CN205404586U - Hand -held type chrysanthemum ester class pesticide immunochromatographic test strip - Google Patents
Hand -held type chrysanthemum ester class pesticide immunochromatographic test strip Download PDFInfo
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- CN205404586U CN205404586U CN201620151305.7U CN201620151305U CN205404586U CN 205404586 U CN205404586 U CN 205404586U CN 201620151305 U CN201620151305 U CN 201620151305U CN 205404586 U CN205404586 U CN 205404586U
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Abstract
The utility model discloses a hand -held type chrysanthemum ester class pesticide immunochromatographic test strip, including bar PVC bottom plate, its characterized in that: bar PVC bottom plate from top to bottom in proper order the overlap paste gold mark pad, the nitrocellulose membranes and the pad that absorbs water that has two anti complexes of gold mark goat -anti mouse, the overlap zone length is 1mm between them, nitrocellulose membranes on be equipped with detection line and matter accuse line, wherein, the immunogen of spraying on the detection line is hapten and carrier protein's conjugate, the anti goat igG antibody of spraying rabbit on the matter accuse line, the utility model discloses can detect the first cyanogen chrysanthemum ester, bromine cyanogen chrysanthemum ester, cypermethrin, trifluoro cypermethrin pesticide residue simultaneously. Its testing result repeatability tradition gold mark test paper strip better, more stable, adopt handheld handle to has got rid of the sample pad, has made the structure simpler, operate safer, swift, to first of the ten heavenly stems cyanogen chrysanthemum ester, bromine cyanogen chrysanthemum ester, cypermethrin, the sensitivity of trifluoro cypermethrin residue detection can reach 0.5, 5.0, 5.0, 5.0mgL.
Description
Technical field
This utility model relates to a kind of test strips, in particular for the hand-held test strips of the rapid screening detection of pyrethrin pesticide in vegetable and fruit, belonging to gold-marking immunity detection field.
Background technology
Pyrethrin pesticide as a kind of Insecticidal and acaricidal agent, has efficiently, feature that low toxicity, action of contace poison are strong, is widely used in the production estimation such as veterinary antibiotics, Folium Camelliae sinensis.But its residual period is long, be there is harm risk in Environment-Ecosystem and human health.Pyrethroid pesticide residual on vegetables and fruits has been formulated strict limit standard by the international bodies such as the World Health Organization (WHO) and FAO (Food and Agriculture Organization of the United Nation) (FAO), and limit standard (GB2763-2014) also formulated by such pesticide by China.
The detection method of the pyrethrin pesticide of report, mainly includes the method such as instrument analytical method, immunochemistry at present.But, although these detection techniques have higher detection sensitivity, length consuming time, complicated operation, even need the training of specialty, need at experiment in-house operation, it is impossible to meet on-the-spot quick, high throughput testing, particularly when a large amount of samples are carried out low cost rapid screening, advantage is not enough.
Colloidal gold immunochromatographimethod technology is the fast simple immunoassay technology of one grown up in the 80-90 age in 20th century, has the advantages such as quick, easy, easy judgement.nullIt is to be fixed on nitrocellulose filter by specific antigen，Colloidal gold labeled monoclonal antibody is adsorbed on gold mark pad，When sample liquid to be checked touches test strips gold mark pad end，Moved up by capillarity，Detection thing and the colloidal gold labeled monoclonal antibody reaction bonded on gold mark pad，Move again to detecting band (immunogenic region that spraying is fixing)，Now fixing immunogen and thing competition binding antibody to be checked，The golden labeling antibody not being combined with determinand and immunogen occur specific binding and are trapped，It is gathered on detection band，Thus producing color change，When antibody arrives quality control band (the second antibody region that spraying is fixing) with determinand in conjunction with complex or free antibody，Second antibody and first antibody occur specific binding and are trapped within detection band，Colloidal gold aggregation presents redness，The color change of perusal detection band can be passed through subsequently，Carry out positive judgement.This utility model adopts the method that colloid gold label sheep anti mouse two is anti-, and traditional colloidal gold immunochromatographimethod technology has been improved, and structure is simpler, and improves repeatability and the stability of testing result.
Utility model content
This utility model provides test strips preparation and the application process of a kind of quick diagnosis and half-quantitative detection pyrethrin pesticide residual, can meet the requirement of on-site quick screening.
The purpose of this utility model is achieved in that a kind of hand-held pyrethrin pesticide immunochromatographydetecting detecting test strip.The gold anti-including the sheep anti-Mouse two containing colloid gold label marks pad, nitrocellulose filter, adsorptive pads and PVC base plate；Adsorptive pads, nitrocellulose filter, gold mark pad containing the gold mark anti-complex of sheep anti-Mouse two are from top to bottom overlapping successively to be pasted on PVC base plate；Being sequentially provided with on nitrocellulose filter and detect line and nature controlling line, wherein close gold mark pad is detect line.Sample pad in tradition gold label test strip is removed by this utility model, makes structure more simple and practical, and operates faster, safety.
In this utility model, described gold mark pad long 12mm, wide 3.5mm；The long 25mm of described nitrocellulose filter, wide 3.5mm；The long 40mm of described adsorptive pads, wide 3.5mm；Gold mark pad, nitrocellulose filter, adsorptive pads each overlapping region length be 1mm.
Wherein, the conjugate of detection wire spraying hapten and ovalbumin；Nature controlling line spraying rabbit anti goat igg antibody, the distance of detection line and nature controlling line is 4mm.
The principle of the hand-held pyrethrin pesticide test strips in this utility model is A competitive inhibition method, when in measuring samples without Fenpropathrin, decis, cypermethrin, during Grenade (ICI). pesticide, the monoclonal antibody of pyrethrin pesticide is added after sample liquid mixes, the sheep anti-Mouse two upwards migrating the colloid gold label dissolved on gold mark pad with capillarity resists, formation complex in combination, after continuing up migration arrival detection line, coated immunogen is combined on detection line, the complex that gold mark disome and pyrethrin pesticide monoclonal antibody are formed deposition on detection line, form macroscopic redness, excessive gold mark two complex that is anti-and that formed with monoclonal antibody continues up migration, be combined with the anti-goat antibody of the rabbit being coated on nature controlling line, the macroscopic redness of gold colloidal formation of deposits.Now detection line color will be equal to or be deeper than nature controlling line, testing result be feminine gender.
When in measuring samples containing pyrethrin pesticide, the pyrethrin pesticide monoclonal antibody added is first specific binding with the pesticide in sample liquid, prevent itself and immunogenic combination on detection line, therefore the complex that its sheep anti mouse two anti-binding with colloid gold label is formed cannot form precipitation on detection line, now in sample, pyrethrin pesticide content is more high, detection line color is more shallow even colourless, gold mark two is anti-subsequently and complex continues up and moves to nature controlling line, be combined with the anti-goat antibody of the rabbit being coated on nature controlling line, form macroscopic redness.Now detection line color is shallower than nature controlling line or close to colourless, and testing result is positive.
When nature controlling line does not have color, no matter detect whether line has color, be accordingly to be regarded as null result.
Innovation of the present utility model is in that: the sample pad of traditional gold label test strip removed, and is sprayed at by gold target antibody on golden mark pad, it is easier to standardization control, and the migration of gold mark complex is more clean, thorough.
Hand-held pyrethrin pesticide immuno-chromatographic test paper strip described in the utility model compared with prior art, has the advantage that
(1) the direct labelling sheep anti-Mouse two of gold colloidal is adopted to resist, the method utilizing antibody complex indirect labelling pesticide monoclonal antibody, improve the sensitivity of detection, accuracy rate and stability.
(2) the detection time only needs 8-10min, it is possible to meet the requirement of Site Detection.
(3) adsorptive pads is up to 40mm, makes hand-held safer.
(4) easy to carry, simple to operate, it is not necessary to professional training all can detect immediately.
Hand-held pyrethrin pesticide immunochromatographydetecting detecting test strip described in the utility model is the residual of 4 kinds of pyrethrin pesticides suitable in quickly detection plurality of vegetables and fruit, has broad spectrum activity.
Accompanying drawing explanation
Fig. 1 is the structural representation of this utility model embodiment.
In figure, 1, gold mark pad, 2, nitrocellulose filter, 3, adsorptive pads, 4, detection line, 5, nature controlling line, 6, PVC base plate, 7, the gold mark anti-complex of sheep anti-Mouse two.
Detailed description of the invention
The nonrestrictive concrete structure disclosing this utility model embodiment of accompanying drawing and using method thereof, below in conjunction with accompanying drawing, the utility model will be further described
Embodiment 1
A kind of hand-held pyrethrin pesticide immunochromatographydetecting detecting test strip, as seen from Figure 1, test strips is sized to 75mm × 3.5mm, including containing gold mark the gold mark pad 1 of the anti-complex 7 of sheep anti-Mouse two, nitrocellulose filter 2, adsorptive pads 3, PVC base plate 6.Gold mark pad long 12mm, wide 3.5mm；The long 25mm of nitrocellulose filter, wide 3.5mm；The long 40mm of described adsorptive pads, wide 3.5mm；Gold mark pad, nitrocellulose filter, adsorptive pads each overlapping region length be 1mm, the actual size of test strips finished product is 75mm × 3.5mm.
Detection line 4 and nature controlling line 5 test distance respectively 11mm and 15mm of starting point end with nitrocellulose filter 2, and the distance between detection line 4 and nature controlling line 5 is 4mm.Gold mark pad 1 and nitrocellulose filter 2 containing the gold mark anti-complex 7 of sheep anti-Mouse two are from bottom to top overlapping with on PVC base plate 6 successively, and nitrocellulose filter 2 other end overlaps with adsorptive pads 3.Detecting line 4 on nitrocellulose filter and spray immunogen (conjugate of hapten and carrier protein), concentration is 0.3 μ g/mm, and nature controlling line 5 sprays the anti-goat polyclonal antibodies of rabbit, and concentration is 0.06 μ g/mm.
The each Material texture related in the present embodiment is as follows: base plate is PVC material (SM31-40), and gold mark pad is glass fibre, and model is MilliporeGFCP000800；Nitrocellulose filter model is Millipore135；Adsorptive pads model is SX27.Gold mark pad processes: gold mark pad immerses gold mark pad treatment fluid (20mMNa3PO412H2O, 5%BSA, 0.25%Tween-20,10% sucrose) after 30min, 37 DEG C of drying.
Embodiment 2
The preparation method of hand-held pyrethrin pesticide immunochromatographydetecting detecting test strip, its preparation process is as follows:
The first step, adopts citric acid reducing process to prepare gold colloidal:
(1) 200mL distilled water is joined in 500mL wide mouth glass bottle, add 1mL and cross 2% chlorauric acid solution of 0.22 μm of filter membrane, rock gently to mixing clockwise, be positioned in microwave oven high fire heating 5min to boiling.
(2) vial is taken out, rock vial gently clockwise rapidly, make in bottle, after solution rotating, to rapidly join 4.8mL and cross 1% citric acid three sodium solution of 0.22 μm of filter membrane, continue in the same direction to rock vial 5s gently, being placed again in microwave oven, high fire continues heating 4min.
(3) taking out vial, now in bottle, claret liquid is required colloidal gold solution；After putting room temperature cooling, again it is settled to 200mL with distilled water, lucifuge 4 DEG C preservation after mixing.
Second step, the preparation of gold mark goat anti-mouse antibody:
(1) every milliliter of nano-Au solution adds the K of 7 μ L2CO3(0.1mol/L) solution regulates pH value is 8.0, is slowly added to 5 μ L1mg/mL goat anti-mouse antibody solution, now the final concentration of 5 μ g antibody/mL nano-Au solution of its antibody, stirs 30min under stirring；
(2) adding 10%BSA solution and make BSA final concentration of 1%, after continuing stirring 30min, 1500rpm is centrifuged 15min, leaves and takes supernatant, after supernatant continuation is centrifuged 35min with 12000rpm, and supernatant discarded, add washing and preserve liquid (0.01% boric acid, 0.2%PEG20000, 0.02%NaN3) to original solution volume；
(3) again with after the centrifugal 35min of 12000rpm, supernatant discarded, addition washing preserves liquid dissolution precipitation to volume is original solution volume 1/10, obtains golden mark sheep anti-Mouse solution, and 4 DEG C keep in Dark Place.
3rd step, the preparation of colloidal gold test paper film:
Spray film with the Arijet in Bio-dotXYZ-3050 three-dimensional specking instrument, immunogen and rabbit anti goat igg antibody are sprayed respectively and is coated on NC film, as detection line and nature controlling line；Gold labeling antibody complex is sprayed on processed good gold mark pad by 0.002 μ g/mm；After 37 DEG C of dry 2h, 28 DEG C are stored in drying basin.
4th step, the assembling of test strips:
NC film is affixed on the middle part of PVC base plate, and gold mark pad is affixed on NC film detection line one end, and adsorptive pads is affixed on nature controlling line one end, and overlap each other 1mm compressing.With cutting cutter, the test paper plate assembled is cut into the test strips of 3.5mm, is stored in the drying basin of 28 DEG C.
Embodiment 3
Detection method:
The pyrethrin pesticide solution of 100 μ g/mL is prepared with methanol solution, 20,10,5,2,1,0.5,0.2,0.1 μ g/mL it is diluted to respectively again by the PBS solution containing 20% methanol, set up the PBS solution containing 20% methanol as a control group simultaneously, 100 μ L above-mentioned concentration pyrethrin pesticide standard specimen solution and comparison buffer it is separately added in micropore, add 0.5 μ L pyrethrin pesticide monoclonal antibody (1mg/mL), the gold mark pad end of test strips is inserted in micropore, 8-10min observed result.
Result judges:
Positive findings: detection line 4 does not develop the color or color is substantially shallower than the color of nature controlling line 5, if not developing the color, Fenpropathrin, cypermethrin, decis, Grenade (ICI). pesticide residual concentration be respectively greater than equal to 0.5,5.0,5.0,5.0mg/kg, if detection line color relatively nature controlling line shoals, show that sample is still for the positive, the residual concentration of pesticide is then respectively smaller than 0.5,5.0,5.0,5.0mg/kg；Negative findings: detection line 4 color is basically identical with nature controlling line 5 color or is deeper than nature controlling line 5；When nature controlling line 5 does not develop the color, then test strips lost efficacy.
Embodiment 4
The sample substrate impact on test strips
Test sample: Bulbus Allii Cepae, Herba Alii fistulosi, Rhizoma Zingiberis Recens, Bulbus Allii, Folium Allii tuberosi, Chinese cabbage, Xiaoqinling Nature Reserve, Fructus Capsici, Radix Dauci Sativae, Fructus Lycopersici esculenti, Fructus Cucumidis sativi, Fructus Mali pumilae, Citrullus vulgaris.
Sample pre-treatments: weigh 10g and pulverize (homogenate or chopping) sample, adds 10mL acetonitrile concussion 30min, filters, and filtrate is poured into added with the anhydrous MgSO of 1gNaCl and 4g4Centrifuge tube in, after concussion 1min, take 1mL supernatant nitrogen and dry up, for ELISA test strip after redissolving with 1mL20% methanol-PBS.
Detection: take the treatment fluid of 13 kinds of samples respectively, set up the PBS solution containing 20% methanol as a control group simultaneously, the 100 above-mentioned sample treatment liquid of μ L and comparisons it are separately added in micropore, add 0.5 μ L pyrethrin pesticide monoclonal antibody, the gold mark pad end of test strips is inserted in micropore, stands 8-10min observed result.
Result judges:
(1) positive findings: detection line 4 does not develop the color or color is substantially shallower than the color of nature controlling line 5；Negative findings: detection line 4 color and nature controlling line 5 color is basically identical or is deeper than the color of nature controlling line 5；When nature controlling line 5 does not develop the color, then test strips lost efficacy.
(2) in the implementation case, Bulbus Allii Cepae, Herba Alii fistulosi, Rhizoma Zingiberis Recens, Bulbus Allii, Folium Allii tuberosi, Chinese cabbage, Xiaoqinling Nature Reserve, Radix Dauci Sativae, Fructus Lycopersici esculenti, Fructus Cucumidis sativi, Fructus Mali pumilae, 12 kinds of vegetable and fruit sample liquid of Citrullus vulgaris and the testing result as a control group of the PBS solution containing 20% methanol are feminine gender, and matrix interference effect is few；The sample liquid testing result of Fructus Capsici is false positive, and matrix interference is strong, and this test strips is not suitable for the detection of Fructus Capsici sample.
Embodiment 5
The test strips checking to positive Detection results
Utilize pyrethrin pesticide residual in the ELISA test strip 12 kinds of vegetable and fruits except Fructus Capsici: in 12 kinds of vegetable and fruits, be separately added into 50 μ L, 100 μ L pyrethrin pesticide standard specimens (100 μ g/mL), make interpolation concentration respectively 0.5 μ g/mg, 5 μ g/mg, after carrying out the sample handling processes of same embodiment 4, take 2mL organic facies nitrogen respectively to dry up, wherein 1mL with normal hexane redissolve after in gas chromatographic detection, another 1mL redissolves in ELISA test strip with 20% methanol-PBS, if repeatedly repeating.
The result:
For Folium Allii tuberosi sample: adding concentration is 0.5 μ g/mg, 5 μ g/mg, gas chromatographic detection result: Fenpropathrin is 0.49 μ g/mg, 5.02 μ g/mg, cypermethrin is 0.52 μ g/mg, 5.05 μ g/mg, decis is 0.48 μ g/mg, 5.09 μ g/mg, and Grenade (ICI). is 0.51 μ g/mg, 5.10 μ g/mg；ELISA test strip result is the positive, consistent with gas chromatographic detection result.The detection of 12 kinds of vegetable and fruits is obtained consistent results.
Claims (3)
1. a hand-held pyrethrin pesticide immuno-chromatographic test paper strip, including bar shaped PVC base plate, it is characterized in that: bar shaped PVC base plate is from top to bottom overlapping successively is pasted with the gold mark pad of the gold mark anti-complex of sheep anti-Mouse two, nitrocellulose filter and adsorptive pads, and between them, overlapping region length is 1mm；Described nitrocellulose filter is provided with detection line and nature controlling line, and wherein, on detection line, spraying immunogen is the conjugate of hapten and carrier protein, and nature controlling line sprays rabbit anti goat igg antibody.
2. hand-held pyrethrin pesticide immuno-chromatographic test paper strip according to claim 1, it is characterised in that: the width of bar shaped PVC base plate is 3.5mm, gold mark pad wide with bar shaped PVC base plate, nitrocellulose filter and adsorptive pads long 12mm, 25mm and 40mm respectively.
3. hand-held pyrethrin pesticide immuno-chromatographic test paper strip according to claim 1 and 2, it is characterised in that: the distance between detection line and nature controlling line on nitrocellulose filter is 4mm.
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| CN201620151305.7U CN205404586U (en) | 2016-02-29 | 2016-02-29 | Hand -held type chrysanthemum ester class pesticide immunochromatographic test strip |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470831A (en) * | 2019-07-09 | 2019-11-19 | 华南农业大学 | A kind of chemiluminescence immune analysis method and kit directly detecting tenuazonic acid based on hunchbacked source antibody |
| CN112129956A (en) * | 2020-09-02 | 2020-12-25 | 武汉生之源生物科技股份有限公司 | Colloidal gold labeling solution for colloidal gold immunochromatography, and preparation method and application thereof |
-
2016
- 2016-02-29 CN CN201620151305.7U patent/CN205404586U/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470831A (en) * | 2019-07-09 | 2019-11-19 | 华南农业大学 | A kind of chemiluminescence immune analysis method and kit directly detecting tenuazonic acid based on hunchbacked source antibody |
| CN112129956A (en) * | 2020-09-02 | 2020-12-25 | 武汉生之源生物科技股份有限公司 | Colloidal gold labeling solution for colloidal gold immunochromatography, and preparation method and application thereof |
| CN112129956B (en) * | 2020-09-02 | 2022-05-27 | 武汉生之源生物科技股份有限公司 | Colloidal gold labeling solution for colloidal gold immunochromatography, and preparation method and application thereof |
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