CN202870093U - Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit - Google Patents
Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit Download PDFInfo
- Publication number
- CN202870093U CN202870093U CN 201220482527 CN201220482527U CN202870093U CN 202870093 U CN202870093 U CN 202870093U CN 201220482527 CN201220482527 CN 201220482527 CN 201220482527 U CN201220482527 U CN 201220482527U CN 202870093 U CN202870093 U CN 202870093U
- Authority
- CN
- China
- Prior art keywords
- reagent bottle
- kit
- solution
- chloromycetin
- chloramphenicol
- 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
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The utility model discloses a chloramphenicol chemiluminescent enzyme-linked immunoassay assay (ELISA) kit, comprising a kit body and an ELISA plate which is coated by antigen. Reagent bottles which hold different solution are arranged in the kit body and the reagent bottles are a reagent bottle for ELISA chloramphenicol antibody working solution, a reagent bottle for chloramphenicol series standard solution, a reagent bottle for concentrated phosphate buffered solution, a reagent bottle for concentrated scrubbing solution and a reagent bottle for luminescent solution respectively. The kit has the advantages of simple and convenient operation, short time, no need for long-time coloration and stop, high sensitivity and good specificity. The kit can be used for the detection of chloramphenicol in aquatic products rapidly, conveniently and sensitively. The detectable minimum detection limit of the kit is 0.0125ng/mL. The kit can perform the detection of multiple samples once, thus the time for detecting samples is reduced. The kit is characterized by cost saving, convenient and rapid operation, ability to perform under non-laboratory condition and convenience in carrying and transportation.
Description
Technical field
The utility model relates to the pick-up unit of chloromycetin, refers to particularly a kind of chloromycetin chemical luminescence ELISA detection kit.
Background technology
Chloromycetin (Chloramphenicol, CAP) is to make by extracting in the streptothricial nutrient culture media of Venezuela at first, is a class broad-spectrum antibiotic.Chloromycetin is widespread use in China's animal husbandry once, and vital role has been played in disease control and the treatment of livestock and poultry.Chloromycetin exists serious toxic and side effect, can cause people's alpastic anemia, granular white blood cells deficiency disease, neonate, premature's Synthetic Grey disease etc., the low concentration medicament residue also can bring out the drug resistance of pathogenic bacteria, to the mankind's the huge potential threat of health formation.Only allow chloromycetin to be applied to non-consumption animal in the U.S., be defined in any animal food and must not detect chloromycetin.The European Community forbids that chloromycetin is applied to milking cow and laying hen, and the content in other animal foods also limits, and stipulates that high residue amount is 100ng/kg.The Ministry of Agriculture classifies chloromycetin deletion from 2002 version " Chinese veterinary pharmacopoeia " as banning drugs, in recent years has been defined in the animal food and must not have detected.
As the Hubei aquatic products industry of national fresh water fishery the first province, the freshwater product total amount kept the whole nation first in continuous 13 years.180 of the existing aquatic products processing enterprises of our province, wherein export processing enterprise is 19.Prohibite the veterinary drug of use as country, in the animal derived foods such as aquatic products, residual chloromycetin is listed in one of emphasis essential items for inspection always.
At present, widely used CAP detection technique has liquid phase chromatography (LC), mass spectroscopy (MS), high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA) etc.The large-scale instrument analytical approachs such as LC have higher requirements to equipment and operator, and sample treatment is complicated, and detection time is long.The used equipment operating of detection method is complicated, testing cost is higher but have now, can't carry out under non-lab environment, is not suitable for the screening of Site Detection and great amount of samples, makes to promote the use of to be restricted.
The utility model content
The purpose of this utility model will overcome the existing deficiency of prior art exactly, but a kind of chloromycetin chemical luminescence ELISA detection kit of small volume and less weight Site Detection is provided, and it is simple to operate, detects fast, accurate, highly sensitive, and cost is low.
For achieving the above object, the chloromycetin chemical luminescence ELISA detection kit that the utility model is designed, comprise box body and the ELISA Plate that is coated with antigen, the reagent bottle of depositing different solutions is set in the described box body, and described reagent bottle is respectively enzyme mark chloramphenicol antibody working fluid reagent bottle, chloromycetin series standard solution reagent bottle, concentrated phosphoric acid salt buffer solution reagent bottle, concentrated cleaning solution reagent bottle and luminescent solution reagent bottle.
Preferably, described enzyme mark chloramphenicol antibody working fluid reagent bottle is the white plastic bottle of green valve protection cap, described chloromycetin series standard solution reagent bottle is the Brown Glass Brown glass bottles and jars only of white valve protection cap, described concentrated phosphoric acid salt buffer solution reagent bottle is the translucent plastic bottle of blue valve protection cap, the translucent plastic bottle that described concentrated cleaning solution reagent bottle is transparent valve protection cap, the black plastic bottle that described luminescent solution reagent bottle is red valve protection cap.Like this, required reagent is encapsulated in the box body with the reagent bottle of different colours, is easy to quick differentiation.
In the such scheme, described ELISA Plate is the detachable micropore plate that is provided with 48 or 96 reacting holes.
The mark of enzyme described in the utility model chloramphenicol antibody is to be made by the coupling of sodium periodate method by horseradish peroxidase and chloramphenicol antibody.
Antigen described in the utility model is the comlete antigen of being made by the carbodlimide method coupling by chloromycetin and carrier protein.
The ELISA Plate that is coated with antigen described in the utility model be with antigen with diluted after, every hole adding 50 ~ 300 μ L place 4 ℃ to be coated with in microwell plate, next day, the rear every hole of microwell plate taking-up washing with antigen added 50 ~ 300 μ L confining liquids, place 25 ~ 40 ℃ of water-bath sealings again washing after 0.5 ~ 2 hour, pat dry, with vacuum seal 4 ℃ of lower preservations and get final product.
Concentrated phosphoric acid salt buffer solution in the utility model: every liter contains NaCl 80g, KH
2PO
42.0g, Na
2HPO
412H
2The aqueous solution of O 29.0g, KCl 2.0g is used 10 times of dilutions as antigen and enzyme labelled antibody of distilled water diluting during use.
Concentrated cleaning solution in the utility model: in above-mentioned concentrated phosphoric acid salt buffer, add volume ratio and be 0.05% Tween-20, during use with behind 10 times of the distilled water dilutings as cleansing solution.
Luminescent solution in the utility model: the 2mmol/L luminol, the p-Coumaric Acid of 0.25mmol/L and the sodium perborate of 20mmol/L that contain useful 0.1mol/L pH=8.5 trishydroxymethylaminomethane-HCl damping fluid preparation.
The beneficial effects of the utility model are: the chemical luminescent detecting technology that the utility model will have height sensitivity combines with the immune response of high specific, easy and simple to handle, time spent is short, need not long-time colour developing and termination, and highly sensitive, specificity is good, can be used for quick, easy, delicately the detection of aquatic products chloromycetin.The lowest detection that this kit can detect is limited to 0.0125ng/mL, can disposable mensuration of carrying out a plurality of samples, reduce the needed time of sample of detecting.The utlity model has save cost, easy and simple to handle fast, can under non-laboratory condition, carry out, characteristics easy to carry and transport.
Description of drawings
Fig. 1 is the box body structure schematic diagram of chloromycetin chemical luminescence ELISA detection kit.
Fig. 2 is the ELISA Plate structural representation of chloromycetin chemical luminescence ELISA detection kit.
Embodiment
Below in conjunction with the drawings and specific embodiments the utility model is described in further detail.
Chloromycetin chemical luminescence ELISA detection kit shown in Fig. 1,2 comprises box body 1 and the ELISA Plate 2 that is coated with antigen.The reagent bottle of different solutions is deposited in box body 1 interior setting, and reagent bottle is respectively enzyme mark chloramphenicol antibody working fluid reagent bottle 3, chloromycetin series standard solution reagent bottle 4-11, concentrated phosphoric acid salt buffer solution reagent bottle 12, concentrated cleaning solution reagent bottle 13 and luminescent solution reagent bottle 14.Enzyme mark chloramphenicol antibody working fluid reagent bottle 3 is the white plastic bottle of green valve protection cap, chloromycetin series standard solution reagent bottle 4-11 is the Brown Glass Brown glass bottles and jars only of white valve protection cap, concentrated phosphoric acid salt buffer solution reagent bottle 12 is the translucent plastic bottle of blue valve protection cap, concentrated cleaning solution reagent bottle 13 is the translucent plastic bottle of transparent valve protection cap, and luminescent solution reagent bottle 14 is the black plastic bottle of red valve protection cap.
ELISA Plate 2 is for being provided with the detachable micropore plate of 96 reacting holes as shown in Figure 2.
The concentrated phosphoric acid salt buffer solution: every liter contains NaCl 80g, KH
2PO
42.0g, Na
2HPO
412H
2The aqueous solution of O 29.0g, KCl 2.0g is used 10 times of dilutions as antigen and enzyme labelled antibody of distilled water diluting during use.
Concentrated cleaning solution: in above-mentioned concentrated phosphoric acid salt buffer, add volume ratio and be 0.05% Tween-20, during use with behind 10 times of the distilled water dilutings as cleansing solution.
Luminescent solution: the 2mmol/L luminol, the p-Coumaric Acid of 0.25mmol/L and the sodium perborate of 20mmol/L that contain useful 0.1mol/L pH=8.5 trishydroxymethylaminomethane-HCl damping fluid preparation.
Chloramphenicol concentration in the chloromycetin series standard solution reagent bottle 4-11 is respectively 0ng/ml, 0.0125ng/ml, 0.025ng/ml, 0.05ng/ml, 0.15ng/ml, 0.45ng/ml, 1.35ng/ml, 4.05ng/ml.
1, the preparation of antigen in the kit
Take by weighing bovine serum albumin(BSA) (BSA) with the ultrapure water dissolving, being mixed with concentration is the BSA solution of 5 ~ 30mg/mL.Take by weighing chloromycetin (CAP) with the ultrapure water dissolving, being mixed with concentration is the CAP solution of 1 ~ 5mg/mL.Take by weighing 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC), add the ultrapure water dissolving, being mixed with concentration is the EDC solution of 1 ~ 5mg/mL.In EDC solution, add 0.1 ~ 0.5mL CAP solution, mix 2min, the CAP solution that will activate again joins in the 0.5mL BSA solution, after mixing up and down, the room temperature lucifuge is mixed 2h, collects coupled product and is transferred to the 30kDa super filter tube, in 4 ℃ of centrifugal 5min of lower 3000rpm of refrigerated centrifuge, add equal-volume glycerine mixing ,-20 ℃ frozen.
2, the preparation of enzyme labelled antibody in the kit
Take by weighing horseradish peroxidase (HRP) and be dissolved in the ultrapure water, being mixed with concentration is the HRP solution of 1 ~ 10mg/mL, and 4 ℃ of standing over night make it abundant dissolving; Get above-mentioned HRP solution 0.5mL next day and to the 0.01 ~ 1mol/L NaIO that wherein adds 0.025 ~ 1mL and newly join
4Aqueous solution is placed the refrigerator lucifuge and is stirred taking-up after 5 ~ 20 minutes behind the mixing.Take out reactant liquor and pack in the bag filter 4 ℃ of acetum dialysed overnight to pH=4.0 into.Potpourri after will dialyse next day takes out, and regulates potpourri pH value, makes it near 9.6; Chloromycetin monoclonal antibody with 0.5 ~ 5mg adds immediately, and the room temperature lucifuge stirs.Get after reaction finishes and add the NaBH that 0.05 ~ 0.5mL concentration is 1 ~ 10mg/mL in the mixed liquor
4Then the mixed liquor ultrafiltration being added equivalent glycerine preserves.
3, the making of ELISA Plate
With antigen with diluted after, every hole adding 50 ~ 300 μ L place 4 ℃ to be coated with in microwell plate, and next day, the rear every hole of microwell plate taking-up washing with antigen added 50 ~ 300 μ L confining liquids, placed 25 ~ 40 ℃ of water-baths to seal after 0.5 ~ 2 hour and washed, pat dry, with vacuum seal 4 ℃ of lower preservations and get final product.
Testing process of the present utility model:
1) sample pre-treatments: get the fish sample homogenization that 10g removes fat, the sample of weighing 3g homogenization and 6mL ethyl acetate mixing 10min, the centrifugal 5min of 4000g, transferase 12 mL ethyl acetate upper strata is in clean glass tube, 50 ℃ of gentle nitrogen flow evaporator ethyl acetate, the fat residue is dissolved in 1mL normal hexane mixing 1min, and adds the 1mL sample diluting liquid.Vortex mixed 1min, 4000g centrifuging 5min discards the upper strata, draw 100 μ L lower floor solution for detection of.
2) typical curve is set up: every hole adds first the chloromycetin standard solution that 100 μ L kits provide 8 series in the ELISA Plate 2, add again the enzyme labelled antibody after 50 μ L dilute, the dilutability 1:80K of enzyme labelled antibody, 25 ℃ of incubations 15 minutes, the cleansing solution that adds 300 μ L/ holes leaves standstill after 2 minutes and pats dry, and repeats with after the cleansing solution washing 5 times, every hole adds luminescent solution 180 μ L, detects at chemiluminescence detector after 5 minutes.With the luminous value of gained standard solution, the luminous value corresponding divided by least concentration multiply by 100 again, and the rate that is inhibited take inhibiting rate as ordinate, is made typical curve take the logarithm of chloramphenicol concentration as horizontal ordinate.
3) the detection analysis of testing sample: every hole adds first the testing sample of handling well in the ELISA Plate 2, add again the enzyme labelled antibody after 50 μ L dilute, the dilutability 1:80K of enzyme labelled antibody, 25 ℃ of lower incubations 15 minutes, after cleansing solution washing 5 times, every hole adds luminescent solution 180 μ L, detects at chemiluminescence detector after 5 minutes.Calculate the content of chloromycetin the testing sample with the luminous value that adds testing sample from typical curve.
Claims (3)
1. chloromycetin chemical luminescence ELISA detection kit, comprise box body (1) and be coated with the ELISA Plate (2) of antigen, it is characterized in that: the reagent bottle of depositing different solutions is set in the described box body (1), and described reagent bottle is respectively enzyme mark chloramphenicol antibody working fluid reagent bottle (3), chloromycetin series standard solution reagent bottle (4-11), concentrated phosphoric acid salt buffer solution reagent bottle (12), concentrated cleaning solution reagent bottle (13) and luminescent solution reagent bottle (14).
2. described chloromycetin chemical luminescence ELISA detection kit according to claim 1, it is characterized in that: described enzyme mark chloramphenicol antibody working fluid reagent bottle (3) is the white plastic bottle of green valve protection cap, described chloromycetin series standard solution reagent bottle (4-11) is the Brown Glass Brown glass bottles and jars only of white valve protection cap, described concentrated phosphoric acid salt buffer solution reagent bottle (12) is the translucent plastic bottle of blue valve protection cap, described concentrated cleaning solution reagent bottle (13) is the translucent plastic bottle of transparent valve protection cap, and described luminescent solution reagent bottle (14) is the black plastic bottle of red valve protection cap.
3. described chloromycetin chemical luminescence ELISA detection kit according to claim 1, it is characterized in that: described ELISA Plate (2) is for being provided with the detachable micropore plate of 48 or 96 reacting holes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220482527 CN202870093U (en) | 2012-09-20 | 2012-09-20 | Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220482527 CN202870093U (en) | 2012-09-20 | 2012-09-20 | Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202870093U true CN202870093U (en) | 2013-04-10 |
Family
ID=48036802
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201220482527 Expired - Fee Related CN202870093U (en) | 2012-09-20 | 2012-09-20 | Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN202870093U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105486823A (en) * | 2014-10-13 | 2016-04-13 | 江苏维赛科技生物发展有限公司 | Enzyme linked immunosorbent assay kit for detecting pretilachlor and detection method thereof |
| CN110146694A (en) * | 2019-04-26 | 2019-08-20 | 山东省食品药品检验研究院 | A kind of chloramphenicol direct competitive chemiluminescence high specific immunoassay method |
-
2012
- 2012-09-20 CN CN 201220482527 patent/CN202870093U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105486823A (en) * | 2014-10-13 | 2016-04-13 | 江苏维赛科技生物发展有限公司 | Enzyme linked immunosorbent assay kit for detecting pretilachlor and detection method thereof |
| CN110146694A (en) * | 2019-04-26 | 2019-08-20 | 山东省食品药品检验研究院 | A kind of chloramphenicol direct competitive chemiluminescence high specific immunoassay method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103217414B (en) | Chemiluminescent immunoassay method for 3,5,3'-triiodothyronine in blood serum | |
| CN101788560B (en) | Enzyme-linked immunoassay kit for detecting residual chloramphenicol and detection method thereof | |
| CN101819206A (en) | AFP (Alpha-Fetoprotein) testing kit (time-resolved fluoroimmunoassay) for prenatal screening and preparation method thereof | |
| CN102650642A (en) | Antibody compound for detecting human prepalin retinol binding protein 4, as well as immunochromatography test card and kit | |
| CN101526537A (en) | Elisa reagent for detecting chloramphenicol and method thereof | |
| CN103743908A (en) | Neomycin semi-quantitative gold-labeled rapid detection kit, and detection method and application thereof | |
| CN202870093U (en) | Chloramphenicol chemiluminescent enzyme-linked immunoassay assay kit | |
| CN101349696A (en) | Enzyme-linked immunologic reagent and method for detecting alficetin | |
| CN106370862B (en) | A kind of stabilization, sensitive fibronectin detection reagent | |
| CN202794185U (en) | Test paper strip for detecting aflatoxin M1 | |
| CN101592660A (en) | Brucellosis indirect enzyme-linked immunosorbent assay milk humoral antibody detection kit | |
| CN102608316A (en) | Kit or test strip for detecting quinoxaline compound | |
| CN102818894A (en) | Colloidal gold detection card for detecting staphylococcus aureus and preparation method thereof | |
| CN102661946A (en) | Malachite chemiluminescence ELISA detection method and kit | |
| CN102331500A (en) | Method and enzyme linked immunosorbent assay kit for detecting lemon yellow | |
| CN103575878A (en) | Chemiluminiscence enzyme-linked immunosorbent assay kit for chloramphenicol in aquatic products and detection method of chloramphenicol | |
| CN103323597A (en) | Colloidal gold rapid detecting card for shigella detection and preparation method thereof | |
| CN203929780U (en) | A kind of magnetic microparticle chemiluminescence immunity detection reagent of Clenbuterol | |
| CN202486140U (en) | Kit for detecting tetracyclines antibiotic | |
| CN103383354B (en) | Detection method of magnetic particle chemiluminescence kit for detecting enterotoxin SEA | |
| CN103898191B (en) | Detection Methods of Salmonella and detection substratum | |
| CN113970637A (en) | Canine distemper virus latex microsphere detection test strip and preparation method thereof | |
| CN103424550A (en) | Detection kit for chloramphenicol and detection method thereof | |
| CN103105494A (en) | Kit and method for detecting beta-lactam antibiotic and melamine | |
| CN201438191U (en) | Chloromycetin ELISA detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130410 Termination date: 20130920 |