CN107340397A - A kind of time-resolved fluorescence test strips for detecting cypermethrin and its application - Google Patents
A kind of time-resolved fluorescence test strips for detecting cypermethrin and its application Download PDFInfo
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Abstract
The invention discloses a kind of time-resolved fluorescence test strips for detecting cypermethrin and its application.Test strips include sample absorption pad, reaction film and adsorptive pads, and reaction film is overlapped in reaction film left and right ends respectively positioned at centre, sample absorption pad with adsorptive pads；Sample absorption pad is provided with sample application zone and microballoon area, and microballoon area is loaded with cypermethrin time time-resolved fluorescence microballoon；Reaction film is provided with detection line (T lines) and nature controlling line (C lines), and T lines connect coating cypermethrin antigen (coating antigen), C lines coating anti-rabbit antibody.Present invention also offers a kind of method using Cypermethrin Residues in the samples such as above-mentioned cypermethrin ELISA test strip apple, corn, cucumber, tomato, spinach, Chinese cabbage.Test strips provided by the present invention have the characteristics that simple to operate, high sensitivity, quantitative detection, quick detection, cost are low, are adapted to the examination and on-site supervision of great amount of samples.
Description
Technical field
The present invention relates to a kind of time-resolved fluorescence test strips for detecting cypermethrin and its application, it is glimmering to belong to time resolution
Light immunoassay (TRFIA) technical field, for cypermethrin in the samples such as apple, corn, cucumber, tomato, spinach, Chinese cabbage
Content detection.
Background technology
Cypermethrin, also known as Xing Mianbao, Mie Baike, Sai Bokai, it is a kind of pyrethrosin class of synthesis, is one for insect
The quick-acting neurotoxins of kind.The industrial goods of cypermethrin be yellow to viscous brown solid, be thick liquid at 60 DEG C.Animal is acute
Poisoning manifestations are incoordination, instability of gait, occasionally have and tremble, while have chronic toxicity, also have stimulation to mucocutaneous.
It is increasingly extensive with the application of the agricultural chemicals, the pollution to vegetables and other crops will necessarily be caused.Therefore, China's correlation quality inspection portion
Door has carried out limitation regulation to the material.GB 2763-2016《National food safety standard Pesticide MRL》
Define the MRL of cypermethrin：Cereal (part) 0.3mg/kg, soybean and corn 0.05mg/kg, spinach in vegetables
Dish, common Chinese cabbage, lettuce and Chinese cabbage are 2mg/kg, orange in fruit, lemon, shaddock, apple, pears, kernel approaches (except peach) and
Matrimony vine is 2mg/kg, and citrus, peach and durian are 1mg/kg.
At present, detecting the method for cypermethrin has gas chromatography, gas chromatography-mass spectrography, liquid chromatography-mass spectrometry
Deng.The accuracy of instrument detection method is higher, but expensive equipment is, it is necessary to which technical professional, detecting step are complicated, it is difficult to
Batch detection sample, laboratories batch, quick detection sample are not suitable for it.And immunological detection method is simple to operate, fast
It is fast, sensitive, most samples can be detected simultaneously, be preferable quick screening means.
The content of the invention
It is an object of the invention to provide when a kind of high sensitivity, the cypermethrin that simple to operate, cost is low, detection time is short
Between resolved fluorometric test strips.
A kind of cypermethrin time-resolved fluorescence test strips provided by the present invention, the test strips include sample absorption pad,
Reaction film and adsorptive pads three parts.Reaction film is overlapped in reaction film or so respectively positioned at centre, sample absorption pad with adsorptive pads
Both ends.Wherein, sample absorption pad is provided with sample application zone and microballoon area, and microballoon area is loaded with time-resolved fluorescence microballoon；Reaction film
Detection line (T lines) and nature controlling line (C lines) are provided with, T lines connect coating cypermethrin antigen, C lines coating anti-rabbit antibody.
The cypermethrin time-resolved fluorescence microballoon, including detection microballoon and Quality Control microballoon, detection microballoon wrap for surface
The fluorescent microsphere for being had cypermethrin monoclonal antibody, Quality Control microballoon are the fluorescent microsphere that pan coating has rabbit-anti labelled protein.
Bright-coloured series elements compound is filled with the fluorescent microsphere；Preferably, the bright-coloured system member rope huge legendary turtle compound is europium chelant thing；
Optimal, the europium chelant thing can be Eu (TTA) 3/TOPO or Eu (TTA) 3/Phen.
The albumen can be cow's serum Y- globulin (BGG) or bSA (BSA).
The time-resolved fluorescence microspherulite diameter scope is 100-1000nm.
On the nitrocellulose filter, coated antibody is cypermethrin antigen on T lines, coated anti-rabbit antibody on C lines.
The cypermethrin time-resolved fluoroimmunoassay quantitative testing test paper bar bottom is provided with plastic bottom board.
It is to be obtained by cypermethrin haptens with carrier protein couplet that the T lines, which connect coating cypermethrin antigen, described
Carrier protein can be bovine serum albumin(BSA), ovalbumin, hemocyanin, thyroprotein, human serum albumins.
The cypermethrin monoclonal antibody is prepared using cypermethrin hapten-carrier protein conjugate as immunogene
Obtain, be to be secreted to obtain by the strain of cypermethrin monoclonal antibody hybridoma cell；The anti-rabbit antibody is by rabbit source antibody mediated immunity
Sheep obtains.
The sample absorption pad is Fusion5 films or other function identical films；The conjugate release pad can be glass
Cotton or polyester material；The adsorptive pads are adsorptive pads；The reaction film can be nitrocellulose filter or cellulose acetate film.
It is a further object to provide a kind of method for preparing above-mentioned test strips, it includes step：
(1) prepared by Quality Control microballoon：
1. use biotinylated protein；
2. aldehyde group modified fluorescent microsphere is coated with using above-mentioned labelled protein；
(2) prepared by haptens：Cypermethrin haptens product is obtained by series of chemical；
(3) by cypermethrin haptens and carrier protein couplet, cypermethrin hapten-carrier protein conjugate is obtained；
(4) new zealand white rabbit is immunized with cypermethrin hapten-carrier protein conjugate, new zealand white rabbit spleen is thin
Born of the same parents and myeloma cell obtain cypermethrin monoclonal hybridoma strain by merging, screening；
(5) new zealand white rabbit IgG immune health goats are extracted, obtain anti-rabbit antibody；
(6) microballoon is detected to prepare：Aldehyde group modified fluorescent microsphere is coated with using the monoclonal antibody of cypermethrin；
(7) blank kilocalorie is pasted：Nitrocellulose filter is pasted onto among plastic bottom board, Fusion5 films and adsorptive pads point
Nitrocellulose filter left and right ends are not overlapped in；
(8) film is sprayed：The fluorescent microsphere of rabbit-anti labelled protein and the fluorescent microsphere of cypermethrin monoclonal antibody are used and released
Slow down fliud flushing mixed diluting to finite concentration, be sprayed onto the microballoon area of Fusion5 films；Cypermethrin antigen and the dilution of anti-rabbit antibody
Afterwards, the T lines and C line positions of nitrocellulose filter are sprayed onto respectively；
(9) dry and cut：By the above-mentioned kilocalorie drying for having sprayed reagent, slitting.
The release buffer solution used in spray film step contains 10%-20% sucrose, 3%-10% trehaloses, 0.5%-1%
The double trimethylsilyl acetamides (BSA) of N, 0-, 0.2%-0.5% gentamicins.
The final concentration of 0.5mg/mL-2mg/mL of microballoon, Quality Control microballoon final concentration 0.05mg/ are detected after spraying the dilution of film step
mL-0.2mg/mL；T line antigen final concentration 0.5mg/mL-2mg/mL, C line antigen final concentrations 0.5mg/mL-2mg/mL；C, T line spray
Film liquid amount is 0.5 μ L/cm-1.5 μ L/cm, and microballoon spray film amount is 2 μ L/cm-8 μ L/cm.
The drying dried and cut described in step can be in constant temperature oven or drying room, 37 DEG C of drying 8-12 hours.
It is a further object to provide one kind application above-mentioned ELISA test strip apple, corn, cucumber, tomato,
The method of Cypermethrin Residues in the samples such as spinach, Chinese cabbage, it includes step：
(1) sample pre-treatments；
(2) detected with test strips；
(3) testing result is analyzed.
The cypermethrin time-resolved fluorescence test strips of the present invention are reacted and are immunized using the antibody antigen of high degree of specificity
Chromatographic techniques, cypermethrin monoclonal antibody-time-resolved fluorescence label is fixed on Fusion5 films, in sample
Cypermethrin in flow process, with the cypermethrin monoclonal antibody on Fusion5 films-time-resolved fluorescence label knot
Close, form drug-antibody-time-resolved fluorescence label.Medicine and the cypermethrin in reaction film detection line half in sample
Antigen-carrier protein conjugate competition binding cypermethrin monoclonal antibody-time-resolved fluorescence label, final application are immunized
Chromatographic detector calculates the cypermethrin residue contained in analyte sample fluid.
The test strips of the present invention have that high specificity, high sensitivity, quantitative detection, cost be low, simple to operate, detection time
The advantages of short, suitable various units use, storage is simple, long shelf-life.With ELISA test strip Cypermethrin Residues of the present invention
Method is easy, quick, directly perceived, accurate, applied widely, cost is low, easily promotes the use of.
Brief description of the drawings
Fig. 1 is cypermethrin hapten synthesis route map.
Fig. 2 is cypermethrin haptens hydrogen nuclear magnetic resonance spectrogram.
Embodiment
The present invention is expanded on further with reference to specific embodiment.It should be understood that these embodiments are merely to illustrate this
Invention, and it is not limited to the scope of the present invention.
The preparation of the cypermethrin test strip of embodiment 1
The preparation method of the test strips mainly includes the following steps that：
1) the sample absorption pad for being coated with cypermethrin monoclonal antibody-time-resolved fluorescence label is prepared；
2) prepare with the detection line for being coated with cypermethrin hapten-carrier protein conjugate and be coated with anti-rabbit antibody
Nature controlling line reaction film；
3) test strips 1) will be assembled into sample absorption pad, reaction film and the adsorptive pads 2) prepared.
Substep narration in detail below：
1st, the preparation of cypermethrin haptens
(1) 24.3g 3- (4-nitrophenoxy) benzaldehyde is dissolved in 100ml absolute methanols, dissolving is stirred at room temperature, adds
Enter iodine 0.5g, 11gTMSCN is added dropwise, is added dropwise, react at room temperature 10h, solvent evaporated, column chromatography purifying, obtain 25.65g cyano group
Alcohol object, yield 95%.
(2) the cyano group alcohol 27g obtained in step 1 is added in 100ml chloroforms, dissolving is stirred at room temperature, added
Py8.5g, the 30ml chloroform solns containing 22.6g dichloro chrysanthemum acyl chlorides are added dropwise at room temperature.It is added dropwise, room temperature continues anti-
5h is answered, solvent evaporated, column chromatography purifying, obtains cyano group ester object 42.32g, yield 92%.
(3) by the cyano ester compound 46g obtained by step 2, it is added in 120ml absolute methanols, room-temperature dissolution, pours into
It is hydrogenated with kettle, adds Pd/C3g, add 1 atmospheric pressure, react at room temperature 6h, be filtered to remove Pd/C, decompression removes solvent, obtains amino mesh
Mark thing 40.8g, yield 95%.
(4) amino-compound 43g obtained by step 3 is added in 200ml chloroforms, dissolving is stirred at room temperature.Add 1g
P-methyl benzenesulfonic acid, succinic anhydride 12g is added, reaction is stirred at room temperature overnight.Solvent evaporated, column chromatography purify to obtain carboxylic
Haptens object 45.6g, yield 86%.From cypermethrin haptens hydrogen nuclear magnetic resonance spectrogram, 12.2 carboxyl peak, 7-
8 aromatic ring peak, and 0.97 methyl peak, illustrate hapten synthesis success.
Haptens prepared by this method does not have a structure for changing cypermethrin, but from the stronger two aryl oxide structures of rigidity
Draw linking arm so that whole active structures of cypermethrin can be exposed to outside antigen, cause stronger immune response,
Make it possible the antibody for obtaining high-affinity.
2nd, the preparation of immunogene
133mg BSA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Cypermethrin haptens and BSA ratios can be 1: 1
Adjusted in the range of~60: 1, the application selection is adding 53mg cypermethrins haptens (being 50: 1 with BSA mol ratios), adds
EDC hydrochloride 10mg, are stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.Cypermethrin immunogene is obtained under the ratio can be
Antigenic surface exposes more cypermethrin haptens, so as to preferably cause the immune response for cypermethrin.
3rd, the preparation of coating antigen
86mg OVA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Cypermethrin haptens and OVA ratios can 1: 1~
Adjusted in the range of 40: 1, the application selection is adding 42.4mg cypermethrins haptens (being 40: 1 with OVA mol ratios), adds
EDC hydrochloride 10mg, are stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.Obtained under the ratio cypermethrin coating antigen with sample
, can fully and antibody response in product during antigenic competition antibody, also will not be because of coating antigen exposed sites so as to avoid false positive
Excessively, steric hindrance is formed, so as to avoid false negative.
4th, the preparation of cypermethrin monoclonal antibody
Immunogene is injected in Balb/c new zealand white rabbit bodies, produces antiserum.Take immune Balb/c New Zealand great Bai
Rabbit splenocyte merges with SP2/0 myeloma cell, screens positive hole, obtains the hybridoma of stably excreting monoclonal antibody
Strain.Cell suspension is made with frozen stock solution in hybridoma, it is standby.Hybridoma is placed in cell culture medium, at 37 DEG C
Under the conditions of cultivated, obtained nutrient solution is purified with octanoic acid-saturated ammonium sulfate method, obtains monoclonal antibody, -20 DEG C
Preserve.
The cell culture medium is that calf serum and sodium acid carbonate are added into RPMI1640 culture mediums, calf serum is existed
Final concentration of 20% (mass fraction) in cell culture medium, final concentration of 0.2% (matter of the sodium acid carbonate in cell culture medium
Measure fraction)；The pH of the cell culture medium is 7.4.It is immunogene to pathogen-free domestic using rabbit source antibody using sheep as immune animal
Sheep is immunized, and obtains anti-rabbit antibody.
5th, the preparation of microspheres solution, detection line (T lines) solution and nature controlling line (C lines) solution
(1) configuration of microspheres solution
With release buffer solution (containing 20% sucrose, 5% trehalose, 0.5%BSA, 0.2% gentamicin) by Quality Control microballoon
Time-resolved fluorescence microballoon mixed liquor, the final concentration of 0.1mg/mL-0.3mg/mL of Quality Control microballoon, detection are configured to detection microballoon
The final concentration of 0.8mg/mL-1.2mg/mL of microballoon.
(2) preparation of detection line (T lines) solution
Cypermethrin antigenic compound is diluted to 0.4mg/mL-0.6mg/mL with 0.01M PB solution.
(3) preparation of nature controlling line (C lines) solution
Streptavidin is diluted to 0.4mg/mL-0.6mg/mL with 0.01M PB solution.
6th, the preparation of test strips
Reaction film is overlapped in reaction film left and right ends with blotting paper, is pasted onto and carries respectively positioned at centre, sample absorption pad
On the plastic bottom board of gum.C, T line spray film liquid amount are 0.6 μ L/cm-1.2 μ L/cm, and microspheres solution spray film amount is 2 μ L/cm-6 μ L/
cm.The cypermethrin for having sprayed reagent is stuck in greatly in constant temperature oven into 37 DEG C to dry 12 hours.The cypermethrin kilocalorie of drying is cut
The paper slip of 3mm-5mm width is cut into, that is, obtains cypermethrin test strips.
The detection of Cypermethrin Residues in the sample of embodiment 2
1st, sample treatment
(1) apple pre-treating method.
With homogenizer homogeneous samples；The apple sample after (2.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong, 4mL 0.01M PBS, 2mL methanol is separately added into, with vortex instrument whirling motion 5min；4000r/min centrifuges 5min at room temperature；
The μ L of supernatant 200 are taken, are added in 800 μ L 0.01M PBS, are fully mixed, take its 50 μ L to be used to analyze.
(2) corn pre-treating method.
With homogenizer homogeneous samples；The corn sample after (2.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong, the sodium chloride of 4mL 10% is separately added into, 2mL methanol, vibrates 5min with vortex instrument；4000r/min centrifuges 5min at room temperature；
The μ L of supernatant 100 are taken, are added in 900 μ L0.01M PBS, are fully mixed, take its 50 μ L to be used to analyze.
(3) vegetables pre-treating method.
With homogenizer homogeneous samples；The vegetable sample after (1.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong, 2mL 0.1mol/L sulfuric acid is added, 5mL methanol is added, with vortex instrument whirling motion 5min；, 1000r/min centrifugations at room temperature
5min；The μ L of supernatant 200 are taken, are added in 800 μ L 0.01M PBS, are fully mixed, take 50 μ L to be used to analyze.
2nd, detected with test strips
(1) test strips and extract solution are recovered to room temperature.
(2) 60 μ L measuring samples are accurately drawn with pipettor, (being careful not to suck bubble during sampling) is vertically slowly added dropwise
In well (S), start timing after sample-adding, read result within 8 minutes after sample-adding.
(3) reagent card is inserted in the carrier of immunochromatography detector, presses detection key, instrument will stick into test automatically
Row scanning.
(4) testing result is read from the display screen of immunochromatography detector.
3rd, testing result is analyzed
Test limit scope：1ng/ml-30ng/ml.
The sample detection example of embodiment 3
1st, test limit is tested
The samples such as blank apple/corn/cucumber/tomato/spinach/Chinese cabbage are taken, add cypermethrin respectively to final concentration
For 0.5ng/ml, 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, test strips are taken to carry out
Detection, each sample are repeated three times.
During with samples such as ELISA test strip apple/corn/cucumber/tomato/spinach/Chinese cabbages, shown and tied according to test strips
Fruit determines test limit scope, shows that this ELISA test strip limits scope：1ng/ml-30ng/ml.
2nd, sample preci-sion and accuracy is tested
Using the rate of recovery as accuracy estimating index, the testing result relative standard deviation of a certain concentration samples of replication
(RSD%) it is used as precision evaluation index.Calculation formula is：The rate of recovery (%)=actual measured value/theoretical value × 100%, its
The addition concentration of middle theoretical value sample；Relative standard deviation RSD%=SD/X × 100%, wherein SD are standard deviation, and X is
The average value of determination data.
By five concentration cypermethrins of 1ng/ml, 5ng/ml, 10ng/ml, 20ng/ml, 30ng/ml respectively to apple/jade
The samples such as rice/cucumber/tomato/spinach/Chinese cabbage be added recovery measure, each sample do 4 it is parallel, with three batches of different examinations
Agent is measured, and the average recovery rate and precision result for calculating sample see the table below.
The sample precision of table 1 and accuracy test
With the cypermethrin of five concentration of 1ng/ml, 5ng/ml, 10ng/ml, 20ng/ml, 30ng/ml respectively to apple/
The samples such as corn/cucumber/tomato/spinach/Chinese cabbage are added, and average recovery rate is between 91.1%~109.3%；Batch
It is interior, batch between relative standard deviation be respectively less than 15%.
3rd, cross reacting rate is tested
Selection with cypermethrin there is similar structures and the medicine of similar functions to carry out time-resolved fluorescence test strips measure,
Its 50% inhibition concentration is respectively obtained by the standard curve of various medicines, intersection of the kit to other medicines is calculated with following formula
Reactivity.
The cross reacting rate of table 2 is tested
| Medicine name | Cross reacting rate (%) |
| Cypermethrin | 100 |
| Permethrin | ＜ 1 |
| Decis | ＜ 1 |
| Fenpropathrin | ＜ 1 |
| Biphenthrin | ＜ 1 |
| Cyhalothrin | ＜ 1 |
| Lambda-cyhalothrin | ＜ 1 |
| Fenvalerate | ＜ 1 |
Claims (7)
1. a kind of time-resolved fluorescence test strips for detecting cypermethrin, including sample absorption pad, reaction film and adsorptive pads, its
It is characterised by that the reaction film is overlapped in reaction film left and right ends respectively positioned at centre, sample absorption pad with adsorptive pads；The sample
Product absorption pad is provided with sample application zone and microballoon area, and microballoon area is loaded with cypermethrin time time-resolved fluorescence microballoon；It is described anti-
Film is answered to be provided with detection line (T lines) and nature controlling line (C lines), T lines connect coating cypermethrin antigen (coating antigen), and C lines coating is anti-
Rabbit antibody, the cypermethrin antigen are cypermethrin hapten-carrier protein conjugates, the system of the cypermethrin haptens
Preparation Method is as follows：
(1) 24.3g 3- (4-nitrophenoxy) benzaldehyde is dissolved in 100ml absolute methanols, dissolving is stirred at room temperature, add iodine
0.5g, 11g TMSCN are added dropwise, are added dropwise, react at room temperature 10h, solvent evaporated, column chromatography purifying, obtain 25.65g cyano group alcohol
Object, yield 95%；
(2) the cyano group alcohol 27g obtained in step 1 is added in 100ml chloroforms, dissolving is stirred at room temperature, added
Py8.5g, the 30ml chloroform solns containing 22.6g dichloro chrysanthemum acyl chlorides are added dropwise at room temperature；It is added dropwise, room temperature continues anti-
5h is answered, solvent evaporated, column chromatography purifying, obtains cyano group ester object 42.32g, yield 92%；
(3) by the cyano ester compound 46g obtained by step 2, it is added in 120ml absolute methanols, room-temperature dissolution, pours into hydrogenation
In kettle, Pd/C3g is added, adds 1 atmospheric pressure, reacts at room temperature 6h, is filtered to remove Pd/C, decompression removes solvent, obtains amino object
40.8g, yield 95%；
(4) amino-compound 43g obtained by step 3 is added in 200ml chloroforms, dissolving is stirred at room temperature；1g is added to first
Base benzene sulfonic acid, succinic anhydride 12g is added, reaction is stirred at room temperature overnight；Solvent evaporated, it is anti-that column chromatography purifies to obtain carboxylic half
Former object 45.6g, yield 86%.
2. the time-resolved fluorescence test strips of detection cypermethrin as claimed in claim 1, it is characterised in that the chlorine cyanogen chrysanthemum
The molecular structural formula of ester hapten is：
3. the time-resolved fluorescence test strips of detection cypermethrin as claimed in claim 1, it is characterised in that the chlorine cyanogen chrysanthemum
Ester time-resolved fluorescence microballoon, including detection microballoon and Quality Control microballoon, detection microballoon are that pan coating has cypermethrin monoclonal
The fluorescent microsphere of antibody, Quality Control microballoon are the fluorescent microsphere that pan coating has rabbit-anti labelled protein.
4. the time-resolved fluorescence test strips of detection cypermethrin, cypermethrin monoclonal antibody are as claimed in claim 1
Prepared using cypermethrin hapten-carrier protein conjugate as immunogene, the preparation method of the immunogene is as follows：
133mg BSA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Cypermethrin haptens and BSA ratios can be 1: 1~60:
Adjusted in the range of 1, the application selection is adding 53mg cypermethrins haptens (being 50: 1 with BSA mol ratios), adds EDC salt
Hydrochlorate 10mg, is stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.
5. the time-resolved fluorescence test strips of detection cypermethrin as claimed in claim 1, the preparation method of the coating antigen
It is as follows：
86mg OVA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Cypermethrin haptens and OVA ratios can be 1: 1~40: 1
In the range of adjust, the application selection add 42.4mg cypermethrins haptens (be 40: 1 with OVA mol ratios), addition EDC
Hydrochloride 10mg, is stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.
6. a kind of method for preparing any one of the claim 1-5 test strips, it includes step：
1) the sample absorption pad for being coated with cypermethrin monoclonal antibody-time-resolved fluorescence label is prepared；
2) prepare to have and be coated with the detection line of cypermethrin hapten-carrier protein conjugate and be coated with the matter of anti-rabbit antibody
Control the reaction film of line；
3) test strips 1) will be assembled into sample absorption pad, reaction film and the adsorptive pads 2) prepared.
7. a kind of method for detecting Cypermethrin Residues in the samples such as apple, corn, cucumber, tomato, spinach, Chinese cabbage, it is wrapped
Include step：
1) Sample pretreatment；
2) detected with the test strips described in claim any one of 1-6；
3) testing result is analyzed.
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| CN111269139A (en) * | 2019-12-12 | 2020-06-12 | 中国海洋大学 | Efficient cyhalothrin hapten, activated carrier protein, complete antigen and polyclonal antibody for immunoassay |
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| CN109856091B (en) * | 2018-03-16 | 2021-08-03 | 北方工业大学 | Time-resolved fluorescence test strip for detecting monocrotophos and application thereof |
| CN108469517A (en) * | 2018-03-16 | 2018-08-31 | 北方工业大学 | Time-resolved fluorescent test strip for detecting trichlorfon and application thereof |
| CN108535474A (en) * | 2018-03-16 | 2018-09-14 | 北方工业大学 | Time-resolved fluorescence test strip for detecting permethrin and application thereof |
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| CN109061167A (en) * | 2018-08-29 | 2018-12-21 | 郑州工程技术学院 | A kind of immune chromatography test paper detecting pyrethroid |
| CN111269139A (en) * | 2019-12-12 | 2020-06-12 | 中国海洋大学 | Efficient cyhalothrin hapten, activated carrier protein, complete antigen and polyclonal antibody for immunoassay |
| CN111269139B (en) * | 2019-12-12 | 2022-09-23 | 中国海洋大学 | High-efficiency cyhalothrin hapten, activated carrier protein, complete antigen and polyclonal antibody for immunoassay |
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| CN117384061A (en) * | 2023-12-11 | 2024-01-12 | 深圳市通量检测科技有限公司 | Dioxamine hapten, antigen, antibody, detection device and preparation and application thereof |
| CN117384061B (en) * | 2023-12-11 | 2024-03-15 | 深圳市通量检测科技有限公司 | Dioxamine hapten, antigen, antibody, detection device and preparation and application thereof |
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