CN104897864A - ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine and application of ELISA kit - Google Patents
ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine and application of ELISA kit Download PDFInfo
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Abstract
The invention provides an ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine. The ELISA kit comprises an ELISA plate coated with a coating antigen, an amantadine standard product solution, an enzyme-labeled antibody, a substrate developing solution, a stop solution, a washing solution and a combination solution, wherein the coating antigen is an amantadine coupling antigen, and the enzyme-labeled antibody is an enzyme-labeled amantadine antibody. The invention further discloses a method for applying the ELISA kit to amantadine detection. The method comprises steps as follows: sample pretreatment is performed firstly, then the kit is used for detection, and a detection result is analyzed finally. The ELISA kit can be used for detecting the content of amantadine in an animal tissue sample, is simple and convenient to operate, low in cost and high in sensitivity, can be used for performing on-site monitoring and is suitable for screening a large number of samples.
Description
Technical field
The present invention relates to enzyme linked immunosorbent detection technology, being specifically related to a kind of enzyme linked immunological kit for detecting amantadine, can the residual quantity of amantadine medicine in qualitative and quantitative analysis animal tissue.
Background technology
Amantadine belongs to the antiviral class medicine suppressing influenza virus, be usually used in bird opposing bird flu, thus the favor of numerous raisers is subject to, the current U.S. has determined that its validity for antiparkinsonism drug is uncertain, the data of for want of security and validity, amantadine treats influenza no longer recommended being used as of the U.S., and to nervous system, cardiovascular system has certain toxic and side effect, and domestic having forbidden uses as veterinary drug.
At present, the provincial standard detection Ultra Performance Liquid Chromatography tandem mass spectrometry of amantadine, Rimantadine " in DB21/2394-2014 food security provincial standard chicken gizzard and the chicken ", the mensuration Liquid Chromatography-Tandem Mass Spectrometry of amantadine residual quantity " in the DB32/T 1163-2007 chicken gizzard " all adopts instrumental method, instrumental method has the advantages such as highly sensitive, result is accurate, but the input cost such as fund and personnel is higher.The present invention's application euzymelinked immunosorbent assay (ELISA), measures the residual quantity of amantadine medicine in animal tissue, has the advantages such as detectability is low, high specificity, easy and simple to handle, detection speed is fast, testing cost is low, very easy popularization.
Summary of the invention
The object of the present invention is to provide a kind of enzyme linked immunological kit that can detect amantadine drug residue in animal tissue, and provide a kind of efficient, accurate, easy, be suitable for batch samples screening qualitative and quantitative analysis method.
Kit of the present invention, it comprises: be coated with the ELISA Plate of coating antigen, amantadine standard solution, enzyme labelled antibody, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid, described coating antigen is amantadine coupled antigen, and described enzyme labelled antibody is the amantadine antibody of enzyme labeling.
Described amantadine coupled antigen is obtained by amantadine haptens and carrier protein couplet, described amantadine haptens is obtained by reacting by amantadine and β-chloropropionic acid, and described carrier protein can be mouse haemocyanin, thyroprotein, bovine serum albumin(BSA), rabbit serum proteins, human albumin, ovalbumin, hemocyanin or fibrinogen.
Described amantadine specific antibody prepares using amantadine coupled antigen as immunogene, and described amantadine specific antibody can be amantadine monoclonal antibody or amantadine polyclonal antibody, wherein preferred amantadine monoclonal antibody.
The marker enzyme of described enzyme labelled antibody is that horseradish peroxidase or bacterium extract alkaline phosphatase, wherein preferred horseradish peroxidase; Enzyme labelled antibody is obtained by enzyme and amantadine antibody coupling.
In order to more convenient on-site supervision and great amount of samples examination, described kit also comprises amantadine standard solution, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid.
Described amantadine standard solution 6 bottles, concentration is respectively 0 μ g/L, 0.5 μ g/L, 1.5 μ g/L, 4.5 μ g/L, 13.5 μ g/L, 40.5 μ g/L.
When marker enzyme is horseradish peroxidase, described substrate nitrite ion is made up of substrate solution A liquid and substrate solution B liquid, A is hydrogen peroxide or urea peroxide, and B liquid is o-phenylenediamine or tetramethyl benzidine, and described stop buffer is sulfuric acid or the liquid hydrochloride buffer of 1 ~ 2mol/L; When marker enzyme is bacterium extraction alkaline phosphatase, described substrate nitrite ion is to nitro phosphate buffer, and described stop buffer is 1 ~ 2mol/L sodium hydroxide solution.
It is 7.4 that described cleansing solution is preferably pH value, and the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L, described number percent is percent weight in volume.
Described redissolution liquid is preferably that pH value is 7.0, the phosphate buffer of 0.02mol/L, and described number percent is percent weight in volume.
It is pH value is 9.6 that wherein used in ELISA Plate preparation process bag is buffered liquid, the carbonate buffer solution of 0.05mol/L, confining liquid is pH value is 7.1 ~ 7.5, the phosphate buffer containing 1% ~ 3% casein, 0.1 ~ 0.3mol/L, and described number percent is percent weight in volume.
In the present invention, the preparation process of ELISA Plate is: be buffered liquid with bag and coating antigen is diluted to 20 μ g/mL, every hole adds 100 μ l, 25 DEG C of lucifuges are hatched 2h or 4 DEG C and are spent the night, and liquid in hole of inclining, washs 2 times with cleansing solution, each 30s, pat dry, in every hole, then add 150 ~ 200 μ l confining liquids, 25 DEG C of lucifuges hatch 1 ~ 2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
Cleaning Principle of the present invention is:
This kit adopts direct competive ELISA method, pre-coated coupled antigen on ELISA Plate capillary strip, on amantadine residual in sample and ELISA Plate capillary strip, pre-coated coupled antigen competes the enzyme labelled antibody of anti-amantadine, develop the color with tmb substrate, sample absorbance becomes negative correlation with the content of residue amantadine contained by it, compare with typical curve, then be multiplied by the extension rate of its correspondence, the residual quantity of amantadine in sample can be drawn.
Present invention also offers a kind of method applying above-mentioned enzyme linked immunological kit detection amantadine, it comprises step:
(1) sample pre-treatments;
(2) detect with kit;
(3) testing result is analyzed.
The enzyme linked immunological kit that the present invention detects amantadine mainly adopts the content of the firm alkanamine of the qualitative or quantitative detection Gold Samples of ELISA method; Require low to the pre-treatment of sample, sample pretreatment process is simple, can detect batch samples fast simultaneously; Main agents provides with the form of working fluid, and the method for inspection is convenient and easy, has that specificity is high, highly sensitive, degree of accuracy is high, accuracy high.Enzyme linked immunological kit of the present invention, structure is simple, easy to use, low price, carrying convenience, detection method are efficient, accurate, easy, be suitable for the qualitative, quantitative of batch samples screening.
Accompanying drawing explanation
Fig. 1: amantadine hapten synthesis route map
Fig. 2: amantadine haptens hydrogen nuclear magnetic resonance spectrogram
Fig. 3: kit standard curve map
Embodiment
The present invention is set forth further below in conjunction with specific embodiment.Should be understood that these embodiments are only for illustration of the present invention, and be not used for limiting the scope of the invention.
The preparation of embodiment 1 reagent constituents
1, the haptenic preparation of amantadine
Get 0.8g amantadine hydrochloride, add acetonitrile 40ml, stirring and dissolving, add Anhydrous potassium carbonate 0.41g, stir, add β-chloropropionic acid 0.54g, 80 degree of stirring reaction 6h.Stop reaction, filter, removing sal tartari, evaporate to dryness, upper silicagel column, sherwood oil: ethyl acetate=1:1 wash-out, separation and purification, obtains haptens product 0.43g.
Get above-mentioned product through nuclear magnetic resonance hydrogen spectruming determining, as shown in Figure 2, the signal peak of chemical shift δ 11.0 shows to there is carboxyl hydrogen, and the signal peak of 2.82,2.49 shows the hydrogen existed on spacerarm, therefore proves hapten synthesis success.
2, the preparation of antigen
Prepared by immunogene---and amantadine haptens and bovine serum albumin(BSA) (BSA) coupling obtain immunogene.
Get 12mg haptens, be dissolved in 1mL DMF, obtain solution (1); Get 30mg EDC and NHS 0.2ml water fully dissolve after in adding in (1), stirred at ambient temperature 24h, can obtain reactant liquor A.Take BSA50mg, make it fully to be dissolved in 3.8mL PBS (PH 7.2), reactant liquor A is dropwise slowly added drop-wise in protein solution, and in stirred at ambient temperature 24h, change 3 dislysates every day with 0.01mol/L PBS 4 DEG C dialysis 3d.Packing, saves backup in-20 DEG C.
Prepared by coating antigen---and amantadine haptens and ovalbumin (OVA) coupling obtain immunogene.
Get 12mg haptens, be dissolved in 1mL DMF, obtain solution (1); Get 30mg EDC and NHS 0.2ml water fully dissolve after in adding in (1), stirred at ambient temperature 24h, can obtain reactant liquor A.Take OVA50mg, make it fully to be dissolved in 3.8mL PBS (PH 7.2), reactant liquor A is dropwise slowly added drop-wise in protein solution, and in stirred at ambient temperature 24h, change 3 dislysates every day with 0.01mol/L PBS 4 DEG C dialysis 3d.Packing, saves backup in-20 DEG C.
3, the preparation of amantadine monoclonal antibody
Animal immune: immunogene above-mentioned steps obtained is injected in Balb/c Mice Body, immunizing dose is 150 μ g/, makes it produce antiserum.
Fusion of Cells and cloning: after mice serum measurement result is higher, get its splenocyte, in 8:1 (quantitative proportion) ratio and SP2/0 myeloma cell fusion, adopts indirect competitive ELISA to measure cell conditioned medium liquid, screen positive hole.Limiting dilution assay is utilized to carry out cloning to positive hole, until obtain the hybridoma cell strain secreting amantadine monoclonal antibody.
Cell cryopreservation and recovery: monoclonal hybridoma strain cryopreserving liquid is made 1 × 10
6the cell suspension of individual/mL, preserves for a long time in liquid nitrogen.Take out cryopreservation tube during recovery, put into 37 DEG C of water-bath middling speeds immediately and melt, after centrifugal segregation cryopreserving liquid, move into and cultivate culture in glassware.
The production of monoclonal antibody and purifying: only Balb/c mouse peritoneal is injected sterilizing paraffin oil 0.5mL/, within 7 days, pneumoretroperitoneum injects stable monoclonal hybridoma strain 5 × 10
5individual/only, gather ascites after 7 days.Carry out ascites by sad-saturated ammonium sulfate method to purify ,-20 DEG C of preservations.
4, the preparation of enzyme labelled antibody
Amantadine antibody and horseradish peroxidase (HRP) are carried out coupling obtain.
5, the preparation of ELISA Plate
Be buffered liquid with bag and coating antigen is diluted to 20 μ g/mL, every hole adds 100 μ l, 25 DEG C of lucifuges hatch 2h, and liquid in hole of inclining washs 2 times with cleansing solution, each 30s, pat dry, in every hole, then add 200 μ l confining liquids, 25 DEG C of lucifuges hatch 2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
Embodiment 2 detects the establishment of the enzyme linked immunological kit of amantadine
Set up the enzyme linked immunological kit detecting amantadine, make it comprise following component:
(1) bag is by the ELISA Plate of amantadine coupled antigen;
(2) amantadine standard solution 6 bottles, concentration is respectively 0 μ g/L, 0.5 μ g/L, 1.5 μ g/L, 4.5 μ g/L, 13.5 μ g/L, 40.5 μ g/L;
(3) with the amantadine antibody of horseradish peroxidase-labeled;
(4) substrate nitrite ion is made up of A liquid and B liquid, and A liquid is urea peroxide, and B liquid is tetramethyl benzidine;
(5) stop buffer is 2mol/L sulfuric acid;
(6) cleansing solution is pH value is 7.4, the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L, and described number percent is percent weight in volume;
(7) redissolve liquid to be pH value be 7.0, the phosphate buffer of 0.02mol/L, described number percent is percent weight in volume.
The detection of amantadine in embodiment 3 animal tissue
1, sample pre-treatments
With homogenizer homogeneous structure sample; Take the sample after 2.0 ± 0.05g homogeneous in 10ml polystyrene centrifuge tube, add 6ml acetonitrile, then add 0.5g sodium chloride, use oscillator vibrates 5min, 3000g room temperature (20-25 DEG C/68-77 ℉) centrifugal 5min; Pipette 3ml upper organic phase in 10ml clean dried glass tube, flow down dry up in 40-50 DEG C of (68-86 ℉) water-bath nitrogen; Add 1ml normal hexane, with vortex instrument whirling motion 1min, then add 0.5ml redissolution working fluid, with vortex instrument whirling motion 30s, mixing, 3000g room temperature (20-25 DEG C/68-77 ℉) centrifugal 3min; Removing upper organic phase, takes off layer aqueous phase 100 μ l for analyzing.
2, detect with kit
Add standard items/sample 100 μ l in the micropore of correspondence, then add enzyme conjugates working fluid 50 μ l/ hole, mixing of vibrating gently, react 30min with in cover plate membrane cover plate rearmounted 25 DEG C (77 ℉) light protected environment.Carefully open cover plate film, liquid in hole is dried, adds wash operating solution 250 μ l/ hole, abundant washing 4-5 time, every minor tick 10s, sprinkles cleansing solution in board falling hole, pats dry (bubble be not eliminated after patting dry can be poked with original rifle head) with thieving paper.Add substrate solution A liquid 50 μ l/ hole, then add substrate solution B liquid 50 μ l/ hole, mixing of vibrating gently, react 15min with in cover plate membrane cover plate rearmounted 25 DEG C (77 ℉) light protected environment.Add stop buffer 50 μ l/ hole, mixing of vibrating gently, setting microplate reader, in 450nm place (suggestion dual wavelength 450/630nm detects, and please runs through data in 5min), measures every hole OD value.
3, Analysis of test results
The percentage absorptance of standard items or sample equals the mean value of mean value (diplopore) divided by the absorbance of first standard items (0 standard) of the absorbance of standard items or sample, be multiplied by 100% again, obtain the percentage absorbance of standard items or sample.With standard items percentage absorptance for ordinate, with the logarithm of amantadine standard concentration (μ g/L) for horizontal ordinate, drawing standard curve map.The percentage absorptance of sample substituted in typical curve, read the concentration corresponding to sample from typical curve, the extension rate being multiplied by its correspondence is the actual concentrations of amantadine in sample.
The determination test of embodiment 4 amantadine technical parameter
1, kit sensitivity and detectability
Conventionally measure kit sensitivity, the scope of typical curve is 0.5 ~ 40.5 μ g/L, IC
50(50% inhibition concentration) domain of walker is 2 ~ 4 μ g/L; Detect 20 increment product, find the concentration corresponding to each percentage absorbance from typical curve, add that 3 times of standard deviations represent detectability with the mean value of 20 parts of concentration of specimens, result shows, and the detectability of the method to animal tissue is 0.25 μ g/kg.
2, sample preci-sion and accuracy test
Using the recovery as accuracy estimating index, the testing result relative standard deviation (RSD%) of a certain concentration samples of replication is as precision evaluation index.Computing formula is: the recovery (%)=practical measurement value/theoretical value × 100%, and wherein theoretical value is the interpolation concentration of sample; Relative standard deviation RSD%=SD/X × 100%, wherein SD is standard deviation, and X is the mean value of determination data.
Carry out interpolation to chicken, duck sample respectively by the amantadine of 0.5 μ g/kg, 1.0 μ g/kg, two concentration to reclaim and measure, each sample do 4 parallel, measure with three batches of different reagent, average recovery rate and the precision of calculation sample the results are shown in following table.
Table 1 chicken sample precision and accuracy test
Table 2 duck sample precision and accuracy test
Add chicken, duck respectively with the amantadine of 0.5 μ g/kg, 1.0 μ g/kg, two concentration, average recovery rate is between 82.5% ~ 91.0%; Batch in, batch between relative standard deviation be all less than 10%.
3, stabilization of kit test
Kit preservation condition is 2 ~ 8 DEG C, and through the mensuration of 12 months, the maximum absorbance value (zero standard) of kit, 50% inhibition concentration, amantadine added practical measurement value all within normal range.Consider in transport and use procedure, have improper preservation condition and occur, placed 7 days under 37 DEG C of preservation conditions by kit, carry out accelerated aging tests, result shows that this kit indices meets the requirements completely.Consider that the freezing situation of kit occurs, kit is put into-20 DEG C of refrigerator freezings 7 days, measurement result also shows that kit indices is completely normal.Can show that kit at least can be preserved more than 12 months at 2 ~ 8 DEG C from above result.
Claims (8)
1. one kind is detected the enzyme linked immunological kit of amantadine, it is characterized in that comprising: be coated with the ELISA Plate of coating antigen, amantadine standard solution, enzyme labelled antibody, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid, described coating antigen is amantadine coupled antigen, and described enzyme labelled antibody is the amantadine antibody of enzyme labeling.
2. kit as claimed in claim 1, it is characterized in that described amantadine coupled antigen is obtained by amantadine haptens and carrier protein couplet, described amantadine haptens is obtained by reacting by amantadine and β-chloropropionic acid, and described carrier protein can be mouse haemocyanin, thyroprotein, bovine serum albumin(BSA), rabbit serum proteins, human albumin, ovalbumin, hemocyanin or fibrinogen.
3. kit as claimed in claim 2, it is characterized in that described amantadine haptens is obtained by reacting by amantadine and β-chloropropionic acid, molecular structural formula is:
4. kit as claimed in claim 1, it is characterized in that described amantadine specific antibody prepares using amantadine coupled antigen as immunogene, described amantadine specific antibody can be amantadine monoclonal antibody or amantadine polyclonal antibody.
5. kit as claimed in claim 1, it is characterized in that the marker enzyme of described enzyme marker is that horseradish peroxidase or bacterium extract alkaline phosphatase, when marker enzyme is horseradish peroxidase, substrate nitrite ion A liquid is hydrogen peroxide or urea peroxide, substrate nitrite ion B liquid is o-phenylenediamine or tetramethyl benzidine, and stop buffer is sulfuric acid or the hydrochloride buffer of 1 ~ 2mol/L; When marker enzyme is bacterium extraction alkaline phosphatase, substrate nitrite ion is to nitro phosphate buffer, and stop buffer is 1 ~ 2mol/L NaOH.
6. kit as claimed in claim 1, is characterized in that described cleansing solution be pH value is 7.4, the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L; To be pH value be described redissolution liquid 7.0, the phosphate buffer of 0.02mol/L, and described number percent is percent weight in volume.
7. kit as claimed in claim 1, is characterized in that the concentration of described amantadine standard solution is respectively 0 μ g/L, 0.5 μ g/L, 1.5 μ g/L, 4.5 μ g/L, 13.5 μ g/L, 40.5 μ g/L.
8. detect a method for the firm alkanamine content of Gold Samples, comprise step:
(1) sample pre-treatments;
(2) detect with the kit described in any one of claim 1 ~ 7;
(3) testing result is analyzed.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105906522A (en) * | 2016-04-29 | 2016-08-31 | 北京维德维康生物技术有限公司 | Amantadine artificial antigen and preparation method and application thereof |
| CN106706616A (en) * | 2016-12-03 | 2017-05-24 | 无锡艾科瑞思产品设计与研究有限公司 | Adamantanamine detection method and detection card |
| CN108152499A (en) * | 2017-12-06 | 2018-06-12 | 华南农业大学 | A kind of antigen of amantadine, antibody and its enzyme-linked immunologic detecting kit |
| CN109061171A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | It is a kind of detect flumetralim enzyme linked immunological kit and its application |
| CN111812317A (en) * | 2020-06-17 | 2020-10-23 | 北京勤邦生物技术有限公司 | Application of polychlorinated biphenyl artificial antigen in enzyme linked immunosorbent assay kit |
| CN115819270A (en) * | 2021-09-17 | 2023-03-21 | 洛阳中科生物芯片技术有限公司 | Amantadine hapten, amantadine antigen, preparation method, amantadine monoclonal antibody and amantadine detection kit |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004009116A2 (en) * | 2002-07-18 | 2004-01-29 | Cytos Biotechnology Ag | Hapten-carrier conjugates comprising virus like particles and uses thereof |
| CN102043058A (en) * | 2010-11-23 | 2011-05-04 | 中生北控生物科技股份有限公司 | Detection kit of acetyl amantadine for predicting tumors |
| CN104215780A (en) * | 2013-05-31 | 2014-12-17 | 北京勤邦生物技术有限公司 | Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof |
| CN104402753A (en) * | 2014-11-17 | 2015-03-11 | 浙江省农业科学院 | Amantadine artificial hapten and artificial antigen as well as preparation method and application of amantadine artificial hapten and artificial antigen |
| CN104480072A (en) * | 2014-11-17 | 2015-04-01 | 浙江省农业科学院 | Hybridoma cell strain secreting anti-amantadine monoclonal antibody and application of hybridoma cell strain |
-
2015
- 2015-06-18 CN CN201510342064.4A patent/CN104897864B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004009116A2 (en) * | 2002-07-18 | 2004-01-29 | Cytos Biotechnology Ag | Hapten-carrier conjugates comprising virus like particles and uses thereof |
| CN102043058A (en) * | 2010-11-23 | 2011-05-04 | 中生北控生物科技股份有限公司 | Detection kit of acetyl amantadine for predicting tumors |
| CN104215780A (en) * | 2013-05-31 | 2014-12-17 | 北京勤邦生物技术有限公司 | Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof |
| CN104402753A (en) * | 2014-11-17 | 2015-03-11 | 浙江省农业科学院 | Amantadine artificial hapten and artificial antigen as well as preparation method and application of amantadine artificial hapten and artificial antigen |
| CN104480072A (en) * | 2014-11-17 | 2015-04-01 | 浙江省农业科学院 | Hybridoma cell strain secreting anti-amantadine monoclonal antibody and application of hybridoma cell strain |
Non-Patent Citations (2)
| Title |
|---|
| TETSUHARU IWATA ET AL.: "Determination of Amantadine in Human Plasma by High-Performance Liquid Chromatography with Fluorescence Detection", 《ANALYTICAL SCIENCES》 * |
| 蔡自由等: "微流控芯片测定盐酸金刚烷胺片中的盐酸金刚烷胺", 《分析测试学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105906522A (en) * | 2016-04-29 | 2016-08-31 | 北京维德维康生物技术有限公司 | Amantadine artificial antigen and preparation method and application thereof |
| CN106706616A (en) * | 2016-12-03 | 2017-05-24 | 无锡艾科瑞思产品设计与研究有限公司 | Adamantanamine detection method and detection card |
| CN108152499A (en) * | 2017-12-06 | 2018-06-12 | 华南农业大学 | A kind of antigen of amantadine, antibody and its enzyme-linked immunologic detecting kit |
| CN109061171A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | It is a kind of detect flumetralim enzyme linked immunological kit and its application |
| CN111812317A (en) * | 2020-06-17 | 2020-10-23 | 北京勤邦生物技术有限公司 | Application of polychlorinated biphenyl artificial antigen in enzyme linked immunosorbent assay kit |
| CN115819270A (en) * | 2021-09-17 | 2023-03-21 | 洛阳中科生物芯片技术有限公司 | Amantadine hapten, amantadine antigen, preparation method, amantadine monoclonal antibody and amantadine detection kit |
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