CN105758847A - Chemiluminescence enzyme linked immunoassay kit for detecting sulfonamides - Google Patents
Chemiluminescence enzyme linked immunoassay kit for detecting sulfonamides Download PDFInfo
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Abstract
The invention discloses a chemiluminescence enzyme linked immunoassay kit for detecting sulfonamides. The chemiluminescence enzyme linked immunoassay kit is characterized in that the kit consists of a sulfonamides standard solution, a chemiluminiscence panel, an enzyme labeling antigen concentrated solution, enzyme labeling antigen diluent, a luminescence substrate solution, a scrubbing solution and a combination solution, wherein each pore of the chemiluminiscence panel is coated with an anti-sulfonamides monoclonal antibody. The chemiluminescence established by the invention has the advantages of high flux, high sensitivity, high specificity, low detection cost, high speed, accuracy and the like, and can meet the requirement of the quick screening inspection of large-scale field detection.
Description
Technical field
The invention belongs to field of detection of food safety, be specifically related to a kind of chemical luminescence ELISA detection kit, particularly detect the chemical luminescence ELISA detection kit of residual quantity of sulfonamide in Carnis Sus domestica, Carnis Gallus domesticus, shrimp, fish equal samples.
Background technology
Sulfa drugs refers to the general name of a class medicine with P-aminobenzene-sulfonamide structure, be a class for preventing and treat the chemotherapeutic agent of bacterial infection disease, its antimicrobial spectrum is relatively wide, and most of gram positive bacterias and gram negative bacteria are had inhibitory action.
Sulfonamides (SAs) medicine is applied very extensive in husbandry sector, mainly in animal diseases control, there is significant curative effect, fowl cholera, avian typhoid, avian paratyphoid, pullorum disease, infectious coryza of chicken, turkey Arizona disease etc. can be treated, in addition to the various coccidiosis of poultry, Leucocytozoon Caulleryi etc., also there is better effects.
Although sulfa drugs is widely used, but meanwhile, the significant toxic and side effects of this kind of medicine also result in the extensive concern of people.Clinical research finds that it can affect urinary system function, causes reaction and the carcinogenecitys such as crystalluria, hematuria.Sulfa drugs is distributed in during whole body respectively organizes after absorbing, the highest with blood, liver, kidney content.And, so in vivo hold time length high with plasma protein binding rate.Can also penetrating meninges hydrops and other hydrops, and enter foetal circulation by Placenta Hominis, to anemia of pregnant woman and baby and unfavorable, also easy crystallization in urine, causes that calculus damages kidney.Owing to sulfa drugs exists serious side effects, human body is medium-term and long-term to be existed sulfa drugs and can cause that sulfa drugs is produced drug resistance by many antibacterials, and has potential carcinogenecity, and No. 235 file of the Ministry of Agriculture of China specifies that its residue limits is 100ug/kg.
At present, detect sulfa drug residue, mainly adopt high performance liquid chromatography, thin layer chromatography, immunochromatography, high performance capillary electrophoresis.The instrumental methods such as high performance liquid chromatography, liquid chromatograph, gas chromatography, precision is high, highly sensitive, but large-scale instrument is expensive and needs specialty detection technique personnel, it is difficult to realize large sample field screening.Enzyme-linked immunoassay method detection is cheap, quickly, but insufficient sensitivity, it is adaptable to the detection of trace substance and qualification, the detection in trace materials, it is difficult to make the most of the advantage.
Chemiluminescence immune assay (Chemiluminescenceanalysis, CLlA) high-sensitive chemiluminescence is combined by technology with the immunoreation of high specific, has the feature such as highly sensitive, high specificity, range of linearity width, easy and simple to handle, instrument and equipment that need not be sufficiently expensive.CLIA does not need external light source, there is the signal to noise ratio higher than fluorescence immunoassay, stronger than the anti-background interference ability of conventional enzyme-linked immunosorbent assay method, high 1 to 2 order of magnitude of its remolding sensitivity ELISA, detection range is up to 6 orders of magnitude, automaticity is high, improves the precision of analysis method, and CLIA has become as the trace of a kind of advanced person or the detection technique of ultra trace material.CLIA, in veterinary, medical science, food analysis etc., has a extensive future.
Summary of the invention
The technical problem to be solved is in that to provide a kind of detection sulfa drugs chemiluminescence enzyme linked immunoassay reagent kit, adopt this test kit when carrying out the detection of sulfa drugs, not only to possess higher sensitivity, specificity, and there is response speed feature faster.Further object is that the detection method that a kind of sulfa drugs is provided, the feature that the method specificity is good, highly sensitive, simple to operate.Realizing above-mentioned purpose, the present invention provides a kind of sulfa drugs detection kit, and its main agents comprised has:
It is coated with the polystyrene Chemiluminescent plate of anti-sulfa drugs monoclonal antibody.
Described sulfa drugs standard solution 6 bottles, concentration is 0ug/L, 2ug/L, 4ug/L, 8ug/L, 16ug/L, 32ug/L respectively.
Enzyme-labelled antigen concentrated solution: the artificial antigen being made up of sulfa drugs and bovine serum albumin coupling and horseradish peroxidase prepare.
Enzyme-labelled antigen diluent: 0.01-0.02M phosphate buffer, pH7.0-8.0, the bovine serum albumin of 0.5-1%.
Luminous substrate liquid.Luminous substrate liquid is divided into A, B liquid.A liquid is chemical luminous substrate-luminol and luminescence enhancer-p-cresol solution, and B liquid is hydrogen peroxide urea solution.
Redissolution liquid.The liquid that redissolves is specially 2 times of concentrated phosphoric acid salt buffers, uses front distilled water to use after being diluted to working concentration, for sample pre-treatments.
Cleaning mixture.Wash solution is specially 20 times of concentrated phosphoric acid salt buffers containing tween 20 (Tween-20) buffer, uses front distilled water to use after being diluted to working concentration, for washing chemistry luminous plaque in experimentation.
The preparation of solution of the present invention: the sensitivity impact that test kit of the present invention is detected by the enzyme mark sulfa drugs monoclonal anti original solution, chemiluminescent solution and the wash solution that relate in test kit of the present invention and formula thereof is very big；Wherein the main component of each solution and compound method thereof are as follows.
1, enzyme mark sulfa drugs monoclonal anti original solution: the artificial antigen prepared with sulfa drugs and coupling protein coupling prepares with horseradish peroxidase, and gained enzyme mark sulfa drugs antigen diluent becomes the working concentration of 1:5000.
2, enzyme-labelled antigen diluent: 0.01-0.02M phosphate buffer, pH7.0-8.0, the bovine serum albumin of 0.5-1%.
3, luminous substrate liquid: three (methylol) aminomethane solution of A liquid is luminol content to be 0.01M, p-cresol content be 0.001MpH8.8, B liquid is that every 100mL solution is containing citric acid 2.1g, anhydrous Na2HPO42.82g, the aqueous solution of the carbamide peroxide 0.64mL of 0.75%.
4, redissolution working solution: pH value is 7.5-7.8, and containing 2-4% casein, the phosphate buffer of 0.1-0.2mol/L, described percentage ratio is w/v.
5, cleaning mixture: pH value is 7.2-7.5, containing 0.8-1.2% tween 20,0.3-0.6 ‰ Hydrazoic acid,sodium salt, the phosphate buffer of 0.1-0.2mol/L, described percentage ratio is mass volume ratio.
6, being coated buffer: pH value is the phosphate buffer of 9.2-9.6,0.1-0.2mol/L, described percentage ratio is w/v.
7, lock solution preparation: containing 5-8% defatted milk powder, the phosphate buffer of pH value 7.2-7.6,0.1-0.2mol/L, described percentage ratio is mass volume ratio.
Being coated of Chemiluminescent plate of the present invention:
Being coated Chemiluminescent plate in the present invention adopts what anti-sulfa drugs monoclonal antibody was placed in setting to be coated in solution, with the concentration set, reacts and be coated in 37 DEG C of calorstats.
What the present invention adopted is pH9.2-9.6 phosphate buffer.In the present invention, in microwell plate, coated anti-sulfa drugs can well be combined on microwell plate frosting under alkaline environment, it is possible to stands repeatedly to wash plate, and the antibody of employing is coated concentration can from 10mg/ml-20mg/ml.
The microwell plate being coated can be closed by lock solution, the preferred BSA of inert protein in confining liquid, need to add NaN3Prevent from going bad.
The preparation of enzyme mark sulfa drugs antigenic solution:
In the present invention, enzyme mark sulfa drugs antigen solution concentration is to determine the key factor of sulfa drugs chemical luminescence ELISA detection kit measurement range and sensitivity in the present invention.
The enzyme mark sulfa drugs antigenic solution related in the present invention can become the working concentration of 1:4000 by enzyme mark sulfa drugs diluted.
The test kit prepared according to above-mentioned enzyme mark sulfa drugs antigen solution concentration can reach the good range of linearity (normal line scope can reach 2-32ug/L).
The preparation of chemiluminescent solution: the present invention adopts horseradish peroxidase labeled substrate luminescent system, mainly luminol-hydrogen peroxide system.
Three (methylol) aminomethane solution of described luminous substrate liquid A liquid is luminol content to be 0.01M, p-cresol content be 0.001MpH8.8, B liquid is that 100mL solution is containing citric acid 2.1g, anhydrous Na2HPO42.82g, the aqueous solution of the carbamide peroxide 0.64mL of 0.75%, described percentage ratio is mass percent.Described luminol is chemical luminous substrate, and p-cresol is luminescence enhancer.
Principles of the invention is to be combined with enzymatic high sensitivity by the high degree of specificity that antibody-antigene reacts, and utilizes the chemiluminescence reaction detection production concentration of substrate for enzymatic activity.
The chemical luminescence ELISA detection kit of the present invention has feature fast and accurately highly sensitive, easy, is expected to sulfa drug residue detection is played a significant role.
Accompanying drawing explanation
Fig. 1 sulfonamide hapten synthetic route.
Fig. 2 sulfonamide hapten identifies figure.
Fig. 3 sulfa drugs chemiluminescence enzyme linked immunoassay reagent kit canonical plotting.
Fig. 4 table 1 sulfa drugs test kit accuracy and precision measure.
Fig. 5 table 2 sulfa drugs cross reacting rate.
Detailed description of the invention
The synthesis of embodiment 1 sulfonamide hapten-carrier protein couplet thing and qualification
(1) sulfonamide hapten synthesis
Weigh in the oxolane that 0.5g-2g sulfadiazine is dissolved in after drying; logical nitrogen protection; add 0.24-1 maleic anhydride stirring at normal temperature to dissolve; add 10-50mgDMAP body and carry out catalytic reaction; monitor reaction with TLC lamellae to carry out, until not having raw material or raw material point very shallow, stopped reaction; silicagel column purifies, and concentrates to obtain product.Productivity > 80%.
(2) immunogenic preparation
Weigh 34.8mg hapten and be dissolved in 2mLDMF solution, add each 60mgEDC and 60mgNHS(to be dissolved in 2mL water) carry out activation 30 minutes, join 132-440mg carrier protein BSA and be dissolved in 5mL aqueous solution, adjust solution PH 8-9, room temperature carries out coupling and prepares immunogen, dialysing 3 days with 0.01mol/LPB buffer, every day changes dialysis solution sooner or later, prepares antibody for animal immune after having dialysed.Subpackage, saves backup in-20 DEG C.
(3) preparation of coating antigen
Weigh 34.8mg hapten and be dissolved in 2mLDMF solution, add each 60mgEDC and 60mgNHS(to be dissolved in 2mL water) carry out activation 30 minutes, join 180-520mg carrier protein VOA and be dissolved in 5mL aqueous solution, adjust solution PH 8-9, room temperature carries out coupling and prepares immunogen, dialysing 3 days with 0.01mol/LPB buffer, every day changes dialysis solution sooner or later, prepares antibody for animal immune after having dialysed.Subpackage, saves backup in-20 DEG C.
(4) qualification of sulfonamide hapten-carrier protein couplet thing
By carrier protein, sulfonamide hapten, sulfonamide hapten-carrier protein couplet thing pH7.4 PBS be made into the solution of 0.5mg/mL, return to zero with 0.01mol/LpH7.4PBS, with ultraviolet spectrophotometer in wavelength 200 ~ 750nm scope interscan, obtain the absorption curve of carrier protein, sulfonamide hapten, sulfonamide hapten-carrier protein couplet thing.There is different absorption curves in three, it was shown that sulfonamide hapten and carrier protein couplet success.
(5) preparation of sulfa drugs monoclonal antibody
A. animal immune
Immunogen above-mentioned steps obtained is injected in Balb/c Mice Body, and immunizing dose is 150 μ g/ so that it is produce antiserum.
B. cell fusion and cloning
Take immunity Balb/c mouse boosting cell, in 8:1(quantitative proportion) ratio and SP2/0 myeloma cell fusion, screening obtains the sulfa drugs monoclonal antibody hybridoma cell strain of stably excreting sulfa drugs monoclonal antibody.
C. cell cryopreservation and recovery
Hybridoma frozen stock solution is made 5 × 106The cell suspension of individual/ml, preserves for a long time in liquid nitrogen.Take out cryopreservation tube during recovery, be immediately placed in 37 DEG C of water-bath middling speeds and melt, after centrifugal segregation frozen stock solution, move into and cultivate culture in glassware.
D. the preparation of monoclonal antibody and purification
Increment culture method: be placed in cell culture medium by hybridoma, cultivates under 37 DEG C of conditions, is purified by the culture fluid obtained by sad-saturated ammonium sulfate method, obtains monoclonal antibody, measures titer, and frozen-20 DEG C save backup.
Embodiment 2: the preparation of enzyme-labelled antigen
A. weigh 2mgHRP to be dissolved in 0.5mL distilled water；Add the 0.5mL 0.06mol/LNaIO newly prepared4Solution, 4 DEG C of lucifuge effect 30min；
B. the ethylene glycol 0.5mL of 160mmol/L, room temperature effect 30min are added；
C. adding sulfonamide hapten-carrier protein couplet thing 2mg, in the bag filter that after mixing, loading processed, put in the 0.05mmol/L sodium phosphate buffer of 1000mL and dialyse, 4 DEG C overnight；
D. dialysis solution is drawn in the centrifuge tube of 10mL, adds the 0.25mL 5g/LNaBH newly joined4Liquid, mixes rearmounted 4 DEG C of 2h；Adding isopyknic saturated ammonium sulfate solution, 4 DEG C of effect 30min, at 4 DEG C, the centrifugal 25min of 3000rpm, abandons supernatant；
E. precipitation is dissolved in 1.5mL0.02mol/LpH7.0-7.50.2-0.4%NaN3In 0.2-0.3mol/L borate buffer solution, suck in bag filter, at 0.02mol/LpH7.0-7.50.2-0.4%NaN30.2-0.3mol/L borate buffer solution is dialysed, and 4 DEG C overnight (borate buffer solution is changed 3 times in midway)；
F, is drawn in microcentrifugal tube by liquid in dialysis solution, and at 4 DEG C, the centrifugal 30min of 10000rpm, by supernatant sucking-off, adds equivalent glycerol, and mixing ,-20 DEG C save backup.
The foundation of embodiment 3CLEIA detection method
(1) preferred (the square formation method) of antibody and envelope antigen concentration
Longitudinally it is coated Chemiluminescent plate with every kind of coated antibody by the dilution series of 1,2,4,8,16,32,64,128 μ g/mL, 100 μ L/ holes, after being placed in 37 DEG C of calorstat 2h, pat dry；Closing with 150 μ L/ hole lock solution, 37 DEG C of calorstats are placed 2 hours, wash plate once, pat dry；Adding enzyme mark sulfa drugs antigen (1:1000 to 1:512000) of the 50 a series of dilutions in μ L/ hole, room temperature (20 ~ 25 DEG C) hatches 15min, washes plate five times, pats dry for the last time；It is separately added into chemiluminescence A, B liquid in 50 μ L/ holes, measures luminous intensity values.The envelope antigen concentration of obvious graded and antibody dilution is had to carry out specific assay for optium concentration with luminous intensity values with the concentration of envelope antigen.
(2) mensuration of antibody sensitivity
According to the above-mentioned optimization experiment to coated antibody and enzyme-labelled antigen concentration, selecting and determine that enzyme-labelled antigen concentration is 1:4000, coated antibody concentration is the 8.0 μ g/mL mensuration carrying out the sensitivity of antibody:
A. it is coated: be coated solution with the carbonate of 0.05MpH9.6 and anti-sulfa drugs antibody is made into the solution of 8.0 μ g/mL, each polystyrene board Chemiluminescent plate reacting hole adds 100 μ L, 37 DEG C of calorstat 2h.Discard solution in hole, pat dry.
B. close: closing above-mentioned coated Chemiluminescent plate by lock solution, 150 μ L/ holes, then 37 DEG C of calorstat 2h wash plate once, pat dry.
C. application of sample: add the sulfa drugs standard solution 50 μ L/ hole of variable concentrations, add enzyme mark sulfa drugs antigen (1:4000) of 50 μ L/ hole dilutions in the above-mentioned reacting hole closed, room temperature (20 ~ 25 DEG C) lucifuge hatches 15min, then wash plate five times, pat dry for the last time.
D. luminous: in each reacting hole, to add the chemiluminescent solution 100 μ L/ hole of Extemporaneous, detect with chemical illumination immunity analysis instrument after reaction 3min.
E. testing result calculates with suppression ratio:
Relative luminous intensity (%)=RLU/RLU0, RLU is standard substance or the luminous intensity values of sample solution mensuration, RLU0It it is the luminous intensity values of blank (concentration is the standard solution of 0).
Calculate the concentration of medicine during 50% suppression ratio and be the sensitivity of this antibody.
Embodiment 4 detects the chemiluminescence enzyme linked immunoassay reagent kit of sulfa drugs
(1) composition of the chemiluminescence enzyme linked immunoassay reagent kit of sulfa drugs is detected
A. the solid phase carrier (Chemiluminescent plate) of sulfa drugs monoclonal antibody it is coated with；
B. sulfa drugs standard solution: 0,2,4,8,16,32ug/L.
C. enzyme mark sulfonamide hapten-carrier protein couplet thing solution: prepare with horseradish peroxidase with artificial antigen, by diluted to working concentration during use.
D. enzyme mark sulfonamide hapten-carrier protein couplet thing diluent: sodium phosphate, NaCl buffer solution.
E. luminescent solution: A liquid is luminol, p-cresol solution, B liquid hydrogen peroxide urea solution, during use, A liquid, the mixing of B liquid equal-volume, now with the current.
F. redissolve liquid, during use, be diluted to working concentration with distilled water.
G. cleaning mixture: be diluted to working concentration with distilled water during use.
(2) preparation of Chemiluminescent plate
With being coated liquid, anti-sulfa drugs monoclonal antibody is diluted to 4.0 μ g/mL, every hole adds 100 μ L, 2h placed by 37 DEG C of calorstats, inclines and is coated liquid, pats dry, then every hole adds confining liquid 150 μ L, 37 DEG C of calorstats place 2h, liquid in hole of inclining, and cleaning mixture washs once, pat dry, seal by masking foil vacuum and preserve.
Embodiment 5 detects the application of the chemiluminescence enzyme linked immunoassay reagent kit of sulfa drugs
(1) preparation of reagent
A. cleaning mixture: use after the concentrated cleaning solution deionized water provided in test kit is diluted by 1:19 times.
B. working solution: the concentrated phosphoric acid salt buffer provided in test kit is spent and uses after ionized water dilutes by 1:1 times.
C. chemiluminescent solution: by A liquid and B liquid 1:1 by volume mixing before using.
D. enzyme-labelled antigen working solution: enzyme-labelled antigen diluent and enzyme-labelled antigen concentrated solution are mixed by 10:1 volume ratio and mixes.
(2) sample pre-treatments
A. Carnis Sus domestica, Carnis Gallus domesticus, the flesh of fish
With homogenizer homogenizing sample；Weighing 2.0g ± 0.05g sample to 50ml polystyrene centrifuge tube, add 200 μ l0.1M sodium hydroxide, add 3.8ml acetonitrile, add 2ml ethyl acetate, whirling motion mixes, and more than 3000g, room temperature (20 ~ 25 DEG C) is centrifuged 5min；Pipette 3ml upper organic phase to 10ml clean dried glass tubing, flow down in 50 ~ 60 DEG C of water-bath nitrogen and dry up；Adding 1ml normal hexane, with vortex instrument whirling motion 1min, then add 1ml and redissolve working solution, with vortex instrument whirling motion 1min, more than 3000g, room temperature (20 ~ 25 DEG C) is centrifuged 5min；Remove upper organic phase, take off layer aqueous phase for analyzing.
B. Macrobrachium nipponensis
Weighing 2.0 ± 0.05g sample to 50ml polystyrene centrifuge tube, add 200ul0.1M sodium hydroxide, add 3.8ml acetonitrile, whirling motion mixes, and adds 2ml ethyl acetate, and whirling motion mixes, and more than 3000g, room temperature (20 ~ 25 DEG C) is centrifuged 5min；Pipette 3ml upper organic phase to 10ml clean dried glass tubing, flow down in 50～60 DEG C of water-bath nitrogen and dry up；Adding 1ml normal hexane, with vortex instrument whirling motion 1min, then add 1ml and redissolve working solution, with vortex instrument whirling motion 1min, more than 3000g, room temperature (20 ~ 25 DEG C) is centrifuged 5min；Remove upper organic phase, take off layer aqueous phase for analyzing.
(3) detecting step
A. application of sample: add standard substance/sample 50 μ L in corresponding micropore, adds enzyme-labelled antigen working solution 50 μ L/ hole, and mixing of vibrating gently, with reacting 15min in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate.
B. washing: carefully open cover plate film, dried by liquid in hole, with wash operating solution 250 μ L/ hole, fully washing 7 times, every minor tick 10s, pat dry with absorbent paper；
C. luminescent solution is added: every hole adds the luminous substrate 100 μ L of new preparation, shakes about about 30 seconds, places 3min by room temperature after cover plate membrane cover plate.
D. detection: be directly placed into survey measurements in microwell plate luminescence analyzer.
(4) result judges
The standard substance obtained and the meansigma methods of sample luminous intensity values are multiplied by 100 again divided by the luminous intensity values of first standard (0 standard), with suppression ratio for vertical coordinate, the logarithm of sulfonamides substrate concentration is that abscissa makes standard curve, and the concentration of each sample can read from standard curve.
Relative luminous intensity (%)=RLU/RLU0, RLU is standard substance or the luminous intensity values of sample solution mensuration, and RLU0 is the luminous intensity values of blank (concentration is the standard solution of 0).
Embodiment 6 test kit preci-sion and accuracy is tested
Accuracy refers to the matching degree between measured value and true value, and the conventional response rate of test kit accuracy represents.Precision is also known as repeatability, and the conventional coefficient of variation represents.
Sample extraction method according to embodiment 5, it is added reclaiming to Carnis Sus domestica, Carnis Gallus domesticus, the flesh of fish, Macrobrachium nipponensis sample respectively with the sulfa drugs of tri-concentration of 1ug/kg, 2ug/kg, 4ug/kg, every kind of each concentration of sample each 4 parallel, it is measured with three batches of test kits, calculates average recovery rate and the precision of sample.Experimental result is shown in following table.
Table 1 sulfa drugs test kit accuracy and precision measure
As seen from the table, the average recovery rate scope that in Carnis Sus domestica, Carnis Gallus domesticus, the flesh of fish, Macrobrachium nipponensis sample, three concentration of sulfa drugs are all added between 89.2-101.5%, in batch, batch between all the coefficient of variation less than 10%.
Embodiment 7 test kit specific test
Specificity cross reacting rate represents, cross reacting rate refers to that the ability combined occurs the antibody antigenic determinant different from structure.Cross reacting rate is little, and the specificity of provable antibody is high.Using sulfadiazine as base standard product, cross reacting rate is 100%, selects sulfa drugs, by the sulfa drugs of variable concentrations, substitutes sulfadiazine standard solution, measures its standard curve, and calculate IC50Inhibition concentration, calculates cross reacting rate.
Result of calculation is following table such as.
Table 2 sulfa drugs cross reacting rate
Claims (3)
1. the chemical luminescence ELISA detection kit detecting sulfa drugs, including sulfa drugs standard solution, Chemiluminescent plate, enzyme-labelled antigen concentrated solution, enzyme-labelled antigen diluent, luminous substrate liquid, cleaning mixture, redissolution liquid, each hole of described Chemiluminescent plate is coated with the monoclonal antibody of anti-sulfa drugs.
2. the chemiluminescence enzyme linked immunoassay reagent kit detecting sulfa drugs as claimed in claim 1, it is characterised in that: it is 8.0ug/mL that described anti-sulfa drugs monoclonal antibody is coated concentration.
3. the as claimed in claim 1 chemiluminescence enzyme linked immunoassay reagent kit detecting sulfa drugs, it is characterised in that: described sulfa drugs standard solution concentration is respectively as follows: 0,2,4,8,16,32ug/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107247135A (en) * | 2017-08-11 | 2017-10-13 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfamethazine and preparation method thereof |
| CN107478643A (en) * | 2017-08-24 | 2017-12-15 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfaquinoxaline and preparation method thereof |
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| CN1766631A (en) * | 2005-11-03 | 2006-05-03 | 北京望尔生物技术有限公司 | ELISA kit for detecting sulfanilamides residue in animal derived food |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107247135A (en) * | 2017-08-11 | 2017-10-13 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfamethazine and preparation method thereof |
| CN107478643A (en) * | 2017-08-24 | 2017-12-15 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfaquinoxaline and preparation method thereof |
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Application publication date: 20160713 |