CN103808933A - Chemiluminiscence immunoassay kit for detecting zilpaterol - Google Patents
Chemiluminiscence immunoassay kit for detecting zilpaterol Download PDFInfo
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- CN103808933A CN103808933A CN201210438102.2A CN201210438102A CN103808933A CN 103808933 A CN103808933 A CN 103808933A CN 201210438102 A CN201210438102 A CN 201210438102A CN 103808933 A CN103808933 A CN 103808933A
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- 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
- G01N33/566—Immunoassay; Biospecific binding assay; Materials therefor using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
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- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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Abstract
The invention provides a chemiluminiscence immunoassay kit for detecting zilpaterol, and belongs to the field of immunological detection. The kit comprises a non-transparent white ELISA plate coated with a zilpaterol-carrier protein conjugate, a zilpaterol standard substance, a zilpaterol-peroxidase labeled antibody working solution, a luminescent substrate solution, a concentrated sample diluent and a concentrated washing liquid. The zilpaterol-carrier protein conjugate is obtained by coupling zilpaterol and a carrier protein through a mixed anhydride method or a carbodiimide method, and the concentrated washing liquid contains 0.05% of Twain-20. Compared with a traditional enzyme-linked immunosorbent assay method, the kit has higher sensitivity degree, is short in detection time and low in detection cost, and can be used for detection of the residual amount of zilpaterol in animal urine, blood samples, tissues, viscera and other samples.
Description
Technical field
The present invention relates to a kind of chemiluminescence immune detection reagent kit that detects Zilpaterol, for detection of in animal derived food as Zilpaterol content or residual quantity in animal tissue, internal organ, blood, urine and feed, feedstuff.Belong to immunology detection field.
Background technology
Use β
2-adrenoceptor agonists can increase muscle/fatty ratio, can obviously improve letting animals feed lean meat percentage.But use in a large number or long-term Use out of range, and do not observe safe withdrawal time, can in letting animals feed tissue, accumulate, people has eaten and can cause after this animal tissue poisoning even deadly, and therefore a lot of countries and international organization have forbidden that this compounds raises for food animal.Heavy dose of such medicine that uses of sportsman, can increase muscle/fat compares and improves muscle strength, also respiratory system and nervous system are had to excitation, instantaneous burst in play, obtain the performance of extraordinary level, against the principle of fair play, so prohibiting contestant, the International Olympic Committee uses this type of medicine.
Zilpaterol (Zilpaterol) is a kind of novel β
2-2-adrenergic agonist components, its chemical constitution (seeing formula 1) is diverse with common phenyl amines-adrenaline excitomotor (clenbuterol and salbutamol) and phenol beta-adrenaline excitomotor (Ractopamine), its action effect is equivalent to 1/10th of clenbuterol, but more effective than Ractopamine.Be allowed to use as the feed addictive of slaughtering phase ox in Mexico and South Africa Zilpaterol, but prohibited use in European Union.
The chemical constitution of formula 1 Zilpaterol
At present, European Union member countries relate to GC-MS(gas chromatography-mass spectrography), high performance liquid chromatography-tandem mass method etc. to the more experimental technique of research of residual aspect in Zilpaterol use and animal tissue.Gas chromatography mass spectrometry method all needs sample to derive, and operation steps is many, and experimental period is long, and high performance liquid chromatography-series winding mass spectroscopy is high to instrument and equipment requirement, conventionally need to utilize deuterated internal standard compound matter, and experimental procedure is more complicated.Enzyme linked immunosorbent assay analysis method (ELISA) is one of detection technique that current application is the widest, and major advantage is that detection speed is fast, and sample pre-treatments is simple, simple to operate, and testing cost is low, is convenient to the detection for batch samples simultaneously.
Chemiluminescence immunoassay detection technique 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 present invention has the Zilpaterol antibody of high-affinity, high specific by distinctive immune animal preparation, and adopts enzymic-labelled antibody, sets up a kind of chemiluminescence immunoassay kit that can detect Zilpaterol.The feature such as that this method has is easy and simple to handle, quick, highly sensitive, specificity is good.
Summary of the invention
For problems of the prior art, the present invention utilizes the ultimate principle of the specific immune response of antigen and antibody to realize.Chemiluminescence immune assay 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 whole course of reaction, in sample, Zilpaterol content is higher, and in reaction system, luminous intensity is more weak; Otherwise Zilpaterol content is fewer in sample, luminous intensity is higher.
The present invention is a kind of chemiluminescence immune detection reagent kit that detects Zilpaterol, it is characterized in that containing following composition:
1, be coated with the opaque white color ELISA Plate of Zilpaterol-carrier protein couplet thing; Described Zilpaterol-carrier protein couplet thing is that Zilpaterol and carrier protein are obtained by mixed anhydride method or carbodlimide method coupling, and described carrier protein is human serum albumins, bovine serum albumin(BSA), egg albumin, mouse haemocyanin or rabbit anteserum albumen;
2, Zilpaterol standard items;
3, Zilpaterol-peroxidase labeled antibodies: this composition is the Zilpaterol antibody with peroxidase labelling, and described Zilpaterol antibody is monoclonal antibody or polyclonal antibody;
4, luminous substrate liquid: this luminous substrate liquid is the Chemoluminescent substrate take the different luminol of luminol goods as luminous agent, is divided into A liquid and B liquid and preserves, and presses before use 1:1 and mixes use; Wherein A liquid level luminescence enhancer adds luminol goods luminescence enhancer and adds different luminol, and B liquid is superoxol or urea hydrogen peroxide solution;
5,2 times of concentration and dilution liquid;
6,20 times of concentrated cleaning solutions;
In the present invention, described opaque white color ELISA Plate is the detachable or non-removable opaque white color ELISA Plate in 96 holes.
In the present invention, described Zilpaterol standard items, are made up of the Zilpaterol standard items of a series of variable concentrations, and concentration is the concentration interval of 0.01 ~ 16.0 μ g/L.
In the present invention, described Zilpaterol-peroxidase labeled antibodies is the Zilpaterol antibody with peroxidase labelling, as the Zilpaterol antibody of horseradish peroxidase (HRP) mark.
In the present invention, described luminous substrate liquid is commercial any Chemoluminescent substrate take the different luminol of luminol goods as luminous agent.
In the present invention, described 2 times of concentration and dilution liquid, its composition is 0.01mol/L, phosphate buffer, glycocoll-HCl damping fluid or the Tris-HCl damping fluid of pH7.4 please dilute (1 part of concentrating sample dilution+1 part deionized water) by 1:1 before use.
In the present invention, described 20 times of concentrated cleaning solutions, it comprises 0.05% Tween-20, and the PBST of 0.01mol/L, between pH value scope 7.0-7.5, please dilutes (1 part of concentration and dilution liquid+19 part deionized water) by 1:19 before use.
When the chemiluminescence immune detection reagent kit of Zilpaterol of the present invention is applied to the detection of Zilpaterol, detecting step is:
(1) pre-service testing sample, is fluid sample by sample preparation to be tested, or extracts testing sample with organic solvent, and nitrogen dries up and redissolved in sample diluting liquid working fluid;
(2) required reagent is taken out from cold storage environment, more than being placed in room temperature (20～25 ℃) balance 30 min, note must shaking up before every kind of liquid reagent uses;
(3) get the ELISA Plate that is coated with Zilpaterol antigen, add standard items/testing sample 50 μ L/ holes in corresponding micropore, the each concentration of standard items and sample is done two parallel laboratory tests;
(4) add Zilpaterol antibody working fluid, 50 μ L/ holes, vibration mixes gently, with reacting 45 min in the rearmounted 25 ℃ of lucifuge environment of cover plate membrane cover plate;
(5) carefully open cover plate film, liquid in hole is dried, with wash operating solution 250 μ L/ holes, fully wash 4 ~ 5 times, every minor tick 10 s, pat dry (bubble not being eliminated after patting dry can be poked with original rifle head) with thieving paper;
(6) add luminous substrate liquid mixed liquor (A liquid mixes by 1:1 before use with B liquid) 100 μ L/ holes, vibration mixes gently, detects luminous intensity (RLU) after mixing in chemiluminescence detector;
(7) calculating of testing result: use the ratio of obtained standard solution and sample solution luminous value and blank solution to calculate.See following formula:
Relative luminous intensity=RLU/RULmax
In formula:
The luminous intensity values of RLU=standard (or sample) solution;
The luminous intensity values of RLUmax=blank (standard solution that concentration is 0);
The natural logarithm of the corresponding Zilpaterol of the relative luminous intensity value of calculating (μ g/L) is made to semilog coordinate system curve map.The Zilpaterol concentration of each testing sample is found on typical curve according to its RLU value, or calculates by the corresponding equation of typical curve.As there being dilution in sample preparation, the sample concentration that should draw according to typical curve will be multiplied by its extension rate again.Be the actual concentrations of Zilpaterol in sample.
Kit of the present invention can be used for the residues detection of Zilpaterol in the samples such as animals urine, blood sample, tissue and internal organ.With the comparison of existing other detection Zilpaterol residual quantity, kit of the present invention has following advantage:
(1) kit of the present invention of employing chemiluminescence immunoassay method, more more fast and convenient than chromatographic process (high efficiency liquid phase, LC-MS, gas chromatography mass spectrometry), capillary electrophoresis method, required instrument is more simple, and testing cost is more cheap, has high-throughout feature simultaneously;
(2) kit of the present invention of employing chemiluminescence immunoassay method, more sensitiveer than ELISA method, the Zilpaterol that can detect lower concentration and content is residual, and the range of linearity is wider simultaneously;
(3) adopt the kit of the present invention of chemiluminescence immunoassay method, reduced by two anti-use links, thereby shortened detection time; Opaque white color ELISA Plate has increased chemiluminescence detector sensitivity.In addition, with Zilpaterol-carrier protein couplet thing but not Zilpaterol antibody is coated with opaque white color ELISA Plate, reduce the instability of Zilpaterol antibody, guaranteed the long-term effectiveness of kit.
Embodiment
Below by specific embodiment, the invention will be further described.These embodiment are only for the present invention is described, and are not used for limiting the scope of the invention.
Embodiment 1
1, the preparation of the each component of kit
(1) the haptenic preparation of Zilpaterol: by Zilpaterol acidifying, with sodium nitrite effect, generate the intermediate containing diazo positive ion in 4 ℃ of unglazed low temperature environments.Diazotizing Zilpaterol is as haptens, for synthesizing immunizing antigen and envelope antigen below;
(2) the immunogenic preparation of Zilpaterol-bovine serum albumin(BSA) (BSA): adopt diazotising method to carry out coupling Zilpaterol and bovine serum albumin(BSA) (BSA) and obtain immunizing antigen;
The preparation of Zilpaterol-ovoserum albumin (OVA) envelope antigen: adopt diazotising method to carry out coupling Zilpaterol and ovoserum albumin (OVA) and obtain envelope antigen;
The preparation of Zilpaterol-peroxidase labeled antibodies: (g), heavy dose of immunization protocol is that first immunisation mixes with 160 μ g Zilpaterol-BSA and equivalent Freund's complete adjuvant, hypodermic injection to body weight 18 ~ 20 to the female BALB/c mouse to 6 ~ 8 week age.After 3 weeks, then mix hypodermic injection with 80 μ g Zilpaterol-BSA and equivalent Freund's complete adjuvant.After this mixed lumbar injection with 80 μ g Zilpaterol-BSA and equivalent Freund's complete adjuvant every 3 weeks.Last immunity in the spleen 80 μ g Zilpaterol-BSA are as booster immunization.After three days, put to death mouse, get its spleen, merge with myeloma cell.Screen positive hybridoma cell with indirect ELISA method.Inject hybridoma by mouse peritoneal and prepare in a large number mouse ascites, ascites, after filtration, centrifugal preliminary purification, adopts sad method and affinity chromatography purifying ascites, then obtains the Zilpaterol monoclonal antibody of purifying through dialysis.Zilpaterol monoclonal antibody and horseradish peroxidase, thus Zilpaterol-peroxidase labeled antibodies obtained;
Be coated with the opaque white color ELISA Plate preparation of Zilpaterol-OVA conjugate: with damping fluid by the detection hole for coated opaque white color ELISA Plate after the dilution of Zilpaterol-OVA conjugate, 4 ℃ spend the night after with the washing of PBST damping fluid, then add 180 μ L confining liquids (5% skimmed milk power solution), 37 ℃ of incubation 1.5 h, the liquid in hole that inclines, thieving paper pats dry rear sealing and preserves.
2, detect the establishment of the chemiluminescence immune detection reagent kit of Zilpaterol
The chemiluminescence immune detection reagent kit of the detection Zilpaterol of setting up, has comprised following ingredient:
(1) 96 hole opaque white color ELISA Plate (8 hole × 12) is coated with Zilpaterol-OVA conjugate, uses aluminium foil bag vacuum sealed package.
(2) 6 bottles of Zilpaterol standard solution, concentration is respectively:
0?ng/mL、0.01?ng/mL、0.03?ng/mL、0.09?ng/mL、0.27?ng/mL、0.81?ng/mL
(3) Zilpaterol-Horseradish Peroxidase Conjugates solution.
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide);
(5) 2 times of concentrating sample dilutions.Before use, please diluting (1 part of concentration and dilution liquid+1 part deionized water) by 1:1 becomes working prototype dilution, and the working prototype dilution after its dilution is 0.05 mol/L, the PBST damping fluid of pH 7.4;
(6) 20 times of concentrated cleaning solutions.Before use, please diluting (1 part of concentration and dilution liquid+19 part deionized water) by 1:19 becomes work cleansing solution, and the work cleansing solution after its dilution is between pH value scope 7.0-7.5, containing 0.05% Tween-20, and the PBST damping fluid of 0.01mol/L.
3, the chemiluminescence immune detection reagent kit of Zilpaterol is on probation
1) pre-treatment of sample
I, urine sample (pig urine)
Get urine sample to be checked, if when sample is more muddy, can be more than 5000 r/min, centrifugal 5 min or filtration;
II, serum (pig, ox etc.)
Get serum sample to be checked, if when sample is more muddy, can be more than 5000 r/min, centrifugal 5 min or filtration;
III, pork, pork liver (flesh tissue)
A. accurately take in 2.0 ± 0.05g homogenate sample to 50 mL centrifuge tube;
B. first add 3% trichloroacetic acid 4 mL, with adding again 2 mL acetonitriles after vortex instrument whirling motion extremely evenly and fully vibrating and shake up;
C.4000 centrifugal 10 min of r/min;
D. get supernatant 3 mL to another centrifuge tube, to adding 1 mol/LNa2CO3 solution 400 L in it, and mix (regulating pH value to 9.8 ± 0.2);
E. add 5 mL ethyl acetate, vibration 30 s, centrifugal 5 ~ 10 min of 4000 r/min; Getting 3 mL supernatant liquids flows down and dries up in 50 ~ 60 ℃ of nitrogen;
F. add the sample diluting liquid after 1 mL dilution and fully vibrate and shake up;
G. get 50 μ L for analyzing;
2) chemiluminescence immune detection reagent kit
The hole bar of standard and sample requirement is inserted in microwell plate framework to the position of record standard and sample.In suitable micropore, add respectively 50 μ L/ hole Zilpaterol standard solution and testing samples.Add 50 μ L/ hole Zilpaterol-Horseradish Peroxidase Conjugates in each micropore, after fully mixing, under room temperature, lucifuge leaves standstill incubation 45 min.Liquid in hole is dried, fully wash 4 ~ 5 times with wash operating solution.Remove the liquid in hole completely, pat dry with thieving paper, add luminous substrate liquid mixed liquor (A liquid mixes by 1:1 before use with B liquid) 100 μ L/ holes.After mixing, in chemiluminescence detector, detect immediately luminous intensity (RLU);
3) computational analysis of testing result
Calculate with the ratio of obtained standard solution and sample solution luminous value and blank solution.See following formula:
Relative luminous intensity=RLU/RULmax
In formula:
The luminous intensity values of RLU=standard (or sample) solution;
The luminous intensity values of RLUmax=blank (standard solution that concentration is 0);
The natural logarithm of the corresponding Zilpaterol of the relative luminous intensity value of calculating (μ g/L) is made to semilog coordinate system curve map.The Zilpaterol concentration of each testing sample is found on typical curve according to its RLU value, or calculates by the corresponding equation of typical curve.As sample has passed through beforehand dilution, the sample concentration that should draw according to typical curve will be multiplied by its extension rate again.
Embodiment 2
The chemiluminescence immune detection reagent kit that detects Zilpaterol, has comprised following ingredient:
(1) 96 hole opaque white color ELISA Plate (8 hole × 12) is coated with Zilpaterol-mouse serum albumin conjugate, uses aluminium foil bag vacuum sealed package;
(2) 6 bottles of Zilpaterol standard solution, concentration is respectively:
0?ng/mL、0.01?ng/mL、0.05?ng/mL、0.25?ng/mL、1.25?ng/mL、6.25?ng/mL
(3) Zilpaterol-Horseradish Peroxidase Conjugates solution;
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide);
(5) 2 times of concentrating sample dilutions.Before use, please diluting (1 part of concentration and dilution liquid+1 part deionized water) by 1:1 becomes working prototype dilution, and the working prototype dilution after its dilution is 0.05 mol/L, the Tris-HCl damping fluid of pH 7.4;
(6) 20 times of concentrated cleaning solutions.Before use, please diluting (1 part of concentration and dilution liquid+19 part deionized water) by 1:19 becomes work cleansing solution, and the work cleansing solution after its dilution is between pH value scope 7.0-7.5, containing 0.05% Tween-20, and the PBST damping fluid of 0.01mol/L.
Embodiment 3
The chemiluminescence immune detection reagent kit that detects Zilpaterol, has comprised following ingredient:
(1) 96 hole opaque white color ELISA Plate (8 hole × 12) is coated with Zilpaterol-egg albumin conjugate, uses aluminium foil bag vacuum sealed package;
(2) 6 bottles of Zilpaterol standard solution, concentration is respectively:
0?ng/mL、0.05?ng/mL、0.2ng/mL、0.8?ng/mL、4.0?ng/mL、16.0?ng/mL
(3) Zilpaterol-Horseradish Peroxidase Conjugates solution;
(4) luminous substrate A liquid (luminol and reinforcing agent), luminous substrate B liquid (urea hydrogen peroxide);
(5) 2 times of concentrating sample dilutions.Before use, please diluting (1 part of concentration and dilution liquid+1 part deionized water) by 1:1 becomes working prototype dilution, and the working prototype dilution after its dilution is 0.05 mol/L, glycocoll-HCl damping fluid of pH 7.4;
(6) 20 times of concentrated cleaning solutions.Before use, please diluting (1 part of concentration and dilution liquid+19 part deionized water) by 1:19 becomes work cleansing solution, and the work cleansing solution after its dilution is between pH value scope 7.0-7.5, containing 0.05% Tween-20, and the PBST damping fluid of 0.01mol/L.
Claims (5)
1. detect a chemiluminescence immune detection reagent kit for Zilpaterol, it is characterized in that the following composition in kit:
(1) be coated with the opaque white color ELISA Plate of Zilpaterol-carrier protein couplet thing; Described Zilpaterol-carrier protein couplet thing is that Zilpaterol and carrier protein are obtained by mixed anhydride method or carbodlimide method coupling, and described carrier protein is human serum albumins, bovine serum albumin(BSA), egg albumin, mouse haemocyanin or rabbit anteserum albumen;
(2) Zilpaterol standard items;
(3) Zilpaterol-peroxidase labeled antibodies: this composition is the Zilpaterol antibody with peroxidase labelling, and described Zilpaterol antibody is monoclonal antibody;
(4) luminous substrate liquid: this luminous substrate liquid is the Chemoluminescent substrate take the different luminol of luminol goods as luminous agent, is divided into A liquid and B liquid and preserves, and presses before use 1:1 and mixes use; Wherein A liquid is that luminescence enhancer adds luminol goods luminescence enhancer and adds different luminol, and B liquid is superoxol or urea hydrogen peroxide solution;
(5) 2 times of concentration and dilution liquid;
(6) 20 times of concentrated cleaning solutions;
According to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, described opaque white color ELISA Plate is the detachable or non-removable opaque white color ELISA Plate in 96 holes.
2. according to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, the concentration of described Zilpaterol standard items is the concentration interval of 0.01 ~ 16.0 μ g/L.
3. according to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, described Zilpaterol-peroxidase labeled antibodies working fluid is to be diluted to 1:20000 ratio with antibody diluent;
According to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, described luminous substrate liquid is commercial any Chemoluminescent substrate take the different luminol of luminol goods as luminous agent.
4. according to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, described 2 times of concentration and dilution liquid, its composition is 0.01mol/L, phosphate buffer, glycocoll-HCl damping fluid or the Tris-HCl damping fluid of pH7.4, please dilute (1 part of concentrating sample dilution+1 part deionized water) by 1:1 before use.
5. according to the chemiluminescence immune detection reagent kit of the Zilpaterol of claim 1, wherein, described 20 times of concentrated cleaning solutions, it comprises 0.05% Tween-20, the PBST of 0.01mol/L, between pH value scope 7.0-7.5, before use, please dilute (1 part of concentration and dilution liquid+19 part deionized water) by 1:19.
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| CN201210438102.2A CN103808933A (en) | 2012-11-06 | 2012-11-06 | Chemiluminiscence immunoassay kit for detecting zilpaterol |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105277701A (en) * | 2014-07-22 | 2016-01-27 | 江苏维赛科技生物发展有限公司 | Chemiluminiscence immunodetection kit for detecting adprin |
| CN105334323A (en) * | 2014-08-07 | 2016-02-17 | 暨南大学 | Method and test strip for detecting zilpaterol, and application of test strip |
| CN112679509A (en) * | 2021-01-21 | 2021-04-20 | 新乡学院 | Preparation and application of zilpaterol hapten, complete antigen and monoclonal antibody |
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| WEILIN L. SHELVER ET AL: "Enzyme-Linked Immunosorbent Assay Development for the β-Adrenergic Agonist Zilpaterol", 《AGRICULTURAL AND FOOD CHEMISTRY》, vol. 52, no. 8, 17 March 2004 (2004-03-17), pages 2159 - 2166 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105277701A (en) * | 2014-07-22 | 2016-01-27 | 江苏维赛科技生物发展有限公司 | Chemiluminiscence immunodetection kit for detecting adprin |
| CN105334323A (en) * | 2014-08-07 | 2016-02-17 | 暨南大学 | Method and test strip for detecting zilpaterol, and application of test strip |
| CN112679509A (en) * | 2021-01-21 | 2021-04-20 | 新乡学院 | Preparation and application of zilpaterol hapten, complete antigen and monoclonal antibody |
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Application publication date: 20140521 |