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Group of markers, detection model and application thereof in radiation dose detection

A technology of radiation dose and detection model, which is applied in the measurement/inspection of microorganisms, bioinformatics, recombinant DNA technology, etc., can solve the problems of short window period, complex analysis operation of γ-H2AXfoci, and low detection throughput, and achieve expression Stable level, saving preparation time, rapid detection effect

Inactive Publication Date: 2022-04-19
ACADEMY OF MILITARY MEDICAL SCI
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Problems solved by technology

However, the window period for the change of γ-H2AX foci is very short, it appears a few minutes after irradiation, reaches a peak in about 1 hour, and then gradually decreases, and the analysis operation of γ-H2AX foci is relatively complicated, and the detection throughput is not high, so its application has been widely accepted. Large limitations (Reitsema TJ, Banath JP, MacPhail SH, Olive PL. Hypertonicsaline enhances expression of phosphorylated histone H2AX afterirradiation. Radiat Res. 2004; 161(4):402-8.) (Andrievski A, Wilkins RC. Theresponse of gamma-H2AX in Human lymphocytes and lymphocyte subsets measured in whole blood cultures. Int J Radiat Biol. 2009; 85(4): 369-76.) (Wang Z, Hu H, HuM, Zhang X, Wang Q, Qiao Y, et al. Ratio of gamma-H2AX level in lymphocytes to that in granulocytes detected using flow cytometry as a potential biodosimeter for radiation exposure. Radiat Environ Biophys. 2014; 53(2):283-90.)

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  • Group of markers, detection model and application thereof in radiation dose detection
  • Group of markers, detection model and application thereof in radiation dose detection
  • Group of markers, detection model and application thereof in radiation dose detection

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Screening of markers for detection of acute radiation

[0039] The peripheral blood of 3 healthy adults was irradiated with doses of 0, 0.75Gy, 2Gy and 6Gy for 2 hours, and the NimbleGen genome-wide chip data of the samples were analyzed in depth, and the expression differences among the 3 samples in the same dose group were all greater than 2 times of genes as differentially expressed genes at each dose (such as figure 1 ), the only genes with significant differences in the three samples at 0.75Gy dose were AEN, BBC3, TRAF4 and TRIAP1, and these four genes also had significant differences in the three samples in the 2Gy and 6Gy dose groups at the same time. The numbers of differentially expressed genes at 2Gy and 6Gy were 37 and 36, respectively, among which there were 19 differentially expressed genes (see figure 1 in A). Then we performed functional annotation analysis on the differential expression of each dose, and the results showed that 0.75Gy and ot...

Embodiment 2

[0040] Example 2: Application of Markers in Detecting Radiation Dose

[0041] (1) Irradiation and cultivation of samples

[0042] External irradiation of human peripheral blood: the collected whole blood samples of human peripheral blood were placed in a 37°C water bath to simulate the internal environment. 60 It was irradiated externally with a Coγ radiation source at a dose rate of 1.14Gy / min. The experiment was divided into 7 groups of 0Gy, 0.5Gy, 1Gy, 2Gy, 4Gy, 6Gy and 10Gy. The irradiated samples continued to be cultured in a 37°C incubator, and samples were collected at 0h, 2h, 4h, 8h, 12h, and 24h after irradiation.

[0043] (2) Primer and probe synthesis

[0044] According to the gene information retrieved from GenBank, Shanghai Gemma Company designed and synthesized primer pairs and probes for each gene. The specific sequences are shown in Table 1.

[0045] Table 1 The sequences of primer pairs and probes for each gene

[0046]

[0047] (3) RNA extraction qRT-...

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Abstract

The invention discloses a group of markers, a detection model and application thereof in radiation dose detection, and belongs to the technical field of biological detection. In order to solve the problems existing in an existing radiation biological dose estimation method, based on background levels of DDB2, AEN, TRIAP1 and TRAF4 and sensitivity specificity in the early stage of radiation, a guide threshold value is given for detection of each gene at different detection time points, finally, the four genes are combined according to different change modes of the four genes in the early stage of radiation, and the radiation biological dose estimation method based on DDB2, AEN, TRIAP1 and TRAF4 is obtained. And an effective radiation early dose detection model is constructed. The model can be used for classification and dose estimation of large-batch nuclear radiation irradiated personnel within 24 hours after exposure, and a new thought and method are provided for corresponding scientific research of radiation dose.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a group of markers, a detection model and their application in detecting radiation dose. Background technique [0002] At present, the methods for estimating radiation biological dose mainly include lymphocyte chromosomal aberration analysis, lymphocyte count analysis, lymphocyte γ-H2AX analysis, etc. Among them, the lymphocyte chromosome aberration analysis method is considered to be the "gold standard" for radiation biological dose estimation. This method has good specificity and high accuracy, but its experimental method has high requirements for operators, and requires Cells must be cultured for 48 hours before sample preparation and analysis, and the upper dose limit is usually 5-6Gy (Abe Y, Yoshida MA, Fujioka K, KurosuY, Ujiie R, YanagiA, et al.Dose-response curves for analyzing of dicentric chromosomes and chromosome translocations following dose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6851C12N15/11G16B25/10G16B25/20G16B30/00
CPCC12Q1/6883C12Q1/6851G16B25/10G16B25/20G16B30/00C12Q2600/158C12Q2600/166C12Q2531/113C12Q2521/107C12Q2537/165C12Q2561/101C12Q2545/101
Inventor 王治东李亚琼王琪戚振华厉卫红周仕香
Owner ACADEMY OF MILITARY MEDICAL SCI
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