CN101413952A - Chemiluminescence immune detection reagent kit for detecting ractopamine - Google Patents
Chemiluminescence immune detection reagent kit for detecting ractopamine Download PDFInfo
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- CN101413952A CN101413952A CNA2008102034350A CN200810203435A CN101413952A CN 101413952 A CN101413952 A CN 101413952A CN A2008102034350 A CNA2008102034350 A CN A2008102034350A CN 200810203435 A CN200810203435 A CN 200810203435A CN 101413952 A CN101413952 A CN 101413952A
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Abstract
The invention provides a chemiluminescent immunoassay kit for detecting ractopamine, and the kit belongs to the field of immunoassay. The kit is composed of a non-transparent white enzyme-labeled plate which is coated by a ractopamine-carrier protein cross-linked complex, a ractopamine standard product, a ractopamine-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The ractopamine-carrier protein cross-linked complex is obtained by coupling the ractopamine and a carrier protein through the mixed-anhydride method or the direct protein activation method, and the concentrated buffer solution contains Tween-20 buffer solution. The kit has the advantages of rapidness, simpleness, high sensitivity, low detection cost, good repeatability, high throughput and the like, and the kit can be used for detecting the residual ractopamine in animal urine, blood, tissues, visceral organs and other samples.
Description
Technical field
The present invention relates to a kind of chemiluminescence immune detection reagent kit that detects Ractopamine, be used for detecting Ractopamine content or residual quantity in animal derived food such as animal tissue, internal organ, blood, urine and feed, the feedstuff.Belong to the immunology detection field.
Background technology
Ractopamine (Ractopamin) belongs to beta-stimulants.Beta-stimulants is a kind of of the heavy partitioning agent of nutrition, it is a class formation and function class phenolethanolamine analog derivative like adrenaline and norepinephrine, it can improve the ratio of meat with the fat of lard type animal, reduce the ketoboidies fat content, improve lean meat percentage, and can quicken growth of animal, and be added in animal feed.Common β 2-excitant has Clenbuterol, Ractopamine and salbutamol etc.Along with the increasing of China to Clenbuterol (being commonly called as ＂ clenbuterol hydrochloride ＂) supervision, the use of Clenbuterol reduces gradually, and the anti-depressant use of other β 2-increases gradually.Because Ractopamine has the effect that is similar to Clenbuterol, and is residual in animal tissue, thereby human body is had a negative impact, it uses as feed addictive so China forbids Ractopamine.
Aspect the detection method of Ractopamine, method at present commonly used comprises high performance liquid chromatography (HPLC), gas chromatography-mass spectrography (GC-MS), liquid chromatograph mass spectrography (LC-MS), Capillary Electrophoresis, enzyme linked immunosorbent assay analysis method (enzyme linked immunosorbent assay, method such as ELISA).
1, high performance liquid chromatography (HPLC) is used for the detection of Ractopamine:
The HPLC method has degree of accuracy height, the characteristics that false positive rate is low of detecting.It is more loaded down with trivial details that the major defect of HPLC method is that instrument costs an arm and a leg, operates, length consuming time, testing cost costliness; And the selection of the extraction conditions of sample also has considerable influence to detection sensitivity and accuracy in earlier stage.
2, gas chromatography-mass spectrography (GC-MS) is used for the detection of Ractopamine:
GC-MS sensitivity is very high, and false positive rate is low.The major defect of this method is that sample needs derivatization to handle complicated like this early stage.Though can provide more structural information through the sample of derivatization treatment in mass spectrum, derivatization can produce a plurality of different products and cause the partial loss of sample, causes the deviation of experimental result.
3, liquid chromatograph mass spectrography (LC-MS) is used for the detection of Ractopamine:
LC-MS does not need derivatization treatment to sample, can detect urine, blood, liver, hair and eyeball sample.The MS coupling if LC connects with two can further improve signal to noise ratio (S/N ratio).The shortcoming of LC-MS is the same with GC-MS, and complex operation step needs expensive instrument, so general with the affirmation means of doing positive findings.
4, Capillary Electrophoresis is used for the detection of Ractopamine:
Capillary zone electrophoresis method separation efficiency can reach millions of theoretical cam curves, operates fast and conveniently, and required sample size is few.But the required instrument costliness of this method, and can detect the sensitivity that the minimum residual quantity of Ractopamine does not reach other method.
5, enzyme linked immunosorbent assay analysis method (ELISA) is used for the detection of Ractopamine:
Enzyme linked immunosorbent assay analysis method is one of current application immunoenzyme technics wide, the most with fastest developing speed, and major advantage is to detect rapidly, and sample pre-treatments is simple, and detection system is simple to operate, and cost is low, is convenient to simultaneously detect on a large scale.Its ultimate principle is, antigen (or antibody) is adsorbed on the solid phase carrier, enzymic-labelled antibody (or antigen) and corresponding antigen (or antibody) reaction, be combined in enzyme on the immune complex when running into corresponding substrate, can the catalytic substrate hydrolysis, oxidation or reduction and produce coloring matter, the depth of color is directly proportional with corresponding antibody (or antigen) amount, so can come qualitative or quantitative antibody (or antigen) by the degree of dye-forming reaction.The ELISA detection method mainly comprises double antibody sandwich method, indirect method, competition law etc.Double antibody sandwich method mainly is applicable to the comlete antigen that molecular weight is bigger, and law of competition is applicable to the less small-molecular weight haptens in antigen decision site.Because Ractopamine is micromolecular compound (relative molecular mass is less than 5000), belongs to haptens, competitive enzyme-linked immune absorption method therefore commonly used is measured.
6, the principle of work of chemiluminescence immunoassay detection technique and characteristics
The chemiluminescence immunoassay detection technique is different from enzyme linked immunosorbent assay analysis method, and it is the product of chemoluminescence method and immunoassay combination, therefore has the high sensitivity of chemiluminescence detection technology and the high specific of immuno analytical method simultaneously.
The ultimate principle of chemiluminescence immunoassay is similar to ELISA, and difference is that the reaction system of enzyme labeling thing is a chemiluminescence reaction.HRP (horseradish peroxidase) and luminol or derivatives thereof system are exactly one of reaction system of using always, and adding reinforcing agent in this luminescence system can increase chemiluminescence intensity, and keep stable in the long period, thus the sensitivity that improves immunoassay greatly.As China petty official transmit etc. (petty official is transmitted etc., the analytical chemistry research notes, 2005,33 (5): 699-702) the indirect competition chemiluminescence immunoassay method of setting up detects Clenbuterol in the pig urine sample, the actual detected scope is 0.04～25.8 μ g/kg.The shortcoming that the petty official such as transmits at people's method therefor is to use one anti-and two anti-(wherein two anti-be enzymic-labelled antibody) simultaneously, all needs to carry out incubation reaction after two kinds of antibody addings, has prolonged detection time.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of chemiluminescence immune detection reagent kit that detects Ractopamine is provided, have fast and convenient, highly sensitive, advantages such as the detection cost is low, good reproducibility, high flux, the residual quantity that can be used for Ractopamine in animals urine, blood sample, tissue, internal organ and the samples such as feed, feedstuff detects.
For achieving the above object, the present invention utilizes the ultimate principle of the specific immune response of antigen and antibody to realize.Chemiluminescence immunoassay is the product of chemoluminescence method and immunoassay combination, therefore has the high sensitivity of chemoluminescence method and the high specific of immunoassay simultaneously.In entire reaction course, Ractopamine content is high more in the sample, and luminous intensity is weak more in the reaction system; Otherwise Ractopamine content is few more in the sample, and luminous intensity is high more.
The chemiluminescence immune detection reagent kit of detection Ractopamine of the present invention contains following composition:
1, is coated with the opaque white color ELISA Plate of Ractopamine-carrier protein cross-linked composite.Described Ractopamine-carrier protein cross-linked composite is that Ractopamine and carrier protein are obtained by mixed anhydride method or the coupling of direct activation protein method, and described carrier protein is human serum albumins, bovine serum albumin(BSA), egg albumin, mouse haemocyanin or rabbit anteserum albumen.
2, Ractopamine standard items.
3, Ractopamine-peroxidase labeled antibodies.This composition is the Ractopamine antibody with peroxidase labelling.Described Ractopamine antibody is monoclonal antibody or polyclonal antibody.
4, luminous substrate liquid.This luminous substrate liquid is to be the chemical luminous substrate liquid of luminous agent with luminol or different luminol, need be divided into A liquid and B liquid preserves, facing with using preceding the mixing by 1:1, wherein A liquid (or B liquid) is luminol (or different luminol) and luminescence enhancer, and B liquid (or A liquid) is superoxol or urea hydrogen peroxide solution.
5, concentrate damping fluid.The working concentration (being the working concentration behind the distilled water diluting) that should concentrate damping fluid is 0.01～0.5mol/L, and the pH value is 6.0～9.0, and the content of Tween-20 is 0.01～0.5% (v/v).
Among the present invention, described opaque white color ELISA Plate is the detachable or non-removable opaque white color ELISA Plate in 48 holes or 96 holes.
Among the present invention, described Ractopamine standard items are made up of the Ractopamine standard items of a series of variable concentrations, and concentration range has comprised the concentration interval of 0.02～3 μ g/L at least.
Among the present invention, described Ractopamine-peroxidase labeled antibodies is for the Ractopamine antibody with peroxidase labelling, as the Ractopamine antibody of horseradish peroxidase-labeled.
Luminous substrate liquid of the present invention is that commercially available any is the chemical luminous substrate liquid of luminous agent with luminol or different luminol.
Described concentrated damping fluid, for containing 2～100 times of concentrates of Tween-20 (Tween-20) damping fluid, this damping fluid can be phosphate buffer (PBST), glycocoll-HCl damping fluid or Tris-HCl damping fluid, uses to working concentration with distilled water diluting before using.
When kit of the present invention was applied to the detection of Ractopamine, the detection step was:
1) sample pre-treatments.
2) kit of the present invention (contain ELISA Plate, standard items, concentrate damping fluid, Ractopamine-peroxidase labeled antibodies, luminous substrate solution) is taken out from refrigerator, place under the room temperature, 10-60min rises again.
3) preparation of working concentration damping fluid: will concentrate damping fluid with distilled water and be diluted to working concentration, and promptly can be used as enzyme labeling thing dilution, sample diluting liquid and ELISA Plate cleaning fluid.
4) Ractopamine-peroxidase labeled antibodies preparation: utilize the working concentration damping fluid that Ractopamine-peroxidase labeled antibodies is diluted to working concentration, promptly finish the preparation of this labelled antibody.Labelled antibody after the dilution should use as early as possible, if be stored in-20 ℃, storage life is no more than one month; If be stored in 4 ℃, storage life is no more than a week.
5) add Ractopamine standard solution and sample to be tested respectively in suitable micropore, standard and sample are done two parallel laboratory tests at least.
6) add an amount of dilution good Ractopamine-peroxidase labeled antibodies in each micropore.Make it fully mix back lucifuge under room temperature and leave standstill incubation 30～90 minutes (min).
7) liquid of pouring out in the hole also dries, and the micropore frame is upside down in pats on the thieving paper to guarantee to remove fully the liquid in the hole.
8) the working concentration damping fluid is added in every hole, the liquid of outwelling once more in the micropore dries, and repeats to wash 2～3 times.
9) add luminous substrate liquid, mix the back and in chemiluminescence detector, detect luminous intensity (RLU).
10) calculating of testing result: use the standard solution that obtained and the ratio of sample solution luminous value and blank solution to calculate.Formula as follows:
Relative luminous intensity=RLU/RLUmax
In the formula:
The luminous intensity values of RLU---standard (or sample) solution;
RLUmax---the luminous intensity values of blank (concentration is 0 standard solution).
The natural logarithm of the corresponding Ractopamine concentration of relative luminous intensity value (μ g/L) calculated is made semilog coordinate system curve map.The Ractopamine concentration of each testing sample is found on typical curve according to its RLU value, or draws by the corresponding Equation for Calculating of typical curve.Passed through dilution in advance as sample, should will multiply by its extension rate again according to the sample concentration that typical curve drew.
The residual quantity that kit of the present invention can be used for Ractopamine in the samples such as animals urine, blood sample, tissue and internal organ detects.Compare with the existing experimental technique that other detects the Rct opamine residue amount, kit of the present invention has following advantage: the kit of the present invention that 1) adopts the chemiluminescence immunoassay method, more more fast and convenient than chromatographic process (high performance liquid chromatogram, LC-MS, gas chromatography mass spectrometry), capillary electrophoresis method, required instrument is more simple, it is more cheap to detect cost, has high-throughout characteristics simultaneously.2) kit of the present invention of employing chemiluminescence immunoassay method is more sensitiveer than ELISA method, can detect the Rct opamine residue of lower concentration and content, and the range of linearity is wideer simultaneously.3) kit of the present invention of employing chemiluminescence immunoassay method has reduced by the two use links that resist, thereby has reduced the incubation time, has shortened detection time; The opaque white color ELISA Plate has increased chemiluminescence detector sensitivity.In addition, with Ractopamine-carrier protein cross-linked composite but not Ractopamine antibody wrap by the opaque white color ELISA Plate, reduced the instability of Ractopamine antibody, guaranteed the long-term effectiveness of kit.
Embodiment
Below by specific embodiment the present invention is further described.These embodiment only are used to illustrate the present invention, and are not used for limiting the scope of the invention.
Embodiment 1
1) preparation of each component of kit
The haptenic preparation of Ractopamine:, in 4 ℃ of unglazed low temperature environments,, generate the intermediate that contains the diazo positive ion with the sodium nitrite effect with the Ractopamine acidifying.Diazotizing Ractopamine is used for synthetic immunizing antigen in back and envelope antigen as haptens.
Bovine serum albumin(BSA)-Ractopamine immunizing antigen synthetic: Ractopamine is carried out coupling with bovine serum albumin(BSA) (BSA) employing diazotising method obtain immunizing antigen.
Human serum albumins-Ractopamine envelope antigen synthetic: Ractopamine is carried out coupling with human serum albumins (HSA) employing diazotising method obtain immunizing antigen.
The preparation of Ractopamine-peroxidase labeled antibodies: above-mentioned immunizing antigen is injected in the Balb/C mouse body, behind booster immunization repeatedly, make it produce antibody serum.Take out the splenocyte of immunity back mouse, add mixing in the centrifuge tube.Above-mentioned splenocyte and SP2/0 myeloma cell are merged, in containing 96 well culture plates of HAT solution, carry out CO2 and cultivate, screen positive hole.Utilize limiting dilution assay to obtain the hybridoma cell strain of secrete monoclonal antibody then.Get healthy Balb/C mouse, lumbar injection sterilized liquid paraffin 1mL, behind the 7d to the above-mentioned hybridoma suspension of lumbar injection 1mL.Gather ascites behind the 7d, add equivalent whiteruss dilution, add the SiO 2 powder mixing, centrifugally must clarify ascites.Clarification ascites is carried out purifying with sad-ammonium sulfate method, obtains the Ractopamine monoclonal antibody of purifying again through dialysis.Ractopamine monoclonal antibody and horseradish peroxidase, thus Ractopamine-peroxidase labeled antibodies obtained.
Be coated with the opaque white color ELISA Plate preparation of Ractopamine-HSA cross-linked composite: be used to wrap detection hole after with damping fluid Ractopamine-HSA cross-linked composite being diluted by the opaque white color ELISA Plate, wash with the PBST damping fluid 4 ℃ of backs of spending the night, add 200 μ L confining liquids (PBST that contains 2% (w/v) HSA) then, 37 ℃ of incubation 2h, the liquid in the hole that inclines is finished after the drying.
2) establishment of the chemiluminescence immune detection reagent kit of detection Ractopamine
The chemiluminescence immune detection reagent kit of the detection Ractopamine of setting up has comprised following ingredient:
(1) 96 hole opaque white color ELISA Plate (12 * 8 hole) is coated with Ractopamine-human serum albumins cross-linked composite, with aluminium film vacuum sealed package.
(2) the Ractopamine standard solution is 6 bottles, and concentration is respectively:
0μg/L、0.04μg/L、0.12μg/L、0.36μg/L、1.08μg/L、3.24μg/L
(3) Ractopamine-horseradish peroxidase-labeled antibody-solutions.
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide).
(5) 100 times of concentrated damping fluids.Dilution becomes the working concentration damping fluid for 100 times before using, and the working concentration damping fluid after its dilution is a 0.05mol/L PBST damping fluid, and pH7.4 contains 0.05% (v/v) Tween-20.
3) use of the chemiluminescence immune detection reagent kit of Ractopamine
(1) pre-treatment of sample
A. urine sample
Test after directly urine suitably being diluted (6-10 doubly), ask centrifugal (3000r/min) 10min earlier, get supernatant and suitably dilute post analysis if any muddiness.
B. blood sample
Extract the tested animal blood sample, centrifugal (3000r/min) 10min gets transparent supernatant and uses, or leaves standstill and get its transparent supernatant and use, and notes not having haemolysis.Get 0.1ml serum, add the 0.4ml dilution, whirlpool one minute, mixing.Centrifugal 10 minutes (3000r/min) gets supernatant 100ul, analyzes.
C. muscle, liver and kidney sample
Get the even quality sample of 1g and insert in the centrifuge tube, add 4ml0.01N HCl again, whirlpool one minute.Heated about 3 minutes with 80-100 ℃ of water-bath, centrifugal 10 minutes (5000rpm), supernatant and dilution get 100 μ L and analyze after carrying out the 1:3 dilution.
(2) chemiluminescence immune detection reagent kit detects
The hole bar of standard and sample requirement is inserted in the microwell plate framework position of record standard and sample.In suitable micropore, add Ractopamine standard solution and sample to be tested respectively.Add 50 μ L Ractopamine-horseradish peroxidase-labeled antibody in each micropore, fully mix back lucifuge under room temperature and left standstill incubation 1 hour.Remove the liquid in the hole, use work concentration buffer liquid repeated washing three times.Remove the liquid in the hole fully, add luminous substrate liquid mixed liquor (A liquid and B liquid are pressed the 1:1 mixing before use) 100 μ L/ holes.In chemiluminescence detector, detect luminous intensity after mixing immediately.
(3) computational analysis of testing result
Ratio with the standard solution that is obtained and sample solution luminous value and blank solution calculates.Formula as follows:
Relative luminous intensity=RLU/RLUmax
In the formula:
The luminous intensity values of RLU---standard (or sample) solution;
RLUmax---the luminous intensity values of blank (concentration is 0 standard solution).
The natural logarithm of the corresponding Ractopamine concentration of relative luminous intensity value (μ g/L) calculated is made semilog coordinate system curve map.Passed through dilution in advance as sample, should will multiply by its extension rate again according to the sample concentration that typical curve drew.
Embodiment 2
Detect the chemiluminescence immune detection reagent kit of Ractopamine, comprised following ingredient:
(1) 48 hole opaque white color ELISA Plate (6 * 8 hole) is coated with Ractopamine-mouse serum albumin cross-linked composite, with aluminium film vacuum sealed package.
(2) the Ractopamine standard solution is 5 bottles, and concentration is respectively:
0μg/L、0.04μg/L、0.36μg/L、3.24μg/L、29.16μg/L
(3) Ractopamine-horseradish peroxidase-labeled antibody-solutions.
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide).
(5) 50 times of concentrated damping fluids.Dilution becomes the working concentration damping fluid for 50 times before using, and the working concentration damping fluid after its dilution is a 0.05mol/L Tris-HCl damping fluid, and pH6.9 contains 0.1% (v/v) Tween-20.
Embodiment 3
Detect the chemiluminescence immune detection reagent kit of Ractopamine, comprised following ingredient:
(1) 96 hole opaque white color ELISA Plate (12 * 8 hole) is coated with Ractopamine-egg albumin cross-linked composite, with aluminium film vacuum sealed package.
(2) the Ractopamine standard solution is 6 bottles, and concentration is respectively:
0μg/L、0.05μg/L、0.25μg/L、1.25μg/L、6.25μg/L、31.25μg/L。
(3) Ractopamine-horseradish peroxidase-labeled antibody-solutions.
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide).
(5) 10 times of concentrated damping fluids.Dilution becomes the working concentration damping fluid for 10 times before using, and the working concentration damping fluid after its dilution is 0.05mol/L glycocoll-HCl damping fluid, and pH6.9 contains 0.02% (v/v) Tween-20.
Claims (4)
1. chemiluminescence immune detection reagent kit that detects Ractopamine is characterized in that containing following composition:
1) is coated with the opaque white color ELISA Plate of Ractopamine-carrier protein cross-linked composite; Described Ractopamine-carrier protein cross-linked composite is that Ractopamine and carrier protein are obtained by mixed anhydride method or the coupling of direct activation protein method, and described carrier protein is human serum albumins, bovine serum albumin(BSA), egg albumin, mouse haemocyanin or rabbit anteserum albumen;
2) Ractopamine standard items;
3) Ractopamine-peroxidase labeled antibodies; This composition is the Ractopamine antibody with peroxidase labelling, and described Ractopamine antibody is monoclonal antibody or polyclonal antibody;
4) luminous substrate liquid; This luminous substrate liquid is to be the chemical luminous substrate liquid of luminous agent with luminol or different luminol, is divided into A liquid and B liquid and preserves, and mixes use by 1:1 facing with preceding; Wherein A liquid is that luminescence enhancer adds luminol or luminescence enhancer adds different luminol, and B liquid is superoxol or urea hydrogen peroxide solution;
5) concentrate damping fluid, its working concentration is 0.01～0.5mol/L, and the pH value is 6.0～9.0, and the volume content of Tween-20 is 0.01～0.5%.
2. according to the kit of claim 1, it is characterized in that described opaque white color ELISA Plate is the detachable or non-removable opaque white color ELISA Plate in 48 holes or 96 holes.
3. according to the kit of claim 1, it is characterized in that described luminous substrate liquid is that commercially available any is the chemical luminous substrate liquid of luminous agent with luminol or different luminol.
4. according to the kit of claim 1, it is characterized in that described concentrated damping fluid is 2～100 times of concentrates that contain the Tween-20 damping fluid, this damping fluid is phosphate buffer, glycocoll-HCl damping fluid or Tris-HCl damping fluid.
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| CNA2008102034350A CN101413952A (en) | 2008-11-27 | 2008-11-27 | Chemiluminescence immune detection reagent kit for detecting ractopamine |
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| CNA2008102034350A CN101413952A (en) | 2008-11-27 | 2008-11-27 | Chemiluminescence immune detection reagent kit for detecting ractopamine |
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| CN2012104589124A Division CN102980999A (en) | 2008-11-27 | 2008-11-27 | Chemiluminescence immunodetection kit for detecting ractopamine |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101915845A (en) * | 2010-08-03 | 2010-12-15 | 中国农业大学 | Method for detecting ractopamine and chemiluminescence immunoassay kit special for same |
| CN101949933A (en) * | 2010-08-25 | 2011-01-19 | 南开大学 | Indirect chemiluminescent enzyme linked immunosorbent assay (ELISA) kit for detecting residual ractopamine by using monoclonal antibody |
| CN102286103A (en) * | 2011-09-02 | 2011-12-21 | 安徽缘远博爱生物技术有限公司 | Preparation method and application of monoclonal antibody against ractopamine |
| CN102313810A (en) * | 2011-07-29 | 2012-01-11 | 上海交通大学 | Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin |
| CN101921730B (en) * | 2010-01-19 | 2012-05-09 | 泰州康正生物技术有限公司 | Monoclonal antibody of ractopamine and preparation method and application thereof |
| CN102778564A (en) * | 2012-05-31 | 2012-11-14 | 华中农业大学 | Monoclonal antibody, enzyme linked immunosorbent assay method and kit for detecting ractopamine |
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| CN103808933A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Chemiluminiscence immunoassay kit for detecting zilpaterol |
| CN104655845A (en) * | 2013-11-20 | 2015-05-27 | 南京亿特生物科技有限公司 | Preparation of chemiluminescence immunoassay kit for detecting chloroquine |
| CN107478824A (en) * | 2017-08-11 | 2017-12-15 | 太原瑞盛生物科技有限公司 | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of Ractopamine |
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| CN101921730B (en) * | 2010-01-19 | 2012-05-09 | 泰州康正生物技术有限公司 | Monoclonal antibody of ractopamine and preparation method and application thereof |
| CN101915845A (en) * | 2010-08-03 | 2010-12-15 | 中国农业大学 | Method for detecting ractopamine and chemiluminescence immunoassay kit special for same |
| CN101949933A (en) * | 2010-08-25 | 2011-01-19 | 南开大学 | Indirect chemiluminescent enzyme linked immunosorbent assay (ELISA) kit for detecting residual ractopamine by using monoclonal antibody |
| CN102313810A (en) * | 2011-07-29 | 2012-01-11 | 上海交通大学 | Chemiluminescent immunoassay method for rapidly detecting zearalenone toxin |
| CN102286103A (en) * | 2011-09-02 | 2011-12-21 | 安徽缘远博爱生物技术有限公司 | Preparation method and application of monoclonal antibody against ractopamine |
| CN102778564A (en) * | 2012-05-31 | 2012-11-14 | 华中农业大学 | Monoclonal antibody, enzyme linked immunosorbent assay method and kit for detecting ractopamine |
| CN102778564B (en) * | 2012-05-31 | 2014-06-04 | 华中农业大学 | Monoclonal antibody, enzyme linked immunosorbent assay method and kit for detecting ractopamine |
| CN103792226A (en) * | 2012-11-03 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Chemiluminescence immunodetection kit used for detecting phenylethanolamine A |
| CN103808933A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Chemiluminiscence immunoassay kit for detecting zilpaterol |
| CN104655845A (en) * | 2013-11-20 | 2015-05-27 | 南京亿特生物科技有限公司 | Preparation of chemiluminescence immunoassay kit for detecting chloroquine |
| CN107478824A (en) * | 2017-08-11 | 2017-12-15 | 太原瑞盛生物科技有限公司 | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of Ractopamine |
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Application publication date: 20090422 |