CN105259351A - Test paper for detecting metalaxyl residue and application thereof - Google Patents
Test paper for detecting metalaxyl residue and application thereof Download PDFInfo
- Publication number
- CN105259351A CN105259351A CN201510530296.2A CN201510530296A CN105259351A CN 105259351 A CN105259351 A CN 105259351A CN 201510530296 A CN201510530296 A CN 201510530296A CN 105259351 A CN105259351 A CN 105259351A
- Authority
- CN
- China
- Prior art keywords
- metalaxyl
- test paper
- coated
- pad
- carrier protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- G01N33/577—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
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a test paper for detecting metalaxyl residue and application thereof. The test paper comprises a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3), a water absorption pad (4) and a base plate (7), wherein the reaction membrane comprises a detection line (5) coated with a metalaxyl semiantigen-carrier protein conjugate and a quality control line (6) coated with a goat anti-mouse antiantibody, and the conjugate release pad (2) is coated with a metalaxyl monoclonal antibody-colloidal gold marker. The invention also provides a method for applying the metalaxyl test paper in detection of metalaxyl residue in tea, tobacco and vegetable. The test paper provided by the invention can be used for detecting metalaxyl residue in tea, tobacco and vegetable, has the characteristics of simple operation, high sensitivity, a fast detection speed, low cost, etc., and is applicable to screening and on-site monitoring of a great number of samples.
Description
Technical field
The present invention relates to a kind of test paper and the application thereof that detect metalaxyl residue, be specifically related to a kind of colloid gold test paper for detecting metalaxyl residue, it is specially adapted to the qualitative detection of metalaxyl in tobacco.
Background technology
Metalaxyl (Metalaxyl), D, L-N-(2, 6-3,5-dimethylphenyl)-N-(2'-methoxyl acetyl) methyl lactamine, it is a kind of novel systemic fungicide with protection and therapeutic action, can by the root of plant, stem, leaf absorbs, and each organ of plant is transferred to Dynamic water transport in plant, cauline leaf process can be done, Seed Treatment and soil treatment, to downy mildew, phytophthora, disease caused by pythium spp is effective, agricultural production is commonly used to treat downy mildew of garpe, cucumber downy mildew, angular leaf spot of cucumber, capsicum epidemic disease, black shank, the late blight of potato, tomato late blight, rice seedling blight, paddy rice bacterial wilt, bakanae disease of rice, Causal Organism of Maize Basal Stalk, maize head smut, the diseases such as the downy mildew of millet, in agricultural production and management, of many uses.
Metalaxyl belongs to phenylamide, efficient, low toxicity, less-persistent pesticide, inhale in it and seepage force very strong; dispenser can be conducted up and down for 30 minutes in plant, have protection and therapeutic action, and the drug effect extended period is long to disease plant; therefore the longevity of residure is long, thus cause its remaining on crops.China, for Different Crop, has formulated metalaxyl maximum residue limit standard, and wherein the maximum residue limit of cucumber, capsicum, tomato is 0.5mg/kg, and the maximum residue limit of brown rice is 0.1mg/kg, and other cereal maximum residue limit are 0.05mg/kg.In actual production, using 2mg/kg as the criterion of tobacco maximum residue limit.
From prior art and coherent detection standard, at present for the detection method mainly instrumental method of metalaxyl residue, the most frequently used method is LC-UV, LC-MS and LC-MS/MS, instrumental method possesses the advantages such as detection sensitivity is high, high specificity, but it is loaded down with trivial details, consuming time to detect Sample pretreatment, sample also needs to extract and purified treatment, and instrument detection method needs expensive large-scale instrument and equipment simultaneously, be equipped with the detection technique personnel of specialty, so limit its application; Colloidal gold immunity chromatography has that specificity is good, highly sensitive, easy and simple to handle, testing cost is low, be suitable for the advantages such as the selective mechanisms of batch sample, is desirable rapid screening means, can meets the requirement of field quick detection better.
Summary of the invention
The object of the present invention is to provide a kind of highly sensitive, easy and simple to handle, cost is low, detection time is short metalaxyl residue Test paper.
The test paper of detection metalaxyl residue provided by the present invention, comprises absorption of sample pad (1), bond release pad (2), reaction film (3), adsorptive pads (4) and base plate (7); Described reaction film has the detection line (T line) (5) being coated with metalaxyl hapten-carrier protein conjugate and the nature controlling line (C line) (6) being coated with sheep anti mouse antiantibody, described bond release pad (2) is coated with metalaxyl monoclonal antibody-colloid gold label thing.
Described metalaxyl coupled antigen is obtained by metalaxyl haptens and carrier protein couplet, described metalaxyl haptens is obtained by hydrolysis reaction, and described carrier protein can be mouse haemocyanin, thyroprotein, bovine serum albumin(BSA), rabbit serum proteins, human albumin, ovalbumin, hemocyanin or fibrinogen.
Described metalaxyl monoclonal antibody prepares using metalaxyl medicine hapten-carrier protein conjugate as immunogene; Mouse source antibody mediated immunity sheep obtains by the sheep anti mouse antiantibody in described sheep anti mouse antiantibody-colloid gold label thing.
Described absorption of sample pad (1), bond release pad (2), reaction film (3), adsorptive pads (4) are pasted onto on base plate (7) successively, under described bond release pad 1/3-1/2 is capped on absorption of sample pad.
Described base plate can be the material that PVC base plate or other hard do not absorb water; Described absorption of sample pad can be glass, nonwoven fabrics, hemofiltration film etc.; Described adsorptive pads is thieving paper; Described reaction film can be nitrocellulose filter or cellulose acetate membrane; Described diaphragm is PE material diaphragm.
Another object of the present invention is to provide a kind of method preparing above-mentioned test paper, and it comprises step:
1) preparation is coated with bond release pad (2) of metalaxyl monoclonal antibody-colloid gold label thing;
2) preparation has the reaction film of the detection line being coated with metalaxyl hapten-carrier protein conjugate and the nature controlling line being coated with sheep anti mouse antiantibody;
3) by 1) and 2) bond for preparing release pad, reaction film and absorption of sample pad, adsorptive pads and PVC base plate be assembled into test paper.
Specifically, step comprises:
1) haptens preparation: metalaxyl is dissolved in ethanol, stir under normal temperature and add 10%NaOH aqueous solution, carry out with TLC thin layer plate monitoring reaction, until do not have raw material or raw material point very shallow, stop reaction, purification, recrystallizing methanol, obtain hydrolysate, through nuclear-magnetism, infrared and Mass Spectrometric Identification, the success of metalaxyl hapten synthesis;
2) by metalaxyl haptens and carrier protein couplet, metalaxyl hapten-carrier protein conjugate is obtained;
3) with metalaxyl hapten-carrier protein conjugate immune mouse, mouse boosting cell and myeloma cell are passed through to merge, screen, obtains metalaxyl monoclonal antibody;
4) extract mouse IgG immune health goat, obtain sheep anti mouse antiantibody;
5) react with trisodium citrate and gold chloride and prepare collaurum;
6) metalaxyl monoclonal antibody step 3) prepared adds in collaurum prepared by step 5), obtains metalaxyl monoclonal antibody-colloid gold label thing;
7) metalaxyl monoclonal antibody-colloid gold label thing being coated on bond discharges on pad, takes out after 37 DEG C of baking 60min, seals and be placed in dry environment to save backup;
8) metalaxyl hapten-carrier protein conjugate is coated on reaction film and forms detection line (T line), and is coated on by sheep anti mouse antiantibody on reaction film and forms nature controlling line (C line);
9) by absorption of sample pad be 7.2 containing 0.5% bovine serum albumin(BSA) (volumn concentration), pH, 0.1mol/L phosphate buffer soaks 2h, dries 2h at 37 DEG C;
10) on base plate, paste absorption of sample pad, bond release pad, reaction film, adsorptive pads in order, absorption of sample pad covers bond release pad, is finally cut into the little bar that 3mm is wide, adds plastic casing, pack, can preserve 12 months under 4-30 DEG C of condition.
Another object of the present invention is to provide a kind of method applying metalaxyl residue in above-mentioned detection paper tealeaves, tobacco, vegetables sample, and it comprises step:
(1) sample pre-treatments;
(2) detect with test paper;
(3) testing result is analyzed.
Metalaxyl quick detection test paper of the present invention adopts antibody antigen reaction and the immunochromatographiassays assays technology of high degree of specificity, metalaxyl monoclonal antibody-colloid gold label thing is fixed on bond release pad, metalaxyl in sample is in flow process, first discharge the monoclonal antibody-colloid gold label thing of the metalaxyl on padding with bond, form drug-antibody-colloid gold label thing.Metalaxyl hapten-carrier protein conjugate competition binding metalaxyl monoclonal antibody-colloid gold label thing on medicine in sample and reaction film detection line, according to detection line red stripes with or without or shade judge metalaxyl residue amount in analyte sample fluid, detect be limited to 2mg/kg.
During detection, sample instills in test paper hole clipping after treatment, when metalaxyl concentration in the sample to which lower than detectability or be zero time, metalaxyl monoclonal antibody-colloid gold label thing in chromatography process can be fixed on the metalaxyl hapten-carrier protein conjugate on reaction film and be combined, an each appearance red stripes in detection line (T line) and nature controlling line (C line); If metalaxyl concentration is in the sample to which equal to or higher than detectability, metalaxyl monoclonal antibody-colloid gold label thing all can be combined with metalaxyl, thus does not occur red stripes at T line place because competitive reaction can not be combined with metalaxyl hapten-carrier protein conjugate.As shown in Figure 3.
Negative: when nature controlling line (C line) demonstrates red stripes, detection line (T line) also demonstrates red stripes simultaneously, is judged to feminine gender.
Positive: when nature controlling line (C line) demonstrates red stripes, and detection line (T line) does not develop the color, and is judged to the positive.
Invalid: when nature controlling line (C line) does not demonstrate red stripes, then no matter whether detection line (T line) demonstrates red stripes, and it is invalid that this test paper is judged to.
Test paper of the present invention have highly sensitive, high specificity, cost are low, simple to operate, detection time is short, be applicable to various units uses, stores advantage that is simple, long shelf-life.With the method for detection paper metalaxyl residue of the present invention is easy, quick, directly perceived, accurate, applied widely, cost is low, easily promote the use of.
Accompanying drawing explanation
Fig. 1: metalaxyl hapten synthesis route map;
Fig. 2: test paper cross-sectional view;
Fig. 3: detection paper result process decision chart.
Embodiment
The present invention is set forth further below in conjunction with specific embodiment.Should be understood that these embodiments are only for illustration of the present invention, and be not used for limiting the scope of the invention.
The preparation of embodiment 1 metalaxyl Test paper
The preparation method of this test paper mainly comprises following step:
1) preparation is coated with the bond release pad of metalaxyl monoclonal antibody-colloid gold label thing;
2) preparation has the reaction film of the detection line being coated with metalaxyl hapten-carrier protein conjugate and the nature controlling line being coated with sheep anti mouse antiantibody;
3) by 1) and 2) bond for preparing release pad, reaction film and absorption of sample pad, adsorptive pads and PVC base plate be assembled into test paper.
Substep describes in detail below:
1, the haptenic preparation of metalaxyl
Metalaxyl is dissolved in ethanol, under normal temperature stir add 10%NaOH aqueous solution, with TLC thin layer plate monitoring reaction carry out, until do not have raw material or raw material point very shallow, stop reaction, purification, recrystallizing methanol, obtain hydrolysate, through nuclear-magnetism, infrared and Mass Spectrometric Identification, the success of metalaxyl hapten synthesis.
2, immunogene preparation---metalaxyl haptens and bovine serum albumin(BSA) (BSA) coupling obtain immunogene.
Taking 26.5mg haptens is dissolved in 2mLDMF solution, add each 60mgEDC and NHS(to be dissolved in 2mL water) carry out activation 30 minutes, join 110-264mg carrier protein BSA(to be dissolved in 5mL water) carry out coupling and prepare immunogene, dialyse 3 days with 0.02mol/LPB damping fluid, every day changes dislysate, sooner or later to remove unreacted small-molecule substance; Antibody is prepared for animal immune after having dialysed.Packing, saves backup in-20 DEG C.
3, the preparation of coating antigen
Get 12mg metalaxyl haptens 0.8mlDMF to dissolve, be cooled to 10 DEG C, obtain reactant liquor I; Getting isobutyl chlorocarbonate 10ul adds in I, 10 DEG C of stirring reaction 30min; Get 30mg bovine serum albumin 2.2ml50mmol/LNa
2cO
3dissolve, 10 DEG C of reaction 4h, then 4 DEG C are spent the night; With 0.01mol/LPBS4 DEG C of dialysis 3d, change 3 dislysates every day, to remove unreacted small-molecule substance, namely obtain metalaxyl coating antigen, packing, save backup in-20 DEG C.
4, the preparation method of metalaxyl monoclonal antibody
(1) animal immune
Immunogene step 2 obtained is injected in Balb/c Mice Body, and immunizing dose is 150 μ g/, makes it produce antiserum.
(2) Fusion of Cells and cloning
After mice serum measurement result is higher, get its splenocyte, in 8:1(quantitative proportion) ratio and SP2/0 myeloma cell fusion, adopt indirect competitive ELISA to measure cell conditioned medium liquid, screen positive hole.Limiting dilution assay is utilized to carry out cloning to positive hole, until obtain the hybridoma cell strain secreting metalaxyl monoclonal antibody.
Cell cryopreservation and recovery: cell suspension monoclonal hybridoma strain cryopreserving liquid being made 1 × 106/ml, preserve for a long time in liquid nitrogen.Take out cryopreservation tube during recovery, put into 37 DEG C of water-bath middling speeds immediately and melt, after centrifugal segregation cryopreserving liquid, move into and cultivate culture in glassware.
The production of monoclonal antibody and purifying: Balb/c mouse peritoneal is injected sterilizing paraffin oil 0.5ml/ only, 7 days pneumoretroperitoneums inject stable monoclonal hybridoma strain 5 × 105/only, gather ascites after 7 days.Carry out ascites by sad-saturated ammonium sulfate method to purify ,-20 DEG C of preservations.
(3) cell cryopreservation and recovery
Hybridoma cryopreserving liquid is made the cell suspension of 1 × 106/ml, preserve for a long time in liquid nitrogen.Take out cryopreservation tube during recovery, put into 37 DEG C of water-bath middling speeds immediately and melt, after centrifugal segregation cryopreserving liquid, move into and cultivate culture in glassware.
(4) preparation and purification of monoclonal antibody
Increment cultivation: hybridoma is placed in cell culture medium, cultivates under 37 DEG C of conditions, by sad-saturated ammonium sulfate method, the nutrient solution obtained is carried out purifying, obtains monoclonal antibody ,-20 DEG C of preservations.
Described cell culture medium for add calf serum and sodium bicarbonate in RPMI1640 nutrient culture media, the final concentration of calf serum in cell culture medium is made to be 20%(mass percentage), make the final concentration of sodium bicarbonate in cell culture medium be 0.2%(mass percentage); The pH of described cell culture medium is 7.4.
5, the preparation of sheep anti mouse antiantibody
Using sheep as immune animal, for immunogene, immunity is carried out to pathogen-free domestic sheep with mouse source antibody, obtain sheep anti mouse antiantibody.
The preparation of 6, monoclonal antibody-colloid gold label thing
(1) preparation of collaurum
With two ionized water that boils off, 1% gold chloride is diluted to 0.01%(mass percentage), get 100ml and be placed in conical flask, boiling is heated to thermostatic electromagnetic stirrer, 2.5ml1% trisodium citrate is added under continuous high temperature, Keep agitation, continue at the uniform velocity to be heated with stirring to solution be bright red time stopping, original volume is returned to deionized water, 4 DEG C of preservations after being cooled to room temperature.The collaurum outward appearance prepared is pure, bright, nothing precipitates and floating thing.
(2) preparation of golden labeling antibody
Under magnetic stirring, the pH value to 7.0 of collaurum is adjusted with 0.2mol/L solution of potassium carbonate, in colloidal gold solution, metalaxyl antibody is added by the standard adding 20-50 μ g antibody in every milliliter of colloidal gold solution, continue to stir and evenly mix 30min, add 10%BSA, its final concentration in colloidal gold solution is made to be 1%(volumn concentration), leave standstill 10min.12000r/min, 4 DEG C of centrifugal 40min, abandon supernatant, precipitation with redissolve buffer solution twice, with volume be initial colloid gold volume 1/10 redissolution damping fluid will precipitate resuspended, put 4 DEG C for subsequent use.
Redissolve damping fluid: containing bovine serum albumin(BSA) (BSA) 0.1%-0.3%(volumn concentration), Tween-80 0.05%-0.2%(mass percentage), the 0.02mol/L phosphate buffer of pH7.2.
7, the preparation of bond release pad
Bond is discharged pad to be soaked in containing in bovine serum albumin(BSA) (concentration of bovine serum albumin(BSA) in damping fluid is 0.5%) and pH7.2,0.5mol/L phosphate buffer, evenly soak 1h, 37 DEG C of baking 3h are for subsequent use.ZX1000 type spray film instrument is produced with hundred road companies, the metalaxyl prepared monoclonal antibody-colloid gold label thing is evenly coated on bond release pad, every 1cm bond release pad bag is by after 0.01ml metalaxyl monoclonal antibody-colloid gold label thing, take out after being placed in 37 DEG C of environment (humidity < 20%) 60min, sealing is placed in dry environment (humidity < 20%) and saves backup.
8, the preparation of reaction film
Metalaxyl haptens-bovine serum albumin conjugate bag is formed detection line (T line) to reaction film, sheep anti mouse antiantibody is coated on reaction film and forms nature controlling line (C line).
Bag is by process: with phosphate buffer, metalaxyl haptens-bovine serum albumin conjugate is diluted to 10mg/ml, and produce ZX1000 type point film instrument with hundred road companies, be coated in the detection line (T line) on nitrocellulose filter, package amount is 1.0 μ g/cm
2; With the phosphate buffer of 0.01mol/L, pH7.4 by anti-for rabbit goat-anti antibody dilution to 200 μ g/ml, be coated in the nature controlling line (C line) on nitrocellulose filter with a film instrument, package amount is 1.0 μ g/cm
2.Bag is placed in dry 2h under 37 DEG C of conditions by good reaction film, for subsequent use.
9, the preparation of absorption of sample pad
Be placed in by absorption of sample pad and soak 2h containing 0.5% bovine serum albumin(BSA) (volumn concentration), pH7.20.1mol/L phosphate buffer, 37 DEG C of baking 2h are for subsequent use.
10, the assembling of test paper
Absorption of sample pad, bond release pad, reaction film, adsorptive pads are pasted onto on PVC base plate successively in order; Bond release pad has 1/3 region to be covered by absorption of sample pad from initiating terminal, the end of bond release pad is connected with the top of reaction film, the end of reaction film is connected with the top of adsorptive pads, the top of absorption of sample pad aligns with the top of PVC base plate, and the end of adsorptive pads aligns with the end of PVC base plate; Described reaction film has detection line and nature controlling line, detection line (T line) is the strip tape vertical with the appearance of described test paper with nature controlling line (C line); Detection line is positioned at the side of the end near bond release pad; Nature controlling line is positioned at the side of the end away from bond release pad; Test paper machine is cut into the little bar that 3mm is wide, is contained in special plastics fabrication, form test card, under 4-30 DEG C of condition, can 12 months be preserved.
The detection of metalaxyl residue in embodiment 2 sample
1, the pre-treatment of sample
Get 1.0g ± 0.05g homogeneous sample in 50ml polystyrene centrifuge tube, add 5ml extract, use oscillator vibrates 5min, the centrifugal 5min of 3000g room temperature, gets supernatant to be measured.
2, detect with test paper
Vertically dripping 2-3 with suction pipe absorption measuring samples solution drips in well, and start timing during liquid flow, reaction 5min, result of determination, it is invalid that other times judge.
3, testing result is analyzed
Negative (-): T line and C line all develop the color, represents that in sample, metalaxyl drug concentration is lower than detectability, as Fig. 3.
Positive (+): T line, without the colour developing of colour developing C line, represents that in sample, metalaxyl drug concentration is equal to or higher than detectability, as Fig. 3.
Invalid: not occur C line, show the deterioration failure of incorrect operating process or test paper, as Fig. 3.In the case, again should read over instructions, and retest with new test card.
Embodiment 3 sample detection example
1, detectability test
Get blank tobacco sample, add respectively wherein metalaxyl to final concentration be 1,2,4mg/kg, get test paper and detect, each sample replication three times.
During with detection paper tobacco, tealeaves, vegetables sample, when wherein metalaxyl interpolation concentration is 1mg/kg, test paper demonstrates macroscopic two red lines, become negative; Be 2 when wherein metalaxyl adds concentration, 4mg/kg time, test paper nature controlling line shows, but detection line does not show, and becomes positive; Show that this test paper is 2mg/kg to the detection line of metalaxyl.
2, false positive rate, false negative rate test
Get known metalaxyl content and be greater than the tobacco positive sample 20 parts of 2mg/kg and content is less than 2mg/kg tobacco negative sample 20 parts, detect with three batches of test paper, calculate its yin and yang attribute rate.The results are shown in Table 1, table 2.
Table 1 detects positive sample result
Table 2 detects negative sample result
Result shows: during the detection paper positive sample of producing with 3 batches, and result is positive entirely, and known positive sample coincidence rate is 100%, and false negative rate is 0; When detecting 20 parts of negative sample, have one batch to occur a false positive test results, known negative match-rate is 98%, and false positive rate is less than 5%.Illustrate that the test paper of detection metalaxyl residue of the present invention can detect fast to metalaxyl residue in tobacco.
3, specific test
Specificity is commonly used cross reacting rate and is represented, refers to the ability of the antigenic determinant generation combination that antibody is different from structure.By often examine carbendazim, Triadimenol, chrysanthemum esters medicine 500 μ g/L sample, detect with metalaxyl colloid gold test paper.Result shows, and during with this detection paper carbendazim, Triadimenol, chrysanthemum esters medicine 500 μ g/L, test paper nature controlling line and detection line all develop the color, and present feminine gender, illustrate that this test paper is to these medicine no cross reactions.
Claims (7)
1. one kind is detected the test paper of metalaxyl, comprise absorption of sample pad (1), bond release pad (2), reaction film (3), adsorptive pads (4) and base plate (7), it is characterized in that described reaction film having the detection line (5) being coated with metalaxyl hapten-carrier protein conjugate and the nature controlling line (6) being coated with sheep anti mouse antiantibody, described bond release pad (2) is coated with metalaxyl monoclonal-colloid gold label thing.
2. test paper as claimed in claim 1, is characterized in that described absorption of sample pad (1), bond release pad (2), reaction film (3), adsorptive pads (4) are pasted onto on base plate (7) successively.
3. the test paper as described in any one of claim 1-2, is characterized in that described bond release pad (2) is stacked on bond release pad (3), and bond release pad is under 1/3-1/2 is capped on absorption of sample pad.
4. test paper as claimed in claim 1, it is characterized in that described metalaxyl hapten-carrier protein conjugate is obtained by metalaxyl medicine haptens and carrier protein couplet, described carrier protein can be bovine serum albumin(BSA), ovalbumin, hemocyanin, thyroprotein, human serum albumins.
5. the test paper as described in any one of claim 1 or 4, it is characterized in that described metalaxyl haptens is obtained by metalaxyl hydrolysis reaction, its molecular structural formula is:
。
6. test paper as claimed in claim 1, it is characterized in that described metalaxyl monoclonal antibody prepares using metalaxyl hapten-carrier protein conjugate as immunogene, mouse source antibody mediated immunity sheep obtains by described sheep anti mouse antiantibody.
7. prepare a method for test paper described in any one of claim 1-6, it comprises step:
1) preparation is coated with bond release pad (2) of metalaxyl monoclonal antibody-colloid gold label thing;
2) preparation has the reaction film of the detection line being coated with metalaxyl hapten-carrier protein conjugate and the nature controlling line being coated with sheep anti mouse antiantibody;
3) by 1) and 2) bond for preparing release pad, reaction film and absorption of sample pad, adsorptive pads and PVC base plate be assembled into test paper.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510530296.2A CN105259351A (en) | 2015-08-26 | 2015-08-26 | Test paper for detecting metalaxyl residue and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510530296.2A CN105259351A (en) | 2015-08-26 | 2015-08-26 | Test paper for detecting metalaxyl residue and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105259351A true CN105259351A (en) | 2016-01-20 |
Family
ID=55099117
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510530296.2A Pending CN105259351A (en) | 2015-08-26 | 2015-08-26 | Test paper for detecting metalaxyl residue and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105259351A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572376A (en) * | 2016-03-07 | 2016-05-11 | 中国烟草总公司贵州省公司 | Test paper for detecting residual thiophanate methyl in crops, and application and preparation method thereof |
| CN105807056A (en) * | 2016-03-21 | 2016-07-27 | 贵州勤邦食品安全科学技术有限公司 | Test paper card for detecting efficient cyhalothrin residue |
| CN106918705A (en) * | 2017-01-22 | 2017-07-04 | 贵州勤邦食品安全科学技术有限公司 | Detect test paper and its application of Fenpropathrin |
| CN108931646A (en) * | 2018-05-30 | 2018-12-04 | 中国烟草总公司郑州烟草研究院 | A kind of test strips and its preparation method and application detecting safrole |
| CN108997161A (en) * | 2018-09-21 | 2018-12-14 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method and application of metalaxyl haptens and antigen |
| CN109061154A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of fluorescent micro-ball immune chromatography test paper strip and its preparation method and application detecting metalaxyl |
| CN109061150A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application detecting metalaxyl |
| CN109061155A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of test strips and its preparation method and application detecting metalaxyl |
| CN109336779A (en) * | 2018-09-21 | 2019-02-15 | 中国烟草总公司郑州烟草研究院 | A kind of preparation and application of metalaxyl haptens and antigen |
| CN110887965A (en) * | 2019-12-10 | 2020-03-17 | 中国农业科学院烟草研究所 | Detection card for testing metalaxyl residue |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2833595Y (en) * | 2005-08-24 | 2006-11-01 | 万积成 | Colloid gold test paper for quick detection of metalaxyl residue |
-
2015
- 2015-08-26 CN CN201510530296.2A patent/CN105259351A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2833595Y (en) * | 2005-08-24 | 2006-11-01 | 万积成 | Colloid gold test paper for quick detection of metalaxyl residue |
Non-Patent Citations (1)
| Title |
|---|
| 周洪斌等: "进出口食品中甲霜灵残留量免疫学快速检测方法的研究", 《畜牧与兽医》 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572376A (en) * | 2016-03-07 | 2016-05-11 | 中国烟草总公司贵州省公司 | Test paper for detecting residual thiophanate methyl in crops, and application and preparation method thereof |
| CN105572376B (en) * | 2016-03-07 | 2017-07-11 | 中国烟草总公司贵州省公司 | A kind of test paper and its application, preparation method for detecting that thiophanate-methyl is remained in crops |
| CN105807056A (en) * | 2016-03-21 | 2016-07-27 | 贵州勤邦食品安全科学技术有限公司 | Test paper card for detecting efficient cyhalothrin residue |
| CN106918705A (en) * | 2017-01-22 | 2017-07-04 | 贵州勤邦食品安全科学技术有限公司 | Detect test paper and its application of Fenpropathrin |
| CN108931646B (en) * | 2018-05-30 | 2021-04-27 | 中国烟草总公司郑州烟草研究院 | Test strip for detecting safrole and preparation method and application thereof |
| CN108931646A (en) * | 2018-05-30 | 2018-12-04 | 中国烟草总公司郑州烟草研究院 | A kind of test strips and its preparation method and application detecting safrole |
| CN108997161A (en) * | 2018-09-21 | 2018-12-14 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method and application of metalaxyl haptens and antigen |
| CN109061154A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of fluorescent micro-ball immune chromatography test paper strip and its preparation method and application detecting metalaxyl |
| CN109061150A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application detecting metalaxyl |
| CN109061155A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of test strips and its preparation method and application detecting metalaxyl |
| CN109336779A (en) * | 2018-09-21 | 2019-02-15 | 中国烟草总公司郑州烟草研究院 | A kind of preparation and application of metalaxyl haptens and antigen |
| CN109336779B (en) * | 2018-09-21 | 2021-02-19 | 中国烟草总公司郑州烟草研究院 | Preparation and application of metalaxyl hapten and antigen |
| CN109061155B (en) * | 2018-09-21 | 2021-05-11 | 中国烟草总公司郑州烟草研究院 | Test strip for detecting metalaxyl and preparation method and application thereof |
| CN109061150B (en) * | 2018-09-21 | 2021-07-23 | 中国烟草总公司郑州烟草研究院 | Time-resolved fluorescence immunochromatographic test strip for detecting metalaxyl and preparation method and application thereof |
| CN109061154B (en) * | 2018-09-21 | 2021-08-17 | 中国烟草总公司郑州烟草研究院 | Fluorescent microsphere immunochromatography test strip for detecting metalaxyl and preparation method and application thereof |
| CN110887965A (en) * | 2019-12-10 | 2020-03-17 | 中国农业科学院烟草研究所 | Detection card for testing metalaxyl residue |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105259351A (en) | Test paper for detecting metalaxyl residue and application thereof | |
| CN104569399B (en) | A kind of test strips detecting ochratoxin A and application thereof | |
| CN105759043A (en) | Test paper for simultaneous detection of carbendazim and thiophanate-methyl residues in crops, and application and preparation method thereof | |
| CN103575889B (en) | A kind of test strips and method detecting vancomycin | |
| CN105424939B (en) | A kind of test strips for detecting carbendazim and its preparation method and application | |
| CN103941000B (en) | A kind of detect sulfa drugs and the test strips of FQNS and method | |
| CN103575887B (en) | A kind of test card and application thereof detecting aflatoxin B1 | |
| CN105486872A (en) | Test strip for detecting triadimenol and preparation method and application thereof | |
| CN105785011A (en) | Test strip for detecting maleic hydrazide, as well as preparation method and application thereof | |
| CN105388286A (en) | Test strip for detecting iprodione and preparation method and application of test strip | |
| CN105004844B (en) | A kind of gentamicin residue test strip and application thereof | |
| CN103728449B (en) | A kind of test paper and method detecting florfenicol and thiamphenicol | |
| CN103389373B (en) | A kind of test paper and application thereof detecting sulfa drugs | |
| CN103777015A (en) | Colloidal gold test strip and method for detecting erythromycin | |
| CN103364546B (en) | A kind of kit and method detecting Furaxone metabolite | |
| CN103713133B (en) | Detection spiramycin, streptomycin, gentamycin, the test strips of neomycin and method | |
| CN104345145B (en) | A kind of test strips detecting vomitoxin and application thereof | |
| CN108931640B (en) | Test strip for detecting organophosphorus pesticide and application thereof | |
| CN103389379A (en) | Test strip and method for detecting tylosin and tilmicosin | |
| CN202956385U (en) | Test paper used for detecting florfenicol and thiamphenicol | |
| CN103364555B (en) | A kind of kit and method detecting Furacilin metabolite | |
| CN105158472A (en) | Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method | |
| CN109682961A (en) | A kind of test strips and its application detecting cyphenothrin | |
| CN104215763B (en) | A kind of test strips detecting T-2 toxin and application thereof | |
| CN103293304B (en) | A kind of kit and method detecting sulfa drugs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160120 |
|
| WD01 | Invention patent application deemed withdrawn after publication |