CN101907627A - Clenbuterol hydrochloride magnetic particle separation enzyme-linked immunosorbent assay method - Google Patents
Clenbuterol hydrochloride magnetic particle separation enzyme-linked immunosorbent assay method Download PDFInfo
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Abstract
The invention provides a clenbuterol (CLB) magnetic particle separation enzyme-linked immunosorbent assay (ELISA) method, belonging to the technical field of food safety immunodetection. The method comprises the following steps: the immunodetection principle of the competition method is adopted to connect CLB with bio-enzyme and prepare an enzyme labeled antigen reagent, antibody against fluorescein isothiocyanate (FITC) is absorbed on the surface of magnetic particles to prepare a magnetic separation reagent, FITC is connected with the antibody against CLB to prepare an antibody reagent; CLB in a sample and enzyme-labeled CLB compete to combine with a small quantity of FITC-labeled antibody against CLB and form an antigen-antibody composite; after the magnetic separation reagent is added, the antibody against FITC connected with the surface of magnetic particles captures the composite on the surface of magnetic particles; and washing, and finally adding substrate to detect. The invention has the following advantages: (1) magnetic particles replace the traditional ELISA plate to be used as solid carrier and ensure that the immunoreaction is performed more fully and fast under the near-liquidus condition; compared with the traditional ELISA method, the method is characterized by high specificity and good repeatability; and (2) the one-step competition method principle is adopted, thus the detection time is short.
Description
Technical field
The invention belongs to the immune detection analysis technical field, relate to the relevant immune detection analytical technology of food security, a kind of fast detecting clenobuterol hydrochloride (Clenbuterol is provided especially, CLB) magnetic particle separation enzyme-linked immunoassay is applicable to the detection by quantitative of the clenobuterol hydrochloride in urine, liver, meat and its hetero-organization.
Background technology
(Clenbuterol CLB) is a kind of antiasthmatic to clenobuterol hydrochloride.This medicine is neither veterinary drug, neither feed addictive, but adrenal gland class CNS stimulant.Clenbuterol hydrochloride is a kind of broxaterol, and early 1980s, U.S. a company begins it is added in the feed, increase lean meat percentage, if but as feed addictive, using dosage is more than 10 times of people's dosage, just can reach the effect that improves lean meat percentage.Its consumption is big, service time is long, metabolism is slow, so arrive listing before butchering, the residual quantity in the pig body is all very big.This residual quantity enters human body by food, just makes human body put aside poisoning gradually.If one time intake is excessive, will produce the intoxicating phenomenon of abnormal physiology reaction, so and it is disabled.The regulation of the Ministry of Agriculture is ignored at the domestic family of raising pigs, and in order to make the pork fat that do not flesh up, mixes clenobuterol hydrochloride in feed.Pig feed promotes protein synthesis with the back in metabolic process, quicken the conversion and the decomposition of fat, has improved the lean meat percentage of pork, therefore is called clenbuterol hydrochloride.
Clenobuterol hydrochloride is fast in gastrointestinal absorption, promptly worked in 15-20 minute, 2-3 hour plasma concentration peaking, the edible 20 μ g drug effects of general health people can show to some extent, 5-10 intake doubly then can cause poisoning, and symptom is: heart rate acceleration, face flush, uncomfortable in chest, palpitaition, headache, heating, shiver with cold.Clenobuterol hydrochloride causes human limb, face, neck bone flesh chatter after stimulating human skeletal muscle's beta 2-receptor.Long-time inferior therapeutic dose is taken in, hormone residues is accumulated gradually and is caused that the chronic toxicity effect can make human body produce low responsive phenomenon, be that bronchiectatic activity obviously weakens, acting duration shortens, make asthma incidence and occurring degree increase, also can cause endocrine disturbance simultaneously, cause lactic acid in the blood, pyruvic acid concentration to raise, therefore acidosis of diabetes patient.
The detection of clenbuterol hydrochloride residual quantity is significant in food and the feed.
Magnetic particle separation enzyme-linked immunoassay technology be a kind of be the solid phase carrier of separating with the magnetic particle, immune magnetic particle isolation technics is combined with the enzyme linked immunosorbent detection technology and a kind of novel immunologic detection method set up.Traditional E LISA method, the association reaction of antigen, antibody carries out on solid phase (elisa plate reacting hole) surface, and magnetic particle separation enzyme-linked immunoassay, the association reaction of antigen, antibody also carries out under the condition of approximate liquid phase, thereby reaction is fast, thoroughly.Compare with traditional E LISA and to have highly sensitively, detect few advantage of time spent.
" competition immunodetection " is a kind of haptenic method of micromolecule that is used to detect more, it detects principle: determined antigen combines a spot of insolubilized antibody with the antigenic competition of enzyme labeling, remove unconjugated antigen by washing, add the colour developing or the luminous substrate colour developing of biological enzyme at last.In the finite concentration scope, colour developing or luminous intensity and determined antigen content are inverse ratio.
Summary of the invention
The object of the present invention is to provide a kind of have highly sensitive, specificity good, applicability is wide, easy and simple to handle and save the magnetic particle separation enzyme-linked immunoassay of the detection CLB of detection time.
The reagent of described CLB magnetic particle separation enzyme-linked immunoassay is formed and is comprised: 1. magnetic separation agent [being combined with the magnetic particle suspension of anti-fluorescein isothiocynate (FITC) antibody]; 2. anti-reagent (the CLB antibody-solutions of FITC mark); 3. enzyme-labelled antigen reagent (coupling has the CLB solution of biology enzyme); 4. cleaning fluid; 5. CLB calibration object (solution that contains a certain amount of CLB antigen); 6. quality-control product (solution that contains a certain amount of CLB antigen); 7. substrate solution (colour developing or luminous substrate solution); 8. stop buffer.
The detection method of described CLB magnetic particle separation enzyme-linked immunoassay is as follows:
(1) sample (food, feed etc.) is handled: urine specimen can be directly used in detection.(if sample is very muddy, needs to detect through centrifugal or filtration again); Viscera tissue, muscle etc. need to pulverize evenly, add a certain proportion of 10%Na2CO3, add ethyl acetate, mix, and be centrifugal.Get upper strata liquid and continue to employ, lower sediment is washed once again, and with the upper strata liquid merging of two times centrifugal, 60 ℃ of nitrogen dry up, and add purified water and redissolve, and promptly can be used for detecting.
(2) application of sample and immune response: in test tube, add 20-200 μ l sample, add 20-200 μ l enzyme-labelled antigen reagent and the anti-reagent of 20-200 μ l then successively.Behind the mixing, 37 ℃ incubation 5-30 minute;
(3) add the magnetic separation agent: in test tube, add 20-100 μ l, the magnetic separation agent of 0.1-10mg/ml, behind the mixing, 37 ℃ incubation 5-30 minute;
(4) washing: make magnetic particle sedimentation in magnetic field, remove supernatant, adding 100-500 μ l cleaning fluid is removed magnetic field, shakes to make the abundant suspendible of magnetic particle in 30 seconds, makes magnetic particle sedimentation in magnetic field once more, removes supernatant.So repeated washing is 2-4 time;
(5) add substrate solution: every pipe adds the colour developing or the luminous substrate of 50-300 μ l biological enzyme.Behind the mixing, 37 ℃ incubation 5-30 minute;
(6) color development stopping: every pipe adds 100-300 μ l stop buffer, and is placed on 10 minutes (being applicable to development process) of separation on the magnetic separator;
(7) value of reading: on spectrophotometer or luminous intensity detector, read absorbance or luminous intensity values; The principle of work of described CLB magnetic particle separation enzyme-linked immunoassay is: at first, by the sample process process, CLB in the sample is dissolved in is used for subsequent analysis in the aqueous solution.After sample extraction liquid and enzyme mark CLB antigen reagent, FITC marked anti-reagent mix incubation, the CLB in the sample combined the small amount of FITC labeling antibody with enzyme mark CLB competition, forms antigen antibody complex.After the adding coupling had the magnetic separation agent of goat-anti FITC antibody, the magnetic separation agent can capture immune complex the magnetic particle surface.Through washing, remove unconjugated CLB, enzyme mark CLB and other impurity.Add enzymatic colour developing or luminous substrate at last.In the finite concentration scope, CLB content in the sample to be tested and colour developing or luminous degree are inverse ratio, in view of the above can the quantitative measurement sample in the content of CLB.
Technical solution of the present invention is as follows:
(1) preparation of magnetic separation agent: will resist FITC antibody to be connected the magnetic particle surface.Described magnetic particle diameter has superparamagnetism between 0.1-5.0 μ m, amino (NH2-) or carboxyl (COOH-) reactive group can be contained in the surface.Described goat-anti FITC antibody can pass through chemical cross-linking agent with being connected of magnetic particle, as glutaraldehyde, 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) etc. is with the form of covalent bond, and also can be by the form of physisorption (electrostatic interaction, ionic link or hydrophobic effect etc.).
(2) preparation of enzyme-labelled antigen reagent: CLB derivant and bovine serum albumin(BSA) (BSA) is covalently bound, covalently bound with biology enzyme more afterwards.Described biology enzyme can be horseradish peroxidase (HRP), also can be alkaline phosphatase (ALP).Covalently bound method can be passed through several different methods such as sodium periodate oxidizing process, glutaraldehyde method or Succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC), 2-IminothiolaneHCl (2IT) be crosslinked.
(3) preparation of anti-reagent: with the covalently bound thing solution of CLB antibody and fluorescein isothiocynate (FITC).Described CLB antibody can be monoclonal antibody, also can be polyclonal antibody.
(4) substrate solution: described substrate solution can be the chromogenic substrate methyl biphenyl amine (Tetrabenzidine of HRP catalysis, TMB) or luminous substrate luminol (Luminol) etc., also can be the chromogenic substrate list phosphoric acid phenolphthalein (PMP) or the luminous substrate AMPPD of ALP catalysis, CSPD and CSPD-Star etc.
(5) CLB calibration object and quality-control product: the high-purity C LB antigen back of weighing is made with aqueous solution dissolving packing.
The present invention has the following advantages:
(1) sample processing method is simple.Sample through pulverize, extract, dry up and simple process such as redissolution after promptly can be used for detecting the complicated processes such as handling and cross purification column of need not spending the night.
(2) this method detection sensitivity height, good reproducibility, the easy and simple to handle and advantage of saving detection time.
Description of drawings
The magnetic particle separation enzyme-linked immunization of Fig. 1 detects the CLB synoptic diagram
Embodiment
Embodiment 1: anti-FITC antigen and the coupling of surface amino groups (COOH-) magnetic particle, preparation magnetic separation agent
Get magnetic particle that the 100mg surface contains carboxyl (COOH-) reactive group with 0.1M MES (2-[N-morpholino] ethanesulfonic acid), pH 4.5-5 solution 10ml washing 3 times.Magnetic particle is resuspended with this solution 1ml, adds the anti-FITC antibody of 2mg, mixes.Add 100 μ l 10mg/ml EDC solution, mixed the back room temperature reaction 2 hours.Contain 0.01M phosphate buffer (PBS) the pH7.4 washing magnetic bead 3 times of 1% bovine serum albumin(BSA) (BSA) with 10ml after, be mixed with the magnetic separation agent working fluid of 2.5mg/ml with this solution.
Embodiment 2:CLB-BSA connects alkaline phosphatase (ALP), preparation enzyme-labelled antigen reagent
Get 5mg CLB-BSA antigen, be concentrated into 5mg/ml, add activator 2-IminothiolaneHCl (2IT) the solution 10 μ l of 13.76mg/ml, room temperature is placed and is added glycocoll termination priming reaction after 30 minutes, places 5 minutes under the room temperature.Use Sephadex G25 pillar desalination, collect the albumen eluting peak.
Get 5mg ALP solution, add Succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) the solution 50 μ l of 6.69mg/ml, room temperature was placed 15 minutes, added glycocoll and stopped priming reaction, and room temperature was placed 5 minutes.Use Sephadex G25 pillar desalination, collect the albumen eluting peak.
The CLB-BSA of activation is mixed with the ALP of activation, and room temperature was placed 10 hours, used Supperdex200 gel chromatography column separating purification then, removed the free CLB-BSA and the ALP that do not connect, and connector is stored in 4 ℃.
Enzyme-labelled antigen reagent diluent preparation method is: Tris 12.11g, and sodium chloride 8.77g, Sodium azide 1g adds purified water to 600ml, proofreaies and correct pH value to 7.5.Add Tween-20 2mL, BSA 10g adds purified water and is settled to 1000ml.Filter with 0.22 μ m filtrator, in 4 ℃ of preservations.
Above-mentioned CLB-BSA-ALP connector is mixed with enzyme-labelled antigen reagent working fluid with above-mentioned diluted to 1-5 μ g/ml.
Embodiment 3: anti-CLB antibody connects FITC, prepares anti-reagent
Dialysis surpasses 12h to 20mM pH9.0 carbonic acid buffer to get the anti-CLB antibody-solutions of 5mg, and dialysis back concentration requirement is greater than 1mg/ml.Add the FITC solution (with the preparation of 20mM pH9.0 carbonic acid buffer) of 500 μ l 0.5mg/ml, room temperature reaction surpasses 12h behind the mixing.Use Sephadex G25 pillar to remove, collect the albumen eluting peak not in conjunction with FITC.
Anti-reagent diluent preparation method is identical with enzyme-labelled antigen reagent diluent preparation method, and above-mentioned CLB antibody FITC connector is arrived 1-5 μ g/ml with diluted, is mixed with anti-reagent working fluid.
Embodiment 4:CLB magnetic particle separation enzyme immunodetection detects CLB in the pork sample
Material and instrument
(1) magnetic separation agent: see the preparation of above-mentioned magnetic separation agent;
(2) enzyme-labelled antigen reagent: see the preparation of above-mentioned enzyme-labelled antigen reagent;
(3) anti-reagent: see the preparation of above-mentioned anti-reagent;
(4) cleansing solution: 500mL (Beijing Bei Ai Kang Biotechnology Co., Ltd's production);
(5) the molten thing of substrate (PMP solution): 300mL (Beijing Bei Ai Kang Biotechnology Co., Ltd's production);
(6) stop bath: 600mL (Beijing Bei Ai Kang Biotechnology Co., Ltd's production);
(7) the separation enzyme-linked immunoassays instrument of magnetic: the Beijing Bei Ai Kang Biotechnology Co., Ltd produces;
(8) magnetic separator: the Beijing Bei Ai Kang Biotechnology Co., Ltd produces;
(9) water bath (being used for 37 ℃ of temperature bathes).
(1) the detection step is as follows:
(2) sample process: evenly get 5g the sample from a large amount of, add 3ml 10%Na2CO3, shook 3 minutes; Add 15ml ethyl acetate, mixed 5 minutes, 4000 rev/mins centrifugal 5 minutes; Upper strata liquid is all forwarded in the clean vessel, and lower sediment adds 15ml ethyl acetate again, mixes 5 minutes, and 4000 rev/mins centrifugal 5 minutes; The upper strata liquid of two times centrifugal is respectively got 3ml merge (being equivalent to the 1g sample); 60 ℃ of nitrogen dry up, and add the 1ml purified water and redissolve; Getting 60 μ l is used for detecting.
(3) application of sample and immune response: add 60 μ L sample solutions in flat based tubes, add 60 μ l enzyme-labelled antigen reagent and the anti-reagent of 60 μ l then respectively successively, 37 ℃ of incubations are 15 minutes behind the mixing;
(4) add the magnetic separation agent: add the magnetic separation agent behind the 60 μ l mixings in test tube, 37 ℃ of incubations are 5 minutes behind the mixing;
(5) wash: flat based tubes is placed on the magnetic separator separated 2 minutes, remove supernatant, every pipe adds cleaning fluid 300 μ L, fully behind the mixing, flat based tubes is placed on the magnetic separator again and separates abandoning supernatant 2 minutes.Repeated washing process 2 times;
(6) add substrate solution: every pipe adds single phosphoric acid phenolphthalein (PMP) substrate solution of 100 μ l, behind the mixing, and 37 ℃ of incubations 15 minutes;
(7) color development stopping: every pipe adds 300 μ L stop buffers, and is placed on the magnetic separator and separated 10 minutes;
(8) read absorbance: exempt from analyzer with spectrophotometer or magnetic enzyme and read absorbance, wavelength is 492nm/550nm.
Testing result:
(1) measurement range: 0～10ppb
(2) accuracy: the recovery is 85 ± 15%.
(3) repeatability: variation within batch coefficient: CV≤10%; Interassay coefficient of variation: CV≤15%
(4) sensitivity: sensitivity is not higher than 0.1ppb.
(5) specificity: see the following form with the cross reacting rate of other similar toxin.
| The cross reaction thing | Cross reacting rate (%) |
| Clenbuterol | 100％ |
| Bromine Boot sieve | 66％ |
| Mabuterol | 47％ |
| Salbutamol | 10％ |
| Carbuterol | 3％ |
| Adrenaline | ＜0.01％ |
Claims (3)
1. the magnetic particle separation enzyme-linked immunization method of a clenobuterol hydrochloride (CLB) is characterized in that detecting step and is:
(1) application of sample and immune response: in test tube, add 20-200 μ l sample solution, CLB calibration object or CLB quality-control product, add 20-200 μ l enzyme-labelled antigen reagent and the anti-reagent of 20-200 μ l then.Behind the mixing, 37 ℃ incubation 5-30 minute;
(2) add the magnetic separation agent: in test tube, add 20-100 μ l magnetic separation agent, behind the mixing, 37 ℃ incubation 5-30 minute;
(3) washing: make magnetic particle sedimentation in magnetic field, remove supernatant, add 100-500 μ l cleaning fluid, remove magnetic field, concussion makes the abundant suspendible of magnetic particle 30 seconds, and then makes magnetic particle sedimentation in magnetic field, removes supernatant.So repeat 2-4 time;
(4) add substrate solution: every pipe adds the colour developing or the luminous substrate of 50-300 μ l biological enzyme, behind the mixing, 37 ℃ incubation 5-30 minute;
(5) color development stopping (being only applicable to development process): every pipe adds 100-300 μ l stop buffer, and is placed on the magnetic separator and separated 10 minutes;
(6) value of reading: on spectrophotometer or luminous intensity detector, read absorbance or luminous intensity values;
2. anti-reagent according to claim 1 is characterized in that: the covalently bound thing solution that is anti-CLB antibody and fluorescein isothiocynate (FITC).Described anti-CLB antibody can be monoclonal antibody, also can be polyclonal antibody.Its working concentration is 0.2-10 μ g/ml.
3. enzyme-labelled antigen reagent according to claim 1 is characterized in that: the connector solution that is the further covalently bound biology enzyme formation of covalently bound thing (CLB-BSA) of clenobuterol hydrochloride and bovine serum albumin(BSA) (BSA).Described biology enzyme can be horseradish peroxidase (HRP), also can be alkaline phosphatase (ALP).The method of attachment of biology enzyme and CLB-BSA can be passed through several different methods such as sodium periodate oxidizing process, glutaraldehyde method or Succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC), 2-IminothiolaneHCl (2IT) cross-linking method.Its working concentration is 0.5-5 μ g/ml.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102168072A (en) * | 2010-12-20 | 2011-08-31 | 江苏出入境检验检疫局动植物与食品检测中心 | A monoclonal antibody of anti-clenobuterol hydrochloride and a kit for detecting clenobuterol hydrochloride |
| CN102818892A (en) * | 2012-08-16 | 2012-12-12 | 北京恩济和生物科技有限公司 | Detection kit for prostate specific antigen and preparation method thereof |
| CN103048475A (en) * | 2012-12-18 | 2013-04-17 | 苏州浩欧博生物医药有限公司 | Nano magnetic particle chemiluminiscence assay kit for free HCG (human chorionic gonadotropin) beta subunit, preparation method for Nano magnetic particle chemiluminiscence assay kit and detection method adopting Nano magnetic particle chemiluminiscence assay kit |
| CN103808921A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Enzyme-linked immunosorbent assay kit for detecting residual zilpaterol and use method thereof |
| CN103823064A (en) * | 2014-03-03 | 2014-05-28 | 中华人民共和国张家港出入境检验检疫局 | Vomitoxin quantitative determination kit and use method thereof |
| CN104807993A (en) * | 2015-04-23 | 2015-07-29 | 广东国际旅行卫生保健中心（广东出入境检验检疫局口岸门诊部） | Mycobacterium tuberculosis ESAT-6 protein detection kit, as well as preparation method and use method |
| CN104914080A (en) * | 2015-05-14 | 2015-09-16 | 桂林理工大学 | Rapid detection method of clenbuterol |
| CN105301234A (en) * | 2015-09-21 | 2016-02-03 | 北京勤邦生物技术有限公司 | Clenbuterol immunomagnetic bead separation and enrichment kit and application thereof |
| CN108700584A (en) * | 2017-01-20 | 2018-10-23 | 深圳市新产业生物医学工程股份有限公司 | Labeled complex and preparation method thereof, kit, application and detecting system |
| CN108845125A (en) * | 2018-06-24 | 2018-11-20 | 沭阳康源泰博生物科技有限公司 | A kind of clenobuterol hydrochloride Sample pretreatment kit based on immunomagnetic beads |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101514991A (en) * | 2008-02-19 | 2009-08-26 | 北京科美东雅生物技术有限公司 | Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same |
-
2010
- 2010-08-05 CN CN2010102458919A patent/CN101907627A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101514991A (en) * | 2008-02-19 | 2009-08-26 | 北京科美东雅生物技术有限公司 | Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same |
Non-Patent Citations (3)
| Title |
|---|
| ZHEFEI LI ET AL.: "Flow injection chemiluminescent determination of clenbuterol using GoldMag particles as carrier", 《FOOD ADDITIVES & CONTAMINANTS》 * |
| 宓晓黎等: "盐酸克伦特罗的酶联免疫分析试剂盒的研制-- II克伦特罗酶联免疫检测方法的研究", 《中国卫生检验杂志》 * |
| 汪慧蓉等: "金磁微粒介导的盐酸克伦特罗多克隆抗体的纯化", 《西北农林科技大学学报（自然科学版）》 * |
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| CN102168072A (en) * | 2010-12-20 | 2011-08-31 | 江苏出入境检验检疫局动植物与食品检测中心 | A monoclonal antibody of anti-clenobuterol hydrochloride and a kit for detecting clenobuterol hydrochloride |
| CN102168072B (en) * | 2010-12-20 | 2013-03-06 | 江苏出入境检验检疫局动植物与食品检测中心 | A monoclonal antibody of anti-clenobuterol hydrochloride and a kit for detecting clenobuterol hydrochloride |
| CN102818892A (en) * | 2012-08-16 | 2012-12-12 | 北京恩济和生物科技有限公司 | Detection kit for prostate specific antigen and preparation method thereof |
| CN103808921A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Enzyme-linked immunosorbent assay kit for detecting residual zilpaterol and use method thereof |
| CN103048475A (en) * | 2012-12-18 | 2013-04-17 | 苏州浩欧博生物医药有限公司 | Nano magnetic particle chemiluminiscence assay kit for free HCG (human chorionic gonadotropin) beta subunit, preparation method for Nano magnetic particle chemiluminiscence assay kit and detection method adopting Nano magnetic particle chemiluminiscence assay kit |
| CN103823064B (en) * | 2014-03-03 | 2016-05-25 | 中华人民共和国张家港出入境检验检疫局 | A kind of vomitoxin immue quantitative detection reagent box and using method thereof |
| CN103823064A (en) * | 2014-03-03 | 2014-05-28 | 中华人民共和国张家港出入境检验检疫局 | Vomitoxin quantitative determination kit and use method thereof |
| CN104807993A (en) * | 2015-04-23 | 2015-07-29 | 广东国际旅行卫生保健中心（广东出入境检验检疫局口岸门诊部） | Mycobacterium tuberculosis ESAT-6 protein detection kit, as well as preparation method and use method |
| CN104914080A (en) * | 2015-05-14 | 2015-09-16 | 桂林理工大学 | Rapid detection method of clenbuterol |
| CN104914080B (en) * | 2015-05-14 | 2017-04-26 | 桂林理工大学 | Rapid detection method of clenbuterol |
| CN105301234A (en) * | 2015-09-21 | 2016-02-03 | 北京勤邦生物技术有限公司 | Clenbuterol immunomagnetic bead separation and enrichment kit and application thereof |
| CN108700584A (en) * | 2017-01-20 | 2018-10-23 | 深圳市新产业生物医学工程股份有限公司 | Labeled complex and preparation method thereof, kit, application and detecting system |
| CN108700584B (en) * | 2017-01-20 | 2021-11-30 | 深圳市新产业生物医学工程股份有限公司 | Marker complex, preparation method thereof, kit, application and detection system |
| CN108845125A (en) * | 2018-06-24 | 2018-11-20 | 沭阳康源泰博生物科技有限公司 | A kind of clenobuterol hydrochloride Sample pretreatment kit based on immunomagnetic beads |
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Application publication date: 20101208 |