CN101393210A - Chemiluminescence ELISA detection kit of terbutaline - Google Patents
Chemiluminescence ELISA detection kit of terbutaline Download PDFInfo
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- CN101393210A CN101393210A CNA2008101581406A CN200810158140A CN101393210A CN 101393210 A CN101393210 A CN 101393210A CN A2008101581406 A CNA2008101581406 A CN A2008101581406A CN 200810158140 A CN200810158140 A CN 200810158140A CN 101393210 A CN101393210 A CN 101393210A
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Abstract
The invention discloses a chemiluminescence enzyme immunoassay detection reagent kit for terbutaline, which comprises a kit body, an ELIAS plate arranged in the kit body and a reagent in the kit body. The reagent kit is characterized in that each hole of the ELIAS plate is enveloped with an envelope antigen, wherein the envelope antigen is produced by coupling the terbutaline and ovalbumin; and the reagent comprises a terbutaline series standard solution, an enzyme labeled goat anti-rabbit antibody, a terbutaline antibody, a luminescent solution, a washing solution, an envelope solution, and a confining solution. The chemiluminescence enzyme immunoassay detection reagent kit has the characteristics of high sensitivity, simplicity, convenience, quickness, and accuracy; compared with the prior colorimetric ELISA method, the sensitivity can be improved by one order of magnitude; and the reagent kit is expected to play an important role in residue detection of the terbutaline in animal food such as milk, animal tissue, and urine sample.
Description
Technical field
The present invention relates to a kind of enzyme-linked immunologic detecting kit, relate in particular to a kind of chemical luminescence ELISA detection kit of Terbutaline.
Background technology
Terbutaline belongs to beta-stimulants, and beta-stimulants is the phenolethanolamine analog derivative of a class chemosynthesis.The beta-stimulants mechanism of action and adrenaline, norepinephrine are identical, can influence nutriment in animal body the flow direction and redistribute, effectively promote musculature growth, reduce the trunk fat content, improve lean meat percentage and increase lean meat output, thereby once be extensive use of the America and Europe.The sportsman uses this medicine, can increase muscle and vital capacity, the convalescence after the shortening high pressure training.But then, beta-stimulants has similar adrenergic chemical constitution, and this compounds is generally higher as the consumption of growth accelerator, is generally 5-10 times of treatment consumption.The fact shows that this type of medicine is survivable, if before butchering, do not pass through the off-drug period of certain hour, then muscle and internal organs can residual higher concentration medicine.If the people has eaten the edible tissue that contains these left drugs, these medicines just can produce tangible physiological action with the β-receptors bind of cell surface, as palpitaition, headache, dizzy, feel sick, vomit, tremble, neuroticism, blood vessel dilatation, heart rate quickening etc., especially hypertension, heart disease, diabetes, hyperthyroid patient are endangered bigger, serious caused death.Therefore in order to guarantee consumer's safety, European and American countries is all forbidden its application on herding is produced at present.But have the raiser of some countries and regions in the consumption animal feeding, to add Terbutaline, cause having Terbutaline residual in the relevant animal food, therefore regularly to the residual necessary means that detects the monitoring that becomes many countries of Terbutaline.
At present, the method for detection Terbutaline mainly contains: microbial method, thin-layered chromatography (TLC), radioimmunology, high performance liquid chromatography (HPLC), look/matter coupling analytic approach (LC-MS), liquid/matter coupling analytic approach (LC-MS/MS).The defective of microbial method is: time-consuming and shortage specificity.The defective of thin-layered chromatography is: complicated operating process, and the time is long; Operating personnel need pass through professional training; The disturbing factor of impact analysis is more, as a result poor repeatability.Thin-layered chromatography, radioimmunology, the defective of high performance liquid chromatography, look/matter logotype analytic approach, liquid/these physico-chemical methods of matter coupling analytic approach is instrument and equipment costlinesses, the sample pre-treatments complexity, time-consuming, effort is difficult for popularizing the testing cost height, particularly radioimmunology also needs to be equipped with radioactive source, and certain risk is arranged.Given this, it is significant to set up a kind of method of effective, quick, simple, sensitive detection Terbutaline.
Summary of the invention
The chemical luminescence ELISA detection kit that the purpose of this invention is to provide a kind of Terbutaline.This kit has detection sensitivity height, applying flexible, characteristics easily.
The chemical luminescence ELISA detection kit of Terbutaline of the present invention comprises box body, is located at the ELISA Plate in the box body and is located at reagent in the box body; It is characterized in that each hole of described ELISA Plate is coated with envelope antigen, wherein envelope antigen is made by Terbutaline and ovalbumin coupling; Described reagent comprises: Terbutaline series standard solution, enzyme mark goat anti-rabbit antibody, Terbutaline antibody, luminous solution, wash solution, bag are by solution and lock solution.
In the chemical luminescence ELISA detection kit of above-mentioned Terbutaline:
The preferred 5 μ g/mL of described envelope antigen concentration.
Preferred 6.7KDa～the 6.8KDa of the molecular weight ranges of described ovalbumin.
Described Terbutaline series standard solution is respectively 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL.
Described enzyme mark goat anti-rabbit antibody is horseradish peroxidase-goat anti-rabbit igg stoste, and its working concentration is preferably 1:1000.
Described Terbutaline antibody is to be the polyclonal antibody that artificial immunogen immune animal that the bovine serum albumin coupling of 6.7KDa～6.8KDa is made makes by Terbutaline and molecular weight ranges, and its working concentration is preferably 1: 600.
Described luminescent solution is three (methylol) aminomethane solution (pH=8.8) and 3/10000 (volume ratio) H of 0.01M luminol and 0.001M p-cresol
2O
2Mixed liquor.Described luminol is a luminous substrate, and p-cresol is a luminescence enhancer.
Described wash solution is pH7.5, the 0.1mol/L phosphate buffer that contains volume fraction 0.05% Tween-20.
Described bag is the solution that contains 1.59g sodium carbonate and 2.53g sodium bicarbonate in every premium on currency by solution, and pH is 9.5.
Described lock solution is to contain 10g ovalbumin (OVA, ovalbumin also claim pure albumen of ovum gallinaceum or chicken ovalbumin, are made of the about 43Kd of molecular weight 386aa) in every liter of wash solution and add weight fraction 0.5%NaN
3Solution.
Kit maximum detection range of the present invention is 0.1ng/mL～10ng/mL.
The Terbutaline standard solution that relates in the kit of the present invention, enzyme mark goat anti-rabbit antibody solution, Terbutaline antibody-solutions, luminous solution and wash solution and prescription thereof are very big to the sensitivity influence that kit of the present invention detects; Wherein the principal ingredient of each solution and compound method thereof are:
1, Terbutaline standard solution: 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL.
2, enzyme mark goat anti-rabbit antibody solution: enzyme mark goat anti-rabbit antibody is horseradish peroxidase-goat anti-rabbit igg stoste, is mixed with 1: 1000 working concentration during use with wash solution.
3, Terbutaline antibody-solutions: Terbutaline antibody is the polyclonal antibody that makes with artificial immunizing antigen immune animal, gained Terbutaline antibody is diluted to the working concentration of 1:600 with wash solution.
4, luminous solution: three (methylol) aminomethane solution (pH=8.8) and 3/10000 (volume ratio) H of being 0.01M luminol and 0.001M p-cresol
2O
2Mixed liquor.
5, wash solution: the pH7.5, the 0.1mol/L phosphate buffer that refer to contain volume fraction 0.05% Tween-20.
6, wrap by solution: 1.59g sodium carbonate and 2.53g sodium bicarbonate are dissolved in the 1L water, and regulating pH is 9.5.
7, lock solution preparation: 10g OVA is dissolved in the 1L wash solution, adds weight ratio again and be 0.05% NaN
3
The preparation of microwell plate among the present invention:
Among the present invention the microwell plate of bag quilt adopt Terbutaline-OVA the bag of setting by solution in, with the concentration of setting, reaction overnight bag quilt in 4 ℃.
What the present invention adopted is that pH is sodium carbonate-sodium bicarbonate buffer solution of 9.5.Among the present invention in the microwell plate Terbutaline-OVA of the quilt that wraps under alkaline environment, can well be combined on the microwell plate frosting, can stand repeatedly to wash plate, the coating protein concentration of employing is 5 μ g/mL.
Bag can be with containing the lock solution sealing by good microwell plate, and the preferred OVA of inert protein in the confining liquid needs to add NaN
3Prevent to go bad.
The preparation of Terbutaline antibody-solutions and enzyme mark goat anti-rabbit antibody solution:
Terbutaline antibody-solutions among the present invention, enzyme mark goat anti-rabbit antibody solution concentration are the key factors of Terbutaline enzyme linked immunological test kit measurement range and sensitivity among decision the present invention.
It is 0.1～10ng/mL solution that the Terbutaline antibody-solutions that relates among the present invention can be mixed with concentration with wash solution; Or be diluted to the working concentration of 1:600 with wash solution.
The enzyme mark goat anti-rabbit antibody solution that relates among the present invention is 1:1000 with the concentration of wash solution preparation preferably.
(the standard lines scope can reach 0.1ng/mL～10ng/mL) and the sensitivity of becoming reconciled (0.1ng/mL) can to reach the good range of linearity according to the kit of above-mentioned Terbutaline antibody-solutions concentration and enzyme mark goat anti-rabbit antibody solution concentration preparation.
The preparation of luminous solution:
The present invention adopts horseradish peroxidase labeled substrate luminescent system, mainly is luminol-hydrogen peroxide system.
Described luminescent solution is three (methylol) aminomethane solution (pH=8.8) and 3/10000 (volume ratio) H of 0.01M luminol and 0.001M p-cresol
2O
2Mixed liquor.Described luminol is a luminous substrate, and p-cresol is a luminescence enhancer.
Principle of the present invention is that antibody-antigen reactive high degree of specificity and enzymatic high sensitivity are combined, and utilizes the chemiluminescence reaction of substrate for enzymatic activity to detect production concentration.
The luminescence-producing reaction formula that wherein relates to is:
Chemical luminescence ELISA detection kit of the present invention has highly sensitive, easy characteristics fast and accurately, and with traditional colorimetric ELISA method comparison, sensitivity can improve an order of magnitude; Be expected to play a significant role in the Terbutaline residue detection in animal food (as milk, animal tissue, urine sample).
Description of drawings
Fig. 1 is the uv-spectrogram of antigen of the present invention.
Fig. 2 is the inhibiting rate curve of Terbutaline antibody of the present invention.
Fig. 3 is a working curve of the present invention.
Embodiment
(1) immunogenic synthetic
Terbutaline and bovine serum albumin (BSA) are adopted 1, and 4-fourth diether method is carried out coupling and is obtained immunogene.Specifically may further comprise the steps:
A takes by weighing 50mg bovine serum albumin (BSA) and is dissolved in 3mL50mM carbonate (pH10.7) damping fluid, adds 1 of 13.8 μ L (72.9 μ mol) then in solution, 4-fourth diether (BDE), and room temperature reaction 20 hours gets colorless cleared solution A;
B takes by weighing 30mg (109.4 μ mol) Terbutaline and is dissolved in 1.5mL50mM carbonate (pH10.7) damping fluid, and solution A is fed nitrogen, subsequently Terbutaline solution is dropwise added in the A solution, and room temperature reaction 20 hours gets colorless cleared solution;
C transfers to reactant liquor in the semi-permeable diaphragm, 0-4 ℃ with phosphate buffered solution (PBS) (0.01M, pH7.4) dialysis is 3 days, changes one time dislysate therebetween in every 4-6 hour; Subsequently with high purity water dialysis 3 days, changed one time dislysate therebetween in every 4-6 hour; Dialysis finishes, and uses the freeze drier freeze-drying, and get the white solid powder and be immunogene (conjugate of Terbutaline and bovine serum albumin) ,-20 ℃ of preservations, standby.
(2) envelope antigen is synthetic
Adopt the p-aminobenzoic acid method to carry out coupling Terbutaline and ovalbumin (OVA) and obtain immunogene.Specifically may further comprise the steps:
A, take by weighing 10mg (73 μ mol) p-aminobenzoic acid (ABA) and dissolve in the 1.1mL0.2M hydrochloric acid, take by weighing the sodium nitrite (NaNO of 6mg (87 μ mol) then
2) be dissolved in the distilled water of 0.35mL, 0～4 ℃ of stirring is with sodium nitrite (NaNO
2) solution dropwise joins in the p-aminobenzoic acid solution, lucifuge reaction 1 hour, solution A;
B, take by weighing 20mg (84 μ mol) salbutamol and be dissolved in the ice-cold borate buffer solution of 10mL (0.05M) (pH8.5, the NaCl that contains 0.15M) in, 0～4 ℃ of stirring dropwise joins above-mentioned A solution 2mL in this solution, lucifuge reaction 2 hours obtains orange colour solution;
C, solution is added a spot of boric acid crystal transfer pH to 7.4, add 119.54mg (2.78 μ mol) ovalbumin (OVA) then, add 160mg (834 μ mol) water-soluble carbodiimide (EDC) simultaneously, 48mg (417 μ mol) N-hydroxy-succinamide (NHS), stirring at room 3 hours gets orange colour solution;
D, reactant liquor is transferred in the semi-permeable diaphragm, (pH7.4) dialysis is 3 days for PBS, 0.01M, changes one time dislysate therebetween in every 4-6 hour with phosphate buffered solution at 0～4 ℃; Subsequently with high purity water dialysis 3 days, changed one time dislysate therebetween in every 4-6 hour; Dialysis finishes, and uses the freeze drier freeze-drying, and get the faint yellow solid powder and be envelope antigen (conjugate of Terbutaline and ovalbumin) ,-20 ℃ of preservations, standby.
(3) Terbutaline Polyclonal Antibody Preparation
Adopt new zealand white rabbit as immune animal, conjugate with Terbutaline and bovine serum albumin is an immunogene, first immunisation dosage is 500 μ g/mL, when head exempts from immunogene is dissolved in into isopyknic physiological saline and Freund's complete adjuvant and makes emulsifying agent, nape portion multi-point injection, immunity is later on got the immunogene that dosage reduces by half and is dissolved in isopyknic physiological saline and incomplete Freund mixing and emulsifying, head exempts from and two exempted from 20 days at interval, once be total to immunity five times every two all immunity later on, do not add adjuvant for the last time.Last immunity is the heart blood sampling after 7 days, and the centrifugal antiserum that gets obtains the Terbutaline polyclonal antibody.
The foundation of embodiment 2, CL-ELISA detection method
(1) preferred (square formation) of antibody and envelope antigen concentration
Vertically with the serial dilution degree coated elisa plate of every kind of envelope antigen by 40.0 μ g/mL, 20.0 μ g/mL, 10.0 μ g/mL, 5.0 μ g/mL, 2.5 μ g/mL, 1.25 μ g/mL, 0.625 μ g/mL, 0.3125 μ g/mL, 100 μ L/ holes, 0-4 ℃ of placement spent the night, wash plate three times with cleansing solution, pat dry at every turn; 250 μ L/ hole lock solution sealings, room temperature was placed 3 hours, washed plate three times, patted dry at every turn; The antibody (1:100 to 1:1024000) that adds the 100 a series of dilutions in μ L/ hole, room temperature was placed 2 hours, washed plate three times, patted dry at every turn; Horseradish peroxidase-the goat anti-rabbit igg antibody that adds the 1:1000 in 100 μ L/ holes, room temperature was placed 1 hour, washed plate three times, patted dry at every turn; The luminescent solution that adds 100 μ L/ holes is measured luminous value.With the concentration of envelope antigen the envelope antigen concentration of obvious graded and antibody dilution being arranged with luminous value is that optium concentration is carried out specific assay.
(2) mensuration of antibody sensitivity
According to the optimization experiment of above-mentioned antagonist and envelope antigen concentration, the applicant selects and determines that antibody concentration is 1: 600, and envelope antigen concentration is the mensuration that 5 μ g/mL carry out the sensitivity of antibody:
A, bag quilt: by solution the envelope antigen of Terbutaline is made into the solution of 5 μ g/mL with the carbonate bag of 0.05M pH9.6, adds 100 μ L in the reacting hole of each polystyrene board, 4 ℃ are spent the night.
Discard solution in the hole next day, washes 3 times with lavation buffer solution, and 300 μ L/ holes each 5 minutes, pat dry.(this step is called for short washing, down together).
B, sealing: with the above-mentioned ELISA Plate of having wrapped quilt of lock solution sealing, 250 μ L/ holes, room temperature was incubated 2-4 hour, then washing.
C, application of sample: room temperature 2-4 hour, wash in the above-mentioned reacting hole that has sealed then in the Terbutaline antibody-solutions 50 μ L/ holes that add dilution Terbutaline antibody antibody (1: 600) 50 μ L/ holes and variable concentrations.
D, add enzyme labelled antibody: in each reacting hole, add antibody (1:1000) the 100 μ L/ holes of fresh dilution horseradish peroxidase-goat anti-rabbit igg, 1.5 hours, washing.
E, luminous: in each reacting hole, add the luminous solution 100 μ L/ holes of interim preparation, detect with chemical illumination immunity analysis instrument immediately.
F, testing result are calculated with inhibiting rate:
% inhibiting rate=%B/Bo, B are the luminous value of the medicine of variable concentrations as the rival, and Bo is the luminous value of not dosing.
Drug concentrations is the sensitivity of this antibody when calculating 50% inhibiting rate.
The chemiluminescence enzyme linked immunoassay reagent kit of embodiment 3, detection Terbutaline
(1) composition of the chemiluminescence enzyme linked immunoassay reagent kit of detection Terbutaline
A, be coated with the solid phase carrier (ELISA Plate) of envelope antigen (conjugate of Terbutaline and carrier protein);
B, Terbutaline standard solution: 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL.
C, enzyme mark goat anti-rabbit antibody solution: enzyme mark goat anti-rabbit antibody is horseradish peroxidase-goat anti-rabbit igg stoste, packs into, is mixed with the working concentration of 1:1000 during use with wash solution.
D, Terbutaline antibody-solutions: prepare the polyclonal antibody of gained with artificial immunizing antigen immune animal, gained Terbutaline antibody is diluted to the 1:600 working concentration with wash solution.
E, luminous solution: three (methylol) aminomethane solution of the 0.0001M p-cresol of use pH8.8 is mixed with the luminol solution of 0.01M, again with H
2O
2Volume ratio according to 3:10000 is mixed.
F, wash solution: the pH7.5, the 0.1mol/L phosphate buffer that contain volume fraction 0.05% Tween-20.
G, bag are by solution: 1.59g sodium carbonate and 2.53g sodium bicarbonate are dissolved in the 1L water, regulate pH9.5.
The preparation of H, lock solution: 10g OVA is dissolved in the 1L wash solution, adds weight ratio again and be 0.05% NaN
3
(2) preparation of ELISA Plate
With coating buffer envelope antigen is diluted to 5 μ g/mL, every hole adds 100 μ L, and 4 ℃ are spent the night, and coating buffer inclines, every hole adds 250 μ L cleansing solutions washing 3 times, pat dry, every then hole adds confining liquid 250 μ L, hatches 1h for 37 ℃, liquid in the hole inclines, cleansing solution washing 3 times pats dry, and preserves with masking foil vacuum seal.
The application of the chemiluminescence enzyme linked immunoassay reagent kit of embodiment 4, detection Terbutaline
(1) preparation of reagent
A, sample diluting liquid: the concentrated phosphoric acid salt buffer solution that provides in the kit is used with behind 10 times of the distilled water dilutings.
B, wash solution: the concentrated cleaning solution that provides in the kit is used with behind 10 times of the distilled water dilutings.
C, luminous solution: three (methylol) aminomethane solution (pH8.8)+3/10000 (volume ratio) H of 0.01M luminol+0.001M p-cresol
2O
2
(2) sample pre-treatments
A, milk at first with the plain chocolate sample bought under 4 ℃, 10000 rev/mins conditions, centrifugal 15 minutes, remove fat deposit; The milk of degreasing is diluted to 1:10 with cleansing solution, obtains testing sample.
At 4 ℃, centrifugal 15 minutes of 10000g removes precipitation with the pig urine samples for B, pig urine sample, and remaining pig urine is diluted to 1:10 with cleansing solution, obtains testing sample.
The mixed in hydrochloric acid of C, animal tissue's sample thief and 4mL0.1M, and put together and extract 20min with the excusing from death ripple, the centrifugal 15min of 10000g gets supernatant and transfers pH to 9.5 ± 0.5 with 10M NaOH then, vortex vibration 5min, the centrifugal 15min of 10000g then.Get supernatant and add 5mL isobutyl alcohol vibration 2min, static 15min under the potpourri room temperature, the centrifugal 10min of 3000g then, tell organic phase, water is used isobutyl alcohol (each 10mL) extracting twice again, and the organic phase of three extractions combines 50-60 ℃ of water-bath evaporated under reduced pressure, residue dissolves the solution that is made into 1:10 again with cleansing solution, obtains testing sample.
(3) detect step
A, application of sample: add Terbutaline series standard concentration solution or the molten 50 μ L of sample in the ELISA Plate micropore, add Terbutaline antibody-solutions 50 μ L then, room temperature (25 ℃) constant temperature is hatched 2.5h;
B, washing: the middle liquid that portals that inclines, every hole adds wash solution 250 μ L, washs 3 times, pats dry;
C, enzyme-added mark goat anti-rabbit antibody solution: every hole adds enzyme mark goat anti-rabbit antibody solution 100 μ L, and room temperature constant temperature is hatched 1h;
D, washing: the middle liquid that portals that inclines, every hole adds wash solution 250 μ L, washs 3 times, pats dry;
E, add luminous solution: every hole adds luminous solution 100 μ L;
F, detection: the luminous intensity of measuring every hole with chemical illumination immunity analysis instrument.
(4) result judges
The mean value of standard items that obtained and sample luminous value multiply by 100 again divided by the luminous value of first standard (0 standard), with the inhibiting rate is ordinate, the logarithm of Terbutaline concentration is that horizontal ordinate is made typical curve, and the concentration of each sample can be read from typical curve.
% inhibiting rate=% standard items luminous value (or sample)/0 standard items luminous value.
Get 0.1,0.5,1,5, the Ciprofloxacin standard specimen of 10ppb adds in the milk sample, detects the Ciprofloxacin recovery.The interassay coefficient of variation of each concentration all calculates with 5 repeating datas of different 5 days, and the variation within batch coefficient calculates with 5 repeating datas on the same day.
Carry out the quantitative Analysis of the recovery according to the linear equation of the typical curve of formulating.
The results are shown in following table:
From the said determination result, the coefficient of variation is lower than 19.9%, and the recovery is between 83.8-113%.Show that this kit has good repeatability and accuracy.
Claims (10)
1, a kind of chemical luminescence ELISA detection kit of Terbutaline comprises box body, is located at the ELISA Plate in the box body and is located at reagent in the box body; It is characterized in that each hole of described ELISA Plate is coated with envelope antigen, wherein envelope antigen is made by Terbutaline and ovalbumin coupling; Described reagent comprises: Terbutaline series standard solution, enzyme mark goat anti-rabbit antibody, Terbutaline antibody, luminous solution, wash solution, bag are by solution and lock solution.
2, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described envelope antigen concentration is 5 μ g/mL.
3, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: the molecular weight ranges of described ovalbumin is 6.7KDa～6.8KDa.
4, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described Terbutaline series standard solution is respectively 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL.
5, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described enzyme mark goat anti-rabbit antibody is horseradish peroxidase-goat anti-rabbit igg stoste, and its working concentration is 1:1000.
6, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described Terbutaline antibody is to be the polyclonal antibody that artificial immunogen immune animal that the bovine serum albumin coupling of 6.7KDa～6.8KDa is made makes by Terbutaline and molecular weight ranges, and its working concentration is 1:600.
7, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described luminescent solution is that three (methylol) the aminomethane solution and the volume ratio of 0.01M luminol and 0.001M p-cresol is 3/10000H
2O
2Mixed liquor.
8, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described wash solution is pH7.5, the 0.1mol/L phosphate buffer that contains volume fraction 0.05% Tween-20.
9, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described bag is the solution that contains 1.59g sodium carbonate and 2.53g sodium bicarbonate in every premium on currency by solution, and pH is 9.5.
10, according to the chemical luminescence ELISA detection kit of the described Terbutaline of claim 1, it is characterized in that: described lock solution is to contain the 10g ovalbumin in every liter of wash solution and add weight fraction 0.5% NaN
3Solution.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101581406A CN101393210A (en) | 2008-10-24 | 2008-10-24 | Chemiluminescence ELISA detection kit of terbutaline |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101581406A CN101393210A (en) | 2008-10-24 | 2008-10-24 | Chemiluminescence ELISA detection kit of terbutaline |
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| CN101393210A true CN101393210A (en) | 2009-03-25 |
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ID=40493605
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101915831A (en) * | 2010-08-25 | 2010-12-15 | 南开大学 | Enzyme-linked immunoassay kit for acid orange II |
| CN103698532A (en) * | 2013-12-02 | 2014-04-02 | 镇江出入境检验检疫局检验检疫综合技术中心 | Storage and treatment method for immunoglobulin (IgG)-coated enzyme designation strip |
| CN106770214A (en) * | 2016-12-07 | 2017-05-31 | 无锡艾科瑞思产品设计与研究有限公司 | Terbutaline detection method and detection box in a kind of poultry |
| CN107356746A (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 Terbutaline |
| CN108614114A (en) * | 2016-12-12 | 2018-10-02 | 丹阳亿太生物科技发展有限公司 | A kind of chemiluminescence enzyme-linked immunoassay of detection sarafloxacin |
-
2008
- 2008-10-24 CN CNA2008101581406A patent/CN101393210A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101915831A (en) * | 2010-08-25 | 2010-12-15 | 南开大学 | Enzyme-linked immunoassay kit for acid orange II |
| CN103698532A (en) * | 2013-12-02 | 2014-04-02 | 镇江出入境检验检疫局检验检疫综合技术中心 | Storage and treatment method for immunoglobulin (IgG)-coated enzyme designation strip |
| CN106770214A (en) * | 2016-12-07 | 2017-05-31 | 无锡艾科瑞思产品设计与研究有限公司 | Terbutaline detection method and detection box in a kind of poultry |
| CN108614114A (en) * | 2016-12-12 | 2018-10-02 | 丹阳亿太生物科技发展有限公司 | A kind of chemiluminescence enzyme-linked immunoassay of detection sarafloxacin |
| CN107356746A (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 Terbutaline |
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