CN101173925B - Colloidal gold test paper card for detecting quinolones medicament residue - Google Patents
Colloidal gold test paper card for detecting quinolones medicament residue Download PDFInfo
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Abstract
The invention provides an immune colloidal gold test paper card for testing quinolones, comprising a reaction film, a sample pad, a conjugate release pad, a water absorbing pad, and a back liner, wherein, the reaction film comprises a test zone coated with quinolones-carrier protein conjugate and a quality control zone coated with sheep-anti-mouse IgG; the conjugate release pad coats quinolones monoclonal antibody-colloidal gold marker. The invention also provides the method using the test paper card to test quinolones, which comprises the following steps: firstly, pre-treating sample; secondly, testing the sample with the test paper card; thirdly, analyzing test result. The invention can be used to test residual quantity of quinolones in animal derived food such as chicken, chicken liver, pork, pig liver, urine, and honey, and has the advantages of simple operation, high sensitivity, fast test, low cost, on-site supervision, and satisfaction of test requirement of a large quantity of samples.
Description
Technical field
The present invention relates to the immunochemistry speed survey technology field of residue of veterinary drug, relate more specifically to a kind of colloidal gold test paper card that detects quinolones medicament relict, by the immunochromatography reaction of colloid gold label colour developing, fast detecting quinolones medicament relict.
Background technology
(quinolones, QNs) medicine is the one type of crucial broad-spectrum antibiotic that develops rapidly over nearly 20 years to quinolones.On chemical constitution, this type medicine belongs to the pyridonecarboxylic acid derivant, and (pyridonecarboxylic acids PCAs), is commonly called as " quinolones " (QNs).QNs suppresses the DNA of bacteria helicase, and has a broad antifungal spectrum, efficient, low toxicity, tissue penetration ability are strong, become the animal doctor face examine with aquaculture in one of most important anti-infectives, but because its drug resistance and potential carcinogenicity cause widely pays close attention to.In tissue; The sign residue of Enrofloxacin is Enrofloxacin and Ciprofloxacin; Wherein the highest with the residual concentration in liver organization and the renal tissue, secondly be chicken and the fatty skin histology that adheres to, its metabolic product Ciprofloxacin (CIF) of Enrofloxacin is biologically active still.Ofloxacin (OFL) mainly is present in the tissue with the form of original shape medicine.Metabolic rate is near 100% in vivo for Pefloxacin (PEF), and its metabolic product is the husky star (NOR) of promise oxygen.
The detection method that the QNs medicament residue is commonly used at present has two kinds.A kind of is stratographic analysis, like high performance liquid chromatography (HPLC), because complicated instrument and equipment and loaded down with trivial details process are not suitable for on-site supervision and great amount of samples examination.Another kind is an immunological method, and like enzyme linked immunosorbent assay (ELISA), its shortcoming is that detection time is long, and expense is higher, is unfavorable for promoting the use of in grass-roots unit.And these two kinds of methods all need the professional and technical personnel to operate.
(gold-immunochromatography assay GIGA) is a kind of novel immune standard gold technology that collaurum is applied to antigen-antibody reaction as tracer to colloidal gold immunochromatographimethod.
Summary of the invention
The object of the present invention is to provide the QNs quick detection test paper card that a kind of susceptibility is high, simple to operate, cost is low.
Test card of the present invention comprises reaction film, sample pad, bond releasing cushion, adsorptive pads and backing; On said reaction film, have test section that encapsulates QNS-carrier protein couplet thing and the Quality Control district that encapsulates sheep anti-mouse igg, said bond releasing cushion encapsulates QNS monoclonal antibody-colloid gold label thing.
The present invention is solidified QNs-carrier protein couplet antigen in the reaction film test section; Blended rubber body gold mark QNs monoclonal antibody; Compete QNs monoclonal antibody-colloid gold label thing through the QNs in the sample to be tested jointly with the QNs-carrier protein couplet thing that is coated on the reaction film, according to test section bar shade or have or not red bar to bring to judge whether to contain QNs in the sample to be tested liquid residual.During detection, sample splashes in the reagent card hole, and as QNs when concentration is lower than 10ng/ml in sample, colloidal gold antibody can be fixed on the QNs on the reaction film in the chromatography process conjugate combines, and in test section (T) and Quality Control district (C), a red stripes respectively occurs.If QNs is when concentration is higher than 10ng/ml in sample, colloidal gold antibody all combines with QNs in the sample, thereby red stripes in (T) district, do not occur because competitive reaction does not combine with the quinolone conjugate.Negatives in testing process owing to lack the antibody antigen competitive reaction, will (T) district with in the district red stripes appears (C).As shown in Figure 2.
Positive: demonstrate red stripes when (C) distinguishes, and (T) district does not develop the color, is judged to the positive, expression QNs content surpasses 10ng/ml, representes with "+".
Negative: when (C) district demonstrates red stripes, (T) district demonstrates red stripes simultaneously, and (T) the band color near or when being shallower than (C) band, be judged to feminine gender, expression QNs content is less than 10ng/ml, with "-" expression.
Invalid: do not demonstrate red stripes when (C) distinguishes, then no matter whether (T) district demonstrates red stripes, and it is invalid that this test card is judged to.
The present invention also provides the method for preparing above-mentioned test card, and it comprises step:
1) preparation encapsulates the bond releasing cushion of QNS monoclonal antibody-colloid gold label thing;
2) preparation has test section that encapsulates QNS-carrier protein couplet thing and the reaction film that encapsulates the Quality Control district of sheep anti-mouse igg;
3) with 1) and 2) bond releasing cushion, reaction film and the sample pad, adsorptive pads and the backing that prepare be assembled into test card.
Specifically, its step comprises:
(1), forms QNs hapten-carrier protein conjugate with QNs haptens and carrier protein couplet;
(2) with QNs hapten-carrier protein conjugate immune mouse, mouse boosting cell and murine myeloma cell are passed through to merge, screen, obtain secreting the hybridoma cell strain of QNs monoclonal antibody;
(3) extract mouse IgG immune health goat, obtain sheep anti-mouse igg antibody;
(4) with trisodium citrate and gold chloride prepared in reaction collaurum;
(5) the QNs monoclonal antibody with preparation adds in the collaurum of preparation, obtains QNs monoclonal antibody-colloid gold label thing;
(6) the bond releasing cushion was soaked 30 seconds with containing 0.1～0.5% caseic dipotassium hydrogen phosphate and potassium dihydrogen phosphate cocktail buffer,, then QNs monoclonal antibody-colloid gold label thing is coated on the bond releasing cushion at 37 ℃ of baking 2h;
(7) QNs-human serum albumin conjugate (HSA) is coated on constitutes the test section on the reaction film, and sheep anti-mouse igg is coated on constitutes the Quality Control district on the reaction film, seal with the confining liquid that contains 0.5% skimmed milk power;
(8) sample pad is soaked 2h with dipotassium hydrogen phosphate that contains 0.1～0.5% chicken serum albumen, 0.5～1% Tween-80 and potassium dihydrogen phosphate cocktail buffer, at 37 ℃ of baking 2h;
(9) on backing, stick reaction film, adsorptive pads, bond releasing cushion and sample pad in order, sample pad is covered the bond releasing cushion.Be cut into wide little of 3mm at last, add plastic casing, vacuum packaging.Sample pad is covered the bond releasing cushion can prolong testing result observing time, sample pad is fully absorbed tracer liquid and fully react, thereby reduce error with golden labeling antibody.
That test card of the present invention has is highly sensitive, simple to operate, detection time short, store simple, long shelf-life low cost and other advantages, can on-site supervision, be fit to the great amount of samples examination.
Description of drawings
Fig. 1 detects the assembling synoptic diagram of test card for the present invention;
Fig. 2 is that testing result of the present invention is analyzed synoptic diagram.
Among the figure: 1, sample pad; 2, bond releasing cushion; 3, reaction film; 4, adsorptive pads; 5, test section; 6, Quality Control district; 7, backing.
Embodiment
Come further to set forth the present invention below in conjunction with concrete embodiment.Should be understood that these embodiment only are used to explain the present invention, and be not used for limiting scope of the present invention.In addition, those skilled in the art may carry out various changes or modification to the present invention in the appended claims restricted portion, and these changes or modification should fall into the protection domain of invention equally.
Embodiment 1 QNs detects the preparation of test card
1, the synthetic and evaluation of QNs hapten-carrier protein conjugate
(1) the haptenic preparation of QNs
Get pipemidic acid (8-ethyl-5-oxo-5,8-dihydro-2-(1-piperazinyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid) 500mg and be dissolved in the 30ml water, add EDC 200mg room temperature reaction 1h, add equal amido phenenyl acid 150mg reaction again and spend the night.Extract the QNs haptens through pickling then, the dry 280mg that gets, the QNs haptens is 8-ethyl-5-oxo-5,8-dihydro-2-(1-piperazinyl) pyrido [2,3-d] pyrimidine-6-acid amides phenylacetic acid.
(2) QNs haptens-human albumin's conjugate is synthetic
Take by weighing human serum albumins (HSA) 90mg, add 10ml PBS dissolving, ice bath stirs the N that adds 10ml down again, and N-dimethylformamide (DMF) gets HSA solution.Take by weighing QNs haptens 7.8mg, be dissolved among the 1.5ml DMF, add tri-n-butylamine 8 μ L, add isobutyl chlorocarbonate 4.5 μ L behind the mixing again, stir 25min in 4 ℃ of lower magnetic forces.Under the magnetic agitation, HSA solution poured into begin reaction (this moment reactant liquor can become muddy) in the QNs haptens solution, with the NaOH conditioned reaction liquid pH value to 8.3 (it is limpid that reactant liquor becomes again) of 1mol/L.Under the magnetic agitation, put 4 ℃ of refrigerators reaction 4h, end reaction liquid was dialysed freeze drying ,-6 ℃ of preservations two days with the PBS of 0.01mol/L pH7.4.
(3) QNs haptens-bovine serum albumin is synthetic:
Take by weighing bovine serum albumin (BSA) 70mg, be dissolved among the 25ml 0.05M pH7.4 PBS, add the DMF of 25ml again.Take by weighing QNs haptens 7.8mg, be dissolved among the 2mlDMF.In QNs haptens solution, add tri-n-butylamine 6.0 μ l, add isobutyl chlorocarbonate 3.8 μ l behind the mixing again, stir 25min in 4 ℃ of lower magnetic forces.Under the magnetic agitation, BSA solution poured into begin reaction (this moment reactant liquor can become muddy) in the QNs haptens solution, with the NaOH conditioned reaction liquid pH value to 9.0 (reactant liquor becomes clarification again) of 1mol/L.Under the magnetic agitation, in 4 ℃ of refrigerators, react 4h.End reaction liquid is dialysed freeze drying ,-6 ℃ of preservations with 0.01mol/L pH7.4 PBS.
(4) evaluation of QNs hapten-carrier protein conjugate
Carrier protein, QNs haptens, QNs-carrier protein couplet thing are made into the solution of 0.5mg/ml respectively with the PBS of pH7.4; PBS zeroing with 0.01mol/L pH9; In the interscan of wavelength 200-800nm scope, obtain the absorption curve of carrier protein, QNs haptens, QNs hapten-carrier protein conjugate with ultraviolet spectrophotometer.The differing absorption curve appears in the three, shows the success of QNs and carrier protein couplet.
2, QNs MONOCLONAL ANTIBODIES SPECIFIC FOR
(1) animal immune
Immunogene is injected in the Balb/c mouse body, and dosage is 90 μ g/, makes it produce polyclonal antibody.
(2) Fusion of Cells and cloning
Get immune Balb/c mouse boosting cell, merge, adopt the indirect competitive ELISA method to measure cell conditioned medium liquid, screen positive hole in 5: 1 ratios and SP2/0 myeloma cell.Utilize limiting dilution assay that cloning is carried out in positive hole, up to the hybridoma cell strain that obtains the stably excreting monoclonal antibody.Get wherein the higher hybridoma called after A-4-3 that tires; This cell line has been preserved in (address: Datun Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on 05 28th, 2007; Institute of Microorganism, Academia Sinica), preserving number is CGMCC No.2066.
(3) cell cryopreservation and recovery
The monoclonal hybridoma strain A-4-3 of QNs 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.
(4) MONOCLONAL ANTIBODIES SPECIFIC FOR and purifying
With Balb/c mouse peritoneal injection sterilization paraffinum liquidum, dosage is 0.4ml/, monoclonal hybridoma strain A-4-35 * 10 of 7 days pneumoretroperitoneums injection QNs
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.
3, the preparation of sheep anti mouse antiantibody: as immune animal, is that immunogene to pathogen-free domestic goat carry out immunity with mouse source antibody with goat, obtains the sheep anti mouse antiantibody.
4, the preparation of QNs monoclonal antibody-colloid gold label thing
(1) preparation of collaurum
Boil off ionized water 1% gold chloride is diluted to 0.01% with two; Put to stir on the magnetic force heating rod stirrer and boil, the gold chloride of every 100ml 0.01% adds the trisodium citrate of 2ml 1%, continues to boil; Stop heating when liquid takes on a red color, supply dehydration after being cooled to room temperature.The collaurum outward appearance for preparing should be pure, bright, do not have deposition and floating thing, and the term of validity is one month.
(2) preparation of QNs monoclonal antibody-colloid gold label thing
Under magnetic agitation, the pH value to 8.2 with 0.1M sal tartari is transferred collaurum adds the QNs monoclonal antibody by 1～2 μ g/ml antibody colloidal gold, continues stirring and evenly mixing 30min, adding 10%BSA to final concentration be 1%, leave standstill 30min.12000rpm, 4 ℃ of centrifugal 30min, abandoning supernatant, deposition be with redissolving the damping fluid washed twice, will precipitate with the redissolution damping fluid of initial collaurum volume 1/20 resuspended, put 4 ℃ subsequent use, preserved 60 days.
Redissolution damping fluid: contain bovine serum albumin(BSA) (BSA) 0.02～0.1%, Tween-20 0.05～0.2%, vitamin C (Vit-C) 0.1%, the boric acid redissolution damping fluid of 0.02M pH9.0.
5, the bond releasing cushion encapsulates
The bond releasing cushion put into contain 0.1～0.5% caseic dipotassium hydrogen phosphate and the potassium dihydrogen phosphate cocktail buffer was soaked 30 seconds, 37 ℃ of baking 2h are subsequent use; With Biodot point film appearance the QNs monoclonal antibody-colloid gold label thing for preparing evenly is coated on the bond releasing cushion, every 5cm bond releasing cushion encapsulates 9 μ l QNs monoclonal antibody-colloid gold label things, vacuum drying, Vacuum Package, put 4 ℃ subsequent use.
6, the preparation of reaction film
With encapsulating the Quality Control district that test section that QNs-carrier protein couplet thing constitutes and sheep anti-mouse igg constitute on the nitrocellulose filter respectively, the usefulness confining liquid that contains 0.5% skimmed milk power seals again.
Encapsulate process: with kaliumphosphate buffer (containing 3% methyl alcohol) QNs-human serum albumins (HSA) conjugate is diluted to 15 μ g/mL, with Biodot point film appearance it is encapsulated in nitrocellulose filter as the test section, package amount is 0.7 μ g/cm
2, the test section is near bond releasing cushion end, apart from the about 8mm of bond releasing cushion end; With 0.01M kaliumphosphate buffer (containing 3% methyl alcohol) sheep anti-mouse igg antibody is diluted to 200 μ g/ml, with Biodot point film appearance it is encapsulated in cellulose membrane as the Quality Control district, package amount is 0.7 μ g/ml
2, the Quality Control district is near adsorptive pads, apart from the about 8mm of adsorptive pads, and two linear distances, 5～8mm.37 ℃ of oven dry, encapsulation.
7, the assembling of colloid gold immune test paper card
On the PVC backing, paste nitrocellulose filter, adsorptive pads, bond releasing cushion in order, paste sample pad at last, sample pad is covered the bond releasing cushion.Be cut into wide little of 3mm, add plastic casing, vacuum packaging.Original packing should be stored in 18～25 ℃, is valid for one year.Test card of the present invention covers the bond releasing cushion with sample pad can prolong testing result observing time, and sample pad also can fully absorb tracer liquid and contacts, fully reacts, reduces error fully with golden labeling antibody.
The residual detection of QNs in embodiment 2 samples
1, sample pre-treatment
(1) animal tissue's pre-treatment (chicken, chicken gizzard, pork, pork liver)
Take by weighing tissue samples that the even matter of 2.0 ± 0.05g crosses in centrifuge tube, cover tight bottle cap.With the centrifuge tube that sample is housed water-bath 10min in little water-bath that boils, draw the solution cook more than three and fall in the centrifuge tube of 1.5ml.Please be centrifugal if any obvious yellow muddiness, use supernatant as sample to be tested solution then.
(2) pre-treatment of egg sample
With the homogenate of egg sample, take by weighing 2.0 ± 0.05g in centrifuge tube, cover tight bottle cap.With the centrifuge tube that sample is housed water-bath 10min in little water-bath that boils, draw the solution cook more than three and fall in the centrifuge tube of 1.5ml.Please use supernatant as sample to be tested solution.
(3) pre-treatment of urine specimen
Urine sample: generally can directly detect, after centrifugal 5min of 3000～5000rpm or employing filter paper filtering are handled when urine is muddy state, carry out point sample again and detect.
(4) pre-treatment of honey sample
Take by weighing 2.0 ± 0.05g honey and put in the clean container, the distilled water diluting that adds 4ml gets final product.
2, detect with test card of the present invention
In the reagent hole clipping, drip 3 samples, observations behind 5～8min with dropper.Surpass 10min, then the pattern detection sentence read result is invalid.
3, testing result analysis
QNs is when concentration is higher than 10ng/ml in sample, and colloidal gold antibody all combines with QNs, thereby red stripes do not occur because competitive reaction can not combine with the QNs conjugate in (T) district.Negatives in testing process owing to lack the antibody antigen competitive reaction, will (T) district with in the district red stripes appears (C).Positive: as to demonstrate red stripes when (C) distinguishes, and (T) distinguish when not developing the color, be judged to the positive, with "+" expression.Negative: when (C) district demonstrates red stripes, when (T) district demonstrates red stripes simultaneously, and (T) district's color near or when being shallower than (C) district, be judged to feminine gender, with "-" expression.Invalid: do not demonstrate red stripes when (C) distinguishes, then no matter whether (T) district demonstrates red stripes, and it is invalid that this test card is judged to.As shown in Figure 2.
Embodiment 3 pattern detection instances
Get known QNs concentration greater than each 15 parts of the egg of 10ng/g, chicken, each 15 parts in honey sample and negatives separately, each sample duplicate detection 2 times is calculated its yin and yang attribute rate.The result is shown in table 1, table 2 and table 3.
Table 1 egg sample yin and yang attribute rate detects
Table 2 chicken sample yin and yang attribute rate detects
Table 3 honey sample yin and yang attribute rate detects
The result shows, each 15 parts of egg, chicken, each 15 parts in honey sample and negatives separately, and each sample duplicate detection 2 times, its yin and yang attribute coincidence rate is 100% in the egg, and its yin and yang attribute coincidence rate of chicken is 100%.False positive rate is below 7% in the honey, and its positive coincidence rate is 100%, and negative match-rate is more than 93%.Explain that detection test card of the present invention can be used as conventional method fully and is used for fast detecting residual to QNs on the market.
Experimental example 1 susceptibility and specificity test
Sensitivity tests
With the titer dilution of Norfloxacin is 0,5,6,7,8,9; 10,11,12,13,14; 15,16,17,18,19; 20ng/ml, use testing result of the present invention to be: when dripping the titer of examination 0～8ng/ml, test card demonstrates macroscopic two red stripes, when dripping the titer of examination 9ng/ml; Macroscopic two red stripes also appear in test card, but (T) district's color is very light very fuzzy, and test card (T) district does not develop the color when dripping examination 10ng/ml and the titer more than the 10ng/ml, and test findings shows that the sensitivity of this test card detection QNs is 10ng/ml.
The specificity test
The positive detection concentration that the present invention detects test card is 10ng/ml, detects Norfloxacin medicine and negative standard items, equally Enoxacin, Enrofloxacin, Ofloxacin, Danofloxacin, Pefloxacin, Ciprofloxacin, Lomefloxacin 、 oxolinic acid, flumequine, marbofloxacin, Amifloxacin, two Flucloxacillin, sarafloxacin is pressed 5; 6,7,8; 9,10,11; 12,13,14; 15,16,17; 18,19,20ng/ml dilutes; Detect with test card of the present invention; 90%, 100%, 100%, 85%, 90%, 100%, 90%, 85%, 90%, 85%, 100%, less than 1%, less than 1% the result is: Enoxacin, Enrofloxacin, Ofloxacin, Danofloxacin, Pefloxacin, Ciprofloxacin, Lomefloxacin 、 oxolinic acid, flumequine, marbofloxacin, Amifloxacin, two Flucloxacillin, sarafloxacin be respectively 11,10,10,12,11,10,11,12,11, the test section shows during 12ng/ml concentration, and two Flucloxacillins, sarafloxacin do not show the test section 20000, during 50000ng/ml concentration, can draw cross reacting rate and be through calculating:.As shown in table 4.Cross reaction is big more, explains that this test card is just good more to the specificity that QNs detects.
Table 4 test card specificity of the present invention test findings
Claims (5)
1. colloidal gold test paper card that detects QNS; Comprise reaction film, sample pad, bond releasing cushion, adsorptive pads and backing; It is characterized in that having on the said reaction film test section that encapsulates QNS-carrier protein couplet thing and the Quality Control district that encapsulates sheep anti-mouse igg, said bond releasing cushion encapsulates QNS monoclonal antibody-colloid gold label thing;
Wherein, said QNS-carrier protein couplet thing by pipemidic acid and EDC and equal amido phenenyl acid react 8-ethyl-5-oxo-5,8-dihydro-2-(1-piperazinyl) pyrido [2,3-d] pyrimidine-6-acid amides phenylacetic acid obtains with carrier protein couplet then.
2. colloidal gold test paper card as claimed in claim 1; It is characterized in that the QNS monoclonal antibody in said QNS monoclonal antibody-colloid gold label thing is; With 8-ethyl-5-oxo-5; 8-dihydro-2-(1-piperazinyl) pyrido [2,3-d] pyrimidine-6-acid amides phenylacetic acid and carrier protein couplet obtain coupled complex and obtain as immunogen preparing.
3. claim 1 or 2 said colloid gold test papers are stuck in the application that detects in the QNS.
4. method for preparing claim 1 or 2 said colloidal gold test paper cards, it comprises step:
1) preparation encapsulates the bond releasing cushion of QNS monoclonal antibody-colloid gold label thing;
2) preparation has test section that encapsulates QNS-carrier protein couplet thing and the reaction film that encapsulates the Quality Control district of sheep anti-mouse igg;
3) with 1) and 2) bond releasing cushion, reaction film and the sample pad, adsorptive pads and the backing that prepare be assembled into test card.
5. method that detects quinolones medicament relict in the sample is characterized in that comprising step:
1) sample pre-treatment;
2) detect with claim 1 or 2 described test cards;
3) analyzing and testing result.
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| CN101477124B (en) * | 2008-12-17 | 2013-05-29 | 北京望尔生物技术有限公司 | Method for detecting melamine and its special test paper |
| CN101915844B (en) * | 2010-08-03 | 2013-06-26 | 中国农业大学 | Method and special quantum dot fluorescent immunoassay kit for detecting quinolone compounds |
| CN102478576A (en) * | 2010-11-30 | 2012-05-30 | 内蒙古蒙牛乳业（集团）股份有限公司 | Rapid qualitative detection method for detecting danofloxacin in cream |
| CN102323415B (en) * | 2011-06-13 | 2014-03-19 | 清华大学深圳研究生院 | Immunochromatography test paper for detecting Ciprofloxacin and preparation method thereof |
| CN103323593B (en) * | 2012-03-22 | 2016-03-30 | 北京勤邦生物技术有限公司 | A kind of test paper and application thereof detecting fluoroquinolones |
| CN103454422B (en) * | 2013-08-03 | 2016-02-03 | 河南省农业科学院 | Flumequine quick detection test paper card and preparation method |
| CN104561233A (en) * | 2015-01-24 | 2015-04-29 | 福建出入境检验检疫局检验检疫技术中心 | Quinolone medicine residue detection kit |
| CN105044333B (en) * | 2015-06-24 | 2017-07-11 | 华南农业大学 | A kind of ELISA detection method of pipemidic acid and application |
| CN106282124A (en) * | 2016-08-24 | 2017-01-04 | 江南大学 | The mass selection monoclonal antibody of a kind of monoclonal cell strain C4 and generation thereof and application |
| CN106841600B (en) * | 2016-12-28 | 2018-09-11 | 河南科技学院 | The test card of fluoroquinolones is detected based on pefloxacin class polyclonal antibody |
| CN106771165B (en) * | 2016-12-28 | 2018-09-11 | 河南科技学院 | A kind of fluoroquinolones envelope antigen and preparation method thereof and Test paper card |
| CN110551220A (en) * | 2019-08-29 | 2019-12-10 | 北京勤邦生物技术有限公司 | Preparation and application of DDT monoclonal antibody |
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