CN105158472A - Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method - Google Patents
Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method Download PDFInfo
- Publication number
- CN105158472A CN105158472A CN201510529378.5A CN201510529378A CN105158472A CN 105158472 A CN105158472 A CN 105158472A CN 201510529378 A CN201510529378 A CN 201510529378A CN 105158472 A CN105158472 A CN 105158472A
- Authority
- CN
- China
- Prior art keywords
- metalaxyl
- kit
- enzyme linked
- solution
- linked immunosorbent
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (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)
- Peptides Or Proteins (AREA)
Abstract
The invention provides an enzyme linked immunosorbent assay kit for detecting metalaxyl residues and a preparation method of the enzyme linked immunosorbent assay kit. The enzyme linked immunosorbent assay kit comprises an elisa plate enveloped with metalaxyl coupled antigen, metalaxyl standard solution, an enzyme-labeled second antibody, antibody concentrated solution, substrate developing solution, stop buffer, cleaning solution and reconstitution fluid. The enzyme linked immunosorbent assay kit can be used for quickly detecting metalaxyl residues in tobaccos, tea leaves and vegetables, and has features of quick detection, sensitivity, good specificity, high accuracy and low detection cost; and requirements of quick qualitative and quantitative detection on site can be met.
Description
Technical field
The invention belongs to enzyme linked immunosorbent detection field, being specifically related to a kind of enzyme linked immunological kit for detecting metalaxyl medicament residue, it is specially adapted to the detection of metalaxyl residue in tobacco, tealeaves, vegetables sample.
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, instrument detection method needs expensive large-scale instrument and equipment simultaneously, be equipped with the detection technique personnel of specialty, carry out operating and managing, scene cannot be carried out detect on a large scale, poor in timeliness, be difficult to promote.
Medicament residue immunoassay technology is in Safety of Food Quality detects fast in recent years, be widely used, and gradually approve by people, it is simple that enzyme linked immunosorbent detection technology has sample pre-treatments, detect quick, sensitive, specificity is good, accuracy high, the requirement of field quick detection can be met, thus make up the deficiency of instrument detection.
Summary of the invention
The object of the present invention is to provide a kind of enzyme linked immunological kit that can detect metalaxyl residue, adopt indirect competitive ELISA method, and provide a kind of efficient, accurate, easy, be suitable for batch samples screening qualitative and quantitative analysis method and kit preparation method, the detection of the method to tobacco, tealeaves and vegetables is limited to 100 μ g/kg, and sensitivity is 1 μ g/kg.
Kit of the present invention, it comprises: be coated with the ELISA Plate of coating antigen, metalaxyl standard solution, high standard product, ELIAS secondary antibody, antibody concentrated solution, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid, described coating antigen is metalaxyl coupled antigen, and described ELIAS secondary antibody is enzyme labeling antiantibody.
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 specific antibody prepares using metalaxyl coupled antigen as immunogene, and described metalaxyl specific antibody can be metalaxyl monoclonal antibody or metalaxyl polyclonal antibody, wherein preferred metalaxyl monoclonal antibody.
Described antiantibody is sheep anti mouse antiantibody or goat-anti rabbit antiantibody, wherein preferred sheep anti mouse antiantibody.
The marker enzyme of described ELIAS secondary antibody is that horseradish peroxidase or bacterium extract alkaline phosphatase, wherein preferred horseradish peroxidase; The antiantibody of enzyme labeling adopts glutaraldehyde method or Over-voltage protection that marker enzyme and antiantibody are carried out coupling and obtains.
In order to more convenient on-site supervision and great amount of samples examination, described kit also comprises metalaxyl standard solution, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid.
Described standard solution 6 bottles, concentration is respectively 0 μ g/L, 1 μ g/L, 3 μ g/L, 9 μ g/L, 27 μ g/L, 81 μ g/L.
When marker enzyme is horseradish peroxidase, described nitrite ion is made up of substrate nitrite ion A liquid and substrate nitrite ion B liquid, substrate nitrite ion A liquid is hydrogen peroxide or urea peroxide, substrate nitrite ion B liquid is o-phenylenediamine or tetramethyl benzidine, and described stop buffer is sulfuric acid or the hydrochloride buffer of 1-2mol/L; When marker enzyme is alkaline phosphatase, described nitrite ion is to nitro phosphate buffer, and described stop buffer is 1-2mol/L sodium hydroxide solution.
It is 7.2-7.5 that described cleansing solution is preferably pH value, and the phosphate buffer containing 0.8%-1.0% Tween-20,0.03 ‰-0.05 ‰ sodium azide preservatives, 0.1-0.3mol/L, described number percent is percent weight in volume.
Described redissolution liquid is preferably that pH value is 7.0, the phosphate buffer of 0.02mol/L, and described number percent is percent weight in volume.
It is pH value is 9.2-9.6 that wherein used in ELISA Plate preparation process bag is buffered liquid, the carbonate buffer solution of 0.1-0.2mol/L, confining liquid is pH value is 7.1-7.5, the phosphate buffer containing 1%-3% casein, 0.1-0.3mol/L, and described number percent is percent weight in volume.
In the present invention, the preparation process of ELISA Plate is: be buffered liquid with bag and coating antigen is diluted to 20 μ g/ml, every hole adds 100 μ l, 37 DEG C of lucifuges are hatched 2h or 4 DEG C and are spent the night, and liquid in hole of inclining, washs 1 time with cleansing solution, stop 30s, pat dry, in every hole, then add 150 μ l confining liquids, 37 DEG C of lucifuges hatch 1-2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
Cleaning Principle of the present invention is:
Pre-coated metalaxyl coupled antigen in ELISA Plate, after adding sample solution or standard solution, add metalaxyl specific antibody solution again, metalaxyl coupled antigen metalaxyl medicine residual in sample and ELISA Plate being wrapped quilt competes metalaxyl specific antibody, add ELIAS secondary antibody and carry out amplification, with nitrite ion colour developing, the content of sample absorbance and metalaxyl medicine is negative correlation, compares the residual quantity that can obtain metalaxyl in sample with typical curve; Simultaneously according to the depth of color in ELISA Plate, can the concentration range of metalaxyl residue amount in judgement sample roughly with comparing of the metalaxyl standard solution color of series concentration.
The enzyme linked immunological kit that the present invention detects metalaxyl mainly adopts the content of metalaxyl in the qualitative or quantitative detection sample of competitive ELISA method; Require low to the pre-treatment of sample, sample pretreatment process is simple, can detect batch samples fast simultaneously; Main agents provides with the form of working fluid, and the method for inspection is convenient and easy, has that specificity is high, highly sensitive, degree of accuracy is high, accuracy high.Enzyme linked immunological kit of the present invention, structure is simple, easy to use, low price, carrying convenience, detection method are efficient, accurate, easy, be suitable for the qualitative, quantitative of batch samples screening.
Accompanying drawing explanation
Fig. 1: metalaxyl hapten synthesis route map;
Fig. 2: kit standard curve map.
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 reagent constituents
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, the preparation of antigen
Prepared by immunogene---and 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.
Prepared by immunogene---and metalaxyl haptens and ovalbumin (OVA) 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 OVA(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 metalaxyl monoclonal antibody
Animal immune: immunogene above-mentioned steps obtained is injected in Balb/c Mice Body, immunizing dose is 150 μ g/, makes it produce antiserum.
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: monoclonal hybridoma strain cryopreserving liquid is made 1 × 10
6the cell suspension of individual/ml, preserves 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: only Balb/c mouse peritoneal is injected sterilizing paraffin oil 0.5ml/, within 7 days, pneumoretroperitoneum injects stable monoclonal hybridoma strain 5 × 10
5individual/only, gather ascites after 7 days.Carry out ascites by sad-saturated ammonium sulfate method to purify ,-20 DEG C of preservations.
4, the preparation of sheep anti mouse antiantibody
Take sheep as immune animal, with mouse source antibody for immunogen immune pathogen-free domestic sheep, obtain sheep anti mouse antiantibody.
5, the preparation of ELIAS secondary antibody
The Over-voltage protection after improveing is adopted to carry out coupling sheep anti mouse antiantibody and horseradish peroxidase (HRP).In reaction system after improvement, the molar concentration rate of enzyme and antibody is 1.8-2.4:1.
6, the preparation of ELISA Plate
Be buffered liquid with bag and coating antigen is diluted to 20 μ g/ml, every hole adds 100 μ l, 37 DEG C of lucifuges hatch 2h, and liquid in hole of inclining washs 1 time with cleansing solution, stop 30s, pat dry, in every hole, then add 150 μ l confining liquids, 37 DEG C of lucifuges hatch 2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
Embodiment 2 detects the establishment of the enzyme linked immunological kit of metalaxyl
Set up the enzyme linked immunological kit detecting metalaxyl, make it comprise following component:
(1) bag is by the ELISA Plate of metalaxyl coupled antigen;
(2) metalaxyl standard solution 6 bottles, concentration is respectively 0 μ g/L, 1 μ g/L, 3 μ g/L, 9 μ g/L, 27 μ g/L, 81 μ g/L;
(3) metalaxyl monoclonal antibody working fluid;
(4) with the sheep anti mouse antiantibody of horseradish peroxidase-labeled;
(5) substrate nitrite ion is made up of substrate nitrite ion A liquid and substrate nitrite ion B liquid, and substrate nitrite ion A liquid is urea peroxide, and substrate nitrite ion B liquid is tetramethyl benzidine;
(6) stop buffer is 2mol/L sulfuric acid;
(7) cleansing solution is pH value 7.2-7.5, and the phosphate buffer containing 0.8%-1.0% Tween-20,0.03 ‰-0.05 ‰ sodium azide preservatives, 0.1-0.3mol/L, described number percent is percent weight in volume.(8) redissolve liquid to be pH value be 7.0, the phosphate buffer of 0.02mol/L, described number percent is percent weight in volume.
The detection of metalaxyl in embodiment 3 tobacco, tealeaves, vegetables
1, sample pre-treatments
Tobacco, tealeaves, vegetables Sample pretreatment method
Take 1.0 ± 0.05g sample in 50ml polystyrene centrifuge tube, add 5ml methyl alcohol, dissolve completely to sample with oscillator vibrates, the centrifugal 5min of 3000g under room temperature; Pipetting in supernatant 25 μ l to 2ml clean dried polystyrene centrifuge tube, add 975 μ l redissolution liquid, with vortex instrument whirling motion mixing, getting 50 μ l for analyzing.
2. the configuration of solution
The configuration of redissolution liquid
Undertaken diluting (1 part of redissolution liquid+1 part of deionized water) redissolution for sample by 1:1 volume ratio by redissolution liquid with deionized water, the working fluid that redissolves can preserve one month at 4 DEG C of (39.2 ℉) environment.
The configuration of cleansing solution
Undertaken diluting (1 part of cleansing solution+19 parts of deionized waters) washing for ELISA Plate by 1:19 volume ratio by cleansing solution with deionized water, wash operating solution can preserve one month at 4 DEG C of (39.2 ℉) environment.
Antibody concentrated solution mixes with ELIAS secondary antibody liquid
By antibody concentrated solution and ELIAS secondary antibody working fluid with the volume mixture of 1:10 (namely 1 part of antibody concentrated solution mixes with 10 parts of ELIAS secondary antibody working fluids).Note: now with the current.
3, detect with kit
To in the ELISA Plate micropore being coated with metalaxyl coupled antigen, add metalaxyl standard solution or the sample solution 50 μ l/ hole through pre-treatment, then the mixed liquor 50 μ l/ hole of antibody concentrated solution and ELIAS secondary antibody working fluid is added, to vibrate gently mixing, react 30min with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate; Pour out liquid in hole, every hole adds 250 μ l cleansing solutions and fully washs 4-5 time, and every minor tick 10s, pats dry with thieving paper; Every hole adds substrate nitrite ion A liquid 50 μ l, add substrate nitrite ion B liquid 50 μ l again, to vibrate gently mixing, with cover plate membrane cover plate rearmounted 25 DEG C of lucifuges colour developing 15min, every hole adds stop buffer 2mol/L sulfuric acid 50 μ l, to vibrate gently mixing, with microplate reader dual wavelength 450/630nm place, measure every hole absorbance (OD value).If without microplate reader, then not adding stop buffer ocular estimate can judge.
4, Analysis of test results
The mean absorbance values (diplopore) of standard items or sample divided by the mean absorbance values of first standard items (0 standard), then is multiplied by 100%, obtains the percentage absorbance of standard items or sample.With standard items percentage absorptance for ordinate, with the logarithm of metalaxyl standard concentration (μ g/L) for horizontal ordinate, drawing standard curve map.The percentage absorptance of sample substituted in typical curve, read the concentration corresponding to sample from typical curve, the extension rate being multiplied by its correspondence is metalaxyl actual concentrations in sample.
The determination test of embodiment 4 metalaxyl technical parameter
1, kit sensitivity and detectability
Conventionally measure kit sensitivity, kit standard curve minimum point is 1 μ g/L, and the scope of typical curve is 1-81 μ g/L, IC
50(50% inhibition concentration) domain of walker is 8-16 μ g/L; 20 increment product are detected, the concentration corresponding to each percentage absorbance is found from typical curve, add that 3 times of standard deviations represent detectability with the mean value of 20 parts of concentration of specimens, result obtains the detection of the method to tobacco, tealeaves and vegetables and is limited to 100 μ g/kg, and sensitivity is 1 μ g/kg.
2, sample preci-sion and accuracy test
Using the recovery as accuracy estimating index, the testing result relative standard deviation (RSD%) of a certain concentration samples of replication is as precision evaluation index.Computing formula is: the recovery (%)=practical measurement value/theoretical value × 100%, and wherein theoretical value is the interpolation concentration of sample; Relative standard deviation RSD%=SD/X × 100%, wherein SD is standard deviation, and X is the mean value of determination data.
Respectively by 100 μ g/g, 200 μ g/g, 400 μ g/g, tri-concentration metalaxyls tobacco, tealeaves, vegetable sample are carried out to interpolation and reclaim mensuration, each sample do 4 parallel, measure with three batches of different reagent, average recovery rate and the precision of calculation sample the results are shown in following table.
Table 1 tobacco precision and accuracy test
Table 2 tealeaves precision and accuracy test
Table 3 vegetables precision and accuracy test
Add tobacco, tealeaves and vegetable sample respectively with the metalaxyl of 100,200,400 μ g/g, tri-concentration, average recovery rate is between 84.5%-105.8%; Batch in, batch between relative standard deviation be all less than 10%.
3, stabilization of kit test
Kit preservation condition is 2-8 DEG C, and through the mensuration of 12 months, the maximum absorbance value (zero standard) of kit, 50% inhibition concentration, metalaxyl added practical measurement value all within normal range.Consider in transport and use procedure, have improper preservation condition and occur, placed 7 days under 37 DEG C of preservation conditions by kit, carry out accelerated aging tests, result shows that this kit indices meets the requirements completely.Consider that the freezing situation of kit occurs, kit is put into-20 DEG C of refrigerator freezings 7 days, measurement result also shows that kit indices is completely normal.Can show that kit at least can be preserved more than 12 months at 2-8 DEG C from above result.
Claims (5)
1. one kind is detected the enzyme linked immunological kit of metalaxyl residue, comprise: be coated with the ELISA Plate of metalaxyl coupled antigen, metalaxyl standard solution, ELIAS secondary antibody, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid, it is characterized in that described metalaxyl coupled antigen is obtained by metalaxyl haptens and carrier protein couplet.
2. kit as described in claim 1, it is characterized in that described metalaxyl haptens is obtained by metalaxyl hydrolysis reaction, molecular structural formula is:
。
3. kit as claimed in claim 1, is characterized in that described metalaxyl specific antibody prepares using metalaxyl coupled antigen as immunogene, the preferred metalaxyl monoclonal antibody of described metalaxyl specific antibody.
4. kit as claimed in claim 1, it is characterized in that described cleansing solution is pH value 7.2-7.5, phosphate buffer containing 0.8%-1.0% Tween-20,0.03 ‰-0.05 ‰ sodium azide preservatives, 0.1-0.3mol/L, described number percent is percent weight in volume.
5. kit as claimed in claim 1, is characterized in that the concentration of described metalaxyl standard solution is respectively 0 μ g/L, 1 μ g/L, 3 μ g/L, 9 μ g/L, 27 μ g/L, 81 μ g/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510529378.5A CN105158472A (en) | 2015-08-26 | 2015-08-26 | Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510529378.5A CN105158472A (en) | 2015-08-26 | 2015-08-26 | Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105158472A true CN105158472A (en) | 2015-12-16 |
Family
ID=54799394
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510529378.5A Pending CN105158472A (en) | 2015-08-26 | 2015-08-26 | Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105158472A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN109324187A (en) * | 2018-09-21 | 2019-02-12 | 中国烟草总公司郑州烟草研究院 | It is a kind of detect metalaxyl enzyme linked immunological kit and its application |
| 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 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5601847A (en) * | 1993-04-01 | 1997-02-11 | Ciba-Geigy Corporation | Microbicides |
| 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 CN201510529378.5A patent/CN105158472A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5601847A (en) * | 1993-04-01 | 1997-02-11 | Ciba-Geigy Corporation | Microbicides |
| CN2833595Y (en) * | 2005-08-24 | 2006-11-01 | 万积成 | Colloid gold test paper for quick detection of metalaxyl residue |
Non-Patent Citations (7)
| Title |
|---|
| LEE，KANG-BONG等: "Production of Polycolonal Antibody and Optimum Conditions in ELISA for Metalaxyl", 《KOREAN JOURNAL OF ENVIRONMENTAL AGRICULTURE》 * |
| WILLIAM H.NEWSOME: "An enzyme-linked immunosorbent assay for metalaxyl in foods", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| 周洪斌等: "进出口植物产品中甲霜灵残留量酶联免疫检测试剂盒的研究", 《免疫学杂志》 * |
| 江小雪等: "酶联免疫分析技术在杀菌剂残留检测中的应用", 《浙江农业科学》 * |
| 蒋木庚: "免疫化学技术及其在农化分析中的应用", 《江苏农药》 * |
| 郭乃菲等: "食品中杀菌剂残留的免疫分析技术研究", 《食品工业科技》 * |
| 钱传范等: "免疫检测技术在农药残留分析中的应用", 《农药科学与管理》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN109324187A (en) * | 2018-09-21 | 2019-02-12 | 中国烟草总公司郑州烟草研究院 | It is a kind of detect metalaxyl enzyme linked immunological kit and its application |
| 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 |
| CN109324187B (en) * | 2018-09-21 | 2021-08-24 | 中国烟草总公司郑州烟草研究院 | Enzyme linked immunosorbent assay kit for detecting metalaxyl and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104977406B (en) | Detect enzyme linked immunological kit and its application of FQNS | |
| CN105137009A (en) | ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit | |
| CN106324240B (en) | Detect enzyme linked immunological kit and its application of chlopyrifos | |
| CN101839918B (en) | Method for detecting spiramycin and special enzyme-linked immunoassay kit thereof | |
| CN105044325B (en) | The enzyme linked immunological kit of detection triazophos and application thereof | |
| CN105785012B (en) | Detect enzyme linked immunological kit and its application of Ribavirin | |
| CN102967709A (en) | Enzyme linked immunosorbent assay kit for detecting zearalenone drug and application thereof | |
| CN104076154A (en) | Enzyme linked immunosorbent assay kit detecting folic acid and application thereof | |
| CN105572348B (en) | Detect enzyme linked immunological kit and its application of Triadimenol | |
| CN104897864A (en) | ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine and application of ELISA kit | |
| CN102080067B (en) | Method for detecting deoxynivalenol and special reagent kit thereof | |
| CN105277708A (en) | Enzyme linked immunosorbent assay kit for detecting aflatoxin B1 in chili | |
| CN105021815A (en) | Enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate and application of enzyme linked immunosorbent assay kit | |
| CN105158472A (en) | Enzyme linked immunosorbent assay kit for detecting metalaxyl residues and using method | |
| CN105301233A (en) | Enzyme linked immunosorbent assay kit for detecting imidacloprid residue in tea leaves and application method of enzyme linked immunosorbent assay kit | |
| CN105403703B (en) | Detect enzyme linked immunological kit and its application of carbendazim | |
| CN103364553B (en) | The enzyme linked immunological kit of detection nitroimidazoles medicine and application thereof | |
| CN103018451B (en) | The enzyme linked immunological kit of chlorine detection mycin and application thereof | |
| CN110133306A (en) | Detect enzyme linked immunological kit and its application of Cimaterol | |
| CN105675858A (en) | Enzyme-linked immunosorbent assay kit for detecting quinclorac and application of enzyme-linked immunosorbent assay kit | |
| CN101782579B (en) | Enzyme-linked immunoassay kit for detecting tiamulin medicaments, and application thereof | |
| CN103575885A (en) | Enzyme linked immunoassay kit for detecting T-2 toxin, and application thereof | |
| CN103575914A (en) | Enzyme linked immunosorbent assay kit for detecting vitamin B12 and application thereof | |
| CN103018447B (en) | Detect enzyme linked immunological kit and the method thereof of salbutamol | |
| CN105272866A (en) | Phenylethanolamine A hapten, antigen as well as preparation method and application thereof |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20151216 |