CN103558379A - Rapid detection kit for cucumber green mottle mosaic viruses - Google Patents
Rapid detection kit for cucumber green mottle mosaic viruses Download PDFInfo
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- CN103558379A CN103558379A CN201310548698.6A CN201310548698A CN103558379A CN 103558379 A CN103558379 A CN 103558379A CN 201310548698 A CN201310548698 A CN 201310548698A CN 103558379 A CN103558379 A CN 103558379A
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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Abstract
The invention discloses a rapid detection kit for cucumber green mottle mosaic viruses. The rapid detection kit comprises a kit cover, a kit body and a test paper strip, wherein the box cover is provided with a reaction groove for containing a fluorescent nano particle-cucumber green mottle mosaic virus antibody conjugate; the kit body is internally provided with a test paper strip storing position; the test paper strip is a rapid detection test paper strip for the cucumber green mottle mosaic viruses and is composed of a bottom plate, and a sample cushion, a chromatography film and a water absorption cushion which are attached on the bottom plate and are closely connected in sequence; the chromatography film is provided with a detection region and a quality control region; the detection region is used for fixing a monoclonal antibody or a polyclonal antibody which is subjected to specific reaction with the cucumber green mottle mosaic viruses. The detection kit provided by the invention can be used for finishing the detection of the cucumber green mottle mosaic viruses of plants in very short time, has good specificity and sensitivity and is an idea cucumber green mottle mosaic virus infection detection tool.
Description
Technical field
The present invention relates to field of biological detection, be specifically related to a kind of device that detects the infection of plant cucumber green mottle mosaic virus with immunochromatography technique.
Background technology
Cucumber green mottle mosaic virus (Cucumber green mottle mosaic virus) is one of important plant quarantine venereal disease poison, can cause crop yield loss to reach 15%, and the production of ground family crop is caused to serious threat.Liaoning Province in 2005 Gaizhou City's watermelon has infected cucumber green mottle mosaic virus, suffers heavy losses.After this in Guangxi, the ground such as Liaoning, Hebei, Shandong, Guangdong and Beijing finds epidemic situation successively, the production of the crops such as watermelon, muskmelon, cucumber in serious threat, the detection of therefore strengthening this virus is very important.
Prevent that the seed that carries cucumber green mottle mosaic virus from epidemic-stricken area from entering the territory and propagating is to control the prerequisite that this virus is imported into and spread; Strengthening epidemic monitoring and take the measure of effectively eradicating, is to prevent that disease from spreading popular key.At present, laboratory is to the conventional ELISA of this viral fast detecting and PCR equimolecular biological detection method.ELISA kit exists and detects the shortcoming that reagent is extremely expensive, and PCR method exists complex operation, cannot implement the shortcoming of field large area monitoring.Therefore, simple and easy to do method for quick is significant for the prevention and control of cucumber green mottle virus epidemic situation.
Immunochromatography technique is a kind of easy Fast Detection Technique based on antigen-antibody reaction growing up the nineties in last century, this technology has been widely used in clinical, food safety detection field.Colloidal gold immunochromatographydetection detection test paper bar is one of principal item of extracorporeal fast detection reagent.Quick, easy, inexpensive with it, specimen in use amount is few, can directly measure undressed sample, pollute little, do not need expensive instrument, can intuitive judgment result, be easy to the advantages such as basic unit and on-the-spot use, and be used widely.But the sensitivity of colloidal gold immuno-chromatography test paper strip is not ideal enough, this has limited its widespread use in detection field greatly.
Adopt the more strong mark substance of ratio nano gold grain signal can improve to a certain extent the sensitivity of immuno-chromatographic test paper strip.Novel fluorescent nano particle exactly has a kind of like this advantage.Fluorescent nano particle is that fluorescent material (as FITC, quantum dot, Rare Earth Chelate) is wrapped in macromolecular material, forms nano particle (Nanoparticles, NPs).Package action on the one hand can make more light emitting molecule be connected in biomolecule and play signal amplification, can overcome on the other hand external environment to the impact of luminescence reagent (as cancellation effect etc.), increases the stability of luminescence reagent.The nano particle that is wrapped in fluorescence molecule or other light emitting molecule has much higher luminous intensity and photochemical stability than single fluorescence molecule.The advantages such as fluorescent nano particle also has particle homogeneous, and size is any adjustable; Especially aspect mark, abandoned the mark mode that collaurum relies on passive physisorption completely, adopt directed covalent coupling technology, almost the complete activity that has kept antibody and antigen, reduces the background noise detecting greatly.Adopt identical antigen and antibody reagent, fluorescent nano particle can exceed easily a collaurum 1-2 order of magnitude in sensitivity, is the biomolecular labeling material that the utmost point has application prospect.
Summary of the invention
The object of the present invention is to provide a kind of easy to use, highly sensitive for cucumber green mottle mosaic virus device for fast detecting, the field quick detection infecting for plant cucumber green mottle mosaic virus.
The present invention discloses a kind of cucumber green mottle mosaic virus quick-detecting box, comprises lid, box body and test strips, is provided for taking in the reactive tank of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter on lid, in box body, test strips is set and takes in position; Described test strips is cucumber green mottle mosaic virus Rapid detection test strip, by base plate be attached to that on base plate, the sample pad of close proximity, chromatographic film and adsorptive pads form successively, described chromatographic film is provided with detection zone and Quality Control district, and monoclonal antibody or the polyclonal antibody with cucumber green mottle mosaic virus generation specific reaction fixed in detection zone.The form of described cucumber green mottle mosaic virus quick-detecting box is more of value to preservation and the use of product, during detection, fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter fully contacts with detection sample, more contribute to detecting of positive, and time-proven there is very high sensitivity.
For the cucumber green mottle mosaic virus fast detecting under applicable different situations, cucumber green mottle mosaic virus quick-detecting box of the present invention can be prepared as various concrete embodiments.
For realizing nonexpondable object, one of embodiment of cucumber green mottle mosaic virus quick-detecting box of the present invention is in described box body, reagent bottle to be set to take in position, and described reagent bottle is taken in position and placed the brown reagent bottle of having taken in fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter.In reagent bottle, pour in advance and can supply repeatedly fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter reactant liquor of use amount.During use, the reactant liquor to pouring one-time detection amount in reactive tank into, can carry out subsequent survey.For fear of the inactivation of described fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter, the cucumber green mottle mosaic virus quick-detecting box under this embodiment need to be preserved in 4 ℃ of degree.
For being applied to a plurality of sample detection objects, another embodiment of cucumber green mottle mosaic virus quick-detecting box of the present invention is in described detection box, to configure tool hole reaction plate, and described tool hole reaction plate is provided with for holding the blind hole of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter.As preferably, described tool hole reaction plate matches with box body reactive tank bulk shape, and tool hole reaction plate is provided with the blind hole of a plurality of aperture 0.5～1.0cm, the degree of depth 0.5～1.0cm.Described fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter can adopt the reagent bottle mode of addressing to preserve above, joins in the blind hole of tool hole reaction plate during use.More preferably, be the fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter that adsorbs in advance freeze-drying in blind hole.In testing process, tool hole reaction plate is taken out and to be positioned in reactive tank, and using each blind hole as one independently developing tank carry out application of sample detection.Can realize simultaneously to the detection to a plurality of samples.
Above-mentioned each technical characterictic also can be used in combination in the design of cucumber green mottle mosaic virus quick-detecting box, its technical scheme combining also should the present invention prolong and scope in.
In addition, for realizing the goal of the invention of field quick detection cucumber green mottle mosaic virus of the present invention, it can also be the common test paper form in this area by its design and assembly, the invention provides a kind of concrete test paper mentality of designing is: a kind of cucumber green mottle mosaic virus quick detection test paper, by base plate be attached on base plate the sample pad of close proximity, pad, chromatographic film and adsorptive pads successively and form, it is characterized in that being embedded with fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter on described pad; Described chromatographic film is provided with detection zone and Quality Control district, and monoclonal antibody or the polyclonal antibody with cucumber green mottle mosaic virus generation specific reaction fixed in detection zone.
The clean polybag that cucumber green mottle mosaic virus quick detection test paper described in the invention described above can be used with detection sample extraction is assembled into kit and preserves, and to tackle one or many, detects use.
Detection zone described in cucumber green mottle mosaic virus quick-detecting box described in the arbitrary technical scheme of the invention described above or cucumber green mottle mosaic virus quick detection test paper fix with the preferred monoclonal antibody of cucumber green mottle mosaic virus generation specific reaction antibody.Described monoclonal antibody can, according to the monoclonal antibody preparation method preparation of recording in prior art, also can be bought commodity monoclonal antibody.Described detection zone can adopt the spot printing legal system of recording in prior art standby: the detection zone envelope antigen on nitrocellulose filter using the polyclonal antibody with cucumber green mottle mosaic virus generation specific reaction or monoclonal antibody, by the envelope antigen of debita spissitudo, with some film instrument, the amount with 1 μ L/cm is sprayed on the nitrocellulose filter sticking on PVC base plate.Then be placed in baking 60min in dry 37 ℃ of airtight vacuum drying ovens, after taking out, dry environment saves backup.
Quality Control district described in cucumber green mottle mosaic virus quick-detecting box described in the arbitrary technical scheme of the invention described above or cucumber green mottle mosaic virus quick detection test paper is fixed with sheep anti-mouse igg.Described Quality Control district can adopt the spot printing legal system of recording in prior art standby: by the sheep anti-mouse igg of debita spissitudo, with some film instrument, the amount with 1 μ L/cm is sprayed on the nitrocellulose filter sticking on PVC base plate.Then be placed in baking 60min in dry 37 ℃ of airtight vacuum drying ovens, after taking out, dry environment saves backup.
Use detection box of the present invention or Test paper can complete in the utmost point short time detection of plant cucumber green mottle mosaic virus.Guaranteeing, under good specificity and sensitivity prerequisite, can within 10min, to draw testing result, greatly faster than traditional PCR(1～2 hour), ELISA(1～2 hour) etc. detection method, improved detection efficiency.The present invention had both been applicable to the detection of single duplicate samples, was also applicable to the fast detecting of batch samples; And less demanding to operator and instrument and equipment, be that a kind of desirable cucumber green mottle mosaic virus infects fast detecting tool.
Accompanying drawing explanation
Accompanying drawing 4 width of the present invention, wherein,
Fig. 1 is cucumber green mottle mosaic virus quick-detecting box structural representation of the present invention, and 1 (a) is lid structural representation, and 1 (b) is box body structure schematic diagram;
Fig. 2 is the structural representation of the test strips in cucumber green mottle mosaic virus quick-detecting box of the present invention;
Fig. 3 is the tool hole reaction plate structural representation in cucumber green mottle mosaic virus quick-detecting box of the present invention;
Fig. 4 is cucumber green mottle mosaic virus quick detection test paper structural representation of the present invention;
In accompanying drawing: 1, lid; 2, box body; 3, test strips is taken in position; 4, reactive tank; 5, rare-earth fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter; 6, base plate; 7, sample pad; 8, chromatographic film; 9: adsorptive pads; 81, detection zone; 82, Quality Control district; 10, pad; 11, tool hole reaction plate; 12, blind hole.
Embodiment
Following non-limiting example can make the present invention of those of ordinary skill in the art's comprehend, but does not limit the present invention in any way.If no special instructions, of the present invention and
Bovine serum albumin(BSA) (BSA) is purchased from the raw work in Shanghai.Nitrocellulose filter, glass fibre element film are purchased from U.S. Millipore company.It is pure that agents useful for same is analysis.40nm collaurum for contrast test is prepared ([J] .Nat Phys.Sci, 1973,241 (105): 20) preparation with reference to Frens method.Cucumber green mottle mosaic virus infects positive to be provided by Liaoning Province Entry-Exit Inspection and Quarantine Bureau, through RT-PCR(standard GB/T/T28071-2011) method identifies and is defined as positive.
Embodiment 1. prepares cucumber green mottle mosaic virus quick-detecting box and cucumber green mottle mosaic virus quick detection test paper
1, the preparation of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter reactant liquor
Get fluorescent grain (purchased from Shenzhen City Yirui Bioisystech Co., Ltd) 50 μ L(10mg/ml), add 550 μ L50mM MES(pH6.0), the sulfo-NHS of 100mg/mL and each 200 μ L of EDC, room temperature is shaken 30min; Centrifugal, abandon supernatant; MES(pH6.0 with 1mL50mM) resuspended particle; Add appropriate cucumber green mottle mosaic virus monoclonal antibody (purchased from Beijing blue Bloomsbury Bioisystech Co., Ltd), room temperature jolting 1h; Add isopyknic PBS-BSA(BSA content 2%, pH7.4), continue reaction 1h; Centrifugal, abandon supernatant; Use 50mM MES(pH6.0) washing granule 3 times, 1mL/ time; With the resuspended liquid of 300 μ L (10mM Tris-HCl(pH8.0), 10% sucrose) resuspended particle, 4 ℃ of preservations, standby.
Fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter reactant liquor of preparing gained is processed one of in the following manner:
A. be independently stored in brown reagent bottle, and be assembled in cucumber green mottle mosaic virus quick-detecting box;
B. on the tool hole reaction plate of preparing at polystyrene, adsorb fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter, the method for absorption can be used freeze-drying absorption very ripe in prior art.As a reference, the freeze drying technology parameter that the present invention uses mainly comprises (pilot scale type freeze dryer, German CHRIST, EPSILON2-6D):
1.5 ℃/min of chilling rate (Cooling Rate);
-1 ℃ of early stage cold point (Incipient Freezing Point), 15min;
-40 ℃ of chilling temperatures (Cooling temperature);
-27 ℃ of eutectic points (Melting Point Eutectic temperature);
20 ℃ of heating-up temperatures (Heating temperature), 120min.
C. be carried on the pad of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter of having prepared load on nitrocellulose filter.
2, the preparation of chromatographic film
1) detection zone: take the detection zone envelope antigen of cucumber green mottle mosaic virus polyclonal antibody (purchased from Beijing blue Bloomsbury Bioisystech Co., Ltd) on nitrocellulose filter.With some film instrument, the amount with 1 μ L/cm is sprayed on the nitrocellulose filter sticking on PVC base plate the envelope antigen that is 1mg/mL by concentration.Be placed in baking 60min in dry 37 ℃ of airtight vacuum drying ovens, after taking-up, be put in drying basin stand-by.
2) Quality Control district: with the coated nitrocellulose filter of sheep anti-mouse igg (purchased from Shenzhen City Yirui Bioisystech Co., Ltd), as Quality Control district.With some film instrument, the amount with 1 μ L/cm is sprayed on the nitrocellulose filter sticking on PVC base plate the sheep anti-mouse igg that is 0.5mg/mL by concentration.Be placed in baking 60min in dry 37 ℃ of airtight vacuum drying ovens, after taking-up, be put in drying basin stand-by.
3, the assembling of cucumber green mottle mosaic virus quick-detecting box
As accompanying drawing 1 shows, cucumber green mottle mosaic virus quick-detecting box comprises lid 1, box body 2 and test strips 3, on lid 1, be provided for taking in the reactive tank 4 of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter 5, box body 2 is interior to be arranged test strips and takes in position 3, and test strips is taken in position 3 can take in one or multiple test strips.
Described test strips 3 is cucumber green mottle mosaic virus Rapid detection test strips, and individual test strips is shown as accompanying drawing 2: closely sample pad 7, chromatographic film 8 and the adsorptive pads 9 of overlap joint successively on base plate 6; Wherein, described chromatographic film 8 is provided with 82， detection zone, 81He Quality Control district, detection zone 81 and is fixed with the monoclonal antibody with cucumber green mottle mosaic virus generation specific reaction; Quality Control district 82 is fixed with sheep anti-mouse igg.After assembling, cut into the wide strip of 5mm, can be accommodated in box body 2.
Under non-detection status, described fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter is stored in brown reagent bottle with the form of reactant liquor, and this reagent bottle can be positioned in box body 2.The detection box that assembling obtains needs 4 ℃ of degree to preserve.
Can adopt polystyrene preparation as the tool hole reaction plate 11 of accompanying drawing 3, which is provided with blind hole 12 simultaneously.Described tool hole reaction plate 11 profiles and reactive tank 4 match, blind hole 12 aperture 0.5cm on the reaction plate 11 of tool hole, blind hole 12 degree of depth 0.5cm.Adopt method freeze-drying load fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter in blind hole 12 of the present embodiment 1b.The described tool hole reaction plate 11 of suitable number is also accommodated in the interior appropriate location of box body 2.During detection, test strips can be done to suitable cutting to be applicable to take the testing process that blind hole 12 is developing tank.
4, the assembling of cucumber green mottle mosaic virus quick detection test paper
As accompanying drawing 4 shows, the sample pad 7, pad 10, chromatographic film 8 and the adsorptive pads 9 that on base plate 6, closely overlap successively; Wherein, on described pad 10, be embedded with fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter; Described chromatographic film 8 is provided with 82， detection zone, 81He Quality Control district, detection zone 81 and is fixed with the monoclonal antibody with cucumber green mottle mosaic virus generation specific reaction; Quality Control district 82 is fixed with sheep anti-mouse igg.
After assembling, cut into the wide strip of 5mm, obtain cucumber green mottle mosaic virus quick detection test paper of the present invention, 4 ℃ of kept dry.
1, detect sample preparation
Detect sample preparation: get 1g detected sample blade in clean polybag, add 3ml PBST(1 * PBS, 0.5%Tween20, pH7.4), rub the blade in polybag, taking juice detects for detecting sample.
2, use cucumber green mottle mosaic virus quick-detecting box to detect sample
Prepared detection sample is added in the reactive tank 12 of the prepared cucumber green mottle mosaic virus quick-detecting box of embodiment 1.3, adds fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter reactant liquor, piping and druming mixes; Insert test strips, hatch 3min for 40 ℃, visual inspection tomographic results under ultraviolet light after 5min.
As apparatus hole reaction plate is detected, and directly by detecting sample, being added in the blind hole 12 of tool hole reaction plate 11, piping and druming mixes; Insert test strips, hatch 3min for 40 ℃, visual inspection tomographic results under ultraviolet light after 5min.
3, use cucumber green mottle mosaic virus quick detection test paper to detect sample
The sample pad end of the prepared cucumber green mottle mosaic virus quick detection test paper of embodiment 1.4 is inserted and detects sample, 5min, visual inspection tomographic results under ultraviolet light.
4, result is judged
No matter use cucumber green mottle mosaic virus quick-detecting box or cucumber green mottle mosaic virus quick detection test paper to detect sample, the criterion of result is consistent: all there is fluorescence display in 81He Quality Control district, detection zone 82, testing result is positive, and show sample is infected by cucumber green mottle mosaic virus; Detection zone 81 have fluorescence display and Quality Control district 82 without fluorescence display, testing result is negative, show sample is not infected by cucumber green mottle mosaic virus; 81He Quality Control district, detection zone 82 is all without fluorescence display, and test paper cancels, and need again detect.
5, detection specificity and stability test
Use cucumber green mottle mosaic virus quick-detecting box to detect little cucurbita pepo mosaic virus (ZYMV), cucumber mosaic virus (CMV), tobacco mosaic virus (TMV) (TMV), PRSV (PRSV), pumpkin mosaic virus (SqMV), nepovirus (TRSV), muskmelon necrotic spot virus (MNSV) and cucumber green mottle mosaic virus (CGMMV) positive sample (above virus is all purchased from China Inst. of Quarantine Inspection Sciences), equal no cross reaction, cucumber green mottle mosaic virus quick-detecting box application specific of the present invention is good.
By the test strips in cucumber green mottle mosaic virus quick-detecting box, rare-earth fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter, and extract (1 * PBST) be placed on 37 ℃ 7 days, take out the positive that detects variable concentrations, observe its stability.Experimental result confirms, not obviously reduction of sensitivity under test condition.
The contrast test of embodiment 3. and colloidal gold immunity chromatography
Reference literature " development of pumpkin mosaic virus colloidal gold immuno-chromatography test paper strip " ([J]. plant quarantine, 2012,38 (5): method 88-91.) is prepared monoclonal antibody-collaurum conjugate, and with reference to the method for embodiment 1, prepare cucumber green mottle mosaic virus colloidal gold immunochromatographimethod and detect box and Test paper.Method with reference to embodiment 2 is carried out colloidal gold immunity chromatography detection.
1, detection sensitivity comparison test
Use embodiment 2.2(to detect box, reagent bottle) the relatively prepared cucumber green mottle mosaic virus Rapid detection test strip of the embodiment of the present invention 1.3 and the sensitivity of cucumber green mottle mosaic virus colloidal gold immunochromatographydetection detection test paper bar of test form.
With PBST doubling dilution, with a positive sample, by colloidal gold immunochromatographimethod method and method of the present invention, detect respectively.Sentence read result as shown in the table (in table+represent positive ,-represent negative).The sensitivity of fluorescent nano particle test strips will be higher than the sensitivity of colloidal gold strip as can be seen from the table.
| ? | 4× | 8× | 16× | 32× | 64× |
| Colloidal gold strip | + | + | - | - | - |
| Fluorescent nano particle test strips (naked eyes) | + | + | + | + | - |
Claims (10)
1. a cucumber green mottle mosaic virus quick-detecting box, comprises lid, box body and test strips, is provided for taking in the reactive tank of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter on lid, in box body, test strips is set and takes in position;
Described test strips is cucumber green mottle mosaic virus Rapid detection test strip, by base plate be attached to that on base plate, the sample pad of close proximity, chromatographic film and adsorptive pads form successively, described chromatographic film is provided with detection zone and Quality Control district, and monoclonal antibody or the polyclonal antibody with cucumber green mottle mosaic virus generation specific reaction fixed in detection zone.
2. cucumber green mottle mosaic virus quick-detecting box claimed in claim 1, it is characterized in that in described box body, being also provided with reagent bottle takes in position, described reagent bottle is taken in position and is placed the brown reagent bottle of having taken in fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter.
3. cucumber green mottle mosaic virus quick-detecting box claimed in claim 2, is characterized in that detecting 4 ℃ of degree of box and preserves.
4. cucumber green mottle mosaic virus quick-detecting box claimed in claim 1, characterized by further comprising tool hole reaction plate, and described tool hole reaction plate is provided with for taking in the blind hole of fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter.
5. cucumber green mottle mosaic virus quick-detecting box claimed in claim 4, is characterized in that described tool hole reaction plate profile and box body reactive tank match, blind hole aperture 0.5～1.0cm on tool orifice plate, blind hole depth 0.5～1.0cm.
6. cucumber green mottle mosaic virus quick-detecting box claimed in claim 5, is characterized in that described tool hole reaction plate made by polystyrene, takes in fluorescent nano particle-cucumber green mottle mosaic virus antibody coupling matter of freeze-drying in blind hole.
7. the cucumber green mottle mosaic virus quick-detecting box described in arbitrary claim in right 1～6, is characterized in that described detection zone is fixed with the monoclonal antibody with cucumber green mottle mosaic virus generation specific reaction.
8. the cucumber green mottle mosaic virus quick-detecting box described in arbitrary claim in right 1～6, is characterized in that described Quality Control district is fixed with sheep anti-mouse igg.
9. the cucumber green mottle mosaic virus quick-detecting box described in arbitrary claim in right 1～6, it is characterized in that described fluorescent nano particle is rare-earth fluorescent nano particle, the chelate that it contains rare earth ion and sequestrant formation, described chelate can be launched characteristic fluorescence under ultraviolet excitation; Described rare earth ion is Eu
3+, Sm
3+, Dy
3+or Tb
3+.
10. the cucumber green mottle mosaic virus quick-detecting box described in right 9, is characterized in that described rare-earth fluorescent nano particle diameter 10～10000nm.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105181692A (en) * | 2015-09-30 | 2015-12-23 | 山东博科生物产业有限公司 | Colorimetry detection kit for IMA (ischemia modified albumin) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101368217A (en) * | 2008-09-18 | 2009-02-18 | 中国农业科学院植物保护研究所 | Fast detecting method for cucumber green mottle mosaic virus |
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| CN101368217A (en) * | 2008-09-18 | 2009-02-18 | 中国农业科学院植物保护研究所 | Fast detecting method for cucumber green mottle mosaic virus |
| CN202002935U (en) * | 2011-03-21 | 2011-10-05 | 武汉大学 | Kit for detecting staphylococcus aureus infection in blood |
| CN102305854A (en) * | 2011-07-20 | 2012-01-04 | 汤凌霄 | Ultrasensitive and quantitative immunochromatographic device and detection method using same |
| CN102539749A (en) * | 2012-02-16 | 2012-07-04 | 江阴康奈尔生物科技有限公司 | Field high-sensitivity immunoassay method and kit |
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| CN105181692A (en) * | 2015-09-30 | 2015-12-23 | 山东博科生物产业有限公司 | Colorimetry detection kit for IMA (ischemia modified albumin) |
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