CN109355418A - A method of identifying whether fresh food fruit passes through quarantine irradiation - Google Patents
A method of identifying whether fresh food fruit passes through quarantine irradiation Download PDFInfo
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- CN109355418A CN109355418A CN201811364191.4A CN201811364191A CN109355418A CN 109355418 A CN109355418 A CN 109355418A CN 201811364191 A CN201811364191 A CN 201811364191A CN 109355418 A CN109355418 A CN 109355418A
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- C—CHEMISTRY; METALLURGY
- 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
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
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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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
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- C12Q1/6851—Quantitative amplification
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Abstract
The present invention provides a kind of methods for identifying fresh food fruit and whether passing through quarantine irradiation, the described method includes: the seed of the seed of the fruit without radiation treatment and fruit to be measured is carried out DNA Comet Assay detection respectively, if the trailing length of both testing result displays without significant difference, shows that the fruit to be measured is the fruit without radiation treatment；If the trailing length of both testing result displays there are significant difference, shows that the fruit to be measured is the fruit that is irradiated that treated.This method can identify whether fresh food fruit through quarantine irradiation and can quantitative determine the irradiation dose of sample to be tested, while this method is few using sample, measurement is simple, accuracy is good；Furthermore, this method can detect the fruit of (validity period of phytosanilary certificate is 3 weeks) within after radiation treatment 3 weeks and can identify whether the fruit is irradiated processing, so as to easily judge whether Imported Fruits have passed through enough quarantine irradiations and handle using this method at port.
Description
Technical field
The present invention relates to a kind of methods for identifying fresh food fruit and whether passing through quarantine irradiation, belong to fruits and vegetables authentication technique neck
Domain.
Background technique
China is production and the inlet and outlet big country for eating fruit raw, and in about 4,500,000 tons of import volume of year, export volume also has 4,000,000 tons.
Due to the particularity of product, the risk of the harmful organisms such as fresh and alive fruit vector insects is high, easily invasive plants is caused to be asked
Topic, thus be international community and various countries' quarantine departments focus of attention.
It, according to an international practice, must be to may take in international trade to guarantee that harmful organism is not propagated with international trade
Product quarantine treatment measures with harmful organism.The processing technique of tradition plant product fresh and alive for fruit etc. is mainly with smoked
Based on steaming processing.Suffocating treatment refers to that by means of fumigant gas, passing through certain time in space that can be closed will be harmful
The techniques or methods of biocidal.It carries out stifling being processing technique the most general in the world using fumigant methyl bromide, still
Destruction due to bromomethane to ozone layer, according to Montreal protocol, it is necessary to be phased out.For this purpose, international community
Develop substitute technology of the radiation treatment technology as methyl bromide fumigation.
Radiation treatment is the processing technique for leading to harmful insect infertility using high-energy ray.Radiation treatment technology can be used
For gamma ray, high-energy X-ray or the high-power electron beam of cobalt source release as the energy, the dosage range used is generally 100-300GY,
It has the characteristics that cleaning, quickly, be suitable for refrigeration fruit.Currently, radiation treatment technology grinding in the quarantining treatment of trypetid
Study carefully and application is the most mature, International Plant Protection Convention tissue (IPPC) has promulgated the international mark of a variety of trypetid radiation treatment dosage
Standard, and reviewing using 150GY as the general dosage of trypetid radiation treatment.
Radiation treatment is usually to prevent pest development and breeding as control target, therefore, most with other processing methods
Harmful organism continues to survive after big difference is exactly handled, and worm living can be usually found in port quarantine ping procedure.In order to protect
Gateway of a country bio-safety is demonstrate,proved, needs to judge whether this batch of fruit passes through radiation treatment, and whether be processed sufficient dosage, because
This, the detection technique research after needing to carry out fresh food fruit irradiation.
Currently, after Comet Assay is the universal test method of irradiated food, but whether it can be used to detect quarantine irradiation
Eating fruit raw, there is also queries
Irradiation can not only be used to purposes of quarantining, and is more used to food processing purposes at present, goes out for desinsection
Bacterium.Therefore, although eating the detection method of fruit after also lacking irradiation at present raw, for the food after irradiation, have very much
Detection method, including physical method, chemical method and biological method etc., there presently does not exist a certain methods to differentiate certainly
Whether the food of all categories is by irradiation.
Since high-energy ray irradiation can cause genome DNA break, generate a large amount of fragments, thus can be by electrophoresis after,
It detects genomic DNA and determines that product whether by irradiation, passes through measurement " broom with the presence or absence of the method for hangover (" comet " tail portion)
The parameters such as length, density of star " hangover can further progress quantitative analysis, here it is DNA Comet Assay (DNA Comet
assay).It is current studies have shown that DNA Comet Assay can to potato, wheat, soya bean, almond, preserved apricot, freezing chicken,
Whether the numerous foods such as new fresh chicken meat, freezing cod fillet, chilled fillet, Chinese prickly ash, cassia bark, which pass through radiation treatment, is detected.
But detect whether fresh food fruit passes through quarantine irradiation processing also some queries using DNA Comet Assay, mainly
It is that the exposure dose that food irradiation uses is higher, generally 1000GY or more；The dosage that fruit quarantine irradiation uses is lower, generally
No more than 300GY；Dosage is higher, bigger to the damage of DNA, is more able to observe that " comet " trails.
Summary of the invention
In order to solve the above shortcomings and deficiencies, it is an object of the present invention to provide a kind of identifications whether to eat fruit raw
By the method for quarantine irradiation.
In order to reach the goals above, the present invention provides a kind of identifications to eat the method whether fruit passes through quarantine irradiation raw,
Wherein, which comprises the seed of the seed of the fruit without radiation treatment and fruit to be measured is carried out to DNA comet electricity respectively
Swimming method detection, if the trailing length of both testing result displays without significant difference, shows that the fruit to be measured is not to be irradiated
The fruit of processing；
If the trailing length of both testing result displays there are significant difference, shows that the fruit to be measured is to be irradiated place
Fruit after reason.
Specific embodiment according to the present invention, in the described method, " significant difference " of the present invention are that statistics is general
It reads, judgment criteria are as follows: average and standard deviation statistical analysis p < 0.05 of two groups of samples at least three repeated experiments is
There are significant differences for the two, conversely, then the two is without significant difference.
Specific embodiment according to the present invention, in the described method, the irradiation dose for being irradiated treated fruit is
50-500GY。
Specific embodiment according to the present invention, in the described method, the irradiation dose for being irradiated treated fruit is
50-300GY。
Specific embodiment according to the present invention, in the described method, being irradiated treated fruit is to deposit at room temperature
Store up the fruit within 3 weeks.
Specific embodiment according to the present invention, in the described method, the fresh food fruit include apple, navel orange, pear,
Tangerine orange and nectarine.
Specific embodiment according to the present invention, after judging the fruit to be measured to be irradiated treated fruit, the party
Method further includes that the operation of quantitative detection is carried out to the irradiation dose of the fruit to be measured, is specifically included:
The detection of DNA Comet Assay is carried out to the fruit seeds through different irradiation dose radiation treatments, measures the fruit
The trailing length of seed；
Using the different irradiation doses as abscissa, the line of the two is made using the trailing length of the fruit seeds as ordinate
Sexual intercourse curve graph；
The detection of DNA Comet Assay is carried out to the seed of fruit to be measured again, the hangover for measuring the seed of the fruit to be measured is long
The trailing length is substituted into the linear relationship curve, the irradiation dose of the fruit to be measured is calculated by degree.
Specific embodiment according to the present invention, DNA Comet Assay used are conventional method, and those skilled in the art can be with
It is operated according to existing DNA Comet Assay, as long as guaranteeing that the purpose of the present invention may be implemented.
This method provided by the present invention can identify whether fresh food fruit passes through quarantine irradiation and can be to sample to be tested
Irradiation dose is quantitative determined, while this method is few using sample, measurement is simple, accuracy is good；In addition, provided by the present invention
This method the fruit of (validity period of phytosanilary certificate is 3 weeks) within after radiation treatment 3 weeks can be detected and can be reflected
Whether the fruit is not irradiated processing, so as to easily judge whether Imported Fruits have passed through foot using this method at port
The quarantine irradiation of amount is handled.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, embodiment will be described below
Needed in attached drawing be briefly described, it should be apparent that, the accompanying drawings in the following description is some implementations of the invention
Example, for those of ordinary skill in the art, without any creative labor, can also be according to these attached drawings
Obtain other attached drawings.
Figure 1A is to carry out DNA Comet Assay to non-irradiated apple in the embodiment of the present invention 1 to carry out detecting resulting knot
Fruit figure；
Figure 1B is to carry out DNA Comet Assay to the apple through 300GY irradiation dose radiation treatment in the embodiment of the present invention 1
It carries out detecting resulting result figure；
Fig. 2 is the irradiation dose and sample " comet " trailing length provided in the embodiment of the present invention 4 between 0-500GY
Between linear relationship schematic diagram.
Specific embodiment
In order to which technical characteristic of the invention, purpose and beneficial effect are more clearly understood, now in conjunction in detail below
Embodiment carries out following detailed description to technical solution of the present invention, but should not be understood as the limit to enforceable range of the invention
It is fixed.
Embodiment 1
The present embodiment is using DNA Comet Assay respectively to non-irradiated apple and through 300GY irradiation dose radiation treatment
Apple (apple irradiation after storage at room temperature 1 day) detected, both detection gained result figure respectively as illustrated in figures 1A and ib,
The average data of the two trailing length is as shown in table 1 below.
Table 1
| Sample | Trailing length (μm) |
| Non-irradiated apple | 24.8±3.9a |
| The apple of 300GY irradiation | 25.3±3.4a |
Note: the above results are the average value independently analyzed three times, and identical lowercase is indicated without statistics sex differernce.
From table 1 it follows that the hangover of both non-irradiated apple and apple through 300GY irradiation dose radiation treatment
For length without significant difference, showing to detect entire apple progress DNA Comet Assay can not judge whether its is irradiated place
Reason.
Embodiment 2
The present embodiment is irradiated to non-irradiated apple pulp and through 300GY irradiation dose respectively using DNA Comet Assay
The apple pulp (storage at room temperature 1 day after apple irradiation) of processing is detected, the average data of both detection gained trailing length
As shown in table 2 below.
Table 2
| Sample | Trailing length (μm) |
| Non-irradiated apple pulp | 25.1±2.8a |
| The apple pulp of 300GY irradiation | 24.5±3.7a |
Note: the above results are the average value independently analyzed three times, and identical lowercase is indicated without statistics sex differernce.
From Table 2, it can be seen that non-irradiated apple pulp and the apple pulp two through 300GY irradiation dose radiation treatment
For the trailing length of person without significant difference, showing to detect apple pulp progress DNA Comet Assay can not judge that apple is
No irradiated processing.
Embodiment 3
The present embodiment is irradiated to non-irradiated Apple Seeds and through 300GY irradiation dose respectively using DNA Comet Assay
The Apple Seeds (storage at room temperature 1 day after apple irradiation) of processing are detected, the average data of both detection gained trailing length
As shown in table 3 below.
Table 3
| Sample | Trailing length (μm) |
| Non-irradiated Apple Seeds | 4.3±1.1a |
| The Apple Seeds of 300GY irradiation | 23.2±2.9b |
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
From table 3 it is observed that non-irradiated Apple Seeds and the Apple Seeds two through 300GY irradiation dose radiation treatment
The trailing length of person shows that carrying out detection to Apple Seeds progress DNA Comet Assay may determine that apple there are significant difference
Whether processing is irradiated.
Embodiment 4
The present embodiment uses DNA Comet Assay respectively to non-irradiated Apple Seeds and through different irradiation doses first
The Apple Seeds (apple irradiation after storage at room temperature 1 day) of (such as 100GY, 200GY, 300GY, 400GY and 500GY) radiation treatment into
The average data of row detection, the trailing length of those samples of detection gained is as shown in table 4 below.
Table 4
| Sample | Trailing length (μm) |
| Non-irradiated Apple Seeds | 4.3±1.1 |
| The Apple Seeds of 100GY irradiation | 12.5±1.8 |
| The Apple Seeds of 200GY irradiation | 17.7±1.9 |
| The Apple Seeds of 300GY irradiation | 23.2±2.9 |
| The Apple Seeds of 400GY irradiation | 32.9±3.4 |
| The Apple Seeds of 500GY irradiation | 38.4±4.1 |
Note: the above results are the average value independently analyzed three times.
Using different irradiation doses as abscissa, the relation curve of the two is done using the trailing length of Apple Seeds as ordinate
Figure；As shown in Fig. 2, from figure 2 it can be seen that irradiation dose and sample " comet " trailing length between 0-500GY there are lines
Sexual intercourse, the linear relationship are as follows: y=0.0678x+4.5571, this shows that this method provided by the present invention can be used for apple spoke
According to the quantitative detection of dosage.
The detection of DNA Comet Assay is carried out to the seed of apple to be measured again, the hangover for measuring the seed of the apple to be measured is long
Degree is 15.6 ± 2.1 μm, which is substituted into the linear relationship curve y=0.0678x+4.5571, is calculated
The irradiation dose of the apple to be measured is 163GY.
Embodiment 5
The present embodiment is irradiated to non-irradiated Apple Seeds and through 200GY irradiation dose respectively using DNA Comet Assay
The Apple Seeds (storage at room temperature 1 day, 1 week, 2 weeks and 3 weeks after apple irradiation) that different time is stored after processing are detected, and are detected
The average data of the trailing length of gained each sample is as shown in table 5 below.
Table 5
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
As can be seen from Table 5, using this method provided by the present invention to the apple kind for storing different number of days after irradiation
Son is detected, and discovery this method also has good detection effect for storing the apple within 3 weeks after irradiation, that is, uses this hair
This method provided by bright can identify whether the fruit stored within 3 weeks after radiation treatment passes through radiation treatment.
Embodiment 6
The present embodiment is irradiated to non-irradiated navel orange seed and through 200GY irradiation dose respectively using DNA Comet Assay
The navel orange seed (storage at room temperature 1 day after navel orange irradiation) of processing is detected, the average data of both detection gained trailing length
As shown in table 6 below.
Table 6
| Sample | Trailing length (μm) |
| Non-irradiated navel orange seed | 8.2±3.0a |
| The navel orange seed of 200GY irradiation | 18.4±2.5b |
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
As can be seen from Table 6, non-irradiated navel orange seed and the navel orange seed two through 200GY irradiation dose radiation treatment
The trailing length of person shows that carrying out detection to navel orange seed progress DNA Comet Assay may determine that navel orange there are significant difference
Whether processing is irradiated.
Embodiment 7
The present embodiment is irradiated to non-irradiated pear seed and through 200GY irradiation dose respectively using DNA Comet Assay
The pear seed (storage at room temperature 1 day after pear irradiation) of processing is detected, the average data of both detection gained trailing length
As shown in table 7 below.
Table 7
| Sample | Trailing length (μm) |
| Non-irradiated pear seed | 5.4±2.1a |
| The pear seed of 200GY irradiation | 14.6±1.6b |
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
As can be seen from Table 7, non-irradiated pear seed and the pear seed two through 200GY irradiation dose radiation treatment
The trailing length of person shows that carrying out detection to pear seed progress DNA Comet Assay may determine that pear there are significant difference
Whether processing is irradiated.
Embodiment 8
The present embodiment is using DNA Comet Assay respectively to non-irradiated Nanfeng orange seed and through 200GY irradiation dose
The Nanfeng orange seed (storage at room temperature 1 day after Nanfeng orange irradiation) of radiation treatment is detected, both detection gained hangover length
The average data of degree is as shown in table 8 below.
Table 8
| Sample | Trailing length (μm) |
| Non-irradiated Nanfeng orange seed | 8.7±2.4a |
| The Nanfeng orange seed of 200GY irradiation | 19.5±3.2b |
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
As can be seen from Table 8, non-irradiated Nanfeng orange seed and Nanfeng County's honey through 200GY irradiation dose radiation treatment
The trailing length of both tangerine seeds shows that carrying out DNA Comet Assay to Nanfeng orange seed detects there are significant difference
It may determine that whether Nanfeng orange is irradiated processing.
Embodiment 9
The present embodiment is irradiated to non-irradiated nectarine seed and through 200GY irradiation dose respectively using DNA Comet Assay
The nectarine seed (storage at room temperature 1 day after nectarine irradiation) of processing is detected, the average data of both detection gained trailing length
As shown in table 9 below.
Table 9
| Sample | Trailing length (μm) |
| Non-irradiated nectarine seed | 3.1±2.5a |
| The nectarine seed of 200GY irradiation | 17.7±1.8b |
Note: the above results are the average value independently analyzed three times, and different lowercase a, b indicate there is statistics sex differernce.
As can be seen from Table 9, non-irradiated nectarine seed and the nectarine seed two through 200GY irradiation dose radiation treatment
The trailing length of person shows that carrying out detection to nectarine seed progress DNA Comet Assay may determine that nectarine there are significant difference
Whether processing is irradiated.
Claims (6)
1. a kind of method for identifying fresh food fruit and whether passing through quarantine irradiation, which is characterized in that the described method includes: will be without spoke
The seed of seed and fruit to be measured according to the fruit of processing carries out DNA Comet Assay detection respectively, if testing result shows two
The trailing length of person then shows that the fruit to be measured is the fruit without radiation treatment without significant difference；
If the trailing length of both testing result displays there are significant difference, shows that the fruit to be measured is after being irradiated processing
Fruit.
2. the method according to claim 1, wherein the irradiation dose for being irradiated treated fruit is 50-
500GY。
3. according to the method described in claim 2, it is characterized in that, the irradiation dose for being irradiated treated fruit is 50-
300GY。
4. according to the method described in claim 2, it is characterized in that, being irradiated treated fruit is to store 3 weeks at room temperature
Within fruit.
5. the method according to claim 1, wherein the fresh food fruit includes apple, navel orange, pear, tangerine orange
And nectarine.
6. method according to claim 1-5, which is characterized in that judging the fruit to be measured for irradiated place
After fruit after reason, this method further includes that the operation of quantitative detection is carried out to the irradiation dose of the fruit to be measured, is specifically included:
The detection of DNA Comet Assay is carried out to the fruit seeds through different irradiation dose radiation treatments, measures the fruit seeds
Trailing length；
Using the different irradiation doses as abscissa, the linear pass of the two is done using the trailing length of the fruit seeds as ordinate
It is curve graph；
The detection of DNA Comet Assay is carried out to the seed of fruit to be measured again, measures the trailing length of the seed of the fruit to be measured, it will
The trailing length substitutes into the linear relationship curve, and the irradiation dose of the fruit to be measured is calculated.
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| CN201811364191.4A CN109355418A (en) | 2018-11-16 | 2018-11-16 | A method of identifying whether fresh food fruit passes through quarantine irradiation |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101865877A (en) * | 2010-06-25 | 2010-10-20 | 西北大学 | Gel paving method by using single cell gel electrophoresis |
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2018
- 2018-11-16 CN CN201811364191.4A patent/CN109355418A/en not_active Withdrawn
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101865877A (en) * | 2010-06-25 | 2010-10-20 | 西北大学 | Gel paving method by using single cell gel electrophoresis |
Non-Patent Citations (3)
| Title |
|---|
| AFFAF KHAWAR ET AL.: "Evaluation of irradiation in foods using DNA Comet assay", 《J FOOD SCI TECHNOL》 * |
| H. CERDA ET AL.: "The DNA ‘comet assay’ as a rapid screening technique to control irradiated food", 《MUTATION RESEARCH》 * |
| YUNHEE JO ET AL.: "Characterization of DNA comet and cellulose radical signal in Valencia oranges treated with different forms of ionizing radiation", 《POSTHARVEST BIOLOGY AND TECHNOLOGY》 * |
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