CN105548543A - Kit for detecting yellow spotting virus of peanuts, as well as detection method and application thereof - Google Patents
Kit for detecting yellow spotting virus of peanuts, as well as detection method and application thereof Download PDFInfo
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- CN105548543A CN105548543A CN201610077250.4A CN201610077250A CN105548543A CN 105548543 A CN105548543 A CN 105548543A CN 201610077250 A CN201610077250 A CN 201610077250A CN 105548543 A CN105548543 A CN 105548543A
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- G—PHYSICS
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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Abstract
The invention discloses a kit for detecting yellow spotting virus of peanuts, as well as a detection method and an application thereof. The kit for detecting yellow spotting virus of peanuts comprises a box, a nitrocellulose membrane, a developing liquid reagent bottle, an antibody reagent bottle and a contrast reagent bottle, wherein the developing liquid reagent bottle comprises a developing reagent bottle A and a developing reagent bottle B; the antibody reagent bottle comprises a first antibody reagent bottle and a second antibody reagent bottle; and the contrast reagent bottle comprises a GYSV standard positive contrast reagent bottle and a GYSV standard negative contrast reagent bottle. The application indicates the application of the kit for detecting yellow spotting virus of peanuts in the detection for the yellow spotting virus of peanuts.
Description
Technical field
The invention belongs to plant virus detection technique field, be specifically related to one and cultivate peanut flavimaculatus poison detection kit and detection method and application.
Background technology
Peanut yellow spotting virus (Groundnutyellowspotvirus, GYSV) belongs to bunyaviridae (Bunyaviridae) Tospovirus (Tospovirus) member.GYSV reports on India's peanut, also this virus is found subsequently on the capsicum of Yunnan, the symptom of this virus on peanut and capsicum all shows as on initial stage blade yellow spotting, later stage yellow spotting expands gradually becomes necrotic plaque block, and the symptom main manifestations on pepper fruit is yellow and downright bad patch.GYSV recall rate on the capsicum of Yunnan is not high, main cause can only may be propagated by the yellow hard thrips of tea with this virus, and the yellow hard thrips of tea is still few at Yunnan report, but also can large area break out once possess this virus of suitable condition, cause the yield and quality of the crops such as capsicum to lose, therefore to study and the detection method obtaining a kind of this virus causing disease of Rapid identification has great importance for the Control Technology of this disease.
Summary of the invention
The first object of the present invention is to provide one to cultivate peanut flavimaculatus poison detection kit; Second object is to provide the detection method of described peanut yellow spotting virus detection kit; 3rd object is to provide the application of described peanut yellow spotting virus detection kit.
The first object of the present invention is achieved in that and comprises box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle;
Described nitrite ion reagent bottle comprises chromogenic reagent bottle A and nitrite ion reagent bottle B;
Described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles;
Described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle.
The second object of the present invention is achieved in that and comprises the following steps:
1) cut nitrocellulose filter according to detection sample size, one of nitrocellulose filter jiao is cut off, in order to indicating film direction.
2) detect sample add lavation buffer solution grinding, pipettor gets 2.5 μ l lapping liquid points in nitrocellulose membrane square frame, with the accurate positive control of time point subscript and standard negative control, room temperature place or 37 DEG C be placed to film finish-drying;
3) film is moved to container, add Block buffer, room temperature or 37 DEG C of closed 1h;
4) in Block buffer, add primary antibodie, room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
5) discard primary antibodie and cultivate liquid, lavation buffer solution washes film 3 times, each 5min;
6) discard lavation buffer solution, add Block buffer, then in Block buffer, add two resist, room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
7) discard two anti-cultivation liquid, lavation buffer solution washes film 3 times, each 5min;
8) discard lavation buffer solution, colorbuffer washes film 1 time, each 5min;
9) discard colorbuffer, add the place's colour developing of nitrite ion dark, fully after colour developing, film is moved to color development stopping in clear water, analyze and record testing result.
The third object of the present invention is achieved in that described peanut yellow spotting virus detection kit is detecting the application in peanut yellow spotting virus.
Advantage of the present invention:
New topoviruses separator peanut yellow spotting virus N protein on the capsicum of Yunnan has been prepared by the method for prokaryotic expression, immunizing rabbit obtains the peanut yellow spotting virus rabbit anti-serum of high-titer, optimize and assemble the dot-ELISA detection kit of this new separator, and be applied to the quick detection of field sample, by the development of this kit, the band poison situation of peanut yellow spotting virus is carried, for disease prevention and control provide detection technique guarantee in the doubtful sample in detection field that can be quick, accurate, a large amount of.
Accompanying drawing explanation
Fig. 1 is the check point master drawing of GYSV on pepper plant;
Fig. 2 is the test result figure of GYSV on pepper plant.
Embodiment
Below in conjunction with drawings and Examples, the present invention is further illustrated, but limited the present invention never in any form, and any conversion done based on training centre of the present invention or replacement, all belong to protection scope of the present invention.
Peanut yellow spotting virus peanut yellow spotting virus (Groundnutyellowspotvirus, GYSV) detection kit of the present invention, comprises box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle;
Described nitrite ion reagent bottle comprises chromogenic reagent bottle A and nitrite ion reagent bottle B;
Described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles;
Described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle.
Described nitrocellulose filter is provided with grid, and grid empty place is in order to point sample.
Described nitrite ion reagent bottle A contains NBT nitrite ion; Nitrite ion reagent bottle B contains BCIP nitrite ion.
Described NBT nitrite ion is dimethyl formamide 0.5gNBT being dissolved in 10ml70%; BCIP nitrite ion is dimethylformamide 0.5gBCIP being dissolved in 10ml100%.
Described primary antibodie reagent bottle contains peanut yellow spotting virus specific polyclonal antibody; Two anti-reagent bottles contain specificity goat-anti rabbit.
The described preparation containing peanut yellow spotting virus specificity clonal antibody adopts the method for its N gene prokaryotic to obtain immunogenic, and immunizing rabbit obtains, and uses and tires as 1:500 ~ 20000; The GYSV pimento separator N gene complete sequence that peanut yellow spotting virus N gene PCR amplimer register report according to fourth inscription (Ding.M.) designs (number of registration as: EF528556), primer sequence and annealing temperature thereof specifically see the following form:
Described GYSV standard positive control reagent bottle contains the Ben Shi cigarette plant drymeal infecting GYSV; GYSV standard negative control reagent bottle is containing unsoundness Ben Shi cigarette plant drymeal.
The detection method of peanut yellow spotting virus detection kit of the present invention, comprises the following steps:
1) cut nitrocellulose filter (such as detect sample and have 5, add 1 positive control and 1 negative control, the nitrocellulose filter of 7 grids should be cut) according to detection sample size, one of nitrocellulose filter jiao is cut off, in order to indicating film direction.
2) detect sample add lavation buffer solution grinding (1:10 ~ 50 times w/v), pipettor gets 2.5 μ l lapping liquid points in nitrocellulose membrane square frame, with the accurate positive control of time point subscript and standard negative control, room temperature place or 37 DEG C be placed to film finish-drying;
3) film is moved to appropriate containers, add appropriate Block buffer (proceed to container according to film to determine, such as diameter is that the double dish of 20cm adds 25ml Block buffer), room temperature or 37 DEG C of closed 1h;
4) in Block buffer, add primary antibodie (addition is calculated according to primary antibodie operation instructions), room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
5) discard primary antibodie and cultivate liquid, appropriate lavation buffer solution washes film 3 times, and each 5min(shaking table is even to be shaken);
6) discard lavation buffer solution, add appropriate Block buffer, then in Block buffer, add two anti-(addition is calculated according to two anti-operation instructions), room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
7) discard two anti-cultivation liquid, appropriate lavation buffer solution washes film 3 times, and each 5min(shaking table is even to be shaken);
8) discard lavation buffer solution, appropriate colorbuffer washes film 1 time, and each 5min(shaking table is even to be shaken);
9) discard colorbuffer, add appropriate nitrite ion (by nitrite ion reagent bottle A and the preparation of nitrite ion reagent bottle B instructions) dark place colour developing, fully after colour developing, film is moved to color development stopping in clear water.Analyze and record testing result.
The described peanut yellow spotting virus detection kit that is applied as of peanut yellow spotting virus detection kit of the present invention is detecting the application in peanut yellow spotting virus.
Interpretation of result method:
Standard positive control spot becomes redness, aubergine or black, and the nondiscolouring of standard negative control spot, shows that experimentation is correct.Detection sample point becomes redness, aubergine or black and shows that this sample is with GYSV, and positive made by meter; Detect sample point nondiscolouring and show that this sample is without GYSV, negative sample made by meter.
Washing buffer formula of liquid (1L):
Block buffer is filled a prescription:
Lavation buffer solution adds 5% skimmed milk power mixing by mass volume ratio (W/V).
Colorbuffer formula (100ml):
Nitrite ion compound method:
66 μ l nitrite ion A are added and 33 μ l nitrite ion B mix in every 10ml colorbuffer.
Nitrite ion A fills a prescription: 0.5gNBT is dissolved in the dimethyl formamide of 10ml70%.
Nitrite ion B fills a prescription: 0.5gBCIP is dissolved in the dimethyl formamide of 10ml100%.
Primary antibodie reagent bottle operation instruction: primary antibodie reagent bottle is kept at 4 DEG C, uses and tires as 1:3000.
Two anti-reagent bottle operation instructions: two anti-reagent bottles are kept at 4 DEG C, use and tire as 1:8000.
Contrast agents bottle operation instruction: add the mixing of 2ml lavation buffer solution before use.
Peanut yellow spotting virus detection kit of the present invention is provided with box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions, and described nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions are located in box body.Preparation method: 1) prepare nitrocellulose filter, described nitrocellulose filter is drawn and has grid, grid empty place is in order to point sample; 2) prepare nitrite ion reagent bottle, described nitrite ion reagent bottle comprises nitrite ion reagent bottle A and nitrite ion reagent bottle B; 3) Dispersal risk reagent bottle, described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles, described primary antibodie is peanut yellow spotting virus specific polyclonal antibody, the method of peanut yellow spotting virus N gene prokaryotic is adopted to obtain immunogenic, immunizing rabbit obtains, and uses and tires as 1:500 ~ 20000; Described two resist for specificity goat-anti rabbit; 4) prepare contrast agents bottle, described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle; 5) detection kit instructions is prepared, described detection kit instructions comprises detection method, washing buffer formula of liquid, Block buffer formula, colorbuffer formula, nitrite ion compound method, primary antibodie reagent bottle operation instruction, two anti-reagent bottle operation instructions and contrast agents bottle operation instruction.Described peanut yellow spotting virus detection kit can be applied in detection peanut yellow spotting virus.Specifically comprise:
A, peanut yellow spotting virus detection kit: peanut yellow spotting virus detection kit is provided with box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions, and described nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions are located in box body.
B, peanut yellow spotting virus detection kit preparation method:
1) prepare nitrocellulose filter, described nitrocellulose filter is drawn grid, and grid empty place is in order to point sample;
2) prepare nitrite ion reagent bottle, described nitrite ion reagent bottle comprises nitrite ion reagent bottle A and nitrite ion reagent bottle B;
3) Dispersal risk reagent bottle, described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles, described primary antibodie is peanut yellow spotting virus specific polyclonal antibody, the method of peanut yellow spotting virus N gene prokaryotic is adopted to obtain immunogenic, immunizing rabbit obtains, and uses and tires as 1:500 ~ 20000; Described two resist for specificity goat-anti rabbit;
4) prepare contrast agents bottle, described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle;
5) detection kit instructions is prepared, described detection kit instructions comprises detection method, washing buffer formula of liquid, Block buffer formula, colorbuffer formula, nitrite ion compound method, primary antibodie reagent bottle operation instruction, two anti-reagent bottle operation instructions and contrast agents bottle operation instruction.
Be specially:
Described peanut yellow spotting virus detection kit is provided with box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions, and described nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle and detection kit instructions are located in box body.Specifically comprise:
A, nitrocellulose filter.Film is drawn and has grid, grid empty place is in order to point sample.
B, nitrite ion reagent bottle, described nitrite ion reagent bottle comprises nitrite ion reagent bottle A and nitrite ion reagent bottle B.Nitrite ion reagent bottle A contains 1mlNBT nitrite ion; Nitrite ion reagent bottle B contains 500 μ lBCIP nitrite ions.
C, antibody reagent bottle, described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles.Primary antibodie reagent bottle contains 500 μ l peanut yellow spotting virus specific polyclonal antibodies, and adopt the method for peanut yellow spotting virus N gene prokaryotic to obtain immunogenic, immunizing rabbit obtains, and uses and tires as 1:500 ~ 20000; Two anti-reagent bottles contain 500 μ l specificity goat-anti rabbits.
D, contrast agents bottle, described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle.GYSV standard positive control reagent bottle contains the Ben Shi cigarette plant drymeal that 1g is with GYSV; GYSV standard negative control reagent bottle is containing 1g healthy Ben Shi cigarette plant drymeal.
E, detection kit instructions.Detection kit instructions comprises detection method, washing buffer formula of liquid, Block buffer formula, colorbuffer formula, nitrite ion compound method, primary antibodie reagent bottle operation instruction, two anti-reagent bottle operation instructions and contrast agents bottle operation instruction.
With concrete case study on implementation, the present invention will be further described below:
Embodiment 1
The detection of GYSV on pepper plant
One, by specification prepares following damping fluid and reagent:
1. lavation buffer solution preparation: take NaCl8g, K
2hPO
40.2g, Na
2hPO
412H
2o7.2g, KCl0.2g are dissolved in 1L distilled water, adjust pH to 7.4, add 0.1% polysorbas20 mixing.
2. Block buffer preparation: measure 100ml lavation buffer solution, add 5g skimmed milk power and stir and evenly mix.
3. contrast agents bottle preparation: GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle add the mixing of 2ml lavation buffer solution respectively.
4. prepare 1mol/LTris-HCl(pH9.5): take 12gTris and be dissolved in 100ml distilled water and stir and evenly mix, adjust pH to 9.5.
5. prepare 1mol/LNaCl: take 5.8gNaCl and be dissolved in 100ml distilled water and stir and evenly mix.
6. prepare 1mol/LMgCl
2: take 20.3gMgCl
2be dissolved in 100ml distilled water and stir and evenly mix.
7. colorbuffer preparation: measure the 1mol/LTris-HCl(pH9.5 prepared in advance) 10ml, 1mol/LNaCl10ml, 1mol/LMgCl
2in 0.5ml to 200ml beaker, supply distilled water and mix to 100ml.
Two, getting 22 Pepper Leaves 1 ~ 3g to grind to thickening in plastics valve bag, all adding appropriate lavation buffer solution and mixing for subsequent use.
Three, the nitrocellulose filter cutting 8 grids is in the double dish of 5.5cm as diameter, pipettor gets 2.5 μ l lapping liquid points in nitrocellulose membrane square frame, nextly on same sample put in order, the positive contrast of rearmost point and negative control, 37 DEG C are placed to film finish-drying.
Four, 5ml Block buffer 37 DEG C of closed 1h are added.
Five, in Block buffer, add 1.7 μ l primary antibodies, room temperature shaker shakes up, and cultivates 1h for 37 DEG C.
Six, discard primary antibodie and cultivate liquid, appropriate lavation buffer solution horizontal shaker washes film 3 times, each 5min.
Seven, discard lavation buffer solution, add 5ml Block buffer, then in Block buffer, add 0.7 μ l bis-resist, room temperature shaker shakes up, and cultivates 1h for 37 DEG C.
Eight, discard two anti-cultivation liquid, appropriate lavation buffer solution horizontal shaker washes film 3 times, each 5min.
Nine, discard lavation buffer solution, appropriate colorbuffer horizontal shaker washes film 1 time, each 5min.
Ten, preparation nitrite ion: measure 5ml colorbuffer, adds 33 μ l nitrite ion A and 16.5 μ l nitrite ion B mix.
11, discard colorbuffer, add 5ml nitrite ion in the colour developing of dark place, develop the color to positive control spot variable color, then after time delay 10min, film is transferred to color development stopping in clear water.
12, interpretation of result: standard positive control spot becomes aubergine, the nondiscolouring of standard negative control spot, shows that experimentation is correct.The detection sample point being numbered 3,5,6,9,11,14,20 becomes aubergine, shows that sample is with GYSV, and positive made by meter; All the other sample point nondiscolourings show that this sample is without GYSV, and negative sample made by meter.
Testing result is in table 1, table 2 and accompanying drawing 1, accompanying drawing 2;
The detection sample record table of GYSV on table 1 pepper plant
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 |
| 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 |
| 17 | 18 | 19 | 20 | 21 | 22 | Positive control | Negative control |
| 17 | 18 | 19 | 20 | 21 | 22 | Positive control | Negative control |
The testing result statistical form of GYSV on table 2 pepper plant
| Sample number into spectrum | Testing result |
| 1 | - |
| 2 | - |
| 3 | + |
| 4 | - |
| 5 | + |
| 6 | + |
| 7 | - |
| 8 | - |
| 9 | + |
| 10 | - |
| 11 | + |
| 12 | - |
| 13 | - 6 --> |
| 14 | + |
| 15 | - |
| 16 | - |
| 17 | - |
| 18 | - |
| 19 | - |
| 20 | + |
| 21 | - |
| 22 | - |
| Positive control | + |
| Negative control | - |
Note: in table, "+" represents positive, namely detects in sample to be with and surveys virus to some extent; "-" represents negative, namely detect in sample without survey virus.
SEQUENCELISTING
<110> KUNMING INST OF BOTANY CAS
<120> mono-cultivates peanut flavimaculatus poison detection kit and detection method and application
<130>2016
<160>2
<170>PatentInversion3.3
<210>1
<211>35
<212>DNA
<213> specific reverse primers
<400>1
acgcgtcgactcacagctctccgctgcctatagct35
<210>2
<211>61
<212>DNA
<213> specific forward primer
<400>2
cgcggatccatgtctacaaaaggcgttattaaagacgcgtcgactcagaatccttgattc60
t61
Claims (9)
1. one cultivate peanut flavimaculatus poison detection kit, it is characterized in that comprising box body, nitrocellulose filter, nitrite ion reagent bottle, antibody reagent bottle, contrast agents bottle;
Described nitrite ion reagent bottle comprises chromogenic reagent bottle A and nitrite ion reagent bottle B;
Described antibody reagent bottle comprises primary antibodie reagent bottle and two anti-reagent bottles;
Described contrast agents bottle comprises GYSV standard positive control reagent bottle and GYSV standard negative control reagent bottle.
2. peanut yellow spotting virus detection kit according to claim 1, it is characterized in that described nitrocellulose filter is provided with grid, grid empty place is in order to point sample.
3. peanut yellow spotting virus detection kit according to claim 1, is characterized in that described nitrite ion reagent bottle A contains NBT nitrite ion; Nitrite ion reagent bottle B contains BCIP nitrite ion.
4. peanut yellow spotting virus detection kit according to claim 3, is characterized in that described NBT nitrite ion is dimethyl formamide 0.5gNBT being dissolved in 10ml70%; BCIP nitrite ion is dimethylformamide 0.5gBCIP being dissolved in 10ml100%.
5. peanut yellow spotting virus detection kit according to claim 1, is characterized in that described primary antibodie reagent bottle contains peanut yellow spotting virus specific polyclonal antibody; Two anti-reagent bottles contain specificity goat-anti rabbit.
6. peanut yellow spotting virus detection kit according to claim 5, it is characterized in that the described preparation containing peanut yellow spotting virus specificity clonal antibody adopts the method for its N gene prokaryotic to obtain immunogenic, immunizing rabbit obtains, and uses and tires as 1:500 ~ 20000; The GYSV pimento separator N gene complete sequence that peanut yellow spotting virus N gene PCR amplimer register report according to fourth inscription (Ding.M.) designs (number of registration as: EF528556), primer sequence and annealing temperature thereof specifically see the following form:
。
7. peanut yellow spotting virus detection kit according to claim 1, is characterized in that described GYSV standard positive control reagent bottle contains the Ben Shi cigarette plant drymeal infecting GYSV; GYSV standard negative control reagent bottle is containing unsoundness Ben Shi cigarette plant drymeal.
8. a detection method for claim 1 ~ 7 arbitrary described peanut yellow spotting virus detection kit, is characterized in that comprising the following steps:
1) cut nitrocellulose filter according to detection sample size, one of nitrocellulose filter jiao is cut off, in order to indicating film direction.
2) detect sample add lavation buffer solution grinding, pipettor gets 2.5 μ l lapping liquid points in nitrocellulose membrane square frame, with the accurate positive control of time point subscript and standard negative control, room temperature place or 37 DEG C be placed to film finish-drying;
3) film is moved to container, add Block buffer, room temperature or 37 DEG C of closed 1h;
4) in Block buffer, add primary antibodie, room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
5) discard primary antibodie and cultivate liquid, lavation buffer solution washes film 3 times, each 5min;
6) discard lavation buffer solution, add Block buffer, then in Block buffer, add two resist, room temperature shaker shakes up, and cultivates a 1h or 4 DEG C cultivation for 37 DEG C and spends the night;
7) discard two anti-cultivation liquid, lavation buffer solution washes film 3 times, each 5min;
8) discard lavation buffer solution, colorbuffer washes film 1 time, each 5min;
9) discard colorbuffer, add the place's colour developing of nitrite ion dark, fully after colour developing, film is moved to color development stopping in clear water, analyze and record testing result.
9. an application for claim 1 ~ 7 arbitrary described peanut yellow spotting virus detection kit, is characterized in that described peanut yellow spotting virus detection kit is detecting the application in peanut yellow spotting virus.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610077250.4A CN105548543A (en) | 2016-02-04 | 2016-02-04 | Kit for detecting yellow spotting virus of peanuts, as well as detection method and application thereof |
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|---|---|---|---|
| CN201610077250.4A CN105548543A (en) | 2016-02-04 | 2016-02-04 | Kit for detecting yellow spotting virus of peanuts, as well as detection method and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109929809A (en) * | 2019-02-15 | 2019-06-25 | 云南省农业科学院生物技术与种质资源研究所 | Potato virus S strain and its construction method and application in Potato Cultivars beautiful potato 6 a kind of |
| CN109929810A (en) * | 2019-02-15 | 2019-06-25 | 云南省农业科学院生物技术与种质资源研究所 | Corium solani strain and its construction method and application in a kind of Potato Cultivars meeting -2 |
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