CN101799472A - Diethylstilbestrol detection kit and detection method - Google Patents
Diethylstilbestrol detection kit and detection method Download PDFInfo
- Publication number
- CN101799472A CN101799472A CN201010132778A CN201010132778A CN101799472A CN 101799472 A CN101799472 A CN 101799472A CN 201010132778 A CN201010132778 A CN 201010132778A CN 201010132778 A CN201010132778 A CN 201010132778A CN 101799472 A CN101799472 A CN 101799472A
- Authority
- CN
- China
- Prior art keywords
- diethylstilbestrol
- kit
- antibody
- detection
- reagent
- 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
Images
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a diethylstilbestrol chemiluminescent detection kit, comprising a detection plate and a reagent. The reagent comprises an antibody which is a monoclonal antibody prepared by using a conjugate of diethylstilbestrol and bovine serum albumin as an immunogen. The diethylstilbestrol detection kit can realize quick and large-scale detection and has very high specificity and sensitivity, wherein the detection sensitivity reaches 0.016 ppb.
Description
Technical field
The present invention relates to technical field of bioengineering, relate in particular to a kind of diethylstilbestrol detection kit and detection method thereof.
Background technology
(Diethylstibestrol DES) is the estrogen of synthetic to diethylstilbestrol, because it has the weightening finish of the animal of promotion, increase effect proteins deposited and the minimizing fat deposition, once extensively used, improved the fish meal conversion ratio, promoted fish growth as fish feed additive.Since the seventies in 20th century, people found through experiments, and DES has teratogenesis and carcinogenic toxicity.At present, many in the world countries have been defined in livestock and poultry and the aquaculture and have forbidden DES, and regulation cancellation DES of China Ministry of Agriculture in 2002 and salt thereof, ester formulation are in all purposes of all food animals.Yet, be subjected to ordering about of economic interests, culture the violated situation of adding diethylstilbestrol of merchant and still exist, have a strong impact on China's livestock products, aquatic products reputation in the world, caused heavy economic losses.Unimpeded in order to ensure animal foodstuff safety and China's animal husbandry and fishery product export trade, setting up quick, sensitive detection technique becomes the task of top priority.
Traditional medicament residue analytical approach comprises gas chromatography, liquid chromatography, mass spectrum etc., and the required instrument of these methods costs an arm and a leg, and experimentation is loaded down with trivial details, and requirement is detected at the scene that is difficult to reach quick, easy.At present, that is used widely in the medicament residue fast detecting has microbial method and an euzymelinked immunosorbent assay (ELISA), and still, these two kinds of methods usually do not reach requirement for detecting forbidden drugs in sensitivity.(Chemiluminescence Immunoassay CLIA) for the rapid screening method of representative is the immunology mainstream technology, but does not still have the report that utilizes chemiluminescence immune assay (CLIA) method to detect diethylstilbestrol both at home and abroad at present with chemiluminescence immune assay.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of chemiluminescence immune assay (CLIA) kit of diethylstilbestrol, and this kit not only can be realized fast, mass detection, and detection specificity and sensitivity significantly improve.
In addition, also need to provide a kind of detection method of mentioned reagent box.
In order to solve the problems of the technologies described above, the present invention is achieved through the following technical solutions:
In one aspect of the invention, provide a kind of diethylstilbestrol detection kit, comprised check-out console, reagent, described reagent comprises antibody, and described antibody is the monoclonal antibody that the conjugate with diethylstilbestrol and bovine serum albumin(BSA) goes out as immunogen preparing.
Immunogene adopts the conjugate of diethylstilbestrol and bovine serum albumin(BSA), is because during Antibody Preparation, have only the bigger compound of antigen molecular (＞could the interior generation of induced animal body antibody when 5000D) being expelled in the animal body.If the compound of small-molecular weight (this compounds is haptens), directly induced animal produces antibody, and in order to make antibody, need this micromolecular compound is coupled to macromolecular carrier (as bovine serum albumin(BSA), BSA) make antigen on, use it for immune animal again, preparation antibody.
Each hole of described check-out console is coated with envelope antigen, and this envelope antigen is the conjugate for diethylstilbestrol and oralbumin.
The present invention adopts the conjugate of diethylstilbestrol and oralbumin as each hole of antigen coated check-out console, add testing sample and anti-diethylstilbestrol monoclonal antibody then, allow envelope antigen and testing sample compete antibody with the diethylstilbestrol in the test sample through pre-treatment.
Preferably, described check-out console is the chemiluminescence plate, is applicable to that chemiluminescence immune assay detects.
Preferably, the reagent in the kit of the present invention also comprises ELIAS secondary antibody, and this ELIAS secondary antibody comprises the goat anti-mouse igg concentrate of horseradish peroxidase-labeled.
Preferably, the reagent in the kit of the present invention also comprises substrate reactions liquid, and this substrate reactions liquid comprises Tris-HCl damping fluid, luminol, p-Coumaric Acid and hydrogen peroxide.
Preferably, the reagent in the kit of the present invention also comprises the diethylstilbestrol standard solution.
Preferably, the reagent in the kit of the present invention also comprises concentrated cleaning solution, and this concentrated cleaning solution comprises tween and phosphate buffer.
In another aspect of this invention, also provide a kind of detection method of diethylstilbestrol, may further comprise the steps:
The pre-treatment testing sample; With above-mentioned kit test sample; Analyzing and testing result.
The step of described pre-treatment testing sample comprises: testing sample is smashed and used ethyl acetate extraction, ethyl acetate extract is after nitrogen dries up, remaining residue n-hexane dissolution, the mixed solution vibration that adds phosphate buffer and methyl alcohol again is centrifugal, takes off layer solution and is used for detecting.
Diethylstilbestrol detection kit of the present invention has following advantage:
(1) high specificity, the susceptibility height, security is good.The present invention detects chemiluminescence immune assay (CLIA) kit of diethylstilbestrol in the animal derived food, monoclonal antibody based on diethylstilbestrol, this anti-diethylstilbestrol monoclonal antibody does not contain the antibody at carrier protein, only combine, not with other antibiotic medicine of animal and disinfectant generation cross reaction with the trim of diethylstilbestrol and diethylstilbestrol.Therefore, kit of the present invention has very high specificity and susceptibility, and detection sensitivity reaches 0.016ppb.
(2) easy and simple to handle quick.When using the diethylstilbestrol chemical luminescence immune analysis reagent box to detect diethylstilbestrol, need not to join other reagent in addition, sample need not aseptic process, is the decidable testing result in 3-4 hour by the kit explanation.
(3) result judge accurately, reliable.Diethylstilbestrol chemiluminescence immune assay detection kit of the present invention quantitatively shows testing result with the machine-readable number of multi-functional microplate reader, reduces subjectivity, accurately and reliably.
(4) cost is low, small investment.But diethylstilbestrol chemiluminescence immune assay detection kit batch detection of the present invention settles at one go, and is with low cost, small investment, instant effect.
Description of drawings
The present invention is further detailed explanation below in conjunction with the drawings and specific embodiments.
Fig. 1 is the canonical plotting of the DES test sample of the embodiment of the invention 3.
Embodiment
Diethylstilbestrol detection kit of the present invention comprises check-out console, reagent, and described reagent comprises antibody, and this antibody is the monoclonal antibody that the conjugate with diethylstilbestrol and bovine serum albumin(BSA) goes out as immunogen preparing.Use the diethylstilbestrol in the kit detection animal derived food of the present invention, can significantly improve detection sensitivity, its check and analysis principle is: each Kong Jun on the chemiluminescence plate is coated with the diethylstilbestrol and oralbumin conjugate (DES-OVA) antigen of same amount, after adding diethylstilbestrol sample to be measured and diethylstilbestrol monoclonal antibody, envelope antigen and diethylstilbestrol sample to be measured are competed diethylstilbestrol antibody together, because the solid phase antigen in each hole and the antibody content of adding are all consistent, so when diethylstilbestrol concentration to be measured is high, the antibody that then is bonded on the solid phase antigen is few, the ELIAS secondary antibody that adds is few with the antibodies amount that is fixed, add substrate reactions liquid with cleansing solution washing back, luminous value is low, shows the inhibiting rate height; Otherwise when diethylstilbestrol concentration to be measured was low, the luminous value height of then being surveyed showed that inhibiting rate is low.According to detecting the typical curve of being done, can extrapolate the diethylstilbestrol concentration of testing sample with known diethylstilbestrol concentration.
(1) immunogenic preparation
Diethylstilbestrol (DES) and bovine serum albumin(BSA) are carried out coupling, get the DES-BSA conjugate and be immunogene.
(2) Monoclonal Antibody
With above-mentioned synthetic immunogen immune mouse, Freund's complete adjuvant with immunogene and equivalent when head exempts from is mixed and made into emulsifying agent, at the subcutaneous multi-point injection of mouse web portion, later on every two weeks, add emulsifying agent booster immunization that the equivalent incomplete Freund is mixed and made into once with immunogene, the immunity position is that nape portion is subcutaneous, and from immunity for the third time, each immunity one week of back is detected serum titer from the mouse orbit blood sampling.Immune 5 times altogether, last immunity one week of back is merged the splenocyte and the SP2/0 myeloma cell of immune mouse, again through 4 subclones, screens the hybridoma cell strain of energy stably excreting diethylstilbestrol monoclonal antibody.By extracorporeal culture-ing, the collecting cell culture supernatant is removed cell fragment after centrifugal, and supernatant-20 ℃ preservation is standby.Adopt in the body and induce the ascites method, only inject sterilization paraffin oil 0.5ml/, 7-14 days pneumoretroperitoneum injection hybridomas 5 * 10 to mouse peritoneal
5～10
6Individual/as only, to gather ascites after 7-14 days.Ascites is centrifugal, abandon fat deposit and cellular layer, the clear layer in the middle of collecting ,-20 ℃ of preservations are standby.Saturated ammonium sulfate salting out method with 35% is slightly put forward cells and supernatant and ascites, is further purified with the DE-52 anion exchange chromatography at last, diethylstilbestrol monoclonal antibody that must be purer.
Monoclonal antibody is made working fluid: 100x diethylstilbestrol monoclonal antibody (is anti-) concentrate is that 2 μ l and 50 μ l cow's serums add in 50% glycerol liquor of 148 μ l sterilization.Antibody dilution buffer: the 0.01M phosphate buffer (pH7.4) of 0.05% Tween-20 of 5% cow's serum.Monoclonal antibody (one is anti-) concentrate becomes the monoclonal antibody working fluid for 100 times with the antibody dilution buffer dilution.
The preparation of check-out console and reagent in embodiment 2 diethylstilbestrol detection kit
In this embodiment 2, be provided with check-out console, diethylstilbestrol standard solution, diethylstilbestrol monoclonal antibody, ELIAS secondary antibody, substrate reactions liquid, sample diluting liquid, concentrated cleaning solution in the diethylstilbestrol detection kit, it is prepared as follows:
(1) preparation of diethylstilbestrol micropore check-out console
0.2M carbonate buffer solution with pH9.6 is made coating buffer, and diethylstilbestrol and oralbumin conjugate (DES-OVA) are diluted to 200ng/ml, adds in the chemiluminescence orifice plate by 100 μ l/ holes, and 4 ℃ of bags are spent the night.Dry, add by 200 μ l/ holes and contain 1% gelatin, 1% Sodium azide is used in 37 ℃ of sealings of the phosphate buffer of pH7.4 2 hours, and the phosphate cleansing solution of pH7.4 is washed 3 times, dries, and preserves in the packaging bag that contains drying agent of packing into.
(2) preparation of DES standard solution
Accurately take by weighing DES 1mg, earlier with dissolve with methanol and make 5 times of gradient dilutions, then by PBS: methyl alcohol=3: 2 (v/v) adds the 0.01M phosphate buffer that contains 0.05% Tween-20 and prepares that series concentration is 31.25,6.25,1.25,0.25, the standard DES solution of 0.05ng/ml.
(3) preparation of substrate reactions liquid
Substrate colour developing liquid preparation composition and ratio are 0.1M pH8.5 Tris-HCl damping fluid (Tris 6.057g, pH8.5, constant volume 500ml) 1ml, 2mM luminol (the 0.1758g luminol is dissolved among the 2ml DMSO) 4 μ l, 0.25mM reinforcing agent p-Coumaric Acid (the 0.2g p-Coumaric Acid is dissolved in the 3ml absolute ethyl alcohol) 0.625 μ l, 4mM hydrogen peroxide 1.6 μ l.
(4) sample diluting liquid, concentrated cleaning solution
Sample diluting liquid be 1X PBS-methyl alcohol (3: 2, v/v) promptly get 0.01M pH 7.4 phosphate buffer (KH
2PO
40.2g, KCl 0.2g, Na
2HPO4 12H
2O 2.9g, NaCl8.0g, constant volume 1000mL) 60mL mixes with absolute methanol 40mL; 10 * concentrated cleaning solution is the 0.1M phosphate buffer (KH that contains 0.05% Tween-20
2PO
42g, KCl 2g, Na
2HPO412H
2O 29g, NaCl 80g is settled to 1000ml, pH7.4, Tween-205mL).
(5) ELIAS secondary antibody is the goat anti-mouse igg concentrate of 100x horseradish peroxidase-labeled.
Embodiment 3 utilizations kit of the present invention detects diethylstilbestrol
(1) sample pre-treatments
The pre-treatment of animal tissue (chicken, pig), fish, shrimp sample:
1. get the 3g sample and place centrifuge tube, add 9ml ethyl acetate, under homogenizer, fully smash;
2. add 0.3gCaCl then
2, maximal rate vibration 3min; Under the room temperature condition, the centrifugal 5min of 6000rpm;
3. pipette the 6ml supernatant to another new test tube, 60-70 ℃ of water-bath nitrogen dries up;
4. residue is with the n-hexane dissolution of 2.0ml, add again 1.0ml 1X PBS-methyl alcohol (3: 2, v/v), vortex oscillation 2min;
5. under the room temperature condition, the centrifugal 10min of 6000rpm absorbs the upper strata normal hexane, gets 50 μ l lower floor solution and is used for detecting.
(2) kit detection step is as follows:
1. be cleansing solution for 10 times with 10 * concentrated cleaning solution dilution, with one anti-, two anti-ly carry out 10 times of dilutions with antibody dilution buffer;
2. in the suitable micropore of the chemiluminescence plate that has sealed, add the serial DES standard solution that 50 μ l/ holes prepare (31.25,6.25,1.25,0.25,0.050ppb) respectively, in other micropore, add 50 μ l and finished the sample solution of pre-treatment, also in a hole, add control wells that 100 μ l cleansing solutions the do not add monoclonal antibody control wells when being used to measure luminous value.
3. except that control wells, add the suitably DES monoclonal antibody of dilution of 50 μ l again in each micropore, hatched 90 minutes for 37 ℃, dry, every hole adds cleansing solution 300 μ l, washs 3 times, each 2 minutes at interval, pats dry;
4. every hole adds HRP enzyme mark sheep anti mouse (two the is anti-) working fluid of 100 μ l, hatches 90 minutes for 37 ℃, dries, and every hole adds cleansing solution 300 μ l, washs 3 times, each 2 minutes at interval, pats dry;
5. every hole adds 150 μ l substrate reactions liquid and reads chemiluminescence numerical value in 2 minutes.
(3) criterion as a result
Calculate DES standard model inhibiting rate (B/B earlier
0), be ordinate then with the inhibiting rate, be horizontal ordinate with the logarithm value of DES standard model concentration, do typical curve.B/B according to each test sample
0Value just can be read the concentration of its corresponding DES from typical curve.Multiply by corresponding extension rate again.
1) typical curve and sensitivity
With inhibiting rate I is ordinate, and the logarithm value of DES concentration of standard solution is a horizontal ordinate, draws out the typical curve (see figure 1).Inhibiting rate I=B/B
0(B is standard items values of chemiluminescence or sample chemical luminous value, B
0Be the blank values of chemiluminescence).
As can be seen from Figure 1, the B/B of DES in the 0.05ng/ml-31.25ng/ml scope
0(claiming relative light absorption value again) is linear with the logarithm value of DES concentration, carries out regretional analysis, and regression equation is Y=-0.2632x+0.4505, R
2=0.9904, IC
50Value is used B for 0.649ng/ml
0The detection sensitivity that-3SD extrapolation method calculates kit of the present invention is 0.016ppb (ng/ml).
2) repeatability as a result
Criticize the degree of accuracy that an error is represented this method with batch interior sum of errors.1. criticize interior error: each standard model concentration is done 5 times and is repeated, and represents batch interior error with its variation within batch coefficient.2. criticize an error: the measurement operation that repeats to have set up, make error between representing to criticize with its interassay coefficient of variation continuously 5 times.CV%=combination rate (B/B
0) mean value/(combination rate in 1.414 * average batch) * 100% of SD.Error between batch is asked the mean value of the combination rate of 5 mensuration, and is asked SD, asks its total interassay coefficient of variation (CV) by above-mentioned formula.Variation within batch CV%=4.23%, batch variation CV%=6.60%, good reproducibility (seeing Table 1)
Table 1 typical curve criticize interior and batch between error
3) recovery is calculated
Be taken at adding DES standard specimen in the blank swamp eel meat of the 3g sample, addition is respectively 25ng/g, 10ng/g, 2ng/g, and each adds the concentration sample and does three repetitions, carries out sample pre-treatments, and extract carries out chemical luminescent detecting.Calculate relative light absorption value after measuring luminous value, obtain concentration value, determine its accuracy (seeing the following form 2) with the recovery from typical curve.
Table 2 swamp eel meat sample DES adds the recovery test result
Other kind sample recovery rate sees the following form 3.
Other kind sample recovery rate of table 3
| Sample type | Pork | Liver (pig) | Chicken |
| The recovery | ??85％±15％ | ??81±15％ | ??86±13％ |
4) specificity as a result
Judge that the specific index of kit is a crossing-over rate, therefore carry out specificity cross reaction test at medicine.Utilize chemiluminescence immune assay (CLIA) to determine the cross reacting rate of the common forbidding hormone of part and antibacterials in DES and DES similar compound, animal products, the aquaculture, result such as following table 4.The result shows, the DES monoclonal antibody in the kit of the present invention is except having the cross reaction with diethylstilbestrol list carboxylic propyl ether, with other medicines no cross reaction all.
The cross reaction of table 4DES and DES similar compound and other aquaculture forbidden drugs
The above embodiment has only expressed embodiments of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to claim of the present invention.Should be pointed out that for the person of ordinary skill of the art without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (9)
1. a diethylstilbestrol detection kit comprises check-out console, reagent, and described reagent comprises antibody, it is characterized in that, described antibody is the monoclonal antibody that the conjugate with diethylstilbestrol and bovine serum albumin(BSA) goes out as immunogen preparing.
2. kit according to claim 1 is characterized in that, each hole of described check-out console is coated with envelope antigen, and this envelope antigen is the conjugate of diethylstilbestrol and oralbumin.
3. kit according to claim 1 and 2 is characterized in that, described check-out console is the chemiluminescence plate.
4. kit according to claim 1 is characterized in that described reagent also comprises ELIAS secondary antibody.
5. kit according to claim 1 is characterized in that described reagent also comprises substrate reactions liquid, and this substrate reactions liquid comprises Tris-HCl damping fluid, luminol, p-Coumaric Acid and hydrogen peroxide.
6. kit according to claim 1 is characterized in that described reagent also comprises the diethylstilbestrol standard solution.
7. kit according to claim 1 is characterized in that described reagent also comprises concentrated cleaning solution, and this concentrated cleaning solution comprises tween and phosphate buffer.
8. the detection method of a diethylstilbestrol is characterized in that, may further comprise the steps:
The pre-treatment testing sample;
With the described kit test sample of claim 1;
Analyzing and testing result.
9. detection method according to claim 8, it is characterized in that, the step of described pre-treatment testing sample comprises: testing sample is smashed and used ethyl acetate extraction, ethyl acetate extract is after nitrogen dries up, remaining residue n-hexane dissolution, the mixed solution vibration that adds phosphate buffer and methyl alcohol again is centrifugal, takes off layer solution and is used for detecting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010132778A CN101799472A (en) | 2010-03-25 | 2010-03-25 | Diethylstilbestrol detection kit and detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010132778A CN101799472A (en) | 2010-03-25 | 2010-03-25 | Diethylstilbestrol detection kit and detection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101799472A true CN101799472A (en) | 2010-08-11 |
Family
ID=42595235
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010132778A Pending CN101799472A (en) | 2010-03-25 | 2010-03-25 | Diethylstilbestrol detection kit and detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101799472A (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936985A (en) * | 2010-08-03 | 2011-01-05 | 中国农业大学 | Method for detecting diethylstilbestrol and special chemiluminescent immunoassay kit thereof |
| CN101988923A (en) * | 2010-08-31 | 2011-03-23 | 华南农业大学 | Time resolution immunoassay detection kit of diethylstilbestrol residuals and detection method thereof |
| CN101995460A (en) * | 2010-08-31 | 2011-03-30 | 华南农业大学 | Ractopamine residual time resolution immunoassay kit and detection method thereof |
| CN102253224A (en) * | 2011-06-13 | 2011-11-23 | 清华大学深圳研究生院 | Immunochromatographic test paper for detecting diethylstilbestrol and preparation method thereof |
| CN102288770A (en) * | 2011-07-07 | 2011-12-21 | 清华大学深圳研究生院 | Immunofluorescence diethylstilbestrol detecting method based on quantum dots and special kit |
| CN102661946A (en) * | 2012-04-11 | 2012-09-12 | 广州万联生物科技有限公司 | Malachite chemiluminescence ELISA detection method and kit |
| CN103207272A (en) * | 2012-01-13 | 2013-07-17 | 中华人民共和国上海出入境检验检疫局 | A method of simultaneous detection of medroxyprogesterone acetate and diethylstilbestrol in aquatic products |
| CN103207275A (en) * | 2012-01-13 | 2013-07-17 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | A kit for the simultaneous detection of medroxyprogesterone acetate and diethylstilbestrol in aquatic products |
| CN106124772A (en) * | 2016-06-15 | 2016-11-16 | 金华职业技术学院 | A kind of ELISA detection kit based on OMP18 detection campylobacter jejuni and application thereof |
| CN106442512A (en) * | 2016-11-23 | 2017-02-22 | 百奥森（江苏）食品安全科技有限公司 | Detection kit for detecting diethylstilbestrol in food |
| CN107167614A (en) * | 2017-04-24 | 2017-09-15 | 郑州大学 | A kind of hemoglobin cataluminescence enzyme-linked immune detection method of diethylstilbestrol |
-
2010
- 2010-03-25 CN CN201010132778A patent/CN101799472A/en active Pending
Non-Patent Citations (6)
| Title |
|---|
| 《Analytica Chimica Acta》 20041231 Jian Wang et al Determination of diethylstilbestrol by enhancement of luminol-hydrogen peroxide-tetrasulfonated cobalt phthalocyanine chemiluminescence 171-176 第508卷, * |
| 《中国优秀硕士学位论文全文数据库 农业科技辑》 20090915 常小斌 己烯雌酚单克隆抗体的制备及初步应用 D050-237 , 第9期 * |
| E.H.J.M.JANSEN ET AL: "POSSIBILITIES AND LIMITATIONS OF CHEMILUMINESCENT IMMUNOASSAYS", 《PHARMACEUTISCH WEEKBLAD SCIENTIFIC EDITION》 * |
| JIAN WANG ET AL: "Determination of diethylstilbestrol by enhancement of luminol–hydrogen peroxide–tetrasulfonated cobalt phthalocyanine chemiluminescence", 《ANALYTICA CHIMICA ACTA》 * |
| 常小斌: "己烯雌酚单克隆抗体的制备及初步应用", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 廖敏 等: "酶联免疫法：水产品中己烯雌酚检测新招", 《农业技术与装备》 * |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936985B (en) * | 2010-08-03 | 2013-01-09 | 中国农业大学 | Method for detecting diethylstilbestrol and special chemiluminescent immunoassay kit thereof |
| CN101936985A (en) * | 2010-08-03 | 2011-01-05 | 中国农业大学 | Method for detecting diethylstilbestrol and special chemiluminescent immunoassay kit thereof |
| CN101988923A (en) * | 2010-08-31 | 2011-03-23 | 华南农业大学 | Time resolution immunoassay detection kit of diethylstilbestrol residuals and detection method thereof |
| CN101995460A (en) * | 2010-08-31 | 2011-03-30 | 华南农业大学 | Ractopamine residual time resolution immunoassay kit and detection method thereof |
| CN101995460B (en) * | 2010-08-31 | 2013-12-25 | 华南农业大学 | Ractopamine residual time resolution immunoassay kit and detection method thereof |
| CN102253224B (en) * | 2011-06-13 | 2013-10-30 | 清华大学深圳研究生院 | Immunochromatographic test paper for detecting diethylstilbestrol and preparation method thereof |
| CN102253224A (en) * | 2011-06-13 | 2011-11-23 | 清华大学深圳研究生院 | Immunochromatographic test paper for detecting diethylstilbestrol and preparation method thereof |
| CN102288770A (en) * | 2011-07-07 | 2011-12-21 | 清华大学深圳研究生院 | Immunofluorescence diethylstilbestrol detecting method based on quantum dots and special kit |
| CN102288770B (en) * | 2011-07-07 | 2013-11-27 | 清华大学深圳研究生院 | Immunofluorescence diethylstilbestrol detecting method based on quantum dots and special kit |
| CN103207275A (en) * | 2012-01-13 | 2013-07-17 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | A kit for the simultaneous detection of medroxyprogesterone acetate and diethylstilbestrol in aquatic products |
| CN103207272A (en) * | 2012-01-13 | 2013-07-17 | 中华人民共和国上海出入境检验检疫局 | A method of simultaneous detection of medroxyprogesterone acetate and diethylstilbestrol in aquatic products |
| CN102661946A (en) * | 2012-04-11 | 2012-09-12 | 广州万联生物科技有限公司 | Malachite chemiluminescence ELISA detection method and kit |
| CN102661946B (en) * | 2012-04-11 | 2014-08-20 | 广州万联生物科技有限公司 | Malachite chemiluminescence ELISA detection method and kit |
| CN106124772A (en) * | 2016-06-15 | 2016-11-16 | 金华职业技术学院 | A kind of ELISA detection kit based on OMP18 detection campylobacter jejuni and application thereof |
| CN106124772B (en) * | 2016-06-15 | 2018-05-04 | 金华职业技术学院 | A kind of ELISA detection kit and its application based on OMP18 detection campylobacter jejunis |
| CN106442512A (en) * | 2016-11-23 | 2017-02-22 | 百奥森（江苏）食品安全科技有限公司 | Detection kit for detecting diethylstilbestrol in food |
| CN107167614A (en) * | 2017-04-24 | 2017-09-15 | 郑州大学 | A kind of hemoglobin cataluminescence enzyme-linked immune detection method of diethylstilbestrol |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101799472A (en) | Diethylstilbestrol detection kit and detection method | |
| CN102955031B (en) | Enzyme-linked immunosorbent assay kit for detecting aflatoxin B1-containing medicine and application for same | |
| CN101962358B (en) | Ciprofloxacin hapten, artificial antigen and antibody and preparation method and application thereof | |
| CN101113981A (en) | Sulfonamides direct-competition ELISA detecting reagent kit | |
| CN105137009B (en) | The enzyme linked immunological kit of detection carbofuran and application thereof | |
| CN103901201B (en) | Cefalexin residual fluorescence immunochromatographydetecting detecting test strip and preparation thereof | |
| CN102080066B (en) | Method for detecting T-2 toxin and special reagent kit thereof | |
| CN101021536B (en) | Enzyme-linked immunological kit for detecting neomycin drug and method | |
| CN101021535B (en) | Enzyme-linked immunalogical kit for detecting gentamicin medicine and method | |
| CN105838681B (en) | One plant of anti-dexamethasone monoclonal antibody specific hybridoma cell strain C3 and its application | |
| CN105131121B (en) | Detect monoclonal antibody, ELISA method and the kit of Furaxone metabolite | |
| CN103018440B (en) | A kind of clopidol colloidal gold chromatographic test strip or card | |
| CN101776685A (en) | Enzyme linked immunosorbent assay kit for detecting trimethoprim medicament and application thereof | |
| CN106831498B (en) | Furacilin metabolite SEM derivatizations haptens, artificial antigen preparation method and applications | |
| CN101434655A (en) | Preparation of egg yolk antibody against citrinin | |
| CN101806796A (en) | Detection kit and method of furazolidone metabolin | |
| CN103364553B (en) | The enzyme linked immunological kit of detection nitroimidazoles medicine and application thereof | |
| CN110256298A (en) | 4,4 '-dinitro phenylurea haptens and artificial antigen and the preparation method and application thereof | |
| CN105273021B (en) | Erythromycin haptens, artificial antigen and antibody and its preparation method and application | |
| CN101962359B (en) | Hapten, artifical antigen and antibody of enrofloxacin and preparation method as well as application thereof | |
| CN105758846A (en) | Chemiluminescence enzyme-linked immunosorbent assay reagent kit for detecting clenbuterol | |
| CN100465645C (en) | A kit of enzyme-linked immunity detection for toxin of microcapsule alga | |
| CN103288661A (en) | Preparation method and application of malachite green hapten | |
| CN104311659A (en) | Multi-cluster antigen and wide-spectrum specific antibody of beta-adrenoceptor agonists and application of multi-cluster antigen and wide-spectrum specific antibody | |
| CN104744294A (en) | Preparation method and application of di(alpha-ethyl hexyl) phthalate (DEHP) half antigen, artificial antigen and antibody |
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: 20100811 |