Application of radiation sensitive genes as radiological dosimeter

A radiation-sensitive and radiation-dose technology, applied in radiation dose estimation, in the field of radiation biological dosimeters, can solve the problems of low accuracy, high analysis technology requirements, and long time-consuming, etc., and achieve the effect of high accuracy

Active Publication Date: 2019-09-03
NAT INST FOR RADIOLOGICAL PROTECTION & NUCLEAR SAFETY CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the above problems, the present invention preliminarily screens out hundreds of radiation-responsive genes by detecting the changes in the expression of genes contained in the peripheral blood of normal people after being subjected to ionizing radiation; furthermore, the present invention obtains Further research was carried out on genes that showed a good dose-response relationship after exposure to ionizing radiation and could be plotted for R 2 Higher (R of the fitted curve 2 13 radiosensitive genes (PCNA, CCNG1, CDKN1A, XPC, TNFSF4, DDB2, ASTN2, PHPT1, GDF15, TNFRSF10B, BBC3, BAX and FDXR) of the dose-response curve whose value is more than 0.6); Based on the above-mentioned 13 radiation-sensitive genes, the combination of radiation-sensitive genes used as a radiation biological dosimeter to estimate radiation dose was determined, and the accuracy of estimating radiation dose was confirmed by in vivo and in vitro experiments. In the dose estimation method, there are problems such as high requirements for analysis technology, long time consumption, and low accuracy, thus completing the present invention

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  • Application of radiation sensitive genes as radiological dosimeter
  • Application of radiation sensitive genes as radiological dosimeter
  • Application of radiation sensitive genes as radiological dosimeter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preliminary Screening of Radiation Sensitive Genes

[0032] Using gene chip technology to detect the changes in the expression of genes contained in the peripheral blood of normal people after being exposed to ionizing radiation, and initially screened out hundreds of genes that have radiation responses. Furthermore, the genes obtained from the above preliminary screening were further studied, and a good dose-effect relationship was obtained after exposure to ionizing radiation and R 2 Thirteen radiosensitive genes (ie, PCNA, CCNG1, CDKN1A, XPC, TNFSF4, DDB2, ASTN2, PHPT1, GDF15, TNFRSF10B, BBC3, BAX, and FDXR) with higher dose-response curves.

[0033] The details of the studies performed on the 13 radiosensitivity genes in the above primary screen are as follows:

[0034] 1. Primer design and synthesis

[0035] According to the gene information in the GenBank database, 13 radiation-sensitive genes (PCNA, CCNG1, CDKN1A, XPC, TNFSF4, DDB2, ASTN2, PHPT1, GDF...

Embodiment 2

[0048] Example 2: A combination of radiation-sensitive genes used as a radiation biological dosimeter to estimate radiation dose

[0049] 1. Primer design and synthesis

[0050] The same primers as in Example 1 for 13 radiation-sensitive genes and 2 internal reference genes (as shown in Table 1) were used.

[0051] 2. Blood samples were collected and irradiated in the same manner as in Example 1.

[0052] 3. The extraction of total cellular RNA and the synthesis of cDNA were carried out in the same manner as in Example 1.

[0053] 4. Real-time quantitative PCR detection adopts the same method as in Example 1.

[0054] 5. The PCR data processing adopts the same method as in Example 1.

[0055] 6. Establishment of radiation-sensitive gene combinations

[0056] The basic principle of using the multiple stepwise regression method to establish a radiation-sensitive gene combination is: according to the effect of the independent variable on the dependent variable, the regression e...

Embodiment 3

[0066] Example 3: Verification of radiation-sensitive gene combinations by double-blind in vitro experiments

[0067] 1. Blood sample collection and irradiation

[0068] On the premise of signing the informed consent form, peripheral blood was collected from 10 healthy people, half male and half male, 8ml of venous blood sample was collected from each person, and evenly divided into two 4ml lithium heparin anticoagulated vacuum blood collection tubes for later use, and carried out at room temperature 60 Coγ irradiation (double-blind method), the dose rate was 1Gy / min, and the dose points were 0.8, 1.5, 2.5, 3.5, 4.5 and 5Gy. After irradiation, inoculate 2ml of the blood sample at each dose point into RPMI 1640 culture flasks containing 8ml 10% fetal bovine serum, and place them in a constant temperature incubator at 37°C for 24h; add the other 2ml of whole blood to two flasks containing 4ml RPMI 1640 Shake well in a culture flask containing 20% ​​fetal bovine serum, 100 U / ml ...

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Abstract

The invention relates to an application of a combination of multiple radiation sensitive genes as a radiological dosimeter in radiation dose estimation. Specifically, the invention relates to an application of a combination of 5-13 different radiation-sensitive genes as a radiological dosimeter in radiation dose estimation, a kit and a microarray used for radiation dose estimation and prepared based on the combination of 5-13 different radiation-sensitive genes, and a method for radiation dose estimation by the kit and the microarray. Compared with traditional methods including chromosome aberration analysis, the radiological dosimeter and an application method have the advantages of simplicity, rapidity and high throughput, and have high accuracy of dose estimation, and are especially suitable for radiation dose estimation of large-scale population samples in nuclear accidents and radiation accidents.

Description

technical field [0001] The invention belongs to the field of radiation biological dosimetry and relates to the application of a combination of multiple radiation sensitive genes as a radiation biological dosimeter in radiation dose estimation. Specifically, the present invention relates to the combination of 5-13 different radiation-sensitive genes as the application of radiation biological dosimeter in radiation dose estimation, including the preparation of a combination of 5-13 different radiation-sensitive genes for radiation Dose estimation kits, microarrays, and methods and applications for radiation dose estimation. Background technique [0002] With the wide application of nuclear energy and ionizing radiation, nuclear accidents and radiation accidents occur from time to time. In the early stage of the accident, many people may be exposed. In this case, how to quickly, accurately and high-throughput judge a large number of people It is still one of the important issu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/166C12Q2600/158
Inventor 刘青杰李爽陆雪封江彬田梅
Owner NAT INST FOR RADIOLOGICAL PROTECTION & NUCLEAR SAFETY CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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