CN102539412A - Magnetic particle chemical luminous kit for detecting clenbuterol and application thereof - Google Patents
Magnetic particle chemical luminous kit for detecting clenbuterol and application thereof Download PDFInfo
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- CN102539412A CN102539412A CN201010588988XA CN201010588988A CN102539412A CN 102539412 A CN102539412 A CN 102539412A CN 201010588988X A CN201010588988X A CN 201010588988XA CN 201010588988 A CN201010588988 A CN 201010588988A CN 102539412 A CN102539412 A CN 102539412A
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Abstract
The invention relates to a magnetic particle chemiluminescence kit for detecting clenbuterol and application thereof. The magnetic particle chemiluminescence kit comprises the following reagents: a luminescence marker, a fluorescein marker, a standard product, a quality control product and a separating reagent. The luminescence marker is a clenbuterol hapten marked by an isoluminol luminescence marker; the fluorescein marker is a clenbuterol monoclonal antibody marked by fluorescein or derivates thereof; and the separating reagent is a paramagnetic nanometer micro-bead coated with an anti-goat FITC (fluorescein isothiocyanate) monoclonal antibody. The invention further relates to a method for detecting clenbuterol in animal-derived food by using the magnetic particle chemiluminescence kit, wherein the method has the characteristics of higher sensitivity, higher specificity and faster detection speed on clenbuterol detection.
Description
Technical field
The present invention relates to a kind of chemiluminescence detection kit and application method thereof, particularly detect the magnetic granule chemoluminescence detection kit of Ke Lunluo residual quantity in animal tissue, urine, the feed equal samples.
Technical background
Clenbuterol (Clenbuterol) belongs to beta-stimulants, and illegally added to the lean meat percentage and acceleration growth of animal to improve the lard type animal in the feed in recent years, and extensively made an addition in the animal feed, and can be in animal body residual.Yet this medicine has serious adverse to the people.Gently then cause the heartbeat rhythm of the heart undesired, but heavy then cardiac trigger sick.At present, China and many countries have forbidden that it uses as feed addictive.
At present, mainly adopting high performance liquid chromatography (HPLC), LC/MS (LC-MS), gas chromatography-mass spectrography (GC-MS) and ELISA methods such as (ELISA) to be used for the animal derived food residual of kelengtelu detects.
1, high performance liquid chromatography (HPLC) has accuracy of detection height, characteristics that false positive rate is low, and the lowest detection that detects residual of kelengtelu is limited to 1～15 μ g/kg.The major defect of HPLC method is that instrument costs an arm and a leg, operate more loaded down with trivial details, length consuming time, testing cost is expensive, and testing staff's specialty is had relatively high expectations, the sample pre-treatment is complicated.
2, gas chromatography-mass spectrography (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 in mass spectrum, provide more structural information through the sample of derivatization treatment, derivatization can produce a plurality of different products and cause the partial loss of sample, causes the deviation of experimental result.
3, LC/MS (LC-MS), sample does not need derivatization treatment, can detect urine, blood, liver, hair and eyeball sample.This method reaches 0.1 μ g/kg to the LDL of Clenbuterol, and the LC-MS/MS coupling can further improve signal to noise ratio (S/N ratio), so can be used for the affirmation means to positive findings.But, no matter LC-MS or LC-MS/MS, the instrument detecting method is the same with GC-MS, and fails to solve instrument and involve great expense, complex operation step, the sample complex pretreatment requires problems such as height to operating personnel's specialty attainment.
4, ELISA (ELISA) is to occur the seventies in 20th century; Be used for the detection of micro substance; Be applied to Clinical detection such as infectious disease, tumor markers, hormonal readiness the earliest; Beginning the nineties to apply in China food safety detection field, rely on the kit of elisa technique, the leading products that test strips has become food security fast detecting field at present, also is simultaneously the major product type that my company researches and develops and sells.The enzyme linked immunosorbent detection method is based on the specific reaction of Ag-Ab; Detection sensitivity is higher, specificity is better; Technical operation is simple and easy; Easy master, qualitative, the quantitative test work that have solved a large amount of former insoluble many micro-small-molecule substances such as microbiotic, hormone, residues of pesticides etc. have really been played positive facilitation to the development of food security Fast Detection Technique.But there are many defectives that self can't overcome in the ELISA method, and mainly shows:
1) heterogeneous reaction: be separated free thing and bond in the testing process, need multistep to wash the plate process, time and effort consuming is difficult to improve the automation of operation degree.
2) enzymic catalytic reaction: by substrate for enzymatic activity colour developing, assaying reaction liquid absorbance.There are considerable influence in the time and the temperature of reaction to enzyme activity, and reagent stability is poor.
Summary of the invention
Technical matters to be solved by this invention is to provide a kind of clenbuterol detection reagent kit, not only possesses higher sensitivity when adopting this kit to carry out the detection of Clenbuterol, specificity, and have higher reaction velocity.
Another object of the present invention is to provide a kind of method of testing of Clenbuterol, and this method is simple to operate, and is highly sensitive, and specificity is good.
Realize above-mentioned purpose, the present invention provides a kind of clenbuterol detection reagent kit, and its main agents that comprises has: luminous marker, fluorescein-labelled thing, standard items, quality-control product, separation agent.
Described separation agent is the paramagnetism nano microsphere that is coated with goat-anti FITC monoclonal antibody.
Described paramagnetism nano microsphere, surface contain-OH ,-COOH or-NH
2The microballon of reactive group is used to encapsulate goat-anti FITC monoclonal antibody, and its inner core is Fe
3O
4Or γ-Fe
2O
3, make microballon have paramagnetism.
Described luminous marker is the Clenbuterol haptens of different luminol derivant mark.Described different luminol derivant is ABEI, AHEI or ABEN.
Described kit also comprises standard items, quality-control product and concentrated washing lotion.
The Clenbuterol monoclonal antibody that described fluorescein-labelled thing is the FITC mark.
The present invention also provides a kind of method of utilizing kit to detect Clenbuterol, comprises the following steps:
1) draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min;
2) add the paramagnetism nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min;
3) separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant;
4) above-mentioned cleaning step repeats 2-4 time;
5) 4) compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s, content in the sample and RLU proportion relation can be through the concentration of RLU set calibration curve method calculating Clenbuterol.
The principal ingredient of described luminous substrate is NaOH and H
2O
2
The used analyser of analysis test method is among the present invention: comprise power circuit, automatic syringe pump 1 and 2, measuring chamber, illuminated chamber, photomultiplier counter and output system; Also dispose the Windows Control Software of computing machine and Chinese interface simultaneously; Can carry out that data typing, result gather, quality control, the result stores and function such as result queries; Can accomplish the programming of multiple analytical model, quantitative or qualitative reporting the result generates and storage, update functions automatically; Automatically revise typical curve at 2, the system that is adopted is the CI-2008 system.
Separation agent of the present invention is to encapsulate goat-anti FITC monoclonal antibody, and the content of its surface group is 0.1-0.3eqm/g, and it is stored in and contains 0.1-0.5%BSA, 0.01-0.1%Tween-20,0.02%NaN
3, in the PBS damping fluid of pH7.2-7.6.Described FITC is a fluorescein isothiocynate.Said percentage composition is the quality percentage composition.
Described luminous marker is that hapten-marked its of Clenbuterol of different luminol derivant ABEI, AHEI or ABEN mark is stored in pH7.2-7.6, contains 0.1-0.3%Tween-20,0.01%NaN
3The PBS damping fluid in.Said percentage composition is the quality percentage composition.
The Clenbuterol monoclonal antibody that described fluorescein-labelled thing is the FITC mark, it is stored in pH7.2-7.6, contains 5-8%BSA, 0.02-0.04%NaN
3The PBS damping fluid.Said percentage composition is the quality percentage composition.
The Clenbuterol standard solution (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.4,0.03%NaN
3, 0.05mol/L TRIS damping fluid.Said percentage composition is the quality percentage composition.
Clenbuterol quality-control product solution concentration is respectively 0.02ng/ml, 0.5ng/ml, quality-control product dilution pH7.4,0.03%NaN
3, 0.05mol/L TRIS damping fluid.Said percentage composition is the quality percentage composition.
Described concentrated washing lotion is PH7.2-7.8,0.2-0.4%Tween-20,0.02-0.04%NaN
3, 0.1-0.2mol/L PBS damping fluid.Said percentage composition is the quality percentage composition.
Beneficial effect of the present invention is following:
1) but kit specific detection clenbuterol of the present invention does not have intersection to other drug.
2) sensitivity of kit of the present invention is higher, can reach 0.01ng/ml to the detection sensitivity of clenbuterol.
3) detection of kit of the present invention is quick, is lower than 20min detection time.
Embodiment
The preparation of the concrete component of embodiment 1 kit
1, the preparation of luminous marker
A) Clenbuterol is haptenic synthetic
Clenbuterol is carried out acylation reaction with succinic anhydride, become the alcoholic extract hydroxyl group acidylate on the Clenbuterol molecular structure Clenbuterol haptens of the indirect arm of carboxyl that contains 4 carbon.
B) luminous marker preparation
Get 4.5mmol/L ABEI, be dissolved in the 4ml distilled water, the 5.0mmol/L N-hydroxy-succinamide is dissolved in 0.5ml N, in the dinethylformamide, and room temperature reaction 3-4h behind the two abundant mixing.Get the Clenbuterol haptens 15mg of above-mentioned preparation, to 1.5ml, add the ABEI solution of above-mentioned activation with the pH7.4PBS adjusted volume then, fully room temperature reaction spends the night behind the mixing, crosses G-25 gel column purifying.
2, fluorescent marker preparation
A) immunogenic preparation:
Adopt mixed anhydride method to carry out coupling Clenbuterol haptens and ovalbumin and obtain immunogene.
B) Clenbuterol MONOCLONAL ANTIBODIES SPECIFIC FOR
Animal immune: with immunogene the Balb/c mouse is carried out immunity, immunizing dose is 150 μ g/, makes it produce antiserum.
Fusion of Cells and cloning: get immune Balb/c mouse boosting cell, merge, obtain the hybridoma cell strain of monoclonal antibody in 9: 1 ratios and SP2/0 myeloma cell.
Cell cryopreservation and recovery: hybridoma is processed 1 * 10 with cryopreserving liquid
6The cell suspension of individual/ml is in the medium-term and long-term preservation of liquid nitrogen.Take out frozen pipe during recovery, put into 37 ℃ of water-bath middling speeds immediately and melt, behind the centrifugal removal cryopreserving liquid, move in the culture flask and cultivate.
MONOCLONAL ANTIBODIES SPECIFIC FOR and purifying: the Balb/c mouse peritoneal is only injected sterilization paraffinum liquidum 0.5ml/, 7 days pneumoretroperitoneum injection hybridomas 5 * 10
5Individual/as only, to gather ascites after 7 days.Carry out purifying with sad-saturated ammonium sulfate method, the ascites behind the purifying is put into-20 ℃ of environment and is preserved.
C) fluorescent marker preparation
Use 0.025mol/L, the carbonate buffer solution of pH9.0 is diluted to 1% mass percent concentration with the Clenbuterol monoclonal antibody; Total protein according to desiring mark adds 0.01mg FITC by every milligram of immunoglobulin (Ig), accurately takes by weighing required FITC powder with analytical balance.With the solution that FITC is made into 0.1mg/ml,, sneak into the FITC dilution with same damping fluid by 3-5 times of above-mentioned antibody-solutions volume; Electromagnetic agitation, mark 30-48h under 4～℃ lucifuge condition; With label solution 3000r/min, the centrifugal 20min of room temperature removes wherein a spot of sediment, and in the bag filter of packing into, the PBS damping fluid of pH7.4 dialysis is 2-3 days again, during change dislysate at least 3 times; Get the label of dialysed overnight, through SephadexG-25 or G-50 post, the separated free luciferin is collected the fluorescent marker of mark and is identified that packing is stored in 4 ℃ of refrigerators.
3, separation agent preparation
A) magnetic bead activation
The magnetic bead of surface-COOH group (purchase in DYNAL, particle diameter is 2.8 μ m), its content is 0.15eq/g; Get 100 μ l magnetic beads, with 100 μ l 25mmol/L, pH5.0,0.05%Tween-20MES solution washing twice, magnetic removes supernatant after separating; Before the use, dispose EDC, the NHS solution of 50mmol/L respectively with the 25mmol/L MES solution of 4 ℃ of storages; Add new EDC that disposes of 50 μ l and NHS solution respectively in the centrifuge tube that magnetic bead is housed, vortex mixing, room temperature activation 30min; Centrifuge tube places on the magnetic separator frame magnetic to separate 4min, removes supernatant, adds 100 μ l, 25mmol/L, pH5.0, MES clean get final product after 2-3 time the magnetic bead of surperficial activated carboxylic.
B) magnetic bead coupling goat-anti FITC monoclonal antibody
50-100 μ g goat-anti FITC monoclonal antibody is dissolved into 60 μ l, and 25mmol/L among the pH5.0MES, regulates cumulative volume to 100 μ l with said MES solution, soft mixing magnetic bead and antibody; At least coupling 30min or 4 ℃ of coupling 2h under the room temperature condition, vortex appearance capable of using makes magnetic bead keep the mixing state during this period; Centrifuge tube places magnetic separation 3-5min on the magnetic separator frame, removes supernatant; For cancellation unreacted-COOH, can add 100 μ l, pH7.4, TRIS reaction 15min or 100 μ l, pH8.0 contains the PBS sealing magnetic bead of 50mmol/L monoethanolamine; With 100 μ l, 0.1-0.3%BSA, the PBS of 0.1%Tween-20 or TRIS clean the good magnetic bead of sealing 3-5 time; At last magnetic bead is redissolved in containing 0.1-0.5%BSA, 0.01-0.1%Tween-20,0.02%NaN
3PBS or TRIS damping fluid in, 2-8 ℃ of preservation.
The establishment of embodiment two kits
Set up the magnetic granule chemoluminescence detection kit that detects Clenbuterol, make it contain following component:
The fluorescent marker of the Clenbuterol monoclonal antibody of FITC mark
The haptenic luminous marker of the Clenbuterol of ABEI mark
The separation agent of the paramagnetism Nano microsphere of pan coating goat-anti FITC monoclonal antibody
The Clenbuterol standard solution (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.4, contains 0.03%NaN
3, 0.05mol/L TRIS damping fluid.Said percentage composition is the quality percentage composition.
Clenbuterol quality-control product solution concentration is respectively 0.02ng/ml, 0.5ng/ml, and quality-control product dilution pH7.4 contains 0.03%NaN
3, 0.05mol/L TRIS damping fluid.Said percentage composition is the quality percentage composition.
Concentrated washing lotion is pH7.6,0.4%Tween-20,0.02%NaN
3, 0.1mol/L PBS damping fluid.Said percentage composition is the quality percentage composition.
The detection of Clenbuterol in embodiment three actual samples
1, sample pre-treatment
(1) urine
Get the limpid urine sample of 20 μ l directly measure (as urine sample is muddy must be through filtering or more than the 3000g, 15 ℃ of centrifugal 10min are until limpid), the sample that wouldn't use should freezing preservation, diluted sample multiple: 1
(2) tissue has tissues such as low-fat meat, liver
Take by weighing in 2.0 ± 0.05g homogeneous sample to the 50ml polystyrene centrifuge tube; Add 6ml 4%NaCl-0.1M HCl-methyl alcohol mixed liquor, shake to evenly with the oscillator vibration; Left standstill more than the 3000g centrifugal 5 minutes 10 minutes; Liver samples: get the 1ml supernatant and add 20 μ l 1M sodium hydroxide solution mixings (mixing is measured the pH value later on, is approximately 8); Muscle sample: get the 1ml supernatant and add 30 μ l 1M sodium hydroxide solution mixings (mixing is measured the pH value later on, is approximately 8); Getting 20 μ l is used for analyzing diluted sample multiple: 1
(3) feed
Grind the feed sample with mortar, take by weighing in sample to the 50ml polystyrene centrifuge tube of 2.0 ± 0.05g grinding, add 2ml 1M hydrochloric acid solution, add the 16ml deionized water and carry out homogeneous with homogenizer; With vortex appearance whirling motion 3min, put the 15min that vibrates on the oscillator; More than the 3000g, centrifugal 20min migrates out in whole supernatants to the 10ml polystyrene centrifuge tube, and adds 2ml 1M sodium hydroxide solution and mix, and whether inspection pH value is between 6.5-7.5; (regulating the pH value) with hydrochloric acid or sodium hydroxide solution; More than the 3000g, centrifugal 20min gets 100 μ l supernatants to the clean polystyrene centrifuge tube of 2ml, adds 900 μ l deionized waters and mixes; Getting 20 μ l is used for analyzing diluted sample multiple: 100
(4) milk
Get 100 μ l milk samples, add 400 μ l, 4% sodium chloride solution, the whirling motion mixing is got 20 μ l and is used for analyzing diluted sample multiple: 5.
2, detect and interpretation of result with kit
Draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min; Add the paramagnetism nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min; Separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant; The compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s; The content of Clenbuterol and RLU proportion relation in the sample can be through the concentration of RLU combined standard curve method calculating Clenbuterol.
Exciting substrate 1 is NaOH, and exciting substrate 2 is H
2O
2Before substrate loads, clean the substrate pump 10-20 time, behind the remaining water mark, more corresponding substrate is directly put into instrument in the emptying pipe, pump line is inserted in the substrate bottle with distilled water; With substrate flushing pipe 5 times, and measure the RLU value of substrate, under the normal condition, the RLU value of substrate should be above 1200.If surpass 1200, need with distilled water pipeline and substrate pump to be carried out more times cleaning again, in blank value is reduced to zone of reasonableness.Do not do experiment if surpass three days, need unloading substrate bottle, and cover lid, with vaporization prevention.Use the distilled water pipe blow-through subsequently, and emptying, in order to avoid strong base solution corrosion substrate pump.
The present invention adopt 6 Clenbuterol standard items (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml 0.81ng/ml) carries out plotting curves.Instrument detects according to said method and obtains a RLU value and the concentration dependent calibration curve of Clenbuterol, and in after this measuring, the Clenbuterol concentration in each sample is compared with typical curve and drawn the Clenbuterol content in the sample.
The mensuration of embodiment four kit quality
1, the sensitivity of kit
Being defined as of kit sensitivity: measure 20 times the zero standard article, the mean value of mensuration adds 3 times of standard deviations.The sensitivity of this kit is 0.01ng/ml.
2, the accuracy of sample and precision
Accuracy is meant the matching degree between measured value and true value, and the kit accuracy recovery commonly used is represented.Precision is claimed repeatability again, and the coefficient of variation commonly used is represented.
Sample extraction method according to embodiment three; Clenbuterol with 0.05ng/g (ml), two concentration of 0.1ng/g (ml) adds pig urine, pork liver, pork respectively; With the Clenbuterol of 2ng/g, two concentration of 4ng/g the feed sample is added, with the Clenbuterol of 0.1ng/ml, two concentration of 0.2ng/ml the milk sample added recovery; Every kind of each 4 of each concentration of sample are parallel, measure with three batches of kits, calculate the average recovery rate and the precision of sample.Experimental result sees the following form.
Table 1 accuracy and precision are measured ng/g (ml)
Can know from table, the average recovery rate scope that two concentration of Clenbuterol are all added in pig urine, pork liver, pork, feed, the milk sample between 83.4-99.7%, in batch, batch between all the coefficient of variation less than 15%.
3, specificity
Cross reacting rate is meant the ability that antibody combines with structure different antigens determinant.
Table 2 kit cross reacting rate
| Medicine name | Cross reacting rate |
| Clenbuterol | 100％ |
| Salbutamol | 3％ |
| Arubendol | Less than 1% |
| Fenoterol | Less than 1% |
| Mabuterol | Less than 1% |
| Cimaterol | Less than 1% |
| Bromine Boot sieve | Less than 1% |
| Special sieve of Zeeman | Less than 1% |
| Isoprel | Less than 1% |
4, correlativity
X＝CI-2008，Y＝RIA
Y＝1.23X-2.83
R＝0.9763
X is CI-2008 system measurement result, and Y is that the result is measured in the aspect
Claims (8)
1. magnetic granule chemoluminescence detection kit that detects Clenbuterol, the reagent that comprises has: luminous marker, fluorescein-labelled thing, standard items, quality-control product, separation agent.
2. kit according to claim 1 is characterized in that: described separation agent is the paramagnetism nano microsphere that is coated with goat-anti FITC monoclonal antibody.
3. kit according to claim 2 is characterized in that: described paramagnetism nano microsphere is that the inside is coated with Fe
3O
4Or γ-Fe
2O
3, the surface contains-OH ,-COOH or-NH
2The microballon of reactive group.
4. according to each described kit of claim 1-3, it is characterized in that: described luminous marker is the Clenbuterol haptens of different luminol derivant mark.
5. kit according to claim 4 is characterized in that: described different luminol luminous marker is ABEI, AHEI or ABEN.
6. according to each described kit of claim 1-3, it is characterized in that: described kit also comprises titer, quality-control product and concentrated washing lotion.
7. according to the described kit of one of claim 1-3, it is characterized in that: the Clenbuterol monoclonal antibody that described fluorescein-labelled thing is the FITC mark.
8. a method of utilizing each described kit of claim 1-7 to detect Clenbuterol comprises the following steps:
1) draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min;
2) add the paramagnetism nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min;
3) separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant;
4) above-mentioned cleaning step repeats 2-4 time;
5) 4) compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s; The content of Clenbuterol and RLU proportion relation in the sample can be through the concentration of RLU set calibration curve method calculating Clenbuterol.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104892764A (en) * | 2014-03-07 | 2015-09-09 | 北京维德维康生物技术有限公司 | Quantum dot immunofluorescence kit for detecting clenbuterol or derivatives thereof |
| CN105301254A (en) * | 2015-12-22 | 2016-02-03 | 北京勤邦生物技术有限公司 | Immunomagnetic beads for gathering purification of zearalenone and preparing method and application of immunomagnetic beads |
| CN106153607A (en) * | 2016-08-23 | 2016-11-23 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of clenobuterol hydrochloride chemoluminescence method quantitative determination reagent kit |
| CN107356745A (en) * | 2017-08-23 | 2017-11-17 | 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 Clenbuterol |
| CN108181463A (en) * | 2017-12-11 | 2018-06-19 | 华南农业大学 | A kind of beta-2-agonists haptens and artificial antibody and its preparation method and application |
| 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 (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888901A (en) * | 2006-04-21 | 2007-01-03 | 深圳市新产业生物医学工程有限公司 | Magnetic separating direct chemical illuminating reagent and testing method using the same reagent |
| CN101324578A (en) * | 2007-06-13 | 2008-12-17 | 清华大学 | Immune radiometric analysis reagent kit for detecting tumour sign object CA50 and use method thereof |
| CN101413951A (en) * | 2008-11-27 | 2009-04-22 | 上海交通大学 | Chemiluminescence immune detection reagent kit for detecting Clenbuterol |
-
2010
- 2010-12-15 CN CN201010588988XA patent/CN102539412A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888901A (en) * | 2006-04-21 | 2007-01-03 | 深圳市新产业生物医学工程有限公司 | Magnetic separating direct chemical illuminating reagent and testing method using the same reagent |
| CN101324578A (en) * | 2007-06-13 | 2008-12-17 | 清华大学 | Immune radiometric analysis reagent kit for detecting tumour sign object CA50 and use method thereof |
| CN101413951A (en) * | 2008-11-27 | 2009-04-22 | 上海交通大学 | Chemiluminescence immune detection reagent kit for detecting Clenbuterol |
Non-Patent Citations (2)
| Title |
|---|
| 孙武勇等: "醋酸甲羟孕酮完全抗原制备及ELISA检测", 《食品科学》 * |
| 高红军等: "化学发光免疫分析法检测血清NSE的室间比对结果分析", 《中国卫生检验杂志》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104892764A (en) * | 2014-03-07 | 2015-09-09 | 北京维德维康生物技术有限公司 | Quantum dot immunofluorescence kit for detecting clenbuterol or derivatives thereof |
| CN105301254A (en) * | 2015-12-22 | 2016-02-03 | 北京勤邦生物技术有限公司 | Immunomagnetic beads for gathering purification of zearalenone and preparing method and application of immunomagnetic beads |
| CN106153607A (en) * | 2016-08-23 | 2016-11-23 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of clenobuterol hydrochloride chemoluminescence method quantitative determination reagent kit |
| CN107356745A (en) * | 2017-08-23 | 2017-11-17 | 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 Clenbuterol |
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Application publication date: 20120704 |