CN103018449B - Detect enzyme linked immunological kit and the method thereof of AMOZ - Google Patents
Detect enzyme linked immunological kit and the method thereof of AMOZ Download PDFInfo
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Abstract
The invention provides a kind of detection AMOZ enzyme linked immunological kit and method thereof, enzyme linked immunological kit comprises: be coated with the ELISA Plate of coating antigen, AMOZ specific antibody, enzyme marker, AMOZ standard solution, substrate nitrite ion, stop buffer, concentrated cleaning solution, the concentrated liquid that redissolves, 2-nitrobenzaldehyde.Present invention also offers a kind of AMOZ enzyme linked immunological kit detection method, mainly comprise: first carry out Sample pretreatment, then detect with kit, ultimate analysis testing result.Detection kit of the present invention can detect AMOZ medicine in animal tissue, aquatic products, honey fast, has the features such as easy and simple to handle, quick, accurate, highly sensitive, low cost, is applicable to examination and the on-site supervision of great amount of samples.
Description
Technical field
The present invention relates to enzyme linked immunosorbent detection technology, be specifically related to a kind of enzyme linked immunological kit and the detection method that detect AMOZ in animal tissue's (muscle, liver), aquatic products, honey.
Background technology
Nitrofuran metabolites, because having extraordinary antibacterial action and the dynamic (dynamical) characteristic of medicine, was once widely used, as the somatotrophic adjuvant of bird, aquatic products and pig.But find in long process of experimental, Nitrofuran metabolites and metabolin all can make animal used as test generation canceration and gene mutation, this type of drug withdrawal is just caused to use in treatment and feed Just because of this.
Due to itrofurans prototype medicine, metabolism is rapid in vivo, cannot detect, but its metabolic product is because of with protein bound and quite stable, so the product after often will analyzing its metabolism when analyzing this type of medicine residual, administrative authority just reaches to detect metabolic product for means the object that detection itrofurans remains.Furaltadone is as the one of Nitrofuran metabolites, and metabolic product is AMOZ.The most popular method being used for detecting Nitrofuran metatolites is at present LC-UV, LC-MS and LC-MS/MS, by comparison, enzyme-linked immunoassay method has the feature such as pinpoint accuracy and sensitivity, the requirement of lower operative technique, of short duration detection time, larger detection sample size, can meet China's livestock and poultry cultivation family, slaughterhouse, food enterprise, government function supervision department etc. better and carry out testing.
Summary of the invention
The object of this invention is to provide a kind of enzyme linked immunological kit of AMOZ, and a kind of efficient, accurate, easy, qualitative and quantitative analysis method of being applicable to carry out batch samples examination is provided.
AMOZ enzyme-linked immunologic detecting kit provided by the invention, include: be coated with the ELISA Plate of coating antigen, AMOZ specific antibody, enzyme marker, standard solution, substrate nitrite ion, stop buffer, concentrated cleaning solution, concentrated liquid, the 2-nitrobenzaldehyde of redissolving, described coating antigen is AMOZ antigen, and described enzyme marker is enzyme labeling antiantibody.
AMOZ enzyme linked immunological kit provided by the present invention, described AMOZ antigen is the conjugate of AMOZ haptens and carrier protein, and described carrier protein is bovine serum albumin(BSA), mouse haemocyanin, rabbit serum proteins, thyroprotein, ovalbumin, hemocyanin or human serum albumins.
Described AMOZ specific antibody is AMOZ monoclonal antibody, obtains with AMOZ immunogen immune animal.Described AMOZ monoclonal antibody can be mouse source, Ma Yuan, Yang Yuan, rabbit source or cavy source antibody, and described AMOZ monoclonal antibody is preferably AMOZ mouse monoclonal antibody.
The marker enzyme of described enzyme marker is horseradish peroxidase, and enzyme labeling antiantibody adopts Over-voltage protection horseradish peroxidase and the coupling of sheep anti mouse antiantibody to be obtained.
In order to carry out great amount of samples examination and on-site supervision more easily, described kit also comprises: standard solution, substrate nitrite ion, stop buffer, concentrated cleaning solution, concentrated liquid, the 2-nitrobenzaldehyde of redissolving.
Described standard solution is serial AMOZ standard solution, and concentration is respectively 0 μ g/L, 0.05 μ g/L, 0.15 μ g/L, 0.45 μ g/L, 1.35 μ g/L, 4.05 μ g/L, 1mL/ bottle.Described stop buffer is the sulfuric acid solution of 1 ~ 2mol/L, described substrate nitrite ion A liquid is urea peroxide solution, substrate nitrite ion B liquid is tetramethyl biphenyl amine aqueous solution, described concentrated cleaning solution is the 0.2 ~ 0.4mol/LpH7.6 phosphate buffer of sodium azide preservatives containing 1% ~ 2% Tween-20 and 0.01% ~ 0.05%, described concentrated redissolution liquid is pH value is 7.3 ~ 7.9, the phosphate buffer containing 7% ~ 9% casein, 0.1mol/L.
The bag of wherein said ELISA Plate used by preparation process is buffered the carbonate buffer solution that liquid is the 0.05mol/L of pH9.4 ~ 9.6, and Block buffer is the phosphate buffer of the bovine serum albumin(BSA) (BSA) of 0.05% ~ 0.5% of pH7.4.
The preparation of ELISA Plate of the present invention is mainly: be buffered liquid with bag and coating antigen is diluted to 0.2 ~ 0.3 μ g/ml, every hole adds 150 μ l, 37 DEG C of lucifuges are hatched 2h or 4 DEG C and are spent the night, incline liquid in hole, and cleansing solution washing 1 ~ 2 time, pats dry, 150 μ l confining liquids are added in every hole, 37 DEG C of lucifuges hatch 2h, and in hole of inclining, liquid pats dry, and preserve with the vacuum seal of aluminium film.
In the present invention, coating antigen and immunogenic building-up process are:
1. hapten synthesis (synthetic route is as Fig. 1)
The mixed liquor of 2.01g AMOZ (AMOZ) and 20mlDMF, slowly be added dropwise under room temperature in the 50-100mlDMF solution of 2.68-5.36g terephthalaldehyde, dropwise rear room temperature to 60 DEG C reaction 2-4 hour, except desolventizing, column chromatography purification, obtains faint yellow AMOZ derivant.
2. immunogenic synthesis
(1) get AMOZ haptens 14mg 1mlDMF to dissolve, obtain solution 1.
(2) get the water-soluble solution of BSA40mg 6ml, obtain solution 2.
(3) solution 1 is added dropwise in solution 2, obtains solution 3, room temperature reaction 24h.
(4) NaBH is got
4enter in solution 3 in after 14mg 0.2ml0.1MNaOH dissolves, 4 DEG C of reaction 2h.
(5) 3 dislysates are changed every day, to remove unreacted small-molecule substance with 0.01mol/lPBS4 DEG C of dialysis 3d.
(6) packing, saves backup in-20 DEG C.
3. the synthesis of coating antigen
(1) get AMOZ haptens 12mg 1mlDMF to dissolve, obtain solution 1.
(2) get the water-soluble solution of OVA36mg 7ml, obtain solution 2.
(3) solution 1 is added dropwise in solution 2, obtains solution 3, room temperature reaction 24h.
(4) NaBH is got
4enter in solution 3 in after 16mg 0.2ml0.1MNaOH dissolves, 4 DEG C of reaction 2h.
(5) 3 dislysates are changed every day, to remove unreacted small-molecule substance with 0.01mol/lPBS4 DEG C of dialysis 3d.
(6) packing, saves backup in-20 DEG C.
4. the preparation of monoclonal antibody
Animal immune: AMOZ haptens and carrier protein couplet thing immunity 8-10 Balb/c mouse in age in week.
Fusion of Cells and cloning: get the mice spleen cell after immunity, merge under the effect of fusion agent polyglycol (PEG) 4000 with SP2/0 myeloma cell, and screening obtains the hybridoma cell strain of energy stably excreting monoclonal antibody.
Cell cryopreservation and recovery: hybridoma cryopreserving liquid is made 1 × 10
9the 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.
Present invention also offers a kind of AMOZ enzyme linked immunological kit and detect the method that in sample, AMOZ is residual, this method is adopted to carry out qualitative to the AMOZ in animal tissue, aquatic products, honey sample or quantitatively detect, Sample pretreatment process is simple, batch samples can be detected fast simultaneously, kit has very high degree of accuracy and sensitivity, lower operative technique requires and of short duration detection time, detect the features such as sample size is large, key step comprises:
1) testing sample is carried out pre-treatment, obtain testing sample solution;
2) testing sample solution is detected with enzyme linked immunological kit;
3) testing result is analyzed.
Accompanying drawing explanation
Fig. 1: AMOZ hapten synthesis figure
Fig. 2: AMOZ typical curve
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.
Embodiment one: the preparation of antigen, antibody and enzyme labeling antiantibody
1. the haptenic preparation of AMOZ
The mixed liquor of 2.01g AMOZ (AMOZ) and 20mlDMF, slowly be added dropwise under room temperature in the 50-100mlDMF solution of 2.68-5.36g terephthalaldehyde, dropwise rear room temperature to 60 DEG C reaction 2-4 hour, except desolventizing, column chromatography purification, obtains faint yellow AMOZ derivant.
2. immunogenic synthesis
(1) get AMOZ haptens 14mg 1mlDMF to dissolve, obtain solution 1.
(2) get the water-soluble solution of BSA40mg 6ml, obtain solution 2.
(3) solution 1 is added dropwise in solution 2, obtains solution 3, room temperature reaction 24h.
(4) NaBH is got
4enter in solution 3 in after 14mg 0.2ml0.1MNaOH dissolves, 4 DEG C of reaction 2h.
(5) 3 dislysates are changed every day, to remove unreacted small-molecule substance with 0.01mol/lPBS4 DEG C of dialysis 3d.
(6) packing, saves backup in-20 DEG C.
3. the synthesis of coating antigen
(1) get AMOZ haptens 12mg 1mlDMF to dissolve, obtain solution 1.
(2) get the water-soluble solution of OVA36mg 7ml, obtain solution 2.
(3) solution 1 is added dropwise in solution 2, obtains solution 3, room temperature reaction 24h.
(4) NaBH is got
4enter in solution 3 in after 16mg 0.2ml0.1MNaOH dissolves, 4 DEG C of reaction 2h.
(5) 3 dislysates are changed every day, to remove unreacted small-molecule substance with 0.01mol/lPBS4 DEG C of dialysis 3d.
(6) packing, saves backup in-20 DEG C.
4. the preparation of ELISA Plate
The bag of wherein said ELISA Plate used by preparation process is buffered the carbonate buffer solution that liquid is the 0.05mol/L of pH9.4 ~ 9.6, and Block buffer is the phosphate buffer of the bovine serum albumin(BSA) (BSA) of 0.05% ~ 0.5% of pH7.4.
The preparation of ELISA Plate of the present invention is mainly: be buffered liquid with bag and coating antigen is diluted to 0.2 ~ 0.3 μ g/ml, every hole adds 150 μ l, 37 DEG C of lucifuges are hatched 2h or 4 DEG C and are spent the night, incline liquid in hole, and cleansing solution washing 1 ~ 2 time, pats dry, 150 μ l confining liquids are added in every hole, 37 DEG C of lucifuges hatch 2h, and in hole of inclining, liquid pats dry, and preserve with the vacuum seal of aluminium film.
5. 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.
6. the preparation of enzyme labeling sheep anti mouse antiantibody (enzyme labeling antiantibody)
Adopt the Over-voltage protection after improveing to carry out coupling horseradish peroxidase and antiantibody, eliminate amino closed process, because it is seldom actual to produce self the amino amino connected; Horseradish peroxidase: the volumetric molar concentration ratio of antiantibody is 2: 1, the method after improvement is easier than traditional method, reduces the loss of the activity of enzyme.
7. the preparation method of monoclonal antibody
Animal immune: AMOZ haptens and carrier protein couplet thing immunity 8-10 Balb/c mouse in age in week.
Fusion of Cells and cloning: get the mice spleen cell after immunity, merge under the effect of fusion agent polyglycol (PEG) 4000 with SP2/0 myeloma cell, and screening obtains the hybridoma cell strain of energy stably excreting monoclonal antibody.
The monoclonal hybridoma strain of AMOZ is obtained through screening.The monoclonal hybridoma strain of AMOZ can be endless generation AMOZ specific antibody, this antibody specificity is for AMOZ, and sensitivity reaches 0.05 μ g/L.
Cell cryopreservation and recovery: hybridoma cryopreserving liquid is made 1 × 10
9the 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.
Embodiment two: the establishment of each component of AMOZ enzyme linked immunological kit
Set up AMOZ enzyme linked immunological kit, comprise following each component:
(1) ELISA Plate of coating antigen is coated with.
(2) enzyme labeling antiantibody: horseradish peroxidase-sheep anti mouse antiantibody.
(3) AMOZ monoclonal antibody working fluid.
(4) standard solution: adopt gradient dilution method preparation standard solution, obtain serial standards 6 bottles, concentration is respectively 0 μ g/L, 0.05 μ g/L, 0.15 μ g/L, 0.45 μ g/L, 1.35 μ g/L, 4.05 μ g/L, and high standard product 100 μ g/L, 1mL/ bottle.
(5) substrate nitrite ion A liquid is urea peroxide solution, and substrate nitrite ion B liquid is tetramethyl biphenyl amine aqueous solution.
(6) stop buffer is the sulfuric acid solution of 1 ~ 2mol/L.
(7) concentrated cleaning solution is the 0.2 ~ 0.4mol/LpH7.6 phosphate buffer of sodium azide preservatives containing 1% ~ 2% Tween-20 and 0.01% ~ 0.05%.
(8) concentrated redissolution liquid is pH value is 7.3 ~ 7.9, the phosphate buffer containing 7% ~ 9% casein, 0.1mol/L.
(9) 2-nitrobenzaldehyde.
Embodiment three: detect AMOZ in sample
1. the pre-treatment of sample
(1) tissue (muscle, liver and aquatic products) Sample pretreatment method
Take the equal pledge of 1.0g and add 4ml deionized water, 0.5ml1M hydrochloric acid solution (measure 8.3ml concentrated hydrochloric acid add deionized water be settled to 100ml) and 100 μ l derivatization reagents (in the reagent bottle that 2-nitrobenzaldehyde is housed, add 10ml methyl alcohol dissolve mixing (concentration is 10mM)), fully to vibrate 2min with oscillator; At 37 DEG C of night incubation (about 16h); Add 5ml0.1M dipotassium hydrogen phosphate solution (take 22.8g tri-hypophosphite monohydrate hydrogen dipotassium and add the mixing of 1L deionized water dissolving), 0.4ml1M sodium hydroxide solution (take 4.0g NaOH and add the mixing of 100ml deionized water dissolving) and 5ml ethyl acetate respectively, with oscillator thermal agitation 30s; More than 3000g, room temperature (20-25 DEG C/68-77 °F) centrifugal 10min.
Get 2.5ml ethyl acetate in 10ml dry glass test tube, flow down in 50 ~ 60 DEG C of water-bath nitrogen and dry up; Add 1ml normal hexane (or normal heptane), with vortex instrument whirling motion 30s, then add 1ml redissolution working fluid (being diluted by 1: 1 volume ratio by the 2 × concentrated liquid that redissolves with deionized water), fully mix with vortex instrument whirling motion 1min; More than 3000g, room temperature (20-25 DEG C/68-77 °F) centrifugal 10min; Removing upper organic phase, takes off layer aqueous phase 50 μ l for analyzing.
(2) honey pre-treating method
Take 1.0g honey in 50ml polystyrene centrifuge tube, add 4ml deionized water, all dissolve to honey with oscillator vibrates, add 0.5ml1M hydrochloric acid solution and 100 μ l derivatization reagents, fully vibrate with oscillator, at 37 DEG C of night incubation (about 16h); Add 5ml0.1M dipotassium hydrogen phosphate solution, 0.4ml1M sodium hydroxide solution and 5ml ethyl acetate respectively, with oscillator thermal agitation 30s, more than 3000g, room temperature (20-25 DEG C/68-77 °F) centrifugal 10min;
Get 2.5ml ethyl acetate in 10ml dry glass test tube, flow down in 50 ~ 60 DEG C of water-bath nitrogen and dry up; Add 1ml normal hexane (or normal heptane), with vortex instrument whirling motion 30s, then add 1ml redissolution working fluid, fully mix with vortex instrument whirling motion 1min, more than 3000g, room temperature (20-25 DEG C/68-77 °F) centrifugal 10min; Removing upper organic phase, takes off layer aqueous phase 50 μ l for analyzing.
2. detection method
(1) prepare: by all reagent and need taking out from cold storage environment with lath before use, be placed in room temperature (20-25 DEG C) balance more than 30min, notice that often kind of liquid reagent must shake up before using, capillary strip needed for taking-up, no microwell plate is put into valve bag, is stored in 2-8 DEG C.
(2) number: sample and the corresponding micropore of standard items are numbered according to the order of sequence, it is parallel that each sample and standard items do 2 holes, and the position at record standard hole and sample aperture place.
(3) standard items/sample is added: add standard items/sample 50 μ l in the micropore of correspondence.
(4) mixing of antibody working fluid and enzyme labeling antiantibody concentrate: antibody working fluid and enzyme labeling antiantibody concentrate are mixed by 10: 1 volume ratios and mixes.(note: this mixed liquor can not be preserved, carries out application of sample after mixing at once)
(5) add the mixed liquor of antibody working fluid and enzyme labeling antiantibody concentrate: the mixed liquor 50 μ l/ hole adding antibody working fluid and enzyme labeling antiantibody concentrate, mixing of vibrating gently, react 30min with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate.
(6) plate is washed: carefully open cover plate film, liquid in hole is dried, with wash operating solution (20 × concentrated cleaning solution being diluted by 1: 19 volume ratio with deionized water) 250 μ l/ hole, abundant washing 4-5 time, every minor tick 10s, pats dry (bubble be not eliminated after patting dry can be poked with original rifle head) with thieving paper.
(7) develop the color: add substrate solution A liquid 50 μ l/ hole, substrate solution B liquid 50 μ l/ hole, mixing of vibrating gently, react 15min with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate.
(8) measure: add stop buffer 50 μ l/ hole, mixing of vibrating gently, setting microplate reader, in 450nm place (suggestion dual wavelength 450/630nm detects, and please runs through data in 5min), measures every hole OD value.
3. Analysis of test results
The percentage absorptance of standard items or sample equals the absorbance of mean value (diplopore) divided by first standard (0 standard) of the absorbance of standard items or sample, then is multiplied by 100%, namely obtains percentage absorptance.With standard items percentage absorptance for ordinate, with the logarithm of AMOZ standard concentration for horizontal ordinate, drawing standard curve map (as Fig. 2).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 AMOZ actual concentrations in sample.
Embodiment four: the sensitivity of AMOZ enzyme linked immunological kit, specificity, preci-sion and accuracy, storage life experiment
1. kit sensitivity determination
Conventionally measure kit sensitivity test, kit standard curve minimum point is 0.05 μ g/L, the scope of typical curve is 0.05 μ g/L ~ 4.05 μ g/L, and in chicken, pork, chicken gizzard, shrimp, fish, honey sample, AMOZ detects and is limited to 0.1 μ g/kg.
2. kit accuracy and precision
Accuracy refers to the matching degree between measured value and true value, and the accuracy of kit is commonly used the recovery and represented; Precision is that reaction assay method repeatedly measures the repetition degree of acquired results to a certain specific sample, and the conventional coefficient of variation represents.Being 0.2 μ g/kg, 0.4 μ g/kg respectively to adding AMOZ to final concentration in blank chicken, pork, chicken gizzard, shrimp, fish, honey sample, repeating 5 times, the kit getting three batches respectively calculates the coefficient of variation, the results are shown in following table.
The accuracy of table 1 kit and precision measure
Result shows, when adding blank chicken, pork, chicken gizzard, shrimp, fish with 0.2 μ g/kg AMOZ, sample TIANZHU XINGNAO Capsul scope is 71.5% ~ 100.0%, when adding blank chicken, pork, chicken gizzard, shrimp, fish with 0.4 μ g/kg AMOZ, sample TIANZHU XINGNAO Capsul scope is 70.8% ~ 99.8%; When adding blank honey with 0.2 μ g/kg AMOZ, sample TIANZHU XINGNAO Capsul scope is 78.0% ~ 103.0%, and when adding blank honey with 0.4 μ g/kg AMOZ, sample TIANZHU XINGNAO Capsul scope is 78.8% ~ 103.8%; In batch, interassay coefficient of variation is all less than 20%, meets " Ministry of Agriculture's file " agriculture doctor [2005] No. 17 annex 2 kits and puts on record with reference to the regulation of the 4th preci-sion and accuracy in judgment criteria.
3. cross reacting rate test
Select AMOZ as follows, Furaxone metabolite, Cistofuran metabolite, Furacilin metabolite, furaltadone, furazolidone, furantoin, nitrofurazone 8 kinds of medicines conventionally to measure cross reacting rate respectively, result is as shown in table 2.
The specificity of table 2 kit
4. storage life experiment
Kit preservation condition is 2-8 DEG C, measures, the maximum absorbance value of kit, IC through 12 months
50value, AMOZ add practical measurement value all within normal range.Do accelerated deterioration and refrigeration test, kit to be placed in 37 DEG C ,-20 DEG C 6 days, measurement result also shows that the indices of kit is normal simultaneously.Obtain AMOZ kit from above result to preserve 12 months at 2-8 DEG C.
Claims (8)
1. an AMOZ enzyme-linked immunologic detecting kit, include: the ELISA Plate being coated with coating antigen, AMOZ specific antibody, enzyme marker, standard solution, substrate nitrite ion, stop buffer, concentrated cleaning solution, the concentrated liquid that redissolves, 2-nitrobenzaldehyde, described coating antigen is AMOZ antigen, described enzyme marker is enzyme labeling antiantibody, described AMOZ antigen is the conjugate of AMOZ haptens and carrier protein, described carrier protein is bovine serum albumin(BSA), mouse haemocyanin, rabbit serum proteins, thyroprotein, ovalbumin, hemocyanin or human serum albumins, described AMOZ is haptenic preparation method mainly comprise the steps:
The mixed liquor of 2.01g AMOZ and 20mlDMF, slowly be added dropwise in the 50-100mlDMF solution containing 2.68-5.36g terephthalaldehyde under room temperature, dropwise rear room temperature to 60 DEG C reaction 2-4 hour, except desolventizing, column chromatography purification, obtain faint yellow AMOZ derivant, i.e. AMOZ haptens.
2. kit as claimed in claim 1, is characterized in that: described AMOZ specific antibody is AMOZ monoclonal antibody.
3. kit as claimed in claim 1, is characterized in that: described antiantibody is sheep anti mouse antiantibody.
4. kit as claimed in claim 1, is characterized in that: the marker enzyme of described enzyme marker is horseradish peroxidase; Enzyme labeling antiantibody adopts Over-voltage protection marker enzyme and antiantibody coupling to be obtained.
5. kit as claimed in claim 1, it is characterized in that: described stop buffer is the sulfuric acid solution of 1 ~ 2mol/L, described substrate nitrite ion A liquid is urea peroxide solution, and substrate nitrite ion B liquid is tetramethyl biphenyl amine aqueous solution.
6. kit as claimed in claim 1, it is characterized in that: described concentrated cleaning solution is the 0.2 ~ 0.4mol/LpH7.6 phosphate buffer of sodium azide preservatives containing 1% ~ 2% Tween-20 and 0.01% ~ 0.05%, described concentrated redissolution liquid is pH value is 7.3 ~ 7.9, the phosphate buffer containing 7% ~ 9% caseic 0.1mol/L.
7. kit as claimed in claim 1, is characterized in that: described standard solution concentration is respectively 0 μ g/L, 0.05 μ g/L, 0.15 μ g/L, 0.45 μ g/L, 1.35 μ g/L, 4.05 μ g/L.
8. detect with the AMOZ enzyme linked immunological kit described in any one of claim 1-7 the method that in sample, AMOZ is residual, key step comprises:
1) testing sample is carried out pre-treatment, obtain testing sample solution;
2) testing sample solution is detected with the enzyme linked immunological kit described in any one of claim 1-7;
3) testing result is analyzed.
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| CN106645687A (en) * | 2016-11-09 | 2017-05-10 | 百奥森（江苏）食品安全科技有限公司 | Kit for detecting furaltadone metabolites in food |
| CN106770227A (en) * | 2016-11-30 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | The detection method and test strips of AMOZ in a kind of dairy products |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB727007A (en) * | 1952-09-30 | 1955-03-23 | Bayer Ag | New semicarbazones |
| CN1405563A (en) * | 2001-08-02 | 2003-03-26 | 兰多克斯实验室有限公司 | Method and kit for detecting and determining beta-lactam penicillin |
| CN101013130A (en) * | 2007-02-13 | 2007-08-08 | 北京望尔康泰生物技术有限公司 | Enzyme linked immunosorbent reagent casing for detecting furantoin metabolite and uses thereof |
| CN201852835U (en) * | 2010-10-27 | 2011-06-01 | 北京勤邦生物技术有限公司 | ELISA (enzyme linked immunosorbent assay) kit of furaltadone metabolite |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB727007A (en) * | 1952-09-30 | 1955-03-23 | Bayer Ag | New semicarbazones |
| CN1405563A (en) * | 2001-08-02 | 2003-03-26 | 兰多克斯实验室有限公司 | Method and kit for detecting and determining beta-lactam penicillin |
| CN101013130A (en) * | 2007-02-13 | 2007-08-08 | 北京望尔康泰生物技术有限公司 | Enzyme linked immunosorbent reagent casing for detecting furantoin metabolite and uses thereof |
| CN201852835U (en) * | 2010-10-27 | 2011-06-01 | 北京勤邦生物技术有限公司 | ELISA (enzyme linked immunosorbent assay) kit of furaltadone metabolite |
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