Biomarker for rapidly evaluating radiation exposure dose of human body and application of biomarker

Abstract

The invention discloses a biomarker for rapidly evaluating the radiation exposure dose of a human body and application of the biomarker. The biomarker is zinc finger matrix protein type protein 3 (ZMAT3). Compared with an existing biological dose estimation method, the method has the advantages of being simple, convenient, rapid, accurate and high in throughput, and is particularly suitable for radiation biological dose estimation of large population samples under nuclear and radiation accidents.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to zinc finger matrin-type protein 3 (ZMAT3) as a biomarker for rapidly evaluating the radiation exposure dose of a human body and its applications. Background Art

[0002] With the wide application of nuclear and ionizing radiation, nuclear and radiation accidents occur from time to time. In particular, nuclear terrorism and radioactive terrorism remain one of the most severe challenges faced by international security, and this threat is constantly evolving. In this context, it is very likely that sudden accidents involving large populations being irradiated will occur. Determining the irradiated dose is required for the harm of ionizing radiation to human health, radiation safety assessment, and the classification, diagnosis, and treatment of casualties with acute radiation injury. Since the direct or indirect physical dose monitoring devices in sudden radioactive accidents cannot provide timely and effective records of the irradiated situation, it directly affects the medical classification and treatment of irradiated personnel. Therefore, establishing a rapid, accurate, and high-throughput biodosimeter has important scientific and practical significance.

[0003] Currently, chromosome aberration analysis widely used for estimating radiation biodosimetry is difficult to meet the requirements of rapid dose estimation for large-scale population emergencies due to disadvantages such as a long test cycle, large workload, and high requirements for analysis techniques. The cytokinesis-block micronucleus assay in binucleated cells is also a commonly used method for dose estimation of accident-irradiated personnel. Although this method is simple to operate, has low requirements for analysts' techniques, and can achieve automated analysis, it is limited in practical applications due to many influencing factors of micronuclei and a relatively high background value. The above actual needs and technical status have prompted the domestic and international radiobiological dosimetry communities to explore a stable, rapid, simple, and high-throughput radiation biodosimeter to meet the needs of dose estimation in large-scale population emergencies. Summary of the Invention

[0004] Analysis of gene expression profiles induced by ionizing radiation shows good application prospects in molecular mechanism research, the establishment of biodosimeters, and the individualized treatment of cancer. It has been found that ionizing radiation can induce changes in the expression levels of certain genes, showing a certain dose-effect relationship and time variation pattern. Human peripheral blood lymphocytes are sensitive to radiation and are easily obtained. They are an ideal model for studying radiation biodosimeters. The gene expression level can be measured in batches through techniques such as real-time fluorescence quantitative PCR and gene chips, which is very suitable for rapid and high-throughput dose estimation under nuclear and radiation accidents. The purpose of the present invention is to provide a simple, rapid, accurate, and high-throughput detection method for evaluating the ionizing radiation dose (exposure level / dose) based on the analysis of ZMAT3 gene expression.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A biomarker for rapidly evaluating the radiation exposure dose of a human body, characterized in that the biomarker is zinc finger matrix protein type 3 (ZMAT3).

[0007] According to an embodiment of the present invention, wherein the radiation is X-ray, γ-ray and / or neutron radiation.

[0008] Use of zinc finger matrix protein type 3 (ZMAT3) as a biomarker for rapidly evaluating the radiation exposure dose of a human body.

[0009] A reagent or kit for rapidly evaluating the radiation exposure dose of a human body, characterized in that the reagent or kit can detect the biomarker according to the present invention.

[0010] A detection method for rapidly evaluating the radiation exposure dose of a human body, characterized in that: by detecting the expression level of the ZMAT3 gene in the peripheral blood outside the body after radiation exposure of the human body, and using the linear relationship between the expression level of the ZMAT3 gene and the radiation exposure dose to calculate the radiation dose.

[0011] According to an embodiment of the present invention, wherein the detection method comprises the steps of:

[0012] (1) Drawing peripheral blood of a human body for non-therapeutic purposes and extracting total RNA therefrom;

[0013] (2) Synthesizing cDNA and performing real-time quantitative PCR detection to detect the expression level of the ZMAT3 gene;

[0014] (3) Using the linear relationship between the expression level of the ZMAT3 gene and the radiation exposure dose to determine the radiation exposure dose.

[0015] According to an embodiment of the present invention, wherein the detection method includes using β-actin and B2M as internal reference genes.

[0016] According to an embodiment of the present invention, wherein the detection method can be completed within 6 to 8 hours after blood sample collection.

[0017] By detecting the changes in the expression level of the ZMAT3 gene in human immortalized lymphocyte cell lines and ex vivo human peripheral blood after exposure to ionizing radiation, the present invention determines that it has a good dose-effect relationship and time response range to radiation. Further, a system for detecting the expression level of the ZMAT3 gene is established by real-time fluorescence quantitative PCR method; and its use for rapidly evaluating the radiation exposure level of a subject to be tested and estimating the irradiated dose is verified. Compared with the existing biological dose estimation methods, this method can complete the preliminary dose classification and estimation within 6-8 hours after blood sample collection, and has the advantages of simplicity, rapidity, accuracy, and high throughput, and is particularly suitable for the radiation biological dose estimation of large population samples under nuclear and radiation accidents. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 For 0-8 Gy according to the embodiment of the present invention 60 The dose-effect relationship of the ZMAT3 gene in the AHH-1 cell line irradiated with Coγ rays at 4-48 h;

[0019] Figure 2 For 60 The fitting curve of the relative expression level of ZMAT3 in the AHH-1 cell line irradiated with Coγ rays at 4-48 h and the irradiation dose;

[0020] Figure 3 For 0-8 Gy 60 The dose-effect relationship of the ZMAT3 gene in ex vivo human peripheral blood irradiated with Coγ rays at 4-48 h;

[0021] Figure 4 For 60 The fitting curve of the relative expression level of ZMAT3 in ex vivo human peripheral blood irradiated with Coγ rays at 4-48 h and the irradiation dose;

[0022] Figure 5 For different dose rates 60 The influence of Coγ ray irradiation on the change of the expression level of the ZMAT3 gene in human peripheral blood; and

[0023] Figure 6 For the comparison between the biological dose estimated based on the ZMAT3 gene expression analysis and the true irradiation dose. DETAILED DESCRIPTION OF THE INVENTION

[0024] The following will describe in detail the applications of multiple ZMAT3s provided by the present invention as radiation biomarkers. Unless otherwise specified, the reagents and culture media used in the following examples are commercially available products, and operations such as nucleic acid electrophoresis and Real-time PCR are carried out according to conventional protocols. For example, they can be carried out as described in Molecular Cloning (Cold Spring Harbor Laboratory Press (1989)). The following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention.

[0025] Example 1: 60 Dose-effect relationship of Coγ-ray-induced ZMAT3 gene expression in AHH-1 cell line

[0026] 1) Primer design and synthesis: According to the gene information in the GeneBank database, exon-spanning primer sequences of the ZMAT3 gene and two internal reference genes (β-actin and B2M) were designed and synthesized by Shanghai Sangon Biotech Co., Ltd. The primer sequences are shown in Table 1.

[0027] Table 1. Primer sequences

[0028]

[0029] 2) Cell culture and irradiation: The immortalized normal human lymphocyte cell line (AHH-1) growing in suspension was inoculated into RPMI 1640 medium containing 10% fetal bovine serum and cultured in a constant temperature incubator at 37°C and 5% CO2. The AHH-1 cell line was irradiated with Coγ-rays at a dose rate of 1 Gy / min 60 at dose points of 0, 1, 2, 4, 6, and 8 Gy, and harvested after incubation at 37°C for 4, 12, 24, and 48 h after irradiation.

[0030] 3) Total RNA extraction and quantification of cells: Collect the cells, centrifuge at 1500 rpm for 5 min, discard the supernatant, and wash the cells 3 times with 1×PBS; Add 1 ml of TRIzol to the cell pellet, pipette repeatedly to lyse all the cells, then transfer it to a 1.5 ml centrifuge tube and let it stand at room temperature for 40 min until the cells are fully lysed; Add 200 μl of chloroform to the 1.5 ml centrifuge tube, mix well, let it stand at room temperature for 15 min, centrifuge at 12000 rpm at 4°C for 20 min; Aspirate the upper aqueous phase into another centrifuge tube; Add 500 μl of isopropanol to the centrifuge tube, gently invert to mix, let it stand at room temperature for 20 min, centrifuge at 12000 rpm at 4°C for 20 min; Carefully remove the supernatant to prevent RNA loss; Wash once with 1 ml of 75% ethanol (prepared with DEPC water), centrifuge at 12000 rpm at 4°C for 10 min; Add another 700 μl of ethanol to suspend the RNA pellet and repeat the washing once; Centrifuge at 8000 rpm at room temperature for 10 min; Aspirate the supernatant as thoroughly as possible to prevent loss of the RNA pellet, invert the EP tube on the absorbent paper and let it dry at room temperature for 30 min; Add 30 μl of DEPC water to dissolve the RNA pellet. Use a UV spectrophotometer to detect the OD values of the nucleic acid sample to be measured at wavelengths of 230 nm, 260 nm, and 280 nm. Generally, it is considered that when the ratios of OD260 / OD230 and OD260 / OD280 are both 1.8 - 2.0, the RNA purity and concentration of the sample to be measured meet the experimental requirements.

[0031] 4) cDNA synthesis of cells, the experimental conditions are as follows:

[0032]

[0033] Reaction conditions: Incubate at a constant temperature of 68°C for 5 min, then let it stand on ice for about 5 min;

[0034]

[0035] 48°C for 90 min, 65°C for 5 min, terminate the reaction at 4°C, and store at -20°C.

[0036] 5) Real-time quantitative PCR detection: The reaction system is 20 μl: 10 μl of 2×SYBR green I, 2 μl of cDNA template, a total of 0.5 μl of upstream and downstream primers, and 7.5 μl of double-distilled water. Reaction conditions: After pre-denaturation at 95°C for 10 min, perform the following 40 cycles: Denaturation at 95°C for 15 s, annealing and extension at 60°C for 1 min. Set 3 parallel samples for each sample, perform melting curve analysis after each real-time quantitative PCR reaction to exclude the influence of primer dimers, and at the same time set up a primer-free blank control group, and repeat the experiment 3 times.

[0037] 6) PCR detection data analysis: The results were analyzed using Applied Biosystems 7500 Sequence Detection Software (SDS). The 2-ΔΔCt method was adopted, with β-actin and B2M as internal reference genes, and the relative expression levels of each gene at 4 time points and 6 dose points were relatively quantified. Among them, the internal reference genes

[0038] 7) Statistical analysis: SPSS 25.0 software was used for data analysis, and the relative gene expression levels were expressed as . The comparison between groups was in line with the normal distribution after normality test. One-way ANOVA was used for two-sided test (significance level α = 0.05); the dose-effect curves at each post-irradiation time point were fitted using the linear regression method.

[0039] The results are shown in Figure 1 and 2 (Note: * compared with the control group: *P < 0.05, 0.01, 0.001). It can be seen from Figure 1 that after the AHH-1 cell line was irradiated with γ-rays, compared with the non-irradiated group, the relative expression levels of the ZMAT3 gene at different irradiation time points (4, 12, 24, and 48 h) and different irradiation dose points (1, 2, 4, 6, and 8 Gy) showed an obvious trend of increasing with the increase of dose (P < 0.05, 0.01, 0.001). Among them, in the 8 Gy dose group at 12 - 48 h after irradiation, the relative expression level of the ZMAT3 gene was 3.5 times that of the non-irradiated group( Figure 1 ). The above results indicate that by detecting the change in the expression level of the ZMAT3 gene in cells, a preliminary dose classification can be carried out on whether the test sample has been irradiated with γ-rays. According to the dose-effect relationship of the ZMAT3 gene expression level at each time point, the fitted curve equation is as shown in Figure 2 . Among them, the fitted curve equation at 4 h after irradiation is: y = 0.152x + 1.465, R 2 = 0.707; the fitted curve equation at 12 h after irradiation is: y = 0.246x + 1.257, R 2 = 0.900; the fitted curve equation at 24 h after irradiation is: y = 0.340x + 1.373, R 2 = 0.892; the fitted curve equation at 48 h after irradiation is: y = 0.364x + 1.403, R 2 = 0.880. By substituting the relative expression level of ZMAT3 into the fitted curve equation at each time point, the received radiation dose can be estimated( Figure 2 ).

[0040] Example 2: 60Dose - effect relationship of Coγ - ray in vitro irradiation on ZMAT3 gene expression in human peripheral blood

[0041] 1) Blood sample collection and irradiation conditions: On the premise of signing the informed consent form, peripheral blood of 8 healthy people aged 20 - 30 years old, with equal numbers of males and females, no recent history of acute disease infection, no history of chronic diseases such as hypertension and diabetes, no smoking history, no history of chemical toxicant exposure, and no history of radiation exposure in the past six months was collected. Each person collected 36 ml of venous blood samples at one time, which were evenly divided and stored in 6 heparin - lithium - anticoagulated vacuum blood collection tubes of 6 ml for standby. Coγ irradiation was carried out at room temperature, with a dose rate of 1 Gy / min, and the dose points were 0, 1, 2, 4, 6, and 8 Gy respectively. After irradiation, the blood samples at each dose point were evenly inoculated into four RPMI 1640 culture flasks containing 8 ml of 10% fetal bovine serum (about 1.5 ml of whole blood in each flask), and cultured in a 37℃ constant - temperature incubator for 4 h, 12 h, 24 h, and 48 h. 60 Coγ irradiation was carried out at room temperature, with a dose rate of 1 Gy / min, and the dose points were 0, 1, 2, 4, 6, and 8 Gy respectively. After irradiation, the blood samples at each dose point were evenly inoculated into four RPMI 1640 culture flasks containing 8 ml of 10% fetal bovine serum (about 1.5 ml of whole blood in each flask), and cultured in a 37℃ constant - temperature incubator for 4 h, 12 h, 24 h, and 48 h.

[0042] 2) Extraction of total RNA from human peripheral blood cells: The total RNA of human peripheral blood at each time point and each dose point was extracted using the RNAprep Pure Blood Total RNA Extraction Kit. The method is as follows:

[0043] (1) Preparation of deoxyribonuclease stock solution: Dissolve deoxyribonuclease with 550 μl of RNase - free double - distilled water, mix well by inverting up and down, and after aliquoting, store it at - 20℃.

[0044] (2) Add 10 ml of red blood cell lysate to 2 ml of human peripheral blood, mix well by suspension oscillation to fully mix the cells, and let it stand on ice for 20 min. During this period, mix well by oscillation every 5 min until the solution becomes red and transparent, indicating that the red blood cells have been fully lysed. Centrifuge at 4℃ for 15 min at a speed of 2100 rpm, and then discard the supernatant.

[0045] (3) Add 5 ml of red blood cell lysate to the centrifuge tube, mix well by suspension oscillation to fully resuspend the white blood cell pellet at the bottom.

[0046] (4) Centrifuge at 4℃ for 15 min at a speed of 2100 rpm, and then discard the supernatant. Then add about 600 μl of RLH lysate containing β - mercaptoethanol to the centrifuge tube to fully lyse the white blood cells.

[0047] (5) Transfer the lysed solution to the CS column, about 600 μl, and then centrifuge at 4℃ for 2 min at a speed of 12000 rpm. Retain the filtrate and discard the CS column.

[0048] (6) Dilute absolute ethanol to 70% with DEPC water, then add approximately 600 μl of 70% ethanol to the collection tube containing the filtrate, and mix well with a pipette tip. At this time, flocculent precipitates can be seen. Transfer the solution to the CR4 column in two portions, approximately 600 μl each time. Centrifuge at 12,000 rpm for 2 min at 4°C, retain the filter column, and discard the filtrate.

[0049] (7) Deoxyribonuclease digestion: Add 350 μl of protein removal solution RW1H to the CR4 column, centrifuge at 12,000 rpm for 2 min at 4°C, and discard the filtrate; Take out 10 μl from the prepared deoxyribonuclease stock solution, then add 70 μl of RDD solution, mix well, and add all of it to the CR4 column. Let it stand at room temperature for about 30 min. Then add 350 μl of protein removal solution RW1H to the CR4 column, centrifuge at 12,000 rpm for 2 min at 4°C, retain the filter column, and discard the filtrate.

[0050] (8) Wash the CR4 column with 500 μl of wash solution RW. After adding RW, let it stand at room temperature for 2 min, then centrifuge at 12,000 rpm for 2 min at 4°C, retain the filter column, and discard the filtrate. Repeat the washing once using the same method.

[0051] (9) To ensure no residual liquid, centrifuge the CR4 column again (centrifuge at 12,000 rpm for 2 min at 4°C). Then open the column cap in the operating table and let it stand at room temperature for 20 min to allow the ethanol to fully volatilize.

[0052] (10) Prepare a brand-new RNase-free centrifuge tube, place the CR4 column in the centrifuge tube, drop 30 μl of DEPC water into the center of the column, cover the lid, let it stand at room temperature for 5 min, then centrifuge at 12,000 rpm for 2 min at 4°C, collect the filtrate, which is the RNA solution, and store it at -70°C.

[0053] 3) cDNA synthesis: Use a high-capacity reverse transcription kit to synthesize cDNA. The advantage of this kit is that it only requires a relatively small amount of RNA to synthesize a relatively high-yield cDNA. The reaction system is as follows:

[0054]

[0055] Reaction conditions: After 25°C for 10 min, 37°C for 120 min, 85°C for 5 min, terminate the reaction at 4°C and store it in a -20°C refrigerator.

[0056] 4) Real-time quantitative PCR detection: The reaction system was 20 μl: 10 μl of 2×SYBR green I, 1 μl of cDNA template, a total of 0.5 μl of upstream and downstream primers, and 7.5 μl of double-distilled water. Reaction conditions: After pre-denaturation at 95°C for 10 min, the following 40 cycles were carried out: denaturation at 95°C for 15 s, annealing and extension at 60°C for a total of 1 min. Three parallel samples were set for each sample. After each real-time quantitative PCR reaction, melting curve analysis was performed to exclude the influence of primer dimers. At the same time, a primer-free blank control group was established, and the experiment was repeated 3 times.

[0057] 5) PCR data processing: Applied Biosystems 7500 Sequence Detection Software (SDS) was used for result analysis. Using the 2 -ΔΔCt -method, with β-actin and B2M as internal reference genes, the relative expression change levels of each gene at 4 time points and 6 dose points were relatively quantified. Among them, the internal reference genes

[0058] 6) Statistical analysis: SPSS 25.0 software was used for data analysis, and the relative gene expression was expressed as . Comparison between groups was in line with normal distribution after normality test. One-way ANOVA was used for two-sided test (significance level α = 0.05); the linear regression method was used to fit the dose-effect curve at each post-irradiation time point.

[0059] The results are shown in Appendix Figure 3 and 4 (Note: *Compared with the control group: *P < 0.05, 0.01, 0.001). It can be seen from Figure 3 that after in vitro irradiation of human peripheral blood with 0-8 Gy 60 Coγ-rays for 4-48 h, compared with the non-irradiated group, the relative expression level of ZMAT3 showed a dose-dependent increase at all four time points (P < 0.05, 0.01, 0.001) ( Figure 3 ), and was consistent with the change trend at the cellular level, indicating that the change in the expression level of the ZMAT3 gene induced by radiation has met the necessary conditions to be used as a radiation biomarker. According to the dose-effect relationship of the ZMAT3 gene in human peripheral blood, the curve equations fitted at the four time points showed a linear square model, and the R 2 values were all higher than 0.90 ( Figure 4 ). Among them, the curve equation fitted at 4 h after irradiation was: y = -0.042x 2 + 0.617x + 1.234, R 2 = 0.957; the curve equation fitted at 12 h after irradiation was: y = -0.087x 2 + 1.116x + 1.494, R 2=0.930; the curve equation fitted 24 hours after illumination is: y = -0.090x 2 +1.312x+1.353, R 2 =0.978; the curve equation fitted 48h after illumination is: y = -0.080x 2 +1.047x+1.427, R 2 =0.957. By substituting the relative expression value of ZMAT3 into the above fitting curve equation, the radiation dose received can be estimated, which is helpful to evaluate the radiation exposure level of the exposed personnel in the radiation accident.

[0060] Example 3. Different dose rates 60 Effects of Coγ-ray irradiation on the expression level of ZMAT3 gene in human peripheral blood

[0061] 1) Blood sample collection: Blood samples were collected from three healthy subjects aged 20 to 30 years old, all female, with no history of smoking or drinking, no history of taking medication within six months, and no history of imaging examinations, with informed consent. 24 ml of peripheral blood was collected at a time, and lithium heparin was used for anticoagulation. Each blood sample was divided into 15 equal portions, each of which was about 1.5 ml.

[0062] 2) Blood sample irradiation and culture: Application 60 The blood samples were irradiated with a Coγ-ray source, with an average irradiation field of 30 cm × 30 cm, an absorbed dose rate of 0.2 Gy / min, 1 Gy / min and 2 Gy / min, and irradiation doses of 0, 1, 2, 4 and 6 Gy. After irradiation, the blood samples were inoculated in RPMI 1640 medium containing 10% fetal bovine serum and cultured for 24 hours at 37°C and 5% CO2 saturated humidity.

[0063] 3) The extraction of total blood RNA, synthesis of cDNA, PCR detection and data analysis were carried out in the same manner as in Example 2.

[0064] Results Figure 5 .Depend on Figure 5 It can be seen that different dose rates 60 After Coγ-ray irradiation, the relative expression of ZMAT3 gene induced by radiation increased with the increase of irradiation dose, which was significantly dose-dependent, consistent with the results in Example 2; in addition, at the same dose point (1, 2, 4 and 6Gy), there was no statistically significant difference between the relative expression of genes induced by radiation at different dose rates (P>0.05), indicating that within the dose rate range of 0.1-2Gy / min, the expression level of ZMAT3 was not affected by the dose rate. This result further confirms that the use of changes in the expression level of ZMAT3 gene to estimate radiation biological dose has a good application prospect.

[0065] Example 4. Validation of human peripheral blood ZMAT3 gene expression analysis for estimating biological dose

[0066] 1) Blood sample collection and irradiation conditions: Under the condition of signing informed consent, peripheral blood was collected from 5 healthy subjects, including 3 males and 2 females, aged 20-30 years old. 4 ml of venous blood was collected from each person and evenly divided into two 2 ml heparin lithium anticoagulation vacuum blood collection tubes for standby use. 60 Coγ irradiation, dose rate 1Gy / min, dose points 0.5 and 3Gy. After irradiation, 2ml of blood sample at each dose point was inoculated into RPMI 1640 culture bottle containing 8ml 10% fetal bovine serum and placed in a 37℃ constant temperature incubator for 24h.

[0067] 2) The extraction of total blood RNA, synthesis of cDNA, PCR detection and data analysis were carried out in the same manner as in Example 2.

[0068] The results are shown in the attached Figure 6 .Depend on Figure 6 It can be seen that the dose-effect curve fitting equation established in Example 2 was used to estimate the dose of human peripheral blood samples irradiated in vitro at two dose points (0.5Gy and 3Gy). The results showed that the average estimated dose of the 0.5Gy irradiation group was 0.50±0.11Gy; the average estimated dose of the 3Gy irradiation group was 3.20±0.74Gy, and the relative deviation of the two dose points (|estimated dose-actual irradiation dose| / actual irradiation dose) was <10%, indicating that the detection method based on ZMAT3 gene expression analysis established in this study can quickly and accurately estimate the radiation dose, and has good practical application value.

[0069] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A biomarker for rapid assessment of human radiation exposure dose, characterized in that: The biomarker is zinc finger matrix type protein 3 (ZMAT3).

2. The biomarker according to claim 1, characterized in that: The radiation is X-rays, gamma rays and / or neutron radiation.

3. Application of zinc finger matrix metalloproteinase-type 3 (ZMAT3) as a biomarker for rapid assessment of human radiation exposure dose.

4. A reagent or kit for rapidly assessing human radiation exposure dose, characterized in that: The reagent or kit can detect the biomarker according to claim 1 or 2.

5. A detection method for rapidly evaluating human radiation exposure dose, characterized in that: The radiation dose was calculated by detecting the expression level of ZMAT3 gene in the peripheral blood of human body after radiation exposure and using the linear relationship between ZMAT3 gene expression and radiation exposure dose.

6. The detection method according to claim 5, characterized in that: Included steps (1) Draw peripheral blood from the human body for non-therapeutic purposes and extract total RNA from it; (2) cDNA synthesis and real-time quantitative PCR to detect the expression of ZMAT3 gene; (3) The radiation exposure dose was determined by using the linear relationship between ZMAT3 gene expression and radiation exposure dose.

7. The detection method according to claim 5, characterized in that: Including β-actin and B2M as internal reference genes.

8. The detection method according to claim 5, characterized in that: The detection method can be completed within 6 to 8 hours after blood sample collection.

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