 113151443 A CN113151443 A CN 113151443A
Authority
CN
China
Prior art keywords
schizophrenia
expression level
csf
mig
ifn
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202110409624.9A
Other languages
Chinese (zh)
Other versions
CN113151443B (en
Inventor
程勇
杜洋
陈磊
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Minzu University of China
Original Assignee
Minzu University of China
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Minzu University of China filed Critical Minzu University of China
Priority to CN202110409624.9A priority Critical patent/CN113151443B/en
Publication of CN113151443A publication Critical patent/CN113151443A/en
Application granted granted Critical
Publication of CN113151443B publication Critical patent/CN113151443B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6866—Interferon
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6869—Interleukin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/475—Assays involving growth factors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/475—Assays involving growth factors
    • G01N2333/485—Epidermal growth factor [EGF] (urogastrone)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/521—Chemokines
    • G01N2333/522—Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4 or KC
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/521—Chemokines
    • G01N2333/523—Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1or LDCF-2
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/525—Tumor necrosis factor [TNF]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/53—Colony-stimulating factor [CSF]
    • G01N2333/535—Granulocyte CSF; Granulocyte-macrophage CSF
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G01N2333/5406—IL-4
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G01N2333/55—IL-2
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/555—Interferons [IFN]
    • G01N2333/57—IFN-gamma
    • 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
    • G01N2800/302—Schizophrenia

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention provides a cytokine combined analysis as a schizophrenia marker and application thereof, in particular to application of reagent materials and/or instrument equipment for detecting the level of cytokines in a sample from an individual to be detected in preparing a kit for diagnosing schizophrenia, wherein the cytokines comprise: G-CSF, IL-2, IL-4, MIG and EGF. The cell factor of the invention is adopted for combined analysis, has high sensitivity and high specificity while having high AUC value, and has larger application prospect.

Description

Cytokine combined analysis as schizophrenia marker and application thereof
Technical Field
The invention relates to a biomarker for diagnosing schizophrenia patients and related application, in particular to a technology for diagnosing suspected schizophrenia patients or detecting the effect of schizophrenia patients after medication by using combined analysis of cytokines, belonging to the technical field of biology and medicine.
Background
Schizophrenia (Schizophrenia, SCZ) is one of the most common mental disorders worldwide, is a group of serious psychosis with unknown etiology, mostly develops slowly or subacute in young and strong years, and is clinically manifested as syndromes with various symptoms, which relate to various disorders such as sensory perception, thinking, emotion and behavior, and uncoordinated mental activities. Thought disorder is the core symptom of schizophrenia, and delusions are the most common and important disorder of thought content. It is estimated that up to 80% of schizophrenic patients have a delusion, the severer feels monitored, rejected, worried about being dosed or consummated, etc., and the patient may make serious defensive or aggressive behaviors under the influence of the delusion, which jeopardizes the safety of society, family, and others. Schizophrenic disorders can also occur with sensory disturbances including auditory hallucinations, visual hallucinations, olfactory hallucinations, taste hallucinations, emotional apathy and emotional discordance, and furthermore, emotional symptoms such as discordant excitement, irritability, depression and anxiety are also common. Furthermore, in schizophrenic patients, about 85% of patients suffer from cognitive dysfunction such as information processing and selective attention, working memory, short-term memory and learning, executive function, etc., so that they cannot take care of themselves in life, and a great burden is also imposed on the family society. Schizophrenia is related to various factors such as genetics and environment, and affects tens of millions of people all over the world. The study shows that the recurrence rate of the first-onset schizophrenia patient in 5 years is over 80 percent, and the recurrence risk of the patient with interrupted drug therapy is 5 times that of the patient with continuous drug therapy, so the adherence to medication is the main measure for maintaining stable state of illness. Second generation (atypical) antipsychotics such as risperidone, olanzapine, quetiapine, etc. are generally recommended for use as first line drugs. Clozapine, the first generation and atypical antipsychotic, was used as a second-line drug. However, the side effects and therapeutic effects of all these anti-schizophrenic drugs are still controversial, the most important reason being that the mechanism of SCZ is not well studied, it is a complex disease of different etiologies, and the cellular and molecular basis of the disease is not clear. Although there have been various hypotheses about schizophrenia over the past few decades, including the dopamine hypothesis, the transmethylation hypothesis, etc., these hypotheses have not yet been fully documented. Therefore, there is a need for more research into the etiology of SCZ to determine biomarkers of disease. At present, the diagnosis of SCZ in clinic is mainly based on the detailed medical history and mental symptoms of patients, and comprehensive judgment is made by subjective scores such as PANSS scale, and the diagnosis results are different due to different subjective experiences of doctors. In order to make the diagnosis of SCZ more objective, reduce human factors, and improve the consistency and accuracy of diagnosis, researchers around the world have been dedicated to finding biomarkers of SCZ and establishing effective detection methods in recent years.
CN101454669B discloses that more than 3 cytokines are combined for detecting mental states, but it does not disclose AUC values, specificity and sensitivity of detection, and the specific detection effect is difficult to determine. Therefore, there is a need to develop an SCZ biomarker with high AUC values while having high sensitivity and high specificity.
Disclosure of Invention
It is an object of the present invention to provide the use of reagent materials and/or instrumentation for detecting cytokine levels in a sample from an individual to be tested for the preparation of a kit for diagnosing schizophrenia and/or for assessing the effectiveness of medication for patients with schizophrenia.
In one study, the present inventors extracted serum and total RNA from peripheral blood of subjects, performed multiplex analysis of the samples, and performed statistical analysis of the ROC curve. In the analysis, 6 cytokines were screened for cytokine levels in 49 samples (24 first schizophrenia and 25 healthy controls) that expressed significant differences between healthy and schizophrenic patients. And comparing the change of the cell factors of the peripheral blood level before and after 19 schizophrenia patients take the medicine, and screening 18 cell factors with obvious difference. In further research, comparison between schizophrenic patients and healthy people is carried out, random forest algorithm model training is carried out on single factor data and joint data, and a classifier is screened out and established. It was found that the combination of the five factors of G-CSF, IL-2, IL-4, MIG and EGF gave an AUC of 0.993 and a sensitivity of 100%.
Thus, in one aspect, the present invention provides the use of a reagent material and/or an instrument device for detecting the level of a cytokine in a sample from an individual to be tested in the manufacture of a kit for the diagnosis of schizophrenia, wherein the cytokine comprises:
G-CSF, IL-2, IL-4, MIG and EGF.
According to a particular embodiment of the invention, preferably, when used for diagnosing schizophrenia, the cytokine further comprises IFN- γ.
According to a specific embodiment of the present invention, when the expression level of G-CSF is greater than 2.057 + -0.174 pg/mL, the expression level of MIG is greater than 1.891 + -0.138 pg/mL, the expression level of EGF is less than 2.206 + -0.143 pg/mL, the expression level of IL-2 is greater than 0.581 + -0.187 pg/mL and the expression level of IL-4 is greater than 1.272pg/mL, the subject to be tested is a patient suspected of having schizophrenia.
According to a specific embodiment of the present invention, preferably, when the expression level of G-CSF is greater than 2.057 + -0.174 pg/mL, the expression level of MIG is greater than 1.891 + -0.138 pg/mL, the expression level of EGF is less than 2.206 + -0.143 pg/mL, the expression level of IL-2 is greater than 0.581 + -0.187 pg/mL, the expression level of IL-4 is greater than 1.272pg/mL and the expression level of IFN-gamma is greater than 0.949 + -0.088 pg/mL, the subject to be tested is a suspected patient with schizophrenia.
In another aspect, the present invention provides the use of a reagent material and/or an apparatus for detecting the level of a cytokine in a sample from an individual to be tested in the manufacture of a kit for assessing the efficacy of a drug for a patient with schizophrenia, wherein the cytokine comprises: G-CSF, IFN-gamma and IL-8.
According to a specific embodiment of the present invention, in the assessment of the effect of the drug administration to the schizophrenic patient, the cytokines further comprise one or more of VEGF, HGF, MIP-1 α, IL-4, TNF- α, MIG and IL-1 RA.
According to a specific embodiment of the present invention, preferably, in the evaluation of the effect of the drug administration to the schizophrenia patient, the cytokines include: G-CSF, IFN-gamma, VEGF, HGF, MIP-1 alpha, IL-4, TNF-alpha, IL-8, MIG and IL-1 RA.
According to a specific embodiment of the present invention, when the expression levels of G-CSF, IFN- γ and IL-8 are reduced as compared to the level before administration, the administration effect is good.
According to a specific embodiment of the present invention, it is preferable that the administration is effective when the expression levels of G-CSF, IFN- γ, VEGF, HGF, MIP-1 α, IL-4, TNF- α, IL-8, MIG and IL-1RA are reduced as compared to before administration.
According to a particular embodiment of the invention, the sample in the present invention comprises peripheral blood.
According to a particular embodiment of the invention, the cytokine level in the present invention is its protein level and/or mRNA level.
Furthermore, the invention provides a test system for diagnosing schizophrenia and/or for evaluating the effectiveness of medication for patients with schizophrenia, comprising a test unit and an analysis unit, wherein:
the detection unit comprises a reagent material and/or an instrument device for detecting the cytokine of the invention in a sample from an individual to be detected;
the analysis unit is used for analyzing the detection result of the detection unit and evaluating whether the individual to be detected suffers from schizophrenia and/or the medication effect of the patient suffering from schizophrenia.
The level of expression of the cytokines described in the present invention can be detected by any available technique in the art.
According to a specific embodiment of the present invention, the reagent material and/or the apparatus for detecting the expression level of the cytokine may be any reagent material and/or apparatus and the like used in any feasible technique for detecting the expression level of the cytokine.
The detection system for diagnosing schizophrenia and/or for evaluating the effect of medication for schizophrenia of the present invention may be a virtual device as long as the functions of the detection unit and the evaluation unit are realized. The detection unit can comprise various detection reagent materials and/or detection instrument equipment and the like; the data analysis unit may be any computing instrument, module or virtual device capable of analyzing and processing the detection result of the detection unit to obtain the disease condition of schizophrenia and/or the medication effect of the patient with schizophrenia, for example, a data chart corresponding to each possible detection result and the corresponding disease condition may be prepared in advance, and the disease condition of schizophrenia may be obtained by comparing the detection result of the detection unit with the data chart.
According to a particular embodiment of the invention, in the test system for diagnosing schizophrenia and/or for evaluating the effectiveness of medication for patients with schizophrenia according to the invention:
when the expression level of G-CSF is more than 2.057 +/-0.174 pg/mL, the expression level of MIG is more than 1.891 +/-0.138 pg/mL, the expression level of EGF is less than 2.206 +/-0.143 pg/mL, the expression level of IL-2 is more than 0.581 +/-0.187 pg/mL and the expression level of IL-4 is more than 1.272pg/mL, the individual to be tested is a suspected patient with schizophrenia;
preferably, when the expression level of G-CSF is more than 2.057 +/-0.174 pg/mL, the expression level of MIG is more than 1.891 +/-0.138 pg/mL, the expression level of EGF is less than 2.206 +/-0.143 pg/mL, the expression level of IL-2 is more than 0.581 +/-0.187 pg/mL, the expression level of IL-4 is more than 1.272pg/mL and the expression level of IFN-gamma is more than 0.949 +/-0.088 pg/mL, the individual to be tested is a suspected patient with schizophrenia;
wherein, when the expression levels of G-CSF, IFN-gamma and IL-8 are reduced compared with those before administration, the administration effect is judged to be good;
preferably, administration is effected when the expression levels of G-CSF, IFN- γ, VEGF, HGF, MIP-1 α, IL-4, TNF- α, IL-8, MIG and IL-1RA are reduced compared to pre-administration.
Since the current diagnosis of psychiatric disorders, including but not limited to schizophrenia, is primarily dependent on the clinician's subjective judgment based on the diagnostic and statistical manual for psychiatric disorders, fifth edition (DSM-V) and tenth edition International Classification of diseases (ICD-10). The invention realizes the diagnosis of schizophrenia from objective factors, ensures that the diagnosis is more rigorous and reliable, and can help doctors to objectively judge whether the effect of patients who are diagnosed after taking the medicine is achieved. Compared with the prior art related to the diagnosis of schizophrenia, the method has higher AUC and further has higher sensitivity and specificity.
Drawings
FIG. 1 is a graph showing the expression of six cytokine levels in the schizophrenic patients and healthy controls in example 1.
FIG. 2 is a graph of cytokine changes before and after dosing in schizophrenic patients, as well as changes in PANSS scale scores.
FIG. 3 is a ROC curve for six significantly varying cytokines in example 2 of the present invention, and for two factors with the highest AUC.
FIG. 4 is a ROC curve of the combined analysis of five factors of G-CSF, IL-2, IL-4, MIG and EGF in example 2 of the present invention.
FIG. 5 is a ROC curve showing the diagnostic effect of the combination of six factors, G-CSF, IL-2, IL-4, IFN-. gamma., MIG and EGF, in example 3 of the present invention.
FIG. 6 is a graph showing the results of ROC curves in example 4 of the present invention.
FIG. 7 is a graph showing the results of ROC curves in comparative example 1 of the present invention.
FIG. 8 is a graph showing the results of the ROC curve in comparative example 2 of the present invention.
Detailed Description
The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.
Example 1
Subjects were 24 first schizophrenic patients (FEDF) admitted to the third national hospital in foshan city, and 25 age and gender matched healthy volunteers were recruited as controls (HC), of which 19 schizophrenic patients received a short-term treatment for eight weeks before the collection of peripheral blood samples again. All doctors participating in the diagnosis work have the qualification of psychiatric medical practitioners and have the psychiatric practical experience of more than 10 years, are skilled in mastering diagnosis standards of ICD-10 and DSM-V, adopt positive and negative schizophrenia scale scores (PANSS) to evaluate the psychopathology state of patients, have unified operation specifications, meet the requirement of consistency detection (Kappa is 0.68-0.82), and exclude the schizophrenic patients with complications before grouping. All participants gave written informed consent prior to inclusion in the study. The study protocol was approved by the ethical committee of the third national hospital in foshan city.
1. Serum extraction
Approximately 4mL of peripheral blood was collected from each subject and allowed to clot at room temperature for 1 hour, and then the sample was centrifuged at 3000 xg for 10 minutes to obtain serum. The sera were then stored in a low temperature freezer at-80 ℃ or directly analyzed.
Total RNA was extracted from 200. mu.L of whole blood by Trizol method and stored at-80 ℃.
2. Determination of cytokines:
cytokine levels in serum were measured using a human cytokine magnetic 30 composite plate on a Luminex platform using multiplex cytokine array analysis.
TABLE 1 cytokine expression levels (pg/mL) in schizophrenic patients versus healthy controls
Figure BDA0003023641080000061
The results, as shown in Table 1 and FIG. 1, show that the expression levels of G-CSF, IFN- γ, IL-2, IL-4, MIG were significantly higher in the first schizophrenia patients than in the healthy controls, and EGF was significantly lower in the first schizophrenia patients than in the healthy controls.
In 19 of the 24 first schizophrenic patients, peripheral blood samples were collected again after receiving eight weeks of short-term treatment and the cytokine expression levels before and after administration were measured, and the results are shown in table 2 below:
TABLE 2 cytokine expression levels (pg/mL) before and after short-term dosing in schizophrenic patients
Figure BDA0003023641080000071
The results, as shown in Table 2 and FIG. 2, show that the expression levels of G-CSF, IFN- α, IFN- γ, IL-1RA, IL-1 β, IL-2, IL-4, IL-7, IL-8, IL-10, MIG, MIP-1 α, MCP-1, TNF- α, FGF-2, HGF, VEGF in patients with schizophrenia after short-term administration are significantly lower than before; and by the change in PANSS scale scores (figure 2, table 3), the improvement in schizophrenic patients after short-term dosing can be seen.
TABLE 3 PANSS Scale Scoring results
Figure BDA0003023641080000081
Example 2
On the basis of example 1, the diagnostic effect of six factors, namely G-CSF, IFN-gamma, IL-2, IL-4, MIG and EGF, were observed individually and are shown by using ROC curve. The larger the area under the ROC curve (AUC), the better the diagnosis.
The results are shown in FIG. 3, in which the AUC of six factors, G-CSF, IFN-gamma, IL-2, IL-4, MIG and EGF are 0.945,0.942,0.774,0.750,0.867 and 0.718, respectively. Among them, the diagnostic effects of G-CSF and IFN-gamma are the best, and the sensitivity and specificity of both factors reach 95.8% and 80.0%.
Further, the diagnostic effect of the combination of five factors, G-CSF, IL-2, IL-4, MIG and EGF, was observed and expressed using ROC curve. The larger the area under the ROC curve (AUC), the better the diagnosis.
The results are shown in FIG. 4, and the AUC of the combined analysis of the five factors of G-CSF, IL-2, IL-4, MIG and EGF is 0.993, and the sensitivity and specificity reach 100% and 92%.
Example 3
The diagnostic effect of the combination of six factors, G-CSF, IL-2, IL-4, IFN-. gamma., MIG and EGF, was observed and expressed using the ROC curve. The larger the area under the ROC curve (AUC), the better the diagnosis.
As shown in FIG. 5, the AUC of the combined analysis of six factors, G-CSF, IFN-. gamma., IL-2, IL-4, MIG and EGF, was 0.997. The sensitivity and specificity reach 100% and 96%.
Example 4
The administration effect of the combination of ten factors, G-CSF, VEGF, HGF, MIP-1 alpha, IL-4, TNF-alpha, IL-8, IFN-gamma, MIG and IL-1RA, was evaluated and shown by using the ROC curve. The larger the area under the ROC curve (AUC), the better the detected medication effect.
The results are shown in FIG. 6, in which the AUCs of the ten factors of G-CSF, VEGF, HGF, MIP-1 α, IL-4, TNF- α, IL-8, IFN- γ, MIG, IL-1RA are 0.970,0.787,0.724,0.787,0.778,0.733,0.878,0.812,0.742,0.690, respectively. The administration effect of G-CSF detection is best, and its sensitivity and specificity are up to 100% and 89.5%. The sensitivity of the remaining factors was, in order, 78.9%, 63.2%, 73.7%, 94.7%, 63.2%, 84.2%, 84.2%, 84.2%, 84.2%, 84.2%, and the specificity was, in turn, 78.9%, 89.5%, 63.2%, 84.2%, 100%, 68.4%, 52.6%, 52.6%.
The combination of the first three AUC factors among the individual factors, namely G-CSF, IFN-gamma and IL8, resulted in a new ROC curve with an AUC of 0.983 and sensitivity and specificity of 100% and 94.7%.
The AUC of the ROC curve obtained by analyzing all the factors together is 1.0, and the sensitivity and the specificity reach 100 percent and 100 percent.
Comparative example 1
G-CSF and IFN-gamma were verified at the transcriptional level and a multifactorial combination analysis was performed on both factors at the protein and mRNA levels.
Results as in figure 7, G-CSF and IFN- γ were also significantly upregulated at the transcriptional level in schizophrenic patients. The combined analysis of the protein level and mRNA level of G-CSF and IFN-gamma, and individual factors, resulted in ROC curves showing AUC of 0.981 and 0.972, respectively, the sensitivity and specificity of G-CSF reached 94.7% and 100.0%, and the sensitivity and specificity of IFN-gamma reached 89.5% and 95.0%. Next, the ROC curve obtained by analyzing the two factors in combination at the protein level showed an AUC of 0.975, and both sensitivity and specificity of 95.0%; combined analysis at the mRNA level showed an AUC of 0.936 with sensitivity and specificity of 88.9% and 95.0%, respectively. Finally, the ROC curve obtained by jointly analyzing the two levels of the two factors showed an AUC of 0.986 and sensitivity and specificity of 94.4% and 100.0%, respectively.
Comparative example 2
The results of the combined analysis of IFN-gamma and IL-2, IL-4, MIG and EGF are shown in FIG. 8, and the AUC is 0.967, and the sensitivity and specificity are 95.8% and 94.2%.
The results show that the combined analysis of the cell factors screened by the invention can be used as a biomarker for diagnosing the schizophrenia patients, and the combined analysis of the cell factors can judge whether the illness state of the schizophrenia patients is improved after the drug is taken.

Claims (10)

1. Use of a reagent material and/or an instrument device for detecting the level of a cytokine in a sample from an individual to be tested for the preparation of a kit for the diagnosis of schizophrenia, wherein the cytokine comprises:
G-CSF, IL-2, IL-4, MIG and EGF.
2. The use according to claim 1, wherein the cytokine further comprises IFN- γ when used for the diagnosis of schizophrenia.
3. The use according to claim 1 or 2, wherein the subject is suspected to be a patient with schizophrenia when the expression level of G-CSF is more than 2.057 ± 0.174pg/mL, the expression level of MIG is more than 1.891 ± 0.138pg/mL, the expression level of EGF is less than 2.206 ± 0.143pg/mL, the expression level of IL-2 is more than 0.581 ± 0.187pg/mL and the expression level of IL-4 is more than 1.272 pg/mL;
preferably, when the expression level of G-CSF is more than 2.057 +/-0.174 pg/mL, the expression level of MIG is more than 1.891 +/-0.138 pg/mL, the expression level of EGF is less than 2.206 +/-0.143 pg/mL, the expression level of IL-2 is more than 0.581 +/-0.187 pg/mL, the expression level of IL-4 is more than 1.272pg/mL and the expression level of IFN-gamma is more than 0.949 +/-0.088 pg/mL, the individual to be tested is a suspected patient with schizophrenia.
4. Use of a reagent material and/or an apparatus for detecting the level of a cytokine in a sample from an individual to be tested for the manufacture of a kit for assessing the effectiveness of a drug for a patient with schizophrenia, wherein the cytokine comprises: G-CSF, IFN-gamma and IL-8.
5. The use of claim 4, wherein in assessing the efficacy of a medication administered to a patient with schizophrenia, the cytokines further comprise one or more of VEGF, HGF, MIP-1 α, IL-4, TNF- α, MIG, and IL-1 RA;
preferably, in assessing the efficacy of a drug administered to a schizophrenic patient, the cytokines comprise: G-CSF, IFN-gamma, VEGF, HGF, MIP-1 alpha, IL-4, TNF-alpha, IL-8, MIG and IL-1 RA.
6. The use according to claim 4 or 5, wherein the administration is effected when the expression levels of G-CSF, IFN- γ and IL-8 are reduced compared to the pre-administration;
preferably, administration is effected when the expression levels of G-CSF, IFN- γ, VEGF, HGF, MIP-1 α, IL-4, TNF- α, IL-8, MIG and IL-1RA are reduced compared to pre-administration.
7. The use of any one of claims 1-6, wherein the sample comprises peripheral blood.
8. The use according to any one of claims 1-2 or claims 4-5, wherein the cytokine level is its protein level and/or mRNA level.
9. A test system for diagnosing schizophrenia and/or for assessing the effectiveness of medication for patients with schizophrenia, comprising a test unit and an analysis unit, wherein:
the detection unit comprises a reagent material and/or an instrument device for detecting the cytokine according to any one of claims 1-2 or 4-5 in a sample from an individual to be tested;
the analysis unit is used for analyzing the detection result of the detection unit and evaluating whether the individual to be detected suffers from schizophrenia and/or the medication effect of the patient suffering from schizophrenia.
10. The detection system of claim 9, wherein:
when the expression level of G-CSF is more than 2.057 +/-0.174 pg/mL, the expression level of MIG is more than 1.891 +/-0.138 pg/mL, the expression level of EGF is less than 2.206 +/-0.143 pg/mL, the expression level of IL-2 is more than 0.581 +/-0.187 pg/mL and the expression level of IL-4 is more than 1.272pg/mL, the subject to be tested is diagnosed with schizophrenia;
preferably, the subject is diagnosed with schizophrenia when the expression level of G-CSF is greater than 2.057 + -0.174 pg/mL, MIG is greater than 1.891 + -0.138 pg/mL, EGF is less than 2.206 + -0.143 pg/mL, IL-2 is greater than 0.581 + -0.187 pg/mL, IL-4 is greater than 1.272pg/mL, and IFN- γ is greater than 0.949 + -0.088 pg/mL;
wherein, when the expression levels of G-CSF, IFN-gamma and IL-8 are reduced compared with those before administration, the administration effect is judged to be good;
preferably, administration is effected when the expression levels of G-CSF, IFN- γ, VEGF, HGF, MIP-1 α, IL-4, TNF- α, IL-8, MIG and IL-1RA are reduced compared to pre-administration.
CN202110409624.9A 2021-04-16 2021-04-16 Cytokine combined analysis as schizophrenia marker and application thereof Expired - Fee Related CN113151443B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110409624.9A CN113151443B (en) 2021-04-16 2021-04-16 Cytokine combined analysis as schizophrenia marker and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110409624.9A CN113151443B (en) 2021-04-16 2021-04-16 Cytokine combined analysis as schizophrenia marker and application thereof

Publications (2)

Publication Number Publication Date
CN113151443A true CN113151443A (en) 2021-07-23
CN113151443B CN113151443B (en) 2022-07-01

Family

ID=76868231

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110409624.9A Expired - Fee Related CN113151443B (en) 2021-04-16 2021-04-16 Cytokine combined analysis as schizophrenia marker and application thereof

Country Status (1)

Country Link
CN (1) CN113151443B (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1394282A (en) * 2000-10-31 2003-01-29 新澙大学校长代表的日本国 Diagnostic kit for schizophrenia
US20050079636A1 (en) * 2001-09-25 2005-04-14 White Keith D. Method and apparatus for diagnosing schizophrenia and schizophrenia subtype
CN101454669A (en) * 2006-02-17 2009-06-10 关山敦生 Biological load indicator and method of measuring biological load
CN107345970A (en) * 2017-07-20 2017-11-14 首都医科大学附属北京安定医院 A kind of peripheral blood biomarker available for Schizophrenia diagnosis
CN109996895A (en) * 2016-11-28 2019-07-09 卡斯滕·科思 The method and biomarker of in-vitro diagnosis phrenoblabia

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1394282A (en) * 2000-10-31 2003-01-29 新澙大学校长代表的日本国 Diagnostic kit for schizophrenia
US20050079636A1 (en) * 2001-09-25 2005-04-14 White Keith D. Method and apparatus for diagnosing schizophrenia and schizophrenia subtype
CN101454669A (en) * 2006-02-17 2009-06-10 关山敦生 Biological load indicator and method of measuring biological load
CN109996895A (en) * 2016-11-28 2019-07-09 卡斯滕·科思 The method and biomarker of in-vitro diagnosis phrenoblabia
CN107345970A (en) * 2017-07-20 2017-11-14 首都医科大学附属北京安定医院 A kind of peripheral blood biomarker available for Schizophrenia diagnosis

Also Published As

Publication number Publication date
CN113151443B (en) 2022-07-01

Similar Documents

Publication Publication Date Title
Joppa et al. Sarcopenic obesity, functional outcomes, and systemic inflammation in patients with chronic obstructive pulmonary disease
US10184931B2 (en) Methods and systems for measuring and using the oxidation-reduction potential of a biological sample
Rinaldi et al. Electrical impedance spectroscopy for the characterization of skin barrier in atopic dermatitis
Gattas et al. Procalcitonin as a diagnostic test for sepsis: health technology assessment in the ICU
Boonpiyathad et al. Interleukin-2 levels in exhaled breath condensates, asthma severity, and asthma control in nonallergic asthma.
Carpagnano et al. Looking for airways periostin in severe asthma: could it be useful for clustering type 2 endotype?
EP3786305A1 (en) Biomarker for depression and use thereof
Krupp et al. Increased prevalence of airway reactivity in children with eosinophilic esophagitis
Moodley et al. Exhaled nitric oxide is elevated in patients with progressive systemic sclerosis without interstitial lung disease
Sandroff et al. Comparing two conditions of administering the six-minute walk test in people with multiple sclerosis
Zhang et al. Association between low handgrip strength and 90-day mortality among older Chinese inpatients: a national multicenter prospective cohort study
Xue et al. Role of CD4+ T and CD8+ T Lymphocytes‐Mediated Cellular Immunity in Pathogenesis of Chronic Obstructive Pulmonary Disease
CN108508211B (en) Serum marker FGF9 for first-onset non-drug schizophrenic patients and application thereof
CN113151443B (en) Cytokine combined analysis as schizophrenia marker and application thereof
Trueba et al. Effects of academic exam stress on nasal leukotriene B4 and vascular endothelial growth factor in asthma and health
Zhang et al. Neural respiratory drive measured using surface electromyography of diaphragm as a physiological biomarker to predict hospitalization of acute exacerbation of chronic obstructive pulmonary disease patients
Yanagi et al. Macrophage migration inhibitory factor in zinc-allergic systemic contact dermatitis
Masoumi et al. Diagnostic value of S100B and neuron-specific enolase in distinguishing acute central and peripheral vertigo
CN114015770B (en) Schizophrenia total peripheral blood RNA marker FGFR3 and application thereof
WO2024088066A1 (en) Sarcopenia diagnosis marker and use thereof
Patsiris et al. Exhaled breath condensate: a potential tool to evaluate the relationship between inflammation and dyspnea in patients with chronic obstructive pulmonary disease
Chen et al. Incidence of frailty and construction of prediction model in elderly male patients with chronic obstructive pulmonary disease
WO2012049864A1 (en) Method for providing data to diagnose depression
Yadav et al. Human Saliva Quality and Holistic Practices: A Review on the Effects of Meditation, Dance, and Exercise
Aretz et al. The role of leukocytes in cognitive impairment due to long‐term exposure to fine particulate matter: A large population‐based mediation analysis

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20220701

CF01 Termination of patent right due to non-payment of annual fee