CN115710600A - Primer group, kit and detection method for schizophrenia detection - Google Patents

Abstract

The invention provides a primer group for schizophrenia detection, which comprises a BDNF primer group, a beta-NGF primer group, a CD133 primer group and a D2 primer group, wherein each primer group consists of an upstream primer and a downstream primer. The invention further provides a kit for detecting schizophrenia, which comprises the four primer groups and is suitable for industrial application. Finally, the invention provides a method for detecting schizophrenia, which comprises the following steps: extracting total RNA of a sample, carrying out reverse transcription on the total RNA into cDNA, preparing a PCR mixed system and carrying out PCR amplification. The primer group for detecting schizophrenia improves the reliability of the detection result of schizophrenia through more proper biomarkers, more proper biomarker quantity and more proper primer groups. The kit is specially developed for detecting schizophrenia, and has the advantages of good specificity, high sensitivity, reliable result and good stability.

Description

Primer set, kit and detection method for schizophrenia detection

technical field

The invention relates to the technical field of biological detection, in particular to a primer set for the detection of schizophrenia; at the same time, the invention also relates to a kit for the detection of schizophrenia and a detection method thereof.

Background technique

Schizophrenia (SC) is a chronic disabling disease with complex etiology and high phenotype heterogeneity. It is generally believed that the lifetime prevalence of schizophrenia in the general population is 1%, and most of them have a poor prognosis. More than 50% of patients have long-term intermittent mental problems, and about 20% of patients have residual symptoms and lead to mental disability. The onset characteristics of schizophrenia include positive symptoms (delusions, hallucinations, excitement, etc.), negative symptoms (affective bluntness, emotional withdrawal, emotional communication disorders, etc.) and cognitive dysfunction. There are three systems of diagnostic criteria for schizophrenia currently used by clinical psychiatrists in my country: the Chinese classification and diagnostic criteria of mental illness, the American statistical classification system for the diagnosis of mental illness, and the international disease classification system. And these diagnostic methods are to judge whether it is schizophrenia according to the scale to evaluate the patient's sensory disturbance, perception disturbance, thinking disturbance, will and behavior disturbance. The limitations of this diagnosis are that single symptoms are used to replace clinical phases, multivariate symptoms are evaluated with a linear scale, symptoms of different natures are described as differences in degree, and both excesses and deficiencies can be diseases and omissions Sexuality and closure, as well as exclusivity, this leads to the fact that experienced psychiatrists may not be good raters. The clinical symptoms of schizophrenia are characterized by diversity, but the diagnosis relies only on mental examination, without effective biological markers as indicators, making accurate diagnosis difficult.

CN104330573 discloses a method for detecting serum protein factors for judging schizophrenia, comprising the following steps: 1) extracting blood from a suspected patient, and separating serum protein; 2) detecting nerve growth factor (NGF), leukocyte Interferon (IL-6), calcium binding protein S100B, interferon (IFN-γ), tumor necrosis factor (TNF-α), brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), The content of 8 protein factors of basic myelin protein (MBP); 3) compare the detected protein factor content with the normal reference value, and scientifically classify and judge schizophrenia. SDA and ROC curves were applied to the concentrations of serum protein factors selected as diagnostic markers for schizophrenia. It was found that the concentration of five factors, BDNF, IL-6, S100β, MBP and GFAP, had a greater contribution to assisting in the diagnosis of schizophrenia. And fitting the concentrations of five important variables, it was found that when using the ROC curve combined with multiple protein factor fitting indicators to diagnose schizophrenia, the diagnostic value was higher than that of any single protein factor.

Those skilled in the art know that in the field of biological detection, more indicators do not mean more accurate results, and the two are not simply positively correlated. More detection indicators do not mean that more reliable detection results can be obtained. Therefore, improving the detection accuracy of schizophrenia is an urgent problem to be solved by those skilled in the art.

Contents of the invention

In view of this, the present invention aims to propose a primer set for the diagnosis of schizophrenia, so as to improve the detection accuracy of schizophrenia.

In order to achieve the above object, technical solution of the present invention is achieved in that way:

A primer set for the detection of schizophrenia, the primer set includes a BDNF primer set, a β-NGF primer set, a CD133 primer set and a D2 primer set;

The BDNF primer set includes a BDNF upstream primer and a BDNF downstream primer; the nucleotide sequence of the BDNF upstream primer is shown in SEQ ID No.1, and the nucleotide sequence of the BDNF downstream primer is shown in SEQ ID No.2 Show;

The β-NGF primer set includes β-NGF upstream primers and β-NGF downstream primers; the nucleotide sequence of the β-NGF upstream primers is shown in SEQ ID No.3, and the core of the β-NGF downstream primers The nucleotide sequence is shown in SEQ ID No.4;

The CD133 primer set includes a CD133 upstream primer and a CD133 downstream primer; the nucleotide sequence of the CD133 upstream primer is shown in SEQ ID No.5, and the nucleotide sequence of the CD133 downstream primer is shown in SEQ ID No.6. Show;

The D2 primer set includes a D2 upstream primer and a D2 downstream primer; the nucleotide sequence of the D2 upstream primer is as shown in SEQ ID No.7, and the nucleotide sequence of the D2 downstream primer is as shown in SEQ ID No.8 Show.

Further, the concentration of the primer set includes one or more of the following (i) to (viii): (i) the concentration of the BDNF upstream primer is 5-15 μM; (ii) the concentration of the BDNF downstream primer The concentration is 5-15 μM; (iii) the concentration of the β-NGF upstream primer is 5-15 μM; (iv) the concentration of the β-NGF downstream primer is 5-15 μM; (v) the concentration of the CD133 upstream primer (vi) the concentration of the CD133 downstream primer is 5-15 μM; (vii) the concentration of the D2 upstream primer is 5-15 μM; (viii) the concentration of the D2 downstream primer is 5-15 μM.

Secondly, the present invention provides a kit for detecting schizophrenia, including the aforementioned primer set.

Further, the kit also includes buffer, dNTP mixture, and TaqDNA polymerase.

Further, the buffer solution includes Tris·HCL; preferably, the concentration of the Tris·HCL is 5-15 mM.

Further, the dNTP mixture includes dATP, dGTP, dTTP, dCTP, dUTP, MgCl ₂ and SYBRGreen1.

Further, the concentration of the dATP is 0.2-0.8mM; and/or the concentration of the dGTP is 0.2-0.8mM; and/or the concentration of the dTTP is 0.2-0.8mM; and/or the concentration of the dCTP and/or the concentration of the dUTP is 0.2-0.8mM; and/or the concentration of the MgCl ₂ is 3-7mM; and/or the volume percentage concentration of the SYBRGreen1 is 0.025%.

Further, the concentration of the TaqDNA polymerase is 3-7 U/μL.

Finally, the present invention also provides a method for detecting schizophrenia, comprising the following steps:

S1: extract the total RNA of the sample;

S2: Reverse transcribing the total RNA into cDNA to obtain a sample to be detected;

S3: Using the aforementioned primer set or the aforementioned kit, perform PCR amplification on BDNF, β-NGF, CD133 and D2 in the sample to be detected to obtain the expression levels of BDNF, β-NGF, CD133 and D2.

Further said samples come from blood, hippocampal tissue or prefrontal cortex tissue.

Compared with the prior art, the present invention has the following advantages:

The primer set for schizophrenia detection of the present invention improves the reliability of schizophrenia detection results through more suitable biomarkers, more suitable biomarker quantities and more suitable primer sets. In order to facilitate industrial application, the invention also provides a kit for detecting schizophrenia. In addition, the primer set and kit of the present invention can also be used to evaluate the efficacy of drugs for schizophrenia. The kit of the invention is specially developed for the detection of schizophrenia, has good specificity, high sensitivity, reliable results and good stability, and is especially suitable for detection of negative symptoms and cognitive dysfunction.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only These are some implementations of the present invention. For those skilled in the art, other drawings can also be obtained according to these drawings without creative work. In the attached picture:

Fig. 1 is the electrophoresis result figure of QPCR amplification product and sterile ultrapure water in embodiment 1;

Fig. 2 is the electrophoresis result figure of BDNF primer set sensitivity verification in embodiment 1;

Fig. 3 is the electrophoresis result figure of the sensitivity verification of β-NGF primer set in embodiment 1;

Fig. 4 is the electrophoresis result figure of CD133 primer set sensitivity verification in embodiment 1;

Fig. 5 is the electrophoresis result figure of D2 primer set sensitivity verification in embodiment 1;

Fig. 6 is the detection result figure of IL-6 and TNF-α content in normal rat group offspring and model rat group offspring in embodiment 2.1;

Fig. 7 is the behavioral PPI detection result figure of normal rat group offspring and model rat group offspring in embodiment 2.1;

Fig. 8 is the relative content figure of four kinds of mRNAs of normal rat group offspring and model rat group offspring in embodiment 2.2;

FIG. 9 is a graph showing the relative contents of four mRNAs in the offspring of the control group and the treatment group in Example 3.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts fall within the protection scope of the present invention.

In the present invention, SD rats were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., the quality certificate number is 11400700258932, and the license number is SCXK (Beijing) 2018-0006.

In the present invention, β-NGF is nerve growth factor, BDNF is brain-derived neurotrophic factor, CD133 is hematopoietic stem cell antigen, and D2 is dopamine receptor G protein-coupled receptor.

The present invention will be described in detail below with reference to the accompanying drawings and examples.

Example 1 This example relates to a primer set and a kit for the detection of schizophrenia.

1. Design and screening of primer sets

Primer design was performed using the Primer-BLAST tool. Enter the sequence information of BDNF, β-NGF, CD133, and D2 genes in the PCR Template column. The specific design principles are: select 100 to 250 for PCR product size, fill in 57, 60, and 63 for Primer melting temperatures (T _m ), select Refseq mRNA for Database, select Rattusnorvegicus (taxid: 10116) for Organism, and perform Get Primers. According to the primer gene name, species, length, t _m value of 60, GC% of 40%---60%, Self complementarity, Self 3'complementarity conditions, for BDNF, β-NGF, CD133, D2, each screening Three sets of primers were selected (Table 1).

Table 1 Primer sets screened against BDNF, β-NGF, CD133, D2

After verification, primers with high specificity suitable for the experimental design were screened out. Specifically as follows: the BDNF primer set includes a BDNF upstream primer and a BDNF downstream primer; the nucleotide sequence of the BDNF upstream primer is shown in SEQ ID No.1, and the nucleotide sequence of the BDNF downstream primer is shown in SEQ ID No. 2. The β-NGF primer set includes β-NGF upstream primers and β-NGF downstream primers; the nucleotide sequence of the β-NGF upstream primers is shown in SEQ ID No.3, and the core of the β-NGF downstream primers The nucleotide sequence is shown in SEQ ID No.4. The CD133 primer set includes a CD133 upstream primer and a CD133 downstream primer; the nucleotide sequence of the CD133 upstream primer is shown in SEQ ID No.5, and the nucleotide sequence of the CD133 downstream primer is shown in SEQ ID No.6. Show. The D2 primer set includes a D2 upstream primer and a D2 downstream primer; the nucleotide sequence of the D2 upstream primer is as shown in SEQ ID No.7, and the nucleotide sequence of the D2 downstream primer is as shown in SEQ ID No.8 Show.

The nucleotide sequence of the BDNF primer set is as follows:

SEQ ID NO. 1: TACCTGGATGCCGCAAACAT.

SEQ ID NO. 2: TGGCCTTTTGATACCGGGAC.

The nucleotide sequence of the β-NGF primer set is as follows:

SEQ ID NO. 3: TACCTGGATGCCGCAAACAT.

SEQ ID NO. 4: GAAGACTGGGTGGGTGGATG.

The nucleotide sequence of the CD133 primer set is as follows:

SEQ ID NO. 5: GGGTCTTGTACGCCATCACA.

SEQ ID NO. 6: CAGTATCGAGACGGGTCATCA.

The nucleotide sequence of the D2 primer set is as follows:

SEQ ID NO. 7: TATCATTGCCAAACCCTGCCTT.

SEQ ID NO. 8: GCAGCATCCTTGAGTGGTGT.

The above primer sets are all in powder form and generally not directly used for PCR amplification. Instead, add an appropriate amount of ultrapure water (DEPC water) to dilute the primers to 10 μmol/L, aliquot them and store them at -20°C for future use. When in use, an appropriate amount of primers can be used to prepare a PCR mixed system. It should be noted that in the actual detection process, the primer set of the present invention needs to be used together with other reagents. The selection and dosage of other reagents can be prepared according to the general knowledge of those skilled in the art, and are not limited to buffers, dNTP mixtures, Taq DNA polymerase. The following is a specific composition of the kit, which is only exemplary and not specifically limited.

One of the specific implementations of the kit of the present invention is as follows: it includes the aforementioned primer pair, and in addition, buffer, dNTP mixture, and TaqDNA polymerase. As one of the preferred embodiments of the present invention, the buffer solution includes Tris·HCL, the concentration of the Tris·HCL is 10 mM; the concentration of the TaqDNA polymerase is 5 U/μL. As another preferred embodiment of the present invention, the dNTP mixture includes dATP, dGTP, dTTP, dCTP, dUTP, MgCl ₂ and SYBRGreen1. Wherein, the concentration of the dATP is 0.5mM; the concentration of the dGTP is 0.5mM; the concentration of the dTTP is 0.5mM; the concentration of the dCTP is 0.5mM; the concentration of the dUTP is 0.5mM; The concentration of ₂ is 5.0 mM; the volume percent concentration of SYBRGreen1 is 0.025%.

The buffer, dNTP mixture, and TaqDNA polymerase are also stored separately at -20°C for future use. When in use, an appropriate amount of buffer, dNTP mixture, and TaqDNA polymerase can be used to prepare a PCR mixed system. It should be noted that the buffer, dNTP mixture, and TaqDNA polymerase can be prepared according to the general knowledge of those skilled in the art in the actual application of the detection process, and are not limited to the foregoing specific examples.

2. Specificity verification of primers

Use the QPCR product in Step 4 of Example 2.2 to perform a specificity verification test. The specificity verification method is agarose gel electrophoresis detection (conventional method in the field, including making agarose gel with 1×TAE electrophoresis buffer, spotting and electrophoresis detection). In this example, sterile ultrapure water was used as a negative control template, and the QPCR amplification product in Step 4 of Example 2.2 was detected by agarose gel electrophoresis, and the results are shown in FIG. 1 . According to Figure 1, all four pairs of primer sets can amplify the target band (the expected fragments are 182 for BDNF, 226 for β-NGF, 238 for CD133, and 186 for D2), and no non-target bands were amplified. This shows that the aforementioned BDNF primer set, β-NGF primer set, CD133 primer set, and D2 primer set (SEQ ID No. 1 to SEQ ID No. 8) have strong specificity.

3. Sensitivity verification test of primers

Steps 1 to 4 are the same as Steps 1 to 4 in Example 2.2, the only difference lies in: cDNA sample preparation. Specifically, after the reverse transcription is completed, the reverse-transcribed genomic cDNA of normal rats is serially diluted by 10 times, and the diluted concentrations are 100 ng/μL, 10 ng/μL, 1 ng/μL, 100 pg/μL, 10 pg/μL, 1pg/μL, 100fg/μL, 10fg/μL, 1fg/μL, also use sterile ultrapure water as negative control. Step 5, the QPCR product is subjected to the sensitivity verification test of the primers, and the sensitivity verification method is agarose gel electrophoresis detection (conventional methods in the field, including making agarose gel with 1×TAE electrophoresis buffer, spotting and electrophoresis detection). The results are shown in Figures 2 to 5.

It can be seen from Figure 2 to Figure 5 that the sensitivity of QPCR to detect BDNF is ≥1pg/μL, the sensitivity of QPCR to detect β-NGF is ≥1pg/μL, the sensitivity of QPCR to detect CD133 is ≥1pg/μL, and the sensitivity of QPCR to detect D2 is ≥ 10 pg/μL. This shows that the aforementioned BDNF primer set, β-NGF primer set, CD133 primer set, and D2 primer set (SEQ ID No. 1 to SEQ ID No. 8) have high sensitivity.

Example 2 This example relates to the application of the kit to the detection of schizophrenia

Example 2.1 Constructing an animal model

In this example, rats with schizophrenia in offspring exposed to maternal immune activation prenatally were used as an animal model. The model is to use successfully pregnant female mice, inject polycytidylic acid (Poly I:C) solution into the tail vein on the ninth day, and then measure the inflammation level of the female mice to determine whether the model is successful. The specific animal model construction method is as follows:

In step 1, the rats are raised in a laboratory standard environment, the temperature is controlled at 22±20°C, and the humidity is controlled at 50±10%. The rats were mated and bred when they were 3 months old, and the vaginal smear was checked the next morning. If the sperm was observed under the microscope, it was considered as a successful conception, which was recorded as 0.5 days of pregnancy (GD0.5).

In step 2, the female rats that were successfully conceived were divided into cages and randomly divided into a normal rat group and a model rat group with equal numbers. On the ninth day of pregnancy, the pregnant rats in the model rat group were injected with polycytidylic acid (Poly I:C) solution (10 mg/kg) into the tail vein; the pregnant rats in the normal rat group were injected with sterile normal saline (0.9% ), which is the control group. Three hours after the injection, blood was collected from the tail vein, and the blood collection volume was 1 mL per mouse. After blood collection, the content of IL-6 and TNF-α was detected by the double-antibody sandwich method, and the detection results are shown in Figure 1. Figure 6 shows the levels of inflammatory factors in female mice. It can be seen from Figure 6 that the concentrations of IL-6 and TNF-α in the serum of pregnant mice injected with Poly I:C virus were significantly increased three hours later. It shows that the injection of Poly I:C virus has activated the immune system of pregnant mice, causing them to produce inflammation. It should be noted that the double-antibody sandwich method is a conventional method in the art and will not be repeated here. The method for constructing an animal model by injecting Poly I:C virus to pregnant mice belongs to one of the maternal immune activation model (MIA), and does not constitute a limitation of the present invention. It is also possible to construct the maternal immune activation model (MIA) by other methods. Can.

Step 3: After the offspring rats became adults (60 days), the offspring of the normal rat group and the offspring of the model rat group were subjected to a startle reflex weak stimulus inhibition (PPI) behavioral test, and the results were shown in Figure 7 Show. It can be seen from Figure 7 that after the Poly I:C virus acts on pregnant mice, the inhibition rate of the offspring to PPI is reduced, indicating that the offspring rat model of schizophrenia was successfully prepared, which can be used for the verification of schizophrenia detection in subsequent examples .

Example 2.2 The present invention is applied to the detection of schizophrenia

Step 1, total RNA extraction. The blood, hippocampus tissue or prefrontal lobe tissue of the offspring of the normal rat group and the model rat group were collected, and the total mRNA was extracted directly using the micro tissue cell RNA extraction kit (RaPure Total RNA Micro Kit). The extraction protocol was as follows: Instructions require it. Use NanoDrop ND-2000 to detect the absorbance of total RNA at 260nm to determine the RNA concentration for use in subsequent steps.

Step two, reverse transcription. Using the NovoScript Plus All-in-one ^1st Strand cDNA Synthesis Super Mix (gDNAPurge) kit, the total mRNA was reverse-transcribed into cDNA, and the reverse transcription method can be operated according to the instructions. After the reverse transcription is completed, the sample to be tested is ready.

Step 3: Prepare PCR mixed system, 54 μL in total. Among them, 15 μL of IxPCR buffer solution, 28 μL of dNTP mixture (among them, 4 μL each of dATP, dGTP, dTTP, dCTP, dUTP, _MgCl and SYBRGreen1), 2 μL of TaqDNA polymerase, 4 μL of primer set, and 5 μL of the sample to be tested. Among them, BDNF primer set, β-NGF primer set, CD133 primer set and D2 primer set, all four primer sets are 1 μL, and the upstream primer and downstream primer of each primer set are 0.5 μL. In order not to affect the results, the above operations were performed on ice. Wherein, the nucleotide sequences of the four primer sets correspond to SEQ ID No.1-SEQ ID No.8.

Step 4, QPCR reaction, performing PCR amplification on BDNF, β-NGF, CD133 and D2. The specific steps are as follows: first cycle: preheating at 95°C for 30 s; second cycle (40 cycles): denaturation at 95°C for 30 s, annealing at 54°C for 30 s, extension at 72°C for 45 s; third cycle: followed by melting curve. Among them, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control for normalization. The relative amount of PCR fragments was calculated using the comparative CT method, and the results are shown in FIG. 8 . Comparing CT methods is common knowledge in this field and will not be repeated here.

Fig. 8 shows the relative content figure of four kinds of mRNAs of normal rat group offspring and model rat group offspring, wherein (N+N) is normal rat group offspring, (P+N) is model rat group pups. As can be seen from Figure 8, compared with the test group (P+N) in the control group (N+N), the relative contents of the four mRNAs of BDNF, β-NGF, CD133 and D2 are significantly different, confirming that the primer pair of the present invention or The kit can distinguish normal rats from schizophrenic rats, that is, it can be applied to the detection of schizophrenia.

Example 3 This example relates to the application of the kit to the evaluation of drug efficacy for schizophrenia

Before starting this example, it is first necessary to construct an animal model. The method for constructing the animal model is the same as that in Example 2.1, and will not be repeated here. In this example, only offspring from the model rat group were used. The offspring of the model rat group were equally divided into two groups, one group was the control group (MIA+Saline), and the other group was the treatment group (MIA+MSCs). The treatment group was given tail vein injection of umbilical cord mesenchymal stem cell suspension ( ^MSCs ). Before preparing the umbilical cord mesenchymal stem cell suspension, it is necessary to weigh the weight of the offspring, and prepare the umbilical cord mesenchymal stem cell suspension according to the weight of the offspring. For example, if the weight of the offspring is 300g, the number of cells of the umbilical cord mesenchymal stem cells is 1.2*10 ⁷ . Rats in the control group were injected with sterile saline (Saline) into the tail vein, and the injection volume was 500 μL. At present, it has been experimentally confirmed that umbilical cord mesenchymal stem cells have a therapeutic effect on schizophrenic mice (see the patent application "umbilical cord mesenchymal stem cells, umbilical cord mesenchymal stem cell preparations and their applications", application number 202211452248.2). The preparation method of the umbilical cord mesenchymal stem cell suspension (MSCs) used in this example is detailed in the patent application "Umbilical cord mesenchymal stem cells, umbilical cord mesenchymal stem cell preparations and their application", the application number is 202211452248.2.

Both the control group and the treatment group were injected for 5 consecutive weeks, once a week, 5 injections in total. One week after the completion of the fifth injection, the relative mRNA levels of the two groups were detected. The detection steps are as Step 1 to Step 4 of Example 2.2, and will not be repeated here. The test results are shown in Figure 9. Fig. 9 is the relative content of the four kinds of mRNA in the offspring of the control group and the treatment group. It can be seen from Figure 9 that after injection of umbilical cord mesenchymal stem cells, the expression of BDNF molecules in the treatment group was increased, the expression of β-NGF molecules was slightly decreased, the expression of CD133 molecules was significantly decreased, and the expression of D2 molecules was increased. It can be known that the primer pair or kit of the present invention can be used for evaluating the curative effect of schizophrenia drug treatment.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of the present invention. within the scope of protection.

Claims (10)

1. A primer set for schizophrenia detection, characterized in that: said primer set includes a BDNF primer set, a β-NGF primer set, a CD133 primer set and a D2 primer set; The BDNF primer set includes a BDNF upstream primer and a BDNF downstream primer; the nucleotide sequence of the BDNF upstream primer is shown in SEQ ID No.1, and the nucleotide sequence of the BDNF downstream primer is shown in SEQ ID No.2 Show; The β-NGF primer set includes β-NGF upstream primers and β-NGF downstream primers; the nucleotide sequence of the β-NGF upstream primers is shown in SEQ ID No.3, and the core of the β-NGF downstream primers The nucleotide sequence is shown in SEQ ID No.4; The CD133 primer set includes a CD133 upstream primer and a CD133 downstream primer; the nucleotide sequence of the CD133 upstream primer is shown in SEQ ID No.5, and the nucleotide sequence of the CD133 downstream primer is shown in SEQ ID No.6. Show; The D2 primer set includes a D2 upstream primer and a D2 downstream primer; the nucleotide sequence of the D2 upstream primer is as shown in SEQ ID No.7, and the nucleotide sequence of the D2 downstream primer is as shown in SEQ ID No.8 . 2. The primer set according to claim 1, characterized in that: the concentration of the primer set includes one or more of the following (i) to (viii): (i) the concentration of the BDNF upstream primer is 5-15 μM; (ii) the concentration of the BDNF downstream primer is 5-15 μM; (iii) the concentration of the β-NGF upstream primer is 5-15 μM; (iv) the concentration of the β-NGF downstream primer is 5 ~15 μM; (v) the concentration of the CD133 upstream primer is 5 ~ 15 μM; (vi) the concentration of the CD133 downstream primer is 5 ~ 15 μM; (vii) the concentration of the D2 upstream primer is 5 ~ 15 μM; (viii ) The concentration of the D2 downstream primer is 5-15 μM. 3. A kit for detecting schizophrenia, characterized in that it comprises the primer set according to any one of claims 1 or 2. 4. The kit according to claim 3, characterized in that: the kit also includes a buffer, a dNTP mixture, and TaqDNA polymerase. 5. The kit according to claim 4, characterized in that: the buffer comprises Tris·HCL; preferably, the concentration of the Tris·HCL is 5-15 mM. 6. The kit according to claim 4, wherein the dNTP mixture comprises dATP, dGTP, dTTP, dCTP, dUTP, _MgCl2 and SYBRGreen1. 7. The kit according to claim 6, characterized in that: the concentration of the dATP is 0.2-0.8 mM; and/or the concentration of the dGTP is 0.2-0.8 mM; and/or the concentration of the dTTP is 0.2-0.8mM; and/or the concentration of dCTP is 0.2-0.8mM; and/or the concentration of dUTP is 0.2-0.8mM; and/or the concentration of MgCl ₂ is 3-7mM; and/or The volume percent concentration of the SYBRGreen1 is 0.025%. 8. The kit according to claim 4, characterized in that: the concentration of the TaqDNA polymerase is 3-7 U/μL. 9. A detection method for schizophrenia, comprising the steps of: S1: extract the total RNA of the sample; S2: Reverse transcribing the total RNA into cDNA to obtain a sample to be detected; S3: using the primer set described in any one of claims 1-2 or the kit described in any one of claims 3-8, to detect BDNF, β-NGF, CD133 and D2 in the sample to be detected Perform PCR amplification to obtain the expression levels of BDNF, β-NGF, CD133 and D2. 10. The detection method according to claim 9, characterized in that: the sample originates from blood, hippocampal tissue or prefrontal cortex tissue.

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