CN202903800U - Neomycin detection kit - Google Patents
Neomycin detection kit Download PDFInfo
- Publication number
- CN202903800U CN202903800U CN201220560765.7U CN201220560765U CN202903800U CN 202903800 U CN202903800 U CN 202903800U CN 201220560765 U CN201220560765 U CN 201220560765U CN 202903800 U CN202903800 U CN 202903800U
- Authority
- CN
- China
- Prior art keywords
- neomycin
- reagent bottle
- reagent
- bottle
- kit
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model relates to a neomycin detection kit. The kit comprises a solid phase carrier, a recessed bottle position for containing a reagent, and reagent bottles at the recessed bottle position, wherein the solid phase carrier is provided with a micro-pore bar; the micro-pore bar comprises a micro pore and a coating layer; the coating layer consists of neomycin coupled antigens; and the reagent bottles include a reagent bottle filled with neomycin monoclonal antibody solution and a reagent bottle filled with horseradish peroxidase marked anti-sheep and anti-mouse antibodies. The kit disclosed by the utility model can be used for residue detection of neomycin, and has the characteristics of convenience in use, high sensitivity, large detected sample amount and the like in comparison with an instrument analysis technique.
Description
(1) technical field: the utility model relates to a kind of neomycin detection kit, particularly detects the kit of Neomycin residue in the animal derived food.
(2) background technology: neomycin is aminoglycosides antibiotics, be applied to the treatment of Animal diseases by wide model, because it has neurotoxicity and Toxicity of Kidney, the residual meeting in animal foodstuff affect human health, and American-European countries and China all require its use of limiting the quantity of.
The conventional sense method of Neomycin residue mainly contains thin-layer chromatography (TLC), high performance liquid chromatography (HPLC), Capillary Electrophoresis (CE) etc., because complicated instrument and equipment and loaded down with trivial details process and to reviewer's high professional qualification requirement are not suitable for the examination of on-site supervision and great amount of samples.
(3) utility model content: the utility model provides a kind of neomycin detection kit of small volume and less weight, it is easy and simple to handle, and detection sensitivity is high, and cost is low, can carry out the mensuration that one-time continuous is carried out Neomycin residue in a plurality of samples, reduce the needed time of detection sample.
The neomycin detection kit, comprise solid phase carrier, put reagent recessed bottle position, be positioned at the reagent bottle on the recessed bottle position, solid phase carrier is provided with capillary strip, capillary strip comprises micropore, coated layer; The reagent bottle of the sheep anti mouse antiantibody that reagent bottle comprises reagent bottle that the neomycin monoclonal antibody solution is housed, be equipped with horseradish peroxidase-labeled.
Described coated layer is comprised of the neomycin coupled antigen.
Described kit also comprises the 6 bottled reagent bottles that the neomycin titer is arranged, the 2 bottled reagent bottles that the substrate nitrite ion is arranged, the 1 bottled reagent bottle that stop buffer is arranged, the 1 bottled reagent bottle that cleansing solution is arranged, the 1 bottled reagent bottle that damping fluid is arranged.
Described solid phase carrier is comprised of 96 hole polystyrene agent plate.
Reagent in each kit enough carries out 96 times to be measured, and both can carry out the detection of a plurality of samples of one-time continuous, also can take plate hole apart repeated detection.During detection, neomycin in the sample will be competed the neomycin monoclonal antibody with the neomycin coupled antigen of the coated layer of capillary strip, after adding the sheep anti mouse antiantibody of horseradish peroxidase-labeled, develop the color with the substrate nitrite ion, the residual quantity of neomycin becomes negative correlation in sample absorbance and the sample, multiply by more again its corresponding extension rate with typical curve, can draw Neomycin residue in the sample.
Our experiments show that this kit has very high sensitivity: detection sensitivity is: 0.5 μ g/L.With respect to other neomycin detection methods, the required instrument of this kit is less, and generally all there is outfit in similar laboratory, and required cost is lower.
The beneficial effects of the utility model are: can be used for quick, easy, sensitive, exactly the detection of animal derived food Neomycin residue.
(4) description of drawings:
The solid phase carrier one row capillary strip figure of Fig. 1 the utility model kit.
Fig. 2 the utility model solid phase carrier figure.
Fig. 3 the utility model box body, recessed bottle bit architecture synoptic diagram;
(5) embodiment:
As shown in Figure 1 to Figure 3, the neomycin detection kit, has the recessed bottle position 2 that fixed mould forms in the box body 1, recessed bottle position is used for placing the reagent bottle that solution is housed, the solid phase carrier 3 that 96 hole polystyrene agent plate form, solid phase carrier capillary strip 4 comprise micropore 5 and coated layer 6, coated layer is comprised of the neomycin coupled antigen, the reagent bottle of the sheep anti mouse antiantibody that reagent bottle comprises reagent bottle that the neomycin monoclonal antibody solution is housed, be equipped with horseradish peroxidase-labeled.
In order more to be conducive to detect the detection of Detection of neomycin residues, kit can also include the 6 bottled reagent bottles that the neomycin titer is arranged, the 2 bottled reagent bottles that the substrate nitrite ion is arranged, the 1 bottled reagent bottle that stop buffer is arranged, the 1 bottled reagent bottle that cleansing solution is arranged, the 1 bottled reagent bottle that damping fluid is arranged.
Concrete composition in the reagent bottle of kit is as follows:
(1) 1 bottle/7ml of neomycin monoclonal antibody working fluid;
(2) neomycin standard solution 1ml/ bottle, concentration are 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) the sheep anti mouse antiantibody working fluid 7ml/ bottle of horseradish peroxidase-labeled;
(4) substrate nitrite ion is respectively substrate colour developing A liquid, substrate colour developing B liquid, and substrate nitrite ion A liquid is urea peroxide 7ml/ bottle, substrate nitrite ion B liquid is tetramethyl benzidine 7ml/ bottle;
(5) stop buffer 7ml/ bottle;
(6) damping fluid 50ml/ bottle is for containing 1% methyl alcohol, 0.01-0.05mol/L phosphate buffer.2 times of dilutions during use.
(7) cleansing solution 40ml/ bottle is for containing 0.8%-1.2% Tween-20,0.5% sodium azide, 0.01mol/L phosphate buffer.20 times of dilutions during use.
The preparation method of the main material of using in the kit:
The preparation of neomycin coupled antigen
(1) preparation of neomycin coupled antigen
Neomycin and hemocyanin are obtained the neomycin coupled antigen by the carbodiimide method coupling.
The preparation of neomycin monoclonal antibody
(1) animal immune
Immunogene (neomycin and ovalbumin obtain by the carbodiimide method coupling) is injected in the Balb/c Mice Body, and immunizing dose is 150 μ g/, makes it produce polyclonal antibody.
(2) Fusion of Cells and cloning
After the mice serum measurement result is higher, get its splenocyte, merge in 9:1 ratio and SP2/0 myeloma cell, adopt indirect competitive ELISA to measure cell conditioned medium liquid, screen positive hole.Utilize limiting dilution assay that cloning is carried out in positive hole, until obtain the hybridoma cell strain of secrete monoclonal antibody.
(3) cell cryopreservation and recovery
The strain of neomycin monoclonal hybridoma is made 1 * 10 with cryopreserving liquid
9The cell suspension of individual/ml is in the medium-term and long-term preservation of liquid nitrogen.Take out cryopreservation tube during recovery, put into immediately 37 ℃ of water-bath middling speeds and melt, behind the centrifugal removal cryopreserving liquid, move in the culture flask and cultivate.
(4) production of monoclonal antibody and purifying
The Balb/c mouse peritoneal is only injected sterilization paraffin oil 0.5ml/, 7 days pneumoretroperitoneum injection neomycin monoclonal hybridomas strain 5 * 10
7Individual/as only, to gather ascites after 7 days.Carry out the ascites purifying with sad-saturated ammonium sulfate method ,-20 ℃ of preservations.
(5) preparation of sheep anti mouse antiantibody
As immune animal, as immunogene the pathogen-free domestic sheep is carried out immunity take mouse source antibody with sheep, obtain the sheep anti mouse antiantibody.
(6) preparation of the sheep anti mouse antiantibody of horseradish peroxidase-labeled
Sodium periodate method after sheep anti mouse antiantibody and horseradish peroxidase (HRP) the employing improvement is carried out coupling to be obtained.
(1) use the neomycin detection kit to detect the operation steps of neomycin:
1, takes out the microwell plate that needs quantity.
2, add standard items/sample: add standard items/sample 50 μ l in the micropore of correspondence, the sheep anti mouse antiantibody 50 μ l/ holes that add immediately horseradish peroxidase-labeled, adding neomycin monoclonal antibody working fluid 50 μ l/ holes, the mixing that vibrates is gently put in 25 ℃ of lucifuge environment and is reacted 30min.
3, wash plate: liquid in the hole is dried, with wash operating solution 250 μ l/ holes, fully wash 4-5 time, every minor tick 10s pats dry with thieving paper.
4, colour developing: add each 50 μ l/ hole of substrate solution nitrite ion, mixing is put in 25 ℃ of lucifuge environment and is reacted 15min.
5, measure: add stop buffer 50 μ l/ holes, mixing is set microplate reader and is measured every hole OD value in the 450nm place.
(2) result judges
(1) calculating of percentage absorptance, the percentage absorptance of standard items or sample equal the mean value (diplopore) of percentage absorbance of standard items or sample divided by the absorbance of first standard (0 standard), multiply by 100%, namely again
The mean light absorbency value of B-standard solution or sample solution
B
0The mean light absorbency value of-0 μ g/L standard solution
(2) drafting of typical curve and calculating
Take standard items percentage absorptance as ordinate, take the semilog of neomycin standard items concentration (μ g/L) as horizontal ordinate, the drawing standard curve map.In the percentage absorptance substitution typical curve with sample, read the corresponding concentration of sample from typical curve, multiply by its corresponding extension rate and be neomycin practical residue limit in the sample.
Claims (4)
1. neomycin detection kit, comprise solid phase carrier, put reagent bottle recessed bottle position, be positioned at the reagent bottle on the recessed bottle position, it is characterized in that: solid phase carrier is provided with capillary strip, capillary strip comprises micropore, coated layer; The reagent bottle of the sheep anti mouse antiantibody that reagent bottle comprises reagent bottle that the neomycin monoclonal antibody solution is housed, be equipped with horseradish peroxidase-labeled.
2. neomycin detection kit as claimed in claim 1 is characterized in that: described coated layer is comprised of the neomycin coupled antigen.
3. neomycin detection kit as claimed in claim 1, it is characterized in that: described kit also comprises the 6 bottled reagent bottles that the neomycin titer is arranged, the 2 bottled reagent bottles that nitrite ion is arranged, the 1 bottled reagent bottle that stop buffer is arranged, the 1 bottled reagent bottle that cleansing solution is arranged, the 1 bottled reagent bottle that damping fluid is arranged.
4. neomycin detection kit as claimed in claim 1, it is characterized in that: described solid phase carrier is comprised of 96 hole polystyrene agent plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201220560765.7U CN202903800U (en) | 2012-10-30 | 2012-10-30 | Neomycin detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201220560765.7U CN202903800U (en) | 2012-10-30 | 2012-10-30 | Neomycin detection kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202903800U true CN202903800U (en) | 2013-04-24 |
Family
ID=48124457
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201220560765.7U Expired - Fee Related CN202903800U (en) | 2012-10-30 | 2012-10-30 | Neomycin detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN202903800U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572345A (en) * | 2014-10-16 | 2016-05-11 | 镇江亿特生物科技发展有限公司 | Fluorescence assay method for detecting neomycin in chicken meat tissue |
| CN105572370A (en) * | 2014-10-16 | 2016-05-11 | 镇江先创生物科技有限公司 | Time-resolved fluorescent immunoassay kit for detecting neomycin and detecting method of kit |
-
2012
- 2012-10-30 CN CN201220560765.7U patent/CN202903800U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572345A (en) * | 2014-10-16 | 2016-05-11 | 镇江亿特生物科技发展有限公司 | Fluorescence assay method for detecting neomycin in chicken meat tissue |
| CN105572370A (en) * | 2014-10-16 | 2016-05-11 | 镇江先创生物科技有限公司 | Time-resolved fluorescent immunoassay kit for detecting neomycin and detecting method of kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101226194B (en) | Malachite green vestigial ELISA detection kit and usage method thereof | |
| CN101241134B (en) | ELISA kit for detecting ractopamine residue and method of use thereof | |
| CN103869070B (en) | Detect enzyme linked immunological kit and the application thereof of dibutyl phthalate | |
| CN102127523A (en) | Hybridoma cell line and application thereof | |
| CN104897864B (en) | The enzyme linked immunological kit of detection amantadine and application thereof | |
| CN106093381A (en) | Rod method toxin rod method phenol monomethyl ether colloidal gold immuno-chromatography test paper strip | |
| CN101526537A (en) | Elisa reagent for detecting chloramphenicol and method thereof | |
| CN100501407C (en) | ELISA kit for detecting avermectins and detection method thereof | |
| CN103421072A (en) | Dexamethasone semi-antigen, preparation method and applications thereof | |
| CN101424686A (en) | ELISA reagent for detecting malachite green and method | |
| CN102495211B (en) | Beta-fructofuranosidase enzyme-linked immunoassay detection kit | |
| CN105319354A (en) | Preparation method of chemiluminescence immunoassay detection kit used for detecting methyl parathion | |
| CN202903800U (en) | Neomycin detection kit | |
| CN105403703B (en) | Detect enzyme linked immunological kit and its application of carbendazim | |
| CN103575885B (en) | The enzyme linked immunological kit of detection T-2 toxin and application thereof | |
| CN102081094B (en) | Method for detecting patulin and special enzyme linked immunosorbent assay kit thereof | |
| CN102331500A (en) | Method and enzyme linked immunosorbent assay kit for detecting lemon yellow | |
| CN203178273U (en) | Cyproheptadine ELISA (enzyme-linked immunosorbent assay) test kit | |
| CN203069596U (en) | Flumequine detection kit | |
| CN202903670U (en) | Zeranol assay kit | |
| CN103808931A (en) | Enzyme linked immunosorbent assay kit for detecting sulfadiazine and detection method thereof | |
| CN103018447A (en) | Enzyme-linked immunoassay kit for salbutamol detection, and applications thereof | |
| CN202903804U (en) | Norethindrone detection kit | |
| CN202903799U (en) | Macrodantin metabolin detection kit | |
| CN203069595U (en) | Sulfadimethoxine detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130424 Termination date: 20211030 |