CN107090496A - A kind of detection method of 8Gy ionising radiations - Google Patents
A kind of detection method of 8Gy ionising radiations Download PDFInfo
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- CN107090496A CN107090496A CN201710211155.3A CN201710211155A CN107090496A CN 107090496 A CN107090496 A CN 107090496A CN 201710211155 A CN201710211155 A CN 201710211155A CN 107090496 A CN107090496 A CN 107090496A
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- 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
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- 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
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- 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
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
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Abstract
The invention discloses a kind of detection method of ionising radiation, it comprises the following steps：1）Experimental animal is placed in radiation environment, taken out after the completion of irradiation；2）Irradiation finishes extraction experimental animal serum after 24 hours；3）MiRNA ionising radiation marks in test experience animal blood serum excretion body, when miRNA contents exceed 5 times of non-irradiated sample, that is, illustrate that the environment has ionising radiation；Wherein, miRNA is selected from mmu miR 378d, the 3p of mmu miR 151.This method can whether there is the ionising radiation intensity for jeopardizing organism in Accurate Determining environment, and it is estimated, with fast, easily feature.
Description
Technical field
The present invention relates to detection field, more particularly to a kind of detection method of 8Gy ionising radiations.
Background technology
MicroRNA (miRNA) is the endogenous non-coding RNA that a class length is about 19~25nt, and wide participation gene turns
Regulation and control activity after record, with important life regulatory function.Intercellular interaction can be by discharging protein, nucleic acid, fat
Matter equimolecular is combined so as to mediate intracellular signal transduction to extracellular with acceptor.In addition to these unimolecules, cell can also be released
Excretion body is put, by horizontal transfer functional mRNA, miRNA and protein to recipient cell, changes the function of cell.
It is all that the radiation for causing ionization can be acted on material, it is referred to as ionising radiation.The band electrochondria word of high speed, such as α particles, β
Particle, proton etc., can directly cause the material being pierced to produce ionization, belong to directly ionizing particles；(such as X is penetrated ionizing photon
Line and gamma-rays) and the uncharged particle such as neutron, it is to produce powered secondary when with matter interaction and cause thing
Matter is ionized, and belongs to indirect ionization particle.Ionising radiation is ubiquitous in the life of the mankind, such as natural background radiation, including comes
Cosmic ray from cosmic space, the cosmogenic isotopes produced by the atom nuclear interaction in cosmic ray and air and presence
Natural radioactive element (such as radioactive element uranium, thorium and radon) in the earth's crust.Also from artificial radiation source, such as core is tried
Test the radionuclide of generation；The ionising radiation of the generations such as atomic reactor, nuclear power plant, nuclear power naval vessels；Industrial or agricultural,
The ionising radiation that the radioactive element that contains in the discharge waste of the departments such as medical science, scientific research is produced, and rare earth metal and other
During the exploitation of associated metal ore deposit, refinement, radionuclide such as uranium, thorium, radon contained in its three wastes emission etc..When this
The effects of ionizing radiation that a little materials are produced can trigger some effects biologically, so research ionization spoke when organism
The biological effect penetrated is necessary, and this is of great importance to human health etc..
The content of the invention
It is an object of the invention to invent a kind of detection method of 8Gy ionising radiations.
The technical solution used in the present invention is：
A kind of detection method of ionising radiation, it comprises the following steps：
1) experimental animal is placed in radiation environment, taken out after the completion of irradiation；
2) irradiation finishes extraction experimental animal serum after 24 hours；
3) the miRNA ionising radiation marks in test experience animal blood serum excretion body, are not shone when miRNA contents exceed
When penetrating 5 times of sample, that is, illustrate that the environment has ionising radiation；Wherein, miRNA is selected from mmu-miR-378d, mmu-miR-
151-3p。
It is preferred that, ionizing radiation dose is 8Gy.
It is preferred that, experimental animal is mouse.
Contain quantitative mmu-miR-378d, mmu-miR-151-3p in a kind of kit for detecting ionising radiation, kit
Detection reagent.
The reagent of quantitative mmu-miR-378d and mmu-miR-151-3p contents is preparing detection ionising radiation strength agent
In application.
The beneficial effects of the invention are as follows：The ionization spoke for jeopardizing organism can be whether there is in Accurate Determining environment
Penetrate, the detection method has fast, easily feature.
Brief description of the drawings
Fig. 1 is serum excretion body under Electronic Speculum.
Fig. 2 is expression quantity block diagrams of the mmu-miR-378d under the conditions of different ionising radiations.
Fig. 3 is expression quantity block diagrams of the mmu-miR-151-3p under the conditions of different ionising radiations.
Embodiment
Experimental example
1 animal and electromagnetic radiation
This experiment employs the male C57BL/6 mouse that initial age is 13 weeks, totally 40.It is randomly divided into 4 groups, every group 10
Only.This experiment has 0,2,6.5,8Gy tetra- dosage, and irradiation is sampled after terminating 24 hours.First beginning and end irradiation group is as a control group
It is compared.
2. the collection of serum：
Every mouse is carried out using eyeball rear vein beard blood taking method to take blood.Every mouse is about obtained outside 800~1200ul
All blood.After sample stands 4~5 hours in 4 DEG C of refrigerators, 4000rpm, is centrifuged 15 minutes by 4 DEG C.Each sample draws 200ul
Supernatant, and collect 10 samples of each group, be collected in the centrifuge tube sterilized, common 2ml.
3. excretion body is extracted
Serum sample is taken out from low-temperature storage environment, is placed on ice.At room temperature, 2000g is centrifuged 20 minutes, with
Remove residual cell and fragment.Blood plasma or final proof sheet are shifted to a new pipe, excretion body extracts reagent is added, overturns mixing or liquid relief
Device, until complete even sample.It is put into 4 DEG C of refrigerators and stands 30 minutes.4 DEG C, 15000g is centrifuged 2 minutes, is carefully inhaled with pipettor
Remove supernatant.The precipitation that centrifugation is obtained is exactly excretion body.
4. excretion shape is analyzed
1) particle size and the distribution of excretion body are extracted in the technical appraisement of nano particle trace analysis
2) transmission electron microscope observation excretion volume morphing and size
3) Western Blotting methods identification excretion body film surface marker TSG101, CD63.
5.MicroRNA deep sequencings and bioinformatic analysis
It is sequenced and tests for small RNA, is prepared from total serum IgE to final cDNA library and complete mainly to include following step
Suddenly：Glue method is cut with electrophoresis and obtains RNA fragment of the length range in 18~30nt or so；Fragment carries out end reparation and at its two ends
5 '-adapter and 3 '-adapter are connected respectively；Then amplification, obtains final cDNA library；Machine is sequenced in DNA library.Will
The sequencing data of gained is compared by the biomolecule information database with miRNA, can obtain existing miRNA in database
Expressing information, including structure, length, expression etc..The Clean Data and miRBase databases obtained will be sequenced
The ripe body sequence alignments of all miRNA of the species, calculate its expression quantity in (version 21).Testing result is as follows：
mmu-miR-378d
| Group | 0Gy | 2Gy | 6.5Gy | 8Gy |
| Normalized expression | 7.6514 | 2.4895 | 8.3223 | 275.4312 |
mmu-miR-151-3p
| Group | 0Gy | 2Gy | 6.5Gy | 8Gy |
| Normalized expression | 191.9472 | 230.2758 | 338.4094 | 1544.8525 |
Testing result shows, when under the conditions of 8Gy, and mmu-miR-378d and mmu-miR-151-3p expression quantity have significantly
Raising.
Claims (5)
1. a kind of detection method of ionising radiation, it comprises the following steps：
1）Experimental animal is placed in radiation environment, taken out after the completion of irradiation；
2）Irradiation finishes extraction experimental animal serum after 24 hours；
3）MiRNA ionising radiation marks in test experience animal blood serum excretion body, when miRNA contents exceed non-exposure sample
At 5 times of product, that is, illustrate that the environment has ionising radiation；
Wherein, miRNA is selected from mmu-miR-378d, mmu-miR-151-3p.
2. detection method according to claim 1, it is characterised in that the ionizing radiation dose is 8Gy.
3. detection method according to claim 1, it is characterised in that the experimental animal is mouse.
4. a kind of kit for detecting ionising radiation, it is characterised in that in the kit containing quantitative mmu-miR-378d,
Mmu-miR-151-3p detection reagent.
5. the reagent of quantitative mmu-miR-378d and mmu-miR-151-3p contents is in detection ionising radiation strength agent is prepared
Application.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3719144A1 (en) * | 2019-04-05 | 2020-10-07 | Fundación para la Investigación Biomédica del Hospital Universitario de la Paz (FIBHULP) | Mir-151a-3p as an universal endogenous control for exosome cargo normalization |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1216069A (en) * | 1996-02-15 | 1999-05-05 | 匹兹堡大学 | Determining exposure to ionizing radiation agent with persistent biological markers |
| CN101184998A (en) * | 2005-02-25 | 2008-05-21 | 国立大学法人广岛大学 | Method of measuring exposure dosage of ionizing radiation |
| CN105803049A (en) * | 2014-12-29 | 2016-07-27 | 中国辐射防护研究院 | Method for screening biomarkers of ionizing radiation as well as application of SZT2 protein determined by method |
| WO2016130572A2 (en) * | 2015-02-10 | 2016-08-18 | Dana-Farber Cancer Institute, Inc. | Methods of determining levels of exposure to radiation and uses thereof |
-
2017
- 2017-03-28 CN CN201710211155.3A patent/CN107090496A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1216069A (en) * | 1996-02-15 | 1999-05-05 | 匹兹堡大学 | Determining exposure to ionizing radiation agent with persistent biological markers |
| CN101184998A (en) * | 2005-02-25 | 2008-05-21 | 国立大学法人广岛大学 | Method of measuring exposure dosage of ionizing radiation |
| CN105803049A (en) * | 2014-12-29 | 2016-07-27 | 中国辐射防护研究院 | Method for screening biomarkers of ionizing radiation as well as application of SZT2 protein determined by method |
| WO2016130572A2 (en) * | 2015-02-10 | 2016-08-18 | Dana-Farber Cancer Institute, Inc. | Methods of determining levels of exposure to radiation and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| 危文俊 等: "循环血miR_21在空间辐射监测中的应用潜力", 《载人航天》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3719144A1 (en) * | 2019-04-05 | 2020-10-07 | Fundación para la Investigación Biomédica del Hospital Universitario de la Paz (FIBHULP) | Mir-151a-3p as an universal endogenous control for exosome cargo normalization |
| WO2020201577A1 (en) * | 2019-04-05 | 2020-10-08 | Fundación Para La Investigación Biomédica El Hospital Universitario La Paz (Fibhulp) | Mir-151a-3p as an universal endogenous control for exosome cargo normalization |
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Application publication date: 20170825 |