CN104297470B - A kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card and preparation method thereof and using method - Google Patents
A kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card and preparation method thereof and using method Download PDFInfo
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- CN104297470B CN104297470B CN201410571154.6A CN201410571154A CN104297470B CN 104297470 B CN104297470 B CN 104297470B CN 201410571154 A CN201410571154 A CN 201410571154A CN 104297470 B CN104297470 B CN 104297470B
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- G—PHYSICS
- 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
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Abstract
A kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card and preparation method thereof and using method, this quick measuring card includes that the interior test strips that gets stuck and get stuck, described test strips include base plate, adhere to sample pad, gold mark pad, reaction film and the adsorptive pads being sequentially connected on base plate；It is coated pyrethroid pesticide gold mark wide range antibody on gold mark pad, the detection line of reaction film is coated pyrethroid pesticide antibody, it is coated goat-anti rabbit two on the control line of reaction film to resist, wherein, described pyrethroid pesticide gold mark wide range antibody is to be coated with nanoscale gold chloride colloidal solid and obtain.Quick measuring card prepared by the present invention can detect at least 9 kinds of pyrethroid pesticides simultaneously, detection limit can reach 0.01mg/kg, can stably preserve more than 1.5 years at normal temperatures, high specificity, highly sensitive, good stability, and it is easy and simple to handle, quick, accurate, the detection time is short, can be widely used for the pyrethroid residual quickly detection in agricultural product and water environment and soil.
Description
Technical field
The invention belongs to pyrethroid pesticide remained detection technique field, be specifically related to the deinsectization of a kind of plan
Pyrethrin pesticide colloidal gold immunochromatographimethod quick measuring card and preparation method thereof and using method.
Background technology
Pyrethroid pesticide (Pyrethriod insecticids) is a natural bionical insecticide of class, has
The features such as deinsectization is fast, toxicity is low, use is safe.It is that agricultural products in China Pesticide Residues recall rate is higher
One of pharmacopoeia.Pyrethroid pesticide yield accounts for the 20% of insecticide yield at present, and usable floor area is about
Account for the 25% of whole insecticide usable floor area.Owing to the part pyrethroid pesticide remained phase is longer, right
Non-targeted bio-toxicity is relatively big, and has environmental endocrine disrupting effect, therefore, sets up quickly detection
Technology tool is of great significance.
At present, use for laboratory mostly is chromatographic process or capillary electrophoresis side in the method for detection pyrethroid
Method, the cycle length of this kind of method detection, cost are high, operation is the easiest, sample preparation complexity, nearly all
It is to lay particular emphasis on terminal detection, even if after obtaining testing result, city may have been flowed to by large quantities of detection materials
In field or consumer's hands.Due to supervision dispersion, sample dispersion, laboratory method is difficult in time quickly,
Detect from source and scene all sidedly.
Immune colloidal gold technique (Immune colloidal gold technique) is using gold colloidal as missing
Mark is applied to a kind of novel immunolabelling technique of antigen-antibody.Gold colloidal is by gold chloride
(HAuCl4) under the effects such as reducing agent such as white phosphorus, ascorbic acid, sodium citrate, polymerization becomes specific
The gold grain of size, and owing to electrostatic interaction becomes a kind of stable colloidal state, claim gold colloidal.Colloid
Gold is electronegative under weakly alkaline environment, can form firm combination with the positive charge group of protein molecule,
Owing to this combination is electrostatical binding, so not affecting the biological nature of protein.Colloidal gold-labeled method
Being the core technology of immunochromatography quick diagnosis, it is applied to antigen using gold colloidal as trace labelling thing and resists
Precursor reactant, its essence is that what the macromolecules such as protein were adsorbed to colloid gold particle surface is coated process.Root
According to some physical behaviors of gold colloidal, such as high cloud density, granular size, shape and color reaction,
Plus immunity and the biological nature of conjugate, thus make gold colloidal be widely used in immunology, histology,
The field such as pathology and cytobiology.Existing immunological detection method resists owing to not having preferable wide range
Body, sensitivity and the stability of detection are the highest, it is difficult to meet the needs of field quick detection.Utilize wide range
Antibody sets up the important development side that immune colloid gold quick measuring card is current Detecting Pesticide field
To.
At present, it is applied to existing immune colloidal gold technique detect pesticide residues, however it remains many is asked
Topic, e.g., poor specificity, poor stability, sensitivity is the highest, and cost is high, operates the easiest, therefore originally
Skilled person needs badly and improves existing Immuno gold technology, develop high specificity, highly sensitive,
Good stability, the reproducible new method detecting multiple pyrethroid pesticide.
Summary of the invention
It is an object of the invention to provide a kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card and
Preparation method and using method, overcome prior art and be difficult to field quick detection agricultural product, water ring
A difficult problem pyrethroid pesticide remained in border and soil, can be used for the scene of pyrethroid pesticide
Quick-detecting box examination.
For reaching above-mentioned purpose, technical scheme is as follows:
A kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card, including getting stuck and get stuck interior examination
Paper slip, described test strips includes base plate, adheres to the sample pad, gold mark pad, anti-being sequentially connected on base plate
Answer film and adsorptive pads；Described get stuck on have well and observation window；Described well is located at described in correspondence
Sample pad upper shell, corresponding described reaction film upper shell is located at by described observation window, on described reaction film
It is provided with detection line and nature controlling line, described gold mark pad is coated pyrethroid pesticide gold mark wide range antibody,
The detection line of reaction film is coated pyrethroid pesticide wide range antibody, the nature controlling line of reaction film wraps
Being resisted by goat-anti rabbit two, wherein, described pyrethroid pesticide gold mark wide range antibody is by pyrethroid
Class pesticide wide range antibody coupling is to 20～25nm gold chloride colloidal grain surface gained.
Further, described sample pad, gold mark the overlap padding, being provided with between reaction film and adsorptive pads 1～2mm
District；Described detection line and nature controlling line interval 0.5～1.0cm.
Described base plate is PVC board, and described reaction film is nitrocellulose filter.Described sample pad is by glass fibers
Dimension film or filter paper are made, and described absorbent paper is made up of filter paper.
Also, described pyrethroid pesticide wide range antibody is pyrethroid pesticide wide range Anti-TNF-α
Body.
The standard gold amount of described pyrethroid pesticide gold mark wide range antibody is 14～16 μ g/mL, on gold mark pad
The concentration of coated pyrethroid pesticide gold mark wide range antibody is 14～16 μ g/mL.
When pyrethroid pesticide wide range antibody coupling of the present invention is to gold chloride colloidal grain surface gained,
The pH value of gold chloride colloidal solid solution used is 8.0～8.5.
Again, the concentration detection line of described reaction film being coated pyrethroid pesticide wide range antibody is
0.8～1.4 μ g/mL.
The preparation method of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card of the present invention, including
Following steps:
1) sample pad, gold mark pad, reaction film and adsorptive pads are pasted onto successively on base plate and make test strips；
2) pyrethroid pesticide wide range antibody is coated with 20～25nm gold chloride colloidal solids, prepares
Protein concentration is 14～16 μ g/mL pyrethroid pesticide gold mark wide range antibody, is sprayed on gold mark pad,
It is dried；Again the pyrethroid pesticide wide range antibody that protein concentration is 0.8～1.4 μ g/mL is sprayed at instead
Answering the detection line position on film, 36～37 DEG C are dried；Then by goat-anti rabbit two anti-(affinitive layer purification) spray
The nature controlling line position being applied on reaction film, is prepared as quick measuring card in installing to get stuck by test strips after drying.
Further, the coated concrete grammar of pyrethroid pesticide wide range antibody nanogold particle is:
Take 20～25nm gold chloride colloidal solid solution in small test tube, centrifugal, go out supernatant, use equal-volume
Ultra-pure water suspends, and regulation pH value is 8.0～8.5, is slowly dripped by pyrethroid pesticide wide range antibody subsequently
Entering nanogold particle suspension, room temperature is shaken, then drips BSA, makes the final concentration of 0.1% (volume of BSA
Concentration), room temperature is shaken, to close on nanogold particle as the site combined, it is to avoid non-specific adsorption.
Centrifugal, remove unnecessary pyrethroid pesticide wide range antibody and BSA, ultra-pure water suspends, washing,
Complete the coupling of nanogold particle and pyrethroid pesticide wide range antibody, obtain pyrethroid agriculture
Medicine gold mark wide range antibody, wherein, standard gold amount is 14～16 μ g/mL.
The using method of a kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card of the present invention,
Comprise the steps:
1) pre-treatment of testing sample: if testing sample is water and plant tissue extracting solution or soil water extract
Deng liquid substance time, can directly take 50 μ l testing samples detection.If testing sample be plant leaf blade, stem,
During the solid matters such as seed, then need testing sample is carried out pretreatment, will grind by testing sample, add
Enter sample treatment liquid, after standing, take 50 μ l detections.Wherein, described sample treatment liquid is phosphate buffer
(including sodium phosphate buffer and kaliumphosphate buffer).
2) detection: being instilled by taken sample in well, sample migrates along sample pad under capillary action,
Room temperature effect 5～10min, observed result.
3) result judges:
Positive: nature controlling line, detection line all present red stripes, illustrate there is pyrethroid agriculture in sample
Medicine exists.
Negative: nature controlling line presents red stripes, detection line does not present red stripes, illustrates do not have in sample
Pyrethroid pesticide exists.
Lost efficacy: nature controlling line, detection line the most do not present red stripes or only have detection line present red stripes,
Illustrate that test strips lost efficacy.
Colloidal gold immunochromatographimethod quick measuring card of the present invention is carried out mainly for two type pyrethroid pesticides
Detection, the standard gold amount of pyrethroid pesticide gold mark wide range antibody is 14～16 μ g/mL, the upper bag of gold mark pad
The concentration of the pyrethroid pesticide gold mark wide range antibody of quilt is 14～16 μ g/mL, the inspection of described reaction film
The concentration being coated pyrethroid pesticide wide range antibody on survey line is 0.8～1.4 μ g/mL, at such bar
Under part, the sensitivity of reaction is the highest, and binding characteristic is best, and test strips is the most stable.The whole inspection of the present invention
Survey process completes in 5～10min, simple to operate, quick, and is not required to any instrument.
Beneficial effects of the present invention:
1) present invention select first pyrethroid pesticide wide range antibody for fast detecting pesticide residue,
Pyrethroid pesticide wide range antibody is by utmost highlighting and retaining pyrethroid pesticide general character
The molecule of structure is prepared as, and is greatly improved and is combined with multiple pyrethroid pesticide antigen molecule
Specificity.
2) present invention selects nanoscale gold chloride colloidal solid to be coated pyrethroid pesticide width first
Spectrum antibody, nano-scale particle coated antibody is better than conventional colloid gold particle and is coated, and nano-scale particle is coated and carries
The high electrostatic attraction with antibodies, does not affect function and the titer of antibody, meanwhile, nano-scale particle
The stability that traget antibody is combined with nitrocellulose filter increases substantially.
3) present invention selects goat-anti rabbit two to resist first, and this antibody titer is better than sheep anti-mouse antibody, simultaneously facilitates
A large amount of preparation and purification.
4) quick measuring card prepared by the present invention can detect at least 9 kinds of pyrethroid pesticides, detection limit simultaneously
Can reach 0.01mg/kg, can stably preserve more than 1.5 years at normal temperatures, high specificity, highly sensitive,
Good stability, and easy and simple to handle, quick, accurate, the detection time is short, can be widely used for agricultural product and
Pyrethroid residual quickly detection in water environment and soil.
Accompanying drawing explanation
Fig. 1 is the structural representation of embodiment of the present invention quick measuring card；
Fig. 2 is the side structure schematic diagram of embodiment of the present invention quick measuring card；
Fig. 3 is the structural representation that the embodiment of the present invention gets stuck；
Fig. 4 is that the testing result of the embodiment of the present invention is positive schematic diagram；
Fig. 5 is that the testing result of the embodiment of the present invention is negative schematic diagram；
Fig. 6 be the testing result of the embodiment of the present invention be one of inefficacy schematic diagram；
Fig. 7 be the testing result of the embodiment of the present invention be the two of inefficacy schematic diagram.
Detailed description of the invention:
Below in conjunction with specific embodiment, technical scheme is described in further detail.
Embodiment 1:
Seeing Fig. 1～7, a kind of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card, it includes card
Shell 1 and the test strips 2 got stuck in 1, wherein, described reagent paper 2 includes base plate 3, adheres on base plate 3
Sample pad 4, gold mark pad 5, reaction film 6 and the adsorptive pads 7 being sequentially connected.Described getting stuck 1 is provided with and adds
Sample hole 8 and observation window 9, described well 8 is located at corresponding described sample pad 4 upper shell, described sight
Examine window 9 and be located at corresponding described reaction film 6 upper shell.Described be sequentially connected sample pad 4, gold mark pad 5,
The duplicate block of 1～2mm it is provided with between reaction film 6 and adsorptive pads 7.Described base plate 3 is PVC board, institute
State reaction film 6 for nitrocellulose filter.Described sample pad 4 is made up of glass fibre or filter paper, described suction
Water cushion 7 is made up of filter paper.
In the present embodiment, gold mark pad 5 is coated pyrethroid wide range antibody-nanoscale colloidal gold conjugate.
Described reaction film 6 is provided with and is coated the detection line 61 of pyrethroid wide range antibody and is coated goat-anti
The nature controlling line 62 of rabbit two anti-igg, described detection line 61 and described nature controlling line 62 are spaced 0.5cm.
The preparation process of the present embodiment pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card is as follows:
1) coupling pyrethroid pesticide wide range antibody to the concrete grammar on nanogold particle surface is: this
The nanogold particle that embodiment uses is to be evaluated and testing laboratory by Academy of Agricultural Sciences, Shanghai City bio-safety
Synthesis, measures 10ml 20nm gold chloride colloidal solid solution in small test tube, centrifugal (10000rpm,
10min), going out supernatant, suspend with equal-volume ultra-pure water, regulation pH value is 8.0, subsequently by affine
(wide range antibody is by Shanghai City agricultural section for 0.3ml pyrethroid pesticide wide range antibody after chromatography purification
Prepared by institute's bio-safety evaluation and testing laboratory) slowly instill nanogold particle suspension, room temperature is shaken
2h, then drip 1%BSA, making final concentration of the 0.1% of BSA, room temperature concussion 2h, to close nanometer gold
For the site combined on grain, it is to avoid non-specific adsorption.Centrifugal (8000rpm, 10min), removing is many
Remaining pyrethroid pesticide wide range antibody and BSA, ultra-pure water suspends, and repeated washing 3 times completes to receive
Rice gold grain and the coupling of pyrethroid pesticide wide range antibody, the nanometer gold labeling antibody prepared can be deposited
It is placed on 4 DEG C of refrigerators standby.Wherein, the standard gold amount of nanometer gold labeling antibody is 16 μ g/mL.
2) 16 μ g/mL nanometer gold labeling antibodies of preparation are sprayed on gold mark pad, are dried；Again by 1.4 μ g/mL
The detection line position that pyrethroid pesticide antibody is sprayed on reaction film, 36-37 DEG C is dried；Then will
The 16 anti-nature controlling line positions being sprayed on reaction film of μ g/mL goat-anti rabbit two, install to get stuck by test strips after drying
Inside it is prepared as quick measuring card.
The use of the present embodiment colloidal gold immunochromatographimethod quick measuring card:
(1) pre-treatment of testing sample: if testing sample is water and plant tissue extracting solution or the soil water carries
During the liquid substances such as liquid, can directly take 50 μ l sample detection.If detected sample be plant leaf blade, stem,
During the samples such as seed, then need sample is carried out pretreatment, will grind by sample, add sample treatment liquid
(5Mm PBS pH 7.4), takes 50 μ l detections after standing.
(2) detection: being instilled in well 8 by taken sample, sample solution is under capillary action along sample
Product pad migrates, room temperature effect 5～10min, then observation window 9 observed result.
(3) result judges:
Positive: to detect line 61, nature controlling line 62, be red stripes, illustrate sample has pyrethroid
Pesticide exists, as shown in Figure 4.
Negative: detection line 61 occurs without red stripes, and nature controlling line 62 red stripes occurs, sample is described
In there is no pyrethroid pesticide, as shown in Figure 5.
Lost efficacy: detection line 61, nature controlling line 62 all occur without red stripes or only have detection line appearance redness
Band, illustrates that quick measuring card lost efficacy, as shown in Figure 6, Figure 7.
The sensitivity test of embodiment 2 pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card is prepared according to the method in example 1.Point
Do not test this colloidal gold immunochromatographimethod quick measuring card to cypermethrin, Fenpropathrin, decis, phenylate cyanogen
9 kinds of chrysanthemum esters such as chrysanthemum ester, flucythrinate, cyfloxylate, cycloprothrin, fenvalerate, fluorine amine cyanogen chrysanthemum
The detection sensitivity of class pesticide.Respectively these 9 kinds of pyrethrin pesticide standard substance are diluted to finite concentration, energy
Enough detect the least concentration of positive findings, be the quick measuring card detection sensitivity to this pesticide.
After tested, 9 kinds of pyrethrin pesticide testing results are the positive, and detection limit is shown in Table 1: to 10mg/kg chlorine
Cyano chrysanthemate, Fenpropathrin, decis, cyphenothrin, flucythrinate, cyfloxylate, second cyanogen
Chrysanthemum ester, fenvalerate, the detection positive rate of fluorine amine cyanogen chrysanthemum are less than 5%, regard testing result as negative.Show
This quick measuring card can be used for the examination of multiple pyrethrin pesticide.
The detection limit of table 1 pyrethrin pesticide
| Pesticide name | Detection limit (mg/kg) |
| Cypermethrin | 0.05 |
| Fenpropathrin | 0.01 |
| Decis | 0.01 |
| Cyphenothrin | 0.01 |
| Flucythrinate | 0.01 |
| Cyfloxylate | 0.01 |
| Cycloprothrin | 0.01 |
| Fenvalerate | 0.05 |
| Fluorine amine cyanogen chrysanthemum | 0.05 |
Embodiment 3: the specificity test of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card is prepared according to the method in example 1.Will
Organochlorine pesticide and chlopyrifos, Bayer 71628, Rogor, the enemies such as clofenotane (DDT), Gamma Hexaochlorocyclohexane (HCH)
The carbamates agricultures such as dichlorvos, the organophosphorus pesticide of Malathion and sevin, Furadan, Methomyl
Medicine is diluted to finite concentration respectively, detects with quick measuring card.
Testing result shows, this quick measuring card is 1mg/ml organochlorine, Organophosphorus and carbamate pesticides to concentration
The positive detection rate of class pesticide sample is below 0.5%, i.e. cross reacting rate is less than 0.5%.This speed of surface is surveyed
Card has the strongest detection specificity to pyrethroid pesticide, it is adaptable to multiple pyrethroid agriculture
The specificity residue detection of medicine.
Embodiment 4: the stability test of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card is prepared according to the method in example 1.System
The quick measuring card performed puts into sealing bag, built-in drying sheet, as 45 DEG C of baking ovens, respectively at the 7th, 14,
21, within 28,35,42,49,56 days, take out, the stability of Test paper.
Quick measuring card to 3 batches carries out batch interior and differences between batches test respectively.Result shows, batch interpolation
Be respectively less than 5%, i.e. quick measuring card detection line is coated uniformly with nature controlling line, and material processes uniformly, nanoscale chlorine gold
Granulates coated antibody quality is good.3 batch products colour developing degree concordance are high, detect reproducible,
Repeatability production requirement can be met.
Shown by high temperature accelerated aging test, 45 DEG C keep 7 days, 14 days, 21 days, 28 days, 35
My god, 42 days, testing result is all without substantially changing.45 DEG C keep 49 days, and the antibody on gold mark pad occurs
Redissolution phenomenon, nature controlling line and detection line color all shoal, and sensitivity declines, and by 56 days, detection line was
The faintest.45 DEG C preserve 49 days, are equivalent to room temperature and keep a year and a half.
Claims (10)
1. a pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card, including getting stuck and get stuck interior reagent paper
Bar, described test strips includes base plate, adheres to the sample pad, gold mark pad, anti-being sequentially connected on base plate
Answer film and adsorptive pads；Described get stuck on have well and observation window；Described well is located at corresponding institute
Stating sample pad upper shell, described observation window is located at corresponding described reaction film upper shell, described reaction
Film is provided with detection line and nature controlling line, it is characterised in that be coated pyrethroid on described gold mark pad
Pesticide gold mark wide range antibody, is coated pyrethroid pesticide wide range on the detection line of reaction film and resists
Body, the nature controlling line of reaction film is coated goat-anti rabbit two and resists, wherein, and described pyrethroid pesticide gold
Mark wide range antibody is to 20～25nm gold chloride colloids by pyrethroid pesticide wide range antibody coupling
Particle surface gained.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
Being, described base plate is PVC board, and described reaction film is nitrocellulose filter, described sample pad by
Glass fibre membrane or filter paper are made, and described adsorptive pads is made up of filter paper.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
Being, described sample pad, gold mark pad, are provided with between reaction film and adsorptive pads the overlay region of 1～2mm；
Described detection line and nature controlling line interval 0.5～1.0cm.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
Being, the standard gold amount of described pyrethroid pesticide gold mark wide range antibody is 14～16 μ g/mL, gold
The concentration of the upper coated pyrethroid pesticide gold mark wide range antibody of mark pad is 14～16 μ g/mL.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
It is, during pyrethroid pesticide wide range antibody coupling to gold chloride colloidal grain surface, chlorine used
The pH value of auric acid colloidal solid solution is 8.0～8.5.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
Being, the concentration that the detection line of described reaction film is coated pyrethroid pesticide wide range antibody is
0.8～1.4 μ g/mL.
Pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card the most according to claim 1, its feature
Being, described pyrethroid pesticide wide range antibody is pyrethroid pesticide wide range Anti-TNF-α
Body.
8. the system of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card as described in any one of claim 1～7
Preparation Method, comprises the steps:
1) sample pad, gold mark pad, reaction film and adsorptive pads are pasted onto successively on base plate and make test strips；
2) pyrethroid pesticide wide range antibody is coated with 20～25nm gold chloride colloidal solids, prepares egg
White concentration is the pyrethroid pesticide gold mark wide range antibody of 14～16 μ g/mL, is sprayed at gold mark
On pad, it is dried；Again the pyrethroid pesticide wide range that protein concentration is 0.8～1.4 μ g/mL is resisted
The detection line position that body is sprayed on reaction film, 36～37 DEG C are dried；Then by anti-for goat-anti rabbit two spray
The nature controlling line position being applied on reaction film, is prepared as speed survey in installing to get stuck by test strips after drying
Card.
Preparation method the most according to claim 8, it is characterised in that step 2) in pyrethroid agriculture
The medicine wide range coated concrete grammar of antibody nanogold particle is: take 20～25nm gold chloride colloidal solids
Solution is in small test tube, centrifugal, removes supernatant, suspends with equal-volume ultra-pure water, and regulation pH value is
8.0～8.5, subsequently pyrethroid pesticide wide range antibody is slowly instilled nanogold particle suspension, room
Temperature concussion, then drip BSA, making final concentration of the 0.1% of BSA, room temperature is shaken, to close nanometer gold
Unconjugated site on granule, it is to avoid non-specific adsorption；Centrifugal, remove unnecessary pyrethroid
Class pesticide wide range antibody and BSA, ultra-pure water suspends, and washing completes nanogold particle and pyrethroid
The coupling of ester pesticides wide range antibody, obtains pyrethroid pesticide gold mark wide range antibody, wherein,
Standard gold amount is 14～16 μ g/mL.
10. the user of pyrethroid pesticide colloidal gold immunochromatographimethod quick measuring card as claimed in claim 1
Method, comprises the steps:
1) pre-treatment of testing sample: if testing sample is liquid substance, directly takes testing sample detection；
If testing sample is solid matter, testing sample is ground, add sample treatment liquid, after standing
Detection；Wherein, described sample treatment liquid is phosphate buffer；
2) detection: being instilled by taken sample in well, sample migrates along sample pad under capillary action,
Room temperature effect 5～10min, observed result；
3) result judges:
Positive: nature controlling line, detection line all present red stripes, illustrate to have in sample pyrethroid
Class pesticide exists；
Negative: nature controlling line presents red stripes, detection line does not present red stripes, illustrates in sample
There is no pyrethroid pesticide；
Lost efficacy: nature controlling line, detection line the most do not present red stripes or only have detection line present red bar
Band, illustrates that test strips lost efficacy.
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| CN106226292B (en) * | 2016-04-21 | 2018-07-03 | 中国科学院大连化学物理研究所 | A kind of fast inspection card of cyano-containing pyrethroid pesticide and preparation and application |
| CN106093378A (en) * | 2016-05-30 | 2016-11-09 | 吉林大学 | Detection dog echinococcus granulosus infection colloidal gold immune chromatography test and preparation method |
| CN107356748A (en) * | 2017-07-17 | 2017-11-17 | 广东志道医药科技有限公司 | Biphenthrin colloidal gold immuno-chromatography test paper strip and its preparation and application |
| CN110045115A (en) * | 2019-04-23 | 2019-07-23 | 山东理工大学 | A kind of Determination of Multiple Pesticide Residues in Vegetables test strip based on Broad specificity antibody identification |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5608104A (en) * | 1993-09-28 | 1997-03-04 | Mitsui Toatsu Chemicals, Inc. | Immunological detection method of pyrethroid compound, haptenic compound, intermediate for the haptenic compound, immunogen and antibody |
| US5723306A (en) * | 1993-09-04 | 1998-03-03 | Dragerwerk Aktiengesellschaft | Monoclonal antibodies for the detection of pyrethroids and a process for their production |
| CN101434652A (en) * | 2008-12-05 | 2009-05-20 | 中国农业科学院油料作物研究所 | Pyrethroid pesticide artificial antigen, antibody and preparation thereof |
| CN102507931A (en) * | 2011-11-16 | 2012-06-20 | 厦门出入境检验检疫局检验检疫技术中心 | Colloidal gold immunochromatographic assay test strip for pyrethroid pesticides and preparation and use methods |
| CN103149351A (en) * | 2013-03-21 | 2013-06-12 | 江南大学 | Synthesis and application of envelope antigen universal for pyrethriods pesticide |
| CN103509757A (en) * | 2013-07-03 | 2014-01-15 | 江南大学 | Hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by hybridoma cell strain No. 15 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08136540A (en) * | 1994-11-10 | 1996-05-31 | Mitsui Toatsu Chem Inc | Immunological analytic method |
| WO2008010837A2 (en) * | 2005-12-15 | 2008-01-24 | The Regents Of The University Of California | Noncompetitive immunoassays to detect small molecules |
-
2014
- 2014-10-23 CN CN201410571154.6A patent/CN104297470B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5723306A (en) * | 1993-09-04 | 1998-03-03 | Dragerwerk Aktiengesellschaft | Monoclonal antibodies for the detection of pyrethroids and a process for their production |
| US5608104A (en) * | 1993-09-28 | 1997-03-04 | Mitsui Toatsu Chemicals, Inc. | Immunological detection method of pyrethroid compound, haptenic compound, intermediate for the haptenic compound, immunogen and antibody |
| CN101434652A (en) * | 2008-12-05 | 2009-05-20 | 中国农业科学院油料作物研究所 | Pyrethroid pesticide artificial antigen, antibody and preparation thereof |
| CN102507931A (en) * | 2011-11-16 | 2012-06-20 | 厦门出入境检验检疫局检验检疫技术中心 | Colloidal gold immunochromatographic assay test strip for pyrethroid pesticides and preparation and use methods |
| CN103149351A (en) * | 2013-03-21 | 2013-06-12 | 江南大学 | Synthesis and application of envelope antigen universal for pyrethriods pesticide |
| CN103509757A (en) * | 2013-07-03 | 2014-01-15 | 江南大学 | Hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by hybridoma cell strain No. 15 |
Non-Patent Citations (5)
| Title |
|---|
| Development of a colloidal-gold based lateral-flow immunoassay kit for ‘quality-control’ assessment of pyrethroid and endosulfan formulations in a novel single strip format;K.R. Kranthi,et al;《Crop Protection》;20090531;第28卷;428-434 * |
| Magnetic bead-based phage anti-immunocomplex assay (PHAIA) for the detection of the urinary biomarker 3-phenoxybenzoic acid to assess human exposure to pyrethroid insecticides;Hee-Joo Kim，et al;《Anal Biochem.》;20090301;第386卷(第1期);45-52 * |
| 拟除虫菊酯及其代谢物免疫检测的研究进展;胥传来 等;《食品与生物技术学报》;20121231;第31卷(第3期);238-246 * |
| 拟除虫菊酯类农药免疫快速检测方法及其定量构效模型的建立;陈秀金;《中国博士学位论文全文数据库 农业科技辑》;20140115(第01期);1-65 * |
| 氰戊菊酯残留胶体金免疫层析试纸条研制;肖琛 等;《化学试剂》;20110831;第33卷(第08期);675-679 * |
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