CN1920573A - Method for detecting monoconal antibody mediated furacilinum residue - Google Patents
Method for detecting monoconal antibody mediated furacilinum residue Download PDFInfo
- Publication number
- CN1920573A CN1920573A CN 200610041481 CN200610041481A CN1920573A CN 1920573 A CN1920573 A CN 1920573A CN 200610041481 CN200610041481 CN 200610041481 CN 200610041481 A CN200610041481 A CN 200610041481A CN 1920573 A CN1920573 A CN 1920573A
- Authority
- CN
- China
- Prior art keywords
- antibody
- mediated
- furacilinum
- residue
- monoconal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for detecting the left amifur of single colon antibody medium, wherein it uses amifur single colon antibody as the medium conductivity to build amifur left ELISA, to produce the detecting agent box and detecting testing paper, to treat the amifur left amount with ELISA quick method. The invention, compared with liquid medium couple spectrum device, has low pretreatment demand, simple process, short detecting time, high sensitivity and low cost.
Description
Technical field:
The invention belongs to the medicament residue detection range.
Background technology:
The itrofurans medicine is the broad-spectrum antibiotic of synthetic, it has extraordinary antibacterial action and pharmacological kinetics characteristic, except being applied in animal doctor's medical treatment as a kind of microbiotic, also Ceng Zuowei pig, fowl and the somatotrophic adjuvant of aquatic products are widely used for it.But finding in the laboratory study process for a long time, itrofurans medicine and metabolic product thereof all can make animal used as test that canceration and gene mutation take place, Given this, European Union forbids using furaltadone (Furaltadone), nitrofurazone (Nitrofurazone), Nitrofurantoin (Nitrofurantoin) and furazolidone four kinds of medicines such as (Furazolidone) respectively at 1993 (European Union's decree 2901/93) and nineteen ninety-five (European Union's decree 1442/95) legislation in the animal derived food process of manufacture.Thereafter, the U.S., Canada, Australia are new all completely forbids use itrofurans medicine with state such as Japan.In March, 2002, China also classified the itrofurans medicine as the forbidding veterinary drug.Studies have shown that, itrofurans medicine some hrs in animal body will resolve into nitrofuran and various metabolic product rapidly, these metabolic products form comparatively stable compound with organizing internal protein, this compound can retain the long period in tissue, so should carry out check and analysis to its metabolic product when analyzing this type of medicament residue, states such as European Union are exactly to be that means reach and detect the residual purpose of nitrofuran to detect metabolic product.The metabolic product of four kinds of itrofurans medicines corresponds to respectively: furaltadone-3-amino-5-morpholinyl-2-oxygen oxazolone (AMOZ); Nitrofurazone-semicarbazides (SEM); Nitrofurantoin-1-amino is chewed pyridine (AHD); Furazolidone-3-amino-2-oxygen oxazolone (AOZ).
Since China's entry into the World Trade Organization more than a year, developed country such as American-European-Japanese utilizes the self-technique advantage that China's export food and agricultural byproducts constantly are provided with new technology barriers measure, to reach the outlet of restriction China's food and agricultural byproducts, the trade dispute of relevant this respect is comed one after another, European Union has just begun a FoodBRAND plan (QLK1-1999-00142) that reaches 42 months at the beginning of 2000, its fundamental purpose is that requirement European Union approves that respectively the laboratory to itrofurans method for detecting residue in the food (comprising ELISA and the LC-MS-MS method) research of tackling key problems, thinks that it is provided with technology barriers technical support is provided.At present, European Union must not stipulate that to detect four kinds of metabolic products of itrofurans medicine in food residual, and Given this, European Union has developed the analytical approach that the LC-MS chromatograph detects the itrofurans metabolic product.And the original detection method of China is only at the former medicine of itrofurans, and detection method is very backward, can not satisfy the demand of outlet certainly.
Summary of the invention:
First purpose of the present invention is to provide a kind of monoconal antibody mediated furacilinum residue method for quick for people.
The present invention serves as that monoconal antibody mediated furacilinum residue ELISA method for quick is set up in mediation with anti-Nitrofurantoin monoclonal antibody.
The method for quick of monoconal antibody mediated furacilinum residue may further comprise the steps:
1) will reduce good nitrofurazone and ovalbumin diazotising method coupling synthetic antigen SEM-OVA;
2) synthetic antigen SEM-OVA bag is arrived ELISA Plate;
3) with the full hole sealing of 5% skimmed milk power;
4) phosphate buffer washing pats dry;
5) join the good bag of washing after will resisting nitrofurazone monoclonal antibody and SEM titer outside plate, to act on by on the plate;
6) phosphate buffer washing pats dry;
7) add enzyme labelled antibody, hatch;
8) phosphate buffer washing pats dry;
9) add substrate solution;
10) add stop buffer, cessation reaction;
11) measure OD
490nmValue.
The present invention utilizes the immunology principle preparation at nitrofurazone and the special monoclonal antibody of metabolic product semicarbazides (SEM) thereof, set up enzyme linked immunosorbent assay (ELISA) to detect nitrofurazone and metabolic product semicarbazides (SEM) thereof, this method is with respect to LC-MS chromatograph analytic approach, its pre-treatment to sample requires lower, analytic process is very simple, detection time is shorter, and detection sensitivity is also higher, use cost is low, for a kind of efficient, easy approach that provides is provided Amprolium hydrochloride medicament residue in China's animal products foreign trade.
Second purpose of the present invention is that the monoconal antibody mediated furacilinum residue detection method for said monoclonal antibody mediation provides the application approach that is convenient to operation.
A kind of is to be used to make monoconal antibody mediated furacilinum residue ELISA quick detection kit.
Monoconal antibody mediated furacilinum residue ELISA quick detection kit comprises:
1) is coated with the polystyrene ELISA Plate of synthetic antigen SEM-OVA;
2) concentration and dilution liquid;
3) concentrate washing lotion;
4) anti-nitrofurazone monoclonal antibody;
5) enzyme labelled antibody;
6) substrate buffer solution;
7) substrate;
8) stop buffer.
Another kind is to be used to make monoconal antibody mediated furacilinum residue ELISA quick detection test paper bar.
The method for making of monoconal antibody mediated furacilinum residue ELISA quick detection test paper bar is:
1) colloid gold label antibody: the monoclonal antibody on the collaurum behind the mark purifying;
2) metal spraying: the antibody of colloid gold label is sprayed onto on the glass fibre, makes the collaurum pad;
3) make nitrocellulose filter: T line in the test strips and C line are sprayed onto on the nitrocellulose filter;
4) with sample pad, collaurum pad, nitrocellulose filter, thieving paper assembling, slitting then.
Kit or test strips are respectively two kinds of application forms of the monoconal antibody mediated furacilinum residue detection method of mediated monoclonal antibody, can be people's fast detecting and provide convenience.
Embodiment:
(1) antigen is synthetic
At first, reduction nitrofurazone:, make the furacilin solution of reduction with the nitro in amino substituted furan XiLin.
The synthetic diazotising method that adopts of antigen is carried out coupling.Get the 23mg potassium nitrite and be dissolved in the 1ml distilled water, with ice-water bath temperature adjustment degree to 0-5 ℃; The furacilin solution that reduction is good, 37 ℃ of water-bath effects 20 minutes, supernatant is cooled to about 4 ℃, transfers pH value 1-2, slowly adds potassium nitrite solution again and constantly stirs, and leaves standstill reaction 30min; The bovine serum albumin(BSA) of 100mg is dissolved in the phosphate buffer of 4ml, is cooled to about 4 ℃, the diazotising drug slow is added and constantly stirring, be stablizing of PH in the maintenance adition process, add 0.2mol/L NaOH simultaneously, transfer pH value to 7.4,4 ℃ of reaction overnight; Product 72-96h after 4 ℃ of PBS dialysis coupling then, every 6h changes liquid once, can not detect any small-molecule substance in dislysate; Product after the coupling is with 0.45 μ m membrane filtration, and-20 ℃ of packing are preserved.
(2) animal immune
With the female mouse of synthetic immunizing antigen lumbar injection immunity 6 ~ 8 week Balb/c in age.Immune programme for children is as follows: head exempts from and two exempts from by antigen 50 a μ g/ dosage, mix (0.3ml altogether) immunity respectively with Freund's complete adjuvant and incomplete Freund equal-volume, be 10 days immune interval, afterwards with 100 μ g/ dosage booster immunization only, once altogether immunity 4 times in 10 days, supplementary immunization is once again to merge preceding 3 days.
(3) foundation of hybridoma screening technique
The mouse orbit blood sampling behind the separation of serum, is done serum 1: 50 and dilution in 1: 100.Coating antigen SEM-OVA diluted by 1: 50,1: 100,1: 200,1: 400,1: 800,1: 1000, afterwards doubling dilution to 1: 64000.Set up the noncompetitive indirect ELISA method with the square formation method.The ELISA response procedures is as follows: 1. SEM-OVA is cushioned liquid with bag and is diluted to debita spissitudo, every hole 100ul, and 4 ℃ of bags are spent the night, with PBST washing 3 times, each 3min.2. with the full hole sealing of 5% skimmed milk, hatch 2h for 37 ℃, wash the same.3. positive serum dilution back order is added, every hole 100ul, each dilution positive serum is established negative control simultaneously, hatches 1h for 37 ℃, washs the same.4. with dilution ELIAS secondary antibody is diluted to suitable working concentration, every hole adds 100ul, hatches 1h for 37 ℃, washs the same.5. every hole adds freshly prepared substrate and uses liquid 100ul, hatches 30min for 37 ℃.6. every hole adds 2N H
2SO
4Stop buffer 50ul.7. measure the OD value,,, be judged to the positive to measure hole OD value 2.1 times more than or equal to negative control hole with the blank well zeroing.
(4) Fusion of Cells and screening
The splenocyte and the SP2/0 Fusion of Cells that is in exponential phase of mouse got in last immunity back the 3rd day, sterile working.The PEG 4000 that adds 1ml 50% in 45～60sec adds the DMEM stopped reaction of a large amount of serum-frees immediately, leave standstill centrifugal after, resuspended with the HAT nutrient solution, add an amount of feeder cells, cover plant is in 5 96 porocyte culture plates, in 37 ℃, 5%CO behind the mixing
2Cultivate in the incubator.When treating 1/10 or culture supernatant flavescence at the bottom of cell grows to the hole, with the ELISA method screening of having set up.The rapid enlarged culture of positive cell also adopts limiting dilution assay to carry out subclone, and selects good stable person to set up cell line.Obtain the hybridoma cell strain of the anti-SEM antibody of energy stably excreting, code name is SEM-1F8E9.
Hybridoma cell strain SEM-1F8E9 has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, preserving number: CGMCC No:1790 on August 30th, 2006.
(5) ascites of monoclonal antibody (McAb) preparation
Get 10 age in week BALB/C mice, lumbar injection sterilization paraffin oil 0.3ml/ only, 5～7 days pneumoretroperitoneum injection positive hybridoma cells 10
6Individual/as only, to treat that its belly back of obviously expanding extracts ascites, the centrifuging and taking supernatant is frozen and survey it and tire.
(6) monoclonal antibody titration
Test articles for use and reagent: elisa plate (envelope antigen was by 1: 40 dilution bag quilt)
One is anti-: 1. ascites was carried out 1: 500, and 1: 1000,1: 1500,1: 2000,1: 2500,1: 3000,1: 3500,1: 4000,1: 4500, dilution in 1: 5000
2. cell conditioned medium is done following doubling dilution: stoste, and 1: 2,1: 4 ..., 1: 2048
Two is anti-: sheep anti-mouse igg
Positive criterion: P 〉=2.1N
Testing result shows: it is 1: 20000 that ascites is tired, and it is 1: 256 that cell conditioned medium is tired
(7) determining of monoclonal antibody and envelope antigen ideal concentration:
With the square formation titration measuring, select OD at last
490nmValue equals envelope antigen about 1.0 and the extension rate of monoclonal antibody is an ideal operation concentration.After measured, the best package amount of antigen is the 0.75ug/ hole, and the monoclonal antibody optimum diluting multiple is 1: 5000.
(8) composition of kit and composition thereof preparation
According to serving as that mediation is set up monoconal antibody mediated furacilinum residue ELISA method for quick and carried out the assembling of kit with anti-nitrofurazone monoclonal antibody.
Adopt the diazotising method to carry out coupling synthetic antigen SEM-OVA:
Reduction nitrofurazone:, make the furacilin solution of reduction with the nitro in amino substituted furan XiLin.
Get the 23mg potassium nitrite and be dissolved in the 1ml distilled water, with ice-water bath temperature adjustment degree to 0～5 ℃; The furacilin solution that reduction is good places 37 ℃ of water-bath effects 20 minutes, and supernatant is cooled to about 4 ℃, transfers pH value 1-2, slowly adds potassium nitrite solution and continuous the stirring again, leaves standstill reaction 30min; The oralbumin (OVA) of 100mg is dissolved in the phosphate buffer of 4ml, is cooled to about 4 ℃, the diazotising drug slow is added and constantly stirring, be stablizing of PH in the maintenance adition process, add 0.2mol/L NaOH simultaneously, transfer pH value to 7.4,4 ℃ of reaction overnight; Product 72-96h after 4 ℃ of PBS dialysis coupling then, every 6h changes liquid once, can not detect any small-molecule substance in dislysate; Product after the coupling, promptly synthetic antigen SEM-OVA is with 0.45 μ m membrane filtration, and-20 ℃ of packing are preserved.
Add (bag quilt) 96 hole ELISA Plate, assembling SEM-Kit with synthetic antigen SEM-OVA.It mainly consists of: 8 * 12 hole ELISA Plate; 10 * concentration and dilution liquid (0.1M PBS-T, PH7.2 contains 0.05%Tween-80, the A bottle); 10 * concentrated cleaning solution (0.1M PBS-T, PH7.2 contains 0.05%Tween-80, the B bottle); Monoclonal antibody (anti-SEM monoclonal antibody, C dactylethrae); HRP mark rabbit anti-mouse igg (full molecule) (D dactylethrae); Substrate solution (mixed liquor of the disodium phosphate soln of the citric acid solution of 4.86mL 0.1M and 5.14mL 0.2M, E bottle); Hydrogen peroxide (30% H
2O
2, the G bottle); O-phenylenediamine (OPD tablet, F dactylethrae); Stop buffer (2NH
2SO
4, the H bottle); Titer (the SEM standard items of dissolved in distilled water dilution, N dactylethrae); The shrouding film; Operation instructions.
The preparation of each component in the kit:
1. be coated with the polystyrene ELISA Plate of synthetic antigen SEM-OVA: the carbonate solution with 22mM is done 1: 5000 times of dilution with antigen, and bag is by 96 hole polystyrene ELISA Plate, and every hole 100 μ L put 4 ℃ of refrigerator overnight; With washing lotion (PBST) washing 3 times, with 37 ℃ of wet box sealings of confining liquid (5% skimmed milk power) 60 minutes, washing lotion was washed 3 times, is reaction plate after the drying.
2. concentration and dilution liquid (10 *): 0.1M pH7.2 contains the phosphate buffer (PBST) of 0.5%Tween-80.Method preparation routinely, 4 ℃ of packing are preserved.
3. concentrate washing lotion (10 *): 0.1M pH7.2 contains the phosphate buffer (PBST) of 0.5%Tween-80.Method preparation routinely, 4 ℃ of packing are preserved.
4. antibody: by this chamber preparation, working concentration is 1: 2000 ,-70 ℃ of preservations.
5. enzyme labelled antibody: available from Sigma company.Working concentration is 1: 30000 ,-20 ℃ of preservations.
6. substrate buffer solution: sodium hydrogen phosphate 14.2g, citric acid 10.5g adds deionized water and is settled to 500mL, 4 ℃ of preservations of packing.
7. substrate solution: in substrate buffer solution, add 0.15% 30% H
2O
2
8. stop buffer: 0.2M sulfuric acid solution.
9. substrate tablet: o-phenylenediamine tablet.The substrate tablet 4 ℃ keep in Dark Place should stablize more than 1 year, nondiscolouring, put into substrate buffer solution should 2-3 minute the dissolving.
(9) composition of test strips and composition thereof preparation
According to serving as that the assembling of test strips is carried out in the foundation that monoconal antibody mediated furacilinum residue ELISA method for quick is set up in mediation with anti-nitrofurazone monoclonal antibody.
1. colloid gold label antibody: will carry out according to a conventional method behind the monoclonal antibody purifying.Concrete grammar:
A, get 522nm colloidal gold solution 20ml and return to room temperature, transfer to PH8.2 with 1% sal tartari;
B, get the centrifugal 5min of anti-nitrofurazone monoclonal antibody 12000rpm, add the ratio of the anti-nitrofurazone monoclonal antibody of 1 microgram in every milliliter of collaurum, getting total amount is the anti-nitrofurazone monoclonal antibody of 20 micrograms, is diluted to the 1ml volume with PB;
C, the anti-nitrofurazone monoclonal antibody after will diluting dropwise join in the colloidal gold solution under the magnetic agitation state lentamente, stir 30min under the room temperature;
D, dropwise to add 1ml 10%BSA be 0.5% to final concentration, magnetic agitation 30min;
E, put in 4 ℃ of refrigerators 2 hours, the centrifugal min of 8800rpm then, careful as far as possible the suction abandoned supernatant, with the phosphate buffer suspension collecting precipitation thing that contains 0.5%BSA and 0.2%PEG20000, to last volume be 1ml, 4 ℃ of refrigerators are preserved standby.
2. metal spraying: the antibody of colloid gold label is sprayed onto on the glass fibre, makes the collaurum pad;
3. spray film: T line in the test strips and C line are sprayed onto on the nitrocellulose filter;
4. sample pad, collaurum pad, nitrocellulose filter, thieving paper are assembled according to a conventional method, then slitting.
(10) sensitivity experiment:
The SEM titer with ultrapure water dissolving, gradient dilution, is added in the elisa plate simultaneously with isopyknic monoclonal antibody.
1. envelope antigen is become finite concentration with diluted, 100 μ l/ holes, 4 ℃ are spent the night;
2. phosphate buffer (PBST) washing is 1 time;
3. with the full hole sealing of 5% skimmed milk power, and issuable adherent bubble in the evacuation aperture as far as possible, 37 ℃ of water-bath effect 1h or 4 ℃ spend the night.
4.PBST wash 3 times, each 3 ~ 5 minutes, wash the back and on thieving paper, pat dry;
With the SEM standard items with add in the entering plate 37 ℃ of water-bath effect 1h after the monoclonal antibody equal-volume mixes;
6.PBST wash 3 times, each 3 ~ 5 minutes, wash the back and on thieving paper, pat dry;
7. add HRP mark sheep anti-mouse igg (1: 5000, in PBS), 100 μ l/ holes, 37 ℃ of water-bath effect 1h;
8.PBST wash 4 times, each 3 ~ 5 minutes, wash the back and on thieving paper, pat dry;
9. o-phenylenediamine (OPD) substrate solution that adds fresh configuration, 100 μ l/ holes, 37 ℃ of water-bath effect 30min;
10. add the reaction of 2M concentrated sulphuric acid color development stopping;
11. measure OD with the enzyme linked immunological reading apparatus
490nmValue.
Experimental result shows that this kit sensitivity 10 μ g/kg are following and more stable.
(11) clinical sample detects
Gather clinical sample, detect with kit.
1. the processing of test sample
Gather tissues such as liver, kidney, muscle, yolk, beef fat, add by a certain percentage that water grinds, centrifugal treating.
2. assay method
The method of operating of A, monoconal antibody mediated furacilinum residue quick detection kit and interpretation of result:
Get B bottle liquid (concentrated cleaning solution) before using and be cleansing solution by 1: 10 usefulness distilled water diluting to working concentration; C pipe liquid 100 μ L are stand-by with the dilution in 1: 100 of A liquid; D pipe liquid 100 μ L are stand-by with the dilution in 1: 100 of A liquid; It is stand-by that N pipe liquid is made serial gradient dilution with A liquid.The according to the form below form, elder generation is 37 ℃ of water-bath effect 1hr outside plate with sample and monoclonal antibody, and integral body moves on on the elisa plate again, 37 ℃ of water-bath 45min (A1, A2 hole only add dilution 100 μ L, do not add monoclonal antibody).Discard liquid in the hole, every hole adds cleansing solution to expiring the hole, discards cleansing solution, pats dry repeated washing 5 times on thieving paper.In every hole, add the good D liquid 100 μ L of dilution after the washing immediately, put 37 ℃ of water-bath 1hr.Discard liquid in the hole, every hole adds cleansing solution to expiring the hole, places 2min, discards cleansing solution, pats dry repeated washing 5 times on thieving paper.Tablet in the F pipe (2) is dissolved in 10mL substrate buffer solution (E liquid), and after the dissolving, every hole adds 100 μ L behind the adding 30ul G liquid mixing, 37 ℃ of lucifuges colour developing 30min fully.Colour developing finishes, and every hole adds the H liquid of 50 μ L; With the A1 hole is the zeroing hole, with the OD490 value of microplate reader working sample.
Criterion as a result:
Add behind the H liquid each hole color and become orange redly by yellow, be determined at the absorbance value (OD value) in every hole, 490nm wavelength place, calculate the average OD value of every part of test sample hole, standard items respectively, utilize following formula to obtain OD ratio with microplate reader.
If OD ratio less than 15%, is judged to be "-"; OD ratio is judged to be " ± " at 16-20%; OD ratio is judged to be "+" between 21-40%; OD ratio is judged to be " ++ " between 41-60%; OD ratio is greater than 60%, be judged to be " +++".
If need, concentration that can each standard items hole is horizontal ordinate, is ordinate with separately light absorption value, makes typical curve.Obtain regression equation, bring the light absorption value of each sample into equation and can obtain residual quantity.
B, monoconal antibody mediated furacilinum residue collaurum fast diagnose test paper bar are used and are judged:
Sample preparation liquid and test strips sample pad are fully soaked into, and sample liquid spreads to the direction of film, if T line and C line occur simultaneously, sample then to be checked is judged to feminine gender, if the T line only occurs, sample then to be checked is judged to the positive.
Claims (6)
1, the monoconal antibody mediated furacilinum residue detection method of mediated monoclonal antibody is characterized in that with anti-nitrofurazone monoclonal antibody serving as that monoconal antibody mediated furacilinum residue ELISA method for quick is set up in mediation.
2, according to the monoconal antibody mediated furacilinum residue detection method of the described mediated monoclonal antibody of claim 1, it is characterized in that may further comprise the steps:
1) will reduce good nitrofurazone and ovalbumin diazotising method coupling synthetic antigen SEM-OVA;
2) synthetic antigen SEM-OVA bag is arrived ELISA Plate;
3) with the full hole sealing of 5% skimmed milk power;
4) phosphate buffer washing pats dry;
5) join the good bag of washing after will resisting nitrofurazone monoclonal antibody and SEM titer outside plate, to act on by on the plate;
6) phosphate buffer washing pats dry;
7) add enzyme labelled antibody, hatch;
8) phosphate buffer washing pats dry;
9) add substrate solution;
10) add stop buffer, cessation reaction;
11) measure OD
490nmValue.
3, the application of the monoconal antibody mediated furacilinum residue detection method of mediated monoclonal antibody according to claim 1 is characterized in that being used to make monoconal antibody mediated furacilinum residue ELISA quick detection kit.
4, according to the application of the monoconal antibody mediated furacilinum residue detection method of the described mediated monoclonal antibody of claim 3, it is characterized in that monoconal antibody mediated furacilinum residue ELISA quick detection kit comprises:
1) is coated with the polystyrene ELISA Plate of synthetic antigen SEM-OVA;
2) concentration and dilution liquid;
3) concentrate washing lotion;
4) anti-nitrofurazone monoclonal antibody;
5) enzyme labelled antibody;
6) substrate buffer solution;
7) substrate;
8) stop buffer.
5, the application of the monoconal antibody mediated furacilinum residue detection method of mediated monoclonal antibody according to claim 1 is characterized in that being used to make monoconal antibody mediated furacilinum residue ELISA quick detection test paper bar.
6, according to the application of the monoconal antibody mediated furacilinum residue detection method of the described mediated monoclonal antibody of claim 5, it is characterized in that the method for making of monoconal antibody mediated furacilinum residue ELISA quick detection test paper bar is:
1) colloid gold label antibody: the monoclonal antibody on the collaurum behind the mark purifying;
2) metal spraying: the antibody of colloid gold label is sprayed onto on the glass fibre, makes the collaurum pad;
3) make nitrocellulose filter: T line in the test strips and C line are sprayed onto on the nitrocellulose filter;
4) with sample pad, collaurum pad, nitrocellulose filter, thieving paper assembling, slitting then.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610041481 CN1920573A (en) | 2006-09-08 | 2006-09-08 | Method for detecting monoconal antibody mediated furacilinum residue |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610041481 CN1920573A (en) | 2006-09-08 | 2006-09-08 | Method for detecting monoconal antibody mediated furacilinum residue |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1920573A true CN1920573A (en) | 2007-02-28 |
Family
ID=37778337
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200610041481 Pending CN1920573A (en) | 2006-09-08 | 2006-09-08 | Method for detecting monoconal antibody mediated furacilinum residue |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1920573A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101320037B (en) * | 2008-06-25 | 2013-01-23 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite |
| CN103018448A (en) * | 2011-09-20 | 2013-04-03 | 北京勤邦生物技术有限公司 | Enzyme-linked immunoassay kit and method for nitrofural metabolite detection |
| CN103197058A (en) * | 2013-03-01 | 2013-07-10 | 杭州迪恩科技有限公司 | Kit of rapid detection of nitrofurazone by using one-step enzyme-linked immunosorbent assay technique |
| CN103288962A (en) * | 2012-02-29 | 2013-09-11 | 华中农业大学 | Monoclonal antibody of furacilin residue marker semicarbazide, and preparation method and application thereof |
| CN106771272A (en) * | 2016-12-07 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | The detection method of Furacilin metabolite and detection card in a kind of animal tissue |
-
2006
- 2006-09-08 CN CN 200610041481 patent/CN1920573A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101320037B (en) * | 2008-06-25 | 2013-01-23 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite |
| CN103018448A (en) * | 2011-09-20 | 2013-04-03 | 北京勤邦生物技术有限公司 | Enzyme-linked immunoassay kit and method for nitrofural metabolite detection |
| CN103018448B (en) * | 2011-09-20 | 2016-01-20 | 北京勤邦生物技术有限公司 | Detect enzyme linked immunological kit and the method thereof of Furacilin metabolite |
| CN103288962A (en) * | 2012-02-29 | 2013-09-11 | 华中农业大学 | Monoclonal antibody of furacilin residue marker semicarbazide, and preparation method and application thereof |
| CN103288962B (en) * | 2012-02-29 | 2014-12-17 | 华中农业大学 | Monoclonal antibody of furacilin residue marker semicarbazide, and preparation method and application thereof |
| CN103197058A (en) * | 2013-03-01 | 2013-07-10 | 杭州迪恩科技有限公司 | Kit of rapid detection of nitrofurazone by using one-step enzyme-linked immunosorbent assay technique |
| CN106771272A (en) * | 2016-12-07 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | The detection method of Furacilin metabolite and detection card in a kind of animal tissue |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101865924B (en) | Method for detecting fumonisins based on immuno-magnetic bead combined enzyme-linked immunosorbent assay | |
| CN1916637A (en) | Method for detecting residue of furaltadone through mediated monoclonal antibody and application | |
| CN104897652B (en) | One step homogeneous chemistry luminescence method carries out the method and particulate used of small molecule detection | |
| CN1807601A (en) | Immune colloid gold test paper strip for detecting sulfadiazine residue and its preparation method | |
| CN1920573A (en) | Method for detecting monoconal antibody mediated furacilinum residue | |
| CN106093381B (en) | Rod method toxin rod method phenol monomethyl ether colloidal gold immuno-chromatography test paper strip | |
| CN105044365B (en) | The preparation method of the time resolved fluoro-immunoassay test paper of detection enrofloxacin residual | |
| CN1861632A (en) | Coupling compound of Norfloxacin, preparation process and application thereof | |
| CN1099894C (en) | Vaccine composition for eliciting an immune response against N-glycolylated gangliosides and its use for cancer treatment | |
| CN1677107A (en) | Immune antibody for testing residual of polyether-like antibiotic and use thereof | |
| CN1233828C (en) | Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith | |
| CN1286971C (en) | Monoclonal antibody, testing kit including the monoclonal antibody and application | |
| CN105301246A (en) | Time-resolved fluoroimmunoassay kit for detecting adprin as well as detecting method of time-resolved fluoroimmunoassay kit | |
| CN1916636A (en) | Method for detecting residue of dantafur through mediated monoclonal antibody and application | |
| CN106770210A (en) | A kind of olaquindox method for quick | |
| CN1888056A (en) | Immuno colloidal gold test paper strip for detecting sulfa drug residue | |
| CN101013134A (en) | Monoclonal antibody mediated furazolidone residual detecting method and uses thereof | |
| CN107523554A (en) | One plant of hybridoma cell strain SS0708 for secreting Madumycin monoclonal antibody specific and its application | |
| CN102875512B (en) | Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen | |
| CN200982971Y (en) | Penicillin quick half quantitative detection test paper | |
| CN102690788B (en) | Zearalenone anti-idiotypic antibody, preparation method thereof, and application thereof | |
| CN1790025A (en) | Test paper for lateral flow immunochromatographic semi-quantitative detection of ractopamine | |
| CN1250329C (en) | Endotoxin adsorbing agent and preparing method thereof | |
| CN1840691A (en) | Function regulation of selenium protein Se1k to interactive protein HCLP1 and anti-oncogene LZIP | |
| CN109022366A (en) | One plant of hybridoma cell strain for secreting anti-levamisol monoclonal antibody and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20070228 |