CN101435823B - Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof - Google Patents
Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof Download PDFInfo
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Images
Abstract
The invention discloses a test paper strip for the fluorescent bead immunochromatography of veterinary medicine remained in a detected sample and a preparation method thereof. A filter paper, a sample pad, a fiberglass membrane, a pyroxylin membrane and water sucking paper are sequentially stuck on a base plate in related joint; fluorescent beads are sprayed on the pyroxylin membrane for marking a veterinary medicine resistant molecule monoantibody; the pyroxylin membrane coated with a veterinary medicine molecule holoantigen is used as a detection region; the pyroxylin membrane coated with an anti-mouse antibody is used as a quality control region; and the test paper strip is prepared through the following steps: (1) the preparation of the pyroxylin membrane; (2) the preparation of a fluorescent bead pad; and (3) the assembling of the test paper strip. In the detecting process, emitted spectrums pass through a proper optical filter device; and all the emitted spectrums are collected through CCD scanning technology, are congregated, are multiplied and are analyzed through a fluorescence analyzer and relevant software to obtain a quantized fluorescence signal, thereby realizing quantitative detection. The test paper strip is mainly used for the qualitative detection and the quantitative detection of all the veterinary medicine classes in food safety detection.
Description
Technical field
The invention belongs to food security herbal medicine detection range, be specifically related to fluorescent micro-ball immune chromatography test paper strip of residual animal medicine in a kind of qualitative and the detection by quantitative sample and preparation method thereof.
Background technology
In food safety detection, the main both at home and abroad GC-MS that adopts, LC-MS, technology such as enzyme linked immunosorbent assay (ELISA) and colloid gold label immuno-chromatographic test paper strip, thus realize qualitative and detection by quantitative.In food safety detection, in order to reach efficient and purpose efficiently, often carry out primary dcreening operation with colloidal gold-labeled method or ELISA earlier after, again positive sample is proved conclusively with GC-MS or LC-MS.Because above method all needs by expensive and complicated instrument and equipment, cost is high, simultaneously operating personnel is needed Special Training, and experimental result can't show immediately.Therefore be not suitable for commodity inspection, epidemic prevention, the herding producer quick online detection and monitoring to object of suspicion.
Immunochromatography technique is the immunoassay mode that comes across a kind of uniqueness at the initial stage eighties; It is a solid phase with the fibre strip chromatographic material usually; Make sample solution swimming on chromatography strip through capillary action; And make on determinand and the chromatographic material in the sample immune response that high special high-affinity takes place to the acceptor (like antibody or antigen) of determinand simultaneously; Immune complex is by enrichment or be trapped in certain zone (detect band) of chromatographic material in the chromatography process, through enzyme reaction or directly the label (like collaurum) that can estimate of utilization obtain experimental result (as demonstrating various colors) intuitively.Free label is then crossed and is detected band, reaches the purpose of separating automatically with binding label.The common trace labelling particle of immunochromatography technique has collaurum, latex, and electroselenium, gelatin etc., wherein the most successful label of utilization is a collaurum.
But there is following defective in colloidal gold immuno-chromatography test paper strip:
(1) the colloid gold label process is the Electrostatic Absorption process, is a kind of physisorption, thus in liquid phase less stable, often cause the protein molecular on the mark to come off once more.
(2) testing result is brought judgement through showing single aubergine bar, and color is single, is difficult to realize many inspections and joint inspection.
(3) have only when gold grain and gather when a certain amount of, people's naked eyes just can be observed purplish red band, and this color band and background contrasts are little, thereby have limited detection sensitivity.
(4) the material different matrix effect is obvious, and background interference is very big.
(5) detection sensitivity is lower.
(6) can't realize detection by quantitative.
The detection method of quantum dot mark fast immune chromatographic test paper bar at present also occurred utilizing, but the method can not realize to detecting the detection by quantitative of thing.
Summary of the invention
An object of the present invention is defective, provide a kind of highly sensitive, the fluorescent micro-ball immune chromatography test paper strip of qualitative and detection by quantitative residual animal medicine easy and simple to handle to above-mentioned prior art; Another object of the present invention provides the preparation method of above-mentioned test strips.
A technical scheme taking of the present invention is to achieve these goals:
A kind of fluorescent micro-ball immune chromatography test paper strip that detects residue of veterinary drug is provided; It comprises base plate, and the filter paper of on base plate, pasting successively overlap joint, sample pad; Glass fibre membrane; Nitrocellulose filter and thieving paper are coated with the anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere mark on the wherein said glass fibre membrane, be coated with on the described nitrocellulose filter veterinary drug molecule artificial antigen as detection zone be coated with anti-mouse antibody as the Quality Control district.
Said fluorescent microsphere is that its surface is connected with reactive group with the made microballoon of polymeric material or silicon dioxide parcel fluorescent material, or with the Streptavidin coupling, this diameter of micro ball is 0.01～10 μ m; Said polymeric material is polystyrene, ethene and PVC, and said fluorescent material comprises organic or inorganic fluorescent material or alloy and the quantum dot be made up of multiple fluorescent material.
Described reactive group is-CHO ,-COOH ,-OH ,-NH
2Or-SH; Said fluorescent material is that phenanthroline joins ruthenium organic dyestuff, different thiocyanate luciferin, different thiocyanate rhodamine, 6-Fluoresceincarboxylic acid carboxylic acid amide esters, cumarin or its alloy or quantum dot.
Another technical scheme that the present invention takes provides a kind of method for preparing the fluorescent micro-ball immune chromatography test paper strip of above-mentioned detection residual animal medicine, and it comprises following step:
1. the preparation of nitrocellulose filter;
(1) preparation veterinary drug molecule artificial antigen;
In order to prepare nitrocellulose filter; At first veterinary drug molecular antigen and protein macromolecule are prepared the required veterinary drug molecule artificial antigen of detection zone through e, the veterinary drug molecular antigen can be selected from: Clenbuterol, Ractopamine, chloromycetin, PCs, sulphadiazine, sulfadimidine, furazolidone, nitrofurazone; Described high molecular weight protein is bovine serum albumin or ovalbumin.The veterinary drug molecule artificial antigen of processing for example has: Clenbuterol-BSA conjugate; Ractopamine-BSA conjugate; Chloromycetin-BSA conjugate; PCs-BSA conjugate; Sulphadiazine-BSA conjugate; Sulfadimidine-BSA conjugate; Furazolidone-OVA conjugate; Nitrofurazone-BSA conjugate.When single inspection, the required veterinary drug molecule artificial antigen of preparation detection zone can be selected from one of above-mentioned veterinary drug molecule artificial antigen; When many inspections or joint inspection, the use that combines of an optional majority above-mentioned veterinary drug molecule artificial antigen.
(2) preparation in detection zone and Quality Control district.
Respectively thing veterinary drug molecule artificial antigen and anti-mouse antibody are encapsulated on the nitrocellulose filter, process detection zone and Quality Control district.Regulating encrusting substance concentration respectively with the PBS (PBS) of 0.01～0.1 MpH7.2 is 0.5～8.0mg/mL; Spray film amount is 0.74 μ L/cm; Detection zone spraying veterinary drug molecule artificial antigen; Under many inspections situation, encapsulate many detection zones with multiple veterinary drug molecule artificial antigen at detection zone, no Quality Control district; Quality Control district spraying anti-mouse antibody, the two districts 5mm of being separated by, the 5mm of also being separated by between each detection zone, an end 2mm of Quality Control offset thieving paper, after 37 ℃ of oven dry were handled and spent the night, the environment of drying at room temperature preservation down was subsequent use.
Since the labeling of monoclonal antibody that the different detection material is corresponding launch the fluorescent microsphere of different fluorescence; If encapsulate the Quality Control district, then nature controlling line is understood the antibody because of the multiple fluorescent microsphere of incorporation of markings, and on same band, shows multiple fluorescence color; Make fluorescence color assorted and random; So under the situation of many inspections and joint inspection, only encapsulate many detection zones on the nitrocellulose filter, and do not encapsulate the Quality Control district.
2. the preparation of fluorescent microsphere pad;
(1) preparation of fluorescent microsphere: the fluorescent microsphere of the different fluorescent materials of conventional method preparation parcel.
(2) be sprayed on the glass fibre membrane with the anti-veterinary drug monoclonal antibody of fluorescent microsphere mark, and with it.
With the mode mark anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere through covalent coupling: get microballoon at the centrifugal 10～15min of 1000 * g, centrifugal back collecting precipitation uses the borate buffer solution adjusting microballoon concentration of 0.01M pH4.8 to be OD
450=0.2, add 20～100mg/mL EDC (ethyl-N, N-dimethyl propyl carbodiimide) then respectively; With 2～20mg/mLNHS (nitrogen HOSu NHS), the borate buffer solution with 0.01M pH 4.8 dissolves again, behind the vibration mixing; Hatch 10～30min under the room temperature; At the centrifugal 5～15min of 1000 * g, deposition is with the borate buffer solution dissolving of 0.01M pH 4.8 then, and regulating microballoon concentration is OD
450Be 0.2～1.0.The anti-veterinary drug molecule monoclonal antibody that adds 0.1～100 μ g in the 1mL fluorescent microsphere; After fully mixing, stirring at room reaction 1～4hr, ultrapure water centrifuge washing 2～5 times; After deposition is precipitated to initial volume with the PBS of 0.01M pH7.2 redissolution; Be sprayed on the glass fibre membrane with BIODOT Dispensing System (BIODOT operating platform), 25 ℃ of vacuum drying 1～2hr are put under the environment of drying at room temperature subsequent use.
The anti-veterinary drug molecule monoclonal antibody of described fluorescent microsphere mark is selected from the monoclonal antibody of following material:
Clenbuterol, Ractopamine, chloromycetin, PCs, sulphadiazine, sulfadimidine, furazolidone, nitrofurazone.
3. assembling test strips;
Following material is pasted on overlap joint ground successively on base plate:
(1) filter paper; (2) sample pad; (3) fluorescent microsphere pad; (4) have veterinary drug molecule artificial antigen as detection zone with nitrocellulose filter (5) thieving paper of anti-mouse antibody as the Quality Control district is arranged, promptly be assembled into fluorescent micro-ball immune chromatography test paper strip of the present invention.
Method with residual animal medicine in the above-mentioned fluorescent micro-ball immune chromatography test paper strip qualitative detection sample comprises following step:
(1) handles sample by conventional method, make it to become aqueous sample;
(2) on the sample pad of test strips, add sample, behind the reaction 10min, test strip is put into detection window;
(3) fluorescent microsphere sends strong fluorescence band under the lamp source excitation;
(4) when not containing material to be detected in the sample, a fluorescence band all appears in detection zone and Quality Control district, and it is negative promptly to detect sample; When containing excessive material to be detected in the sample, detection zone does not have the fluorescence band, and there is a fluorescence band in the Quality Control district, and it is promptly positive to detect sample.
Method with above-mentioned fluorescent micro-ball immune chromatography test paper strip detection by quantitative residue of veterinary drug comprises following step:
(1) handles sample by conventional method, make it to become aqueous sample;
(2) on the sample pad of test strips, add sample, behind the reaction 10min, test strip is put into detection window;
(3) fluorescent microsphere that is trapped in detection zone and Quality Control district sends strong fluorescence band under the lamp source excitation;
(4) emitted fluorescence is assembled pipe through photoelectricity after the CCD scanning system converges, and sends into photomultiplier, and light signal is enhanced, and passes through signal conversion element again, and through after the software processes, the power of fluorescence shows on display with the height of numerical value; Reference material with the material to be detected of different concentration known detects, obtain series of values after, draw the typical curve of material concentration to be detected and fluorescence power numerical relation.
(5), can draw the content that detects residual animal medicine in the sample according to above-mentioned typical curve according to the demonstration numerical value that detects sample.
The invention has the beneficial effects as follows:
1. highly sensitive: fluorescent material is because emission spectrum is longer than excitation spectrum, and fluorescence spectrum is emission spectrum, can with the rectangular direction of incident light on detect; Like this; Fluorescence does not receive the interference from the background of exciting light, and sensitivity is much higher than uv-visible absorption spectra, and its sensitivity is 10～1000 times with traditional dyestuff and coloured mark substance detection method; Reach pg/mL, can compare favourably with accurate detecting methods such as gas chromatography mass spectrometry and HPLC.Advantages such as and two kinds of methods of fluorescence labeling detection method and this are compared, and also have easy operating, detect fast, and are cheap.Because the fluorescent material luminous intensity is big, thereby helps improving widely detection sensitivity, expansion can detect the scope and the kind of thing.
2. microsphere surface carries the activity chemistry group, and what antibody labeling used is the chemical coupling method, forms antibody and combines with the firm of microballoon.
3. the quantum dot of different-grain diameter and kind can send the band of multiple color under identical excitation spectrum; Different inorganic or organic fluorescent dyes can be launched various colors during according to the different proportion blending, thereby realize many inspections and joint inspection.
4. after collecting the emission spectrum after filtering through the CCD scanning technique, re-send to fluorescence analysis detector and process software processes, quantize fluorescence signal, thereby realize detection by quantitative.
5. polymkeric substance or silicon dioxide have often been wrapped up in the fluorescent microsphere surface, realize the protection to fluorescent material, reduce the interference of external environment, increase fluorescent microsphere stability and fluorescence lifetime.
Description of drawings
Fig. 1 is the structural drawing that detects the fluorescent micro-ball immune chromatography test paper strip of residue of veterinary drug;
Fig. 2 is the detection schematic diagram of the fluorescent micro-ball immune chromatography test paper strip of qualitative detection residue of veterinary drug;
Fig. 3 is a kind of detection principle and structural representation thereof of simple and easy fluorescence detector.
As shown in Figure 1; Constituting of this fluorescent micro-ball immune chromatography test paper strip: on adhesive base 18, successively overlap joint ground paste filter paper 11, sample pad 12, be coated with the anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere mark glass fibre membrane 13, be coated with the detection zone 15 of veterinary drug molecule artificial antigen and be coated with the nitrocellulose filter 14 and the thieving paper 17 in the Quality Control district 16 of anti-mouse antibody.Quality Control offset thieving paper 2mm, Quality Control district and detection zone be 5mm at interval.
As illustrated in fig. 1 and 2, it is following to detect principle: on sample pad 12, drip sample, test strip is put into detection window 41; In detecting sample, do not comprise veterinary drug and divide the period of the day from 11 p.m. to 1 a.m; The anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere mark can be directly with detection zone 15 on the veterinary drug molecule artificial antigen that encapsulates and the anti-mouse antibody specificity in the Quality Control district 16 combine; Sample 43 to be checked excites down at light source 42; Emitted fluorescence 44 can be observed two fluorescence bands clearly at detection zone 15 with Quality Control district 16 respectively through watch window 36 after filtering through monochromatic light optical filter 35.When comprising veterinary drug molecule and content in the sample 43 to be checked above detectability; Veterinary drug molecule in the sample can combine with the anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere mark specifically; Thereby cause the anti-veterinary drug molecule monoclonal antibody of fluorescent microsphere mark not combine, can only combine with the anti-mouse antibody in Quality Control district, so on simple and easy fluorescence detector, observe with the veterinary drug molecule artificial antigen of detection zone 15; Detection zone does not have the fluorescence band, and a fluorescence band clearly can be seen by the Quality Control district.
As shown in Figure 3, following with the detection step of fluorescent micro-ball immune chromatography test paper strip qualitative detection residual animal medicine:
[1] on test strips, adds sample, behind the reaction 10min, test strip is put into detection window 41;
[2] fluorescent microsphere 43 excites in lamp source 42 down, sends strong fluorescence band;
[3] when not containing material to be detected in the fluorescent microsphere, a fluorescence band all appears in detection zone and Quality Control district, and it is negative promptly to detect sample; When containing excessive material to be detected in the sample, detection zone does not have the fluorescence band, and there is a fluorescence band in the Quality Control district, and it is promptly positive to detect sample.
As shown in Figure 3, following with the detection step of fluorescent micro-ball immune chromatography test paper strip detection by quantitative residual animal medicine:
(1) on sample pad, drip sample, behind the reaction 10min, test strip is put into and is detected detection window 41;
(2) fluorescent microsphere 43 that is trapped of detection zone or Quality Control district excites down at light source 42, sends strong fluorescence band;
(3) emitted fluorescence 44 is assembled pipe 46 through photoelectricity after CCD scanning system 45 converges, and sends into photomultiplier 47; Light signal is enhanced; Pass through signal conversion element 48 again, through after the software processes 49, the power of fluorescence shows on the display of data output 410 with the height of numerical value.
(4) in the testing process, prepare the sample of known series concentration, measure the numerical value of its corresponding fluorescence intensity, thereby set up a typical curve according to this series of values and corresponding concentration with the standard items of tested residual animal medicine.According to the numerical value of the data output 410 that detects sample, look into canonical plotting and draw the content that detects residual animal medicine in the sample at last.
Embodiment
Embodiment one: detect the preparation of the fluorescent micro-ball immune chromatography test paper strip of clenobuterol hydrochloride residue amount in the sample
One, the preparation of chromatograph test strip
1. the preparation of nitrocellulose filter;
(1) preparation Clenbuterol-BSA conjugate (Clen-BSA) artificial antigen;
Adopt diazonium method coupling Clen-BSA.
(2) preparation in detection zone and Quality Control district:
Clenbuterol-BSA conjugate and anti-mouse antibody encapsulate on the nitrocellulose filter: with the PBS (phosphate buffer of 0.01M pH7.2; Wherein comprising 5% sucrose and 0.05% Tween-20 (Tween-20)) concentration of regulating Clenbuterol-BSA conjugate is 0.5mg/ml; The solution of gained sprays on film as detection zone; The concentration of regulating anti-mouse antibody with the PBS (wherein comprising 5% sucrose and 0.05% Tween-20 (Tween-20)) of 0.01M pH7.2 is 0.5mg/mL, and the solution of gained sprays on film as the Quality Control district, and the spray film amount in two districts is 0.74 μ l/cm; The two zone position 5mm of being separated by; Quality Control offset thieving paper one end 2mm after 37 ℃ of oven dry are handled and spent the night, preserves subsequent use down in the environment of drying at room temperature.
2. the preparation of fluorescent microsphere pad;
The preparation of the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark: get the centrifugal 10～15min of fluorescent microsphere 1000 * g that 1mg is enclosed with cumarin, centrifugal back collecting precipitation uses the borate buffer solution adjusting microballoon concentration of 0.01M pH4.8 to be OD
450=0.2, the 20mg/mL that adds 80 μ l respectively then is to ethyl-N, N-dimethyl propyl carbodiimide (EDC); With 2～20mg/mL nitrogen HOSu NHS (NHS) of 130 μ l, with the borate buffer solution dissolving of pH 4.8, behind the vibration mixing; Behind incubated at room 10～30min; Centrifugal 5～the 15min of 1000 * g, deposition is regulated microballoon concentration OD with the borate buffer solution dissolving of pH 4.8
450Be 0.2-1.0, add the anti-clenbuterol monoclonal antibody of 1 μ g in the 1mL fluorescent microsphere, after fully mixing; Stirring at room reaction 3hr, ultrapure water centrifuge washing 3 times is after deposition is precipitated to initial volume with PBS (wherein comprising 5% sucrose and the 0.05%Tween-20) redissolution of 0.01 MpH7.2; With BIODOT operating platform (BIODOTDispensing System); Amount according to 8 μ l/cm is sprayed on the glass fibre membrane of 30 * 0.8cm, 25 ℃ of vacuum drying 1～2hr, and it is subsequent use to be put in dry environment.
3. assembling test strips: overlap joint ground is pasted successively on the PVC of 5.5 * 30cm base plate: (1) filter paper and sample pad; Sample pad is the glass fibre membrane of a kind of 1 * 30cm that handles through 0.01～5%Tween-20; The sample pad disposal route: with the phosphate buffer of the 0.01MpH7.4 that contains 5% sucrose and 0.01～5%Tween-20 soak moistening after; Place 37 ℃ of oven drying 2hr, be positioned over dry environment and preserve subsequent use; (2) be dispersed with the fluorescent microsphere pad of 0.8 * 30cm of fluorescent microsphere mark anti-clenbuterol monoclonal antibody; (3) have the Clenbuterol artificial antigen as detection zone with the nitrocellulose filter of anti-mouse antibody as the 2.5 * 30cm in Quality Control district arranged; The thieving paper of (4) 1.2 * 30cm.Put into ready test paper barrel after assembling, the aluminium foil bag of packing into behind the adding drying agent, seals preservation, under the environment of drying at room temperature, can preserve at least 1 year.
Two, residual Clenbuterol in the qualitative and detection by quantitative sample
1. qualitative detection: the fresh pig urine sample is recovered room temperature, directly adds sample aperture, after ten minutes; Test card is put into simple and easy fluorescence detector detection window, and fluorescent microsphere is launched the fluorescence of particular color under LED lamp source excitation; After the monochromatic filter filtration, selection can only see through the optical filter of bluish-green coloured light, observes from watch window; On simple and easy fluorescence detector, observe, detection zone does not have the fluorescence band, and a blue-green fluorescence band clearly can be seen by the Quality Control district.Explain to detect in the sample and comprise Clenbuterol.
2. detection by quantitative:
(1) on test strips, add sample, behind the normal-temperature reaction 10min, test strip is put into the detection detection window;
(2) fluorescent microsphere that is trapped in detection zone and Quality Control district sends strong fluorescence band under LED lamp source excitation;
(3) emitted fluorescence is assembled pipe through photoelectricity after the CCD scanning system converges, and sends into photomultiplier; Light signal is enhanced; Through signal conversion element, through after the software processes, the power of fluorescence shows on the display of data output with the height of numerical value.
(4) in the testing process, prepare the sample of known series concentration, measure the numerical value of its corresponding fluorescence intensity, thereby set up a typical curve according to this series of values and corresponding concentration with the Clenbuterol standard items.To the urine examination of concentration known pig (this known detection sample is 1.5ng/mL through the GC-MS standard measure), at last the numerical value according to the data output that detects sample is 0.40, and looking into canonical plotting, to draw the content that detects Clenbuterol in the sample be 1.5ng/mL.
Implement row two: with residual clenobuterol hydrochloride in the fluorescent micro-ball immune chromatography test paper strip qualitative detection sample
The fluorescent micro-ball immune chromatography test paper strip preparation method who detects clenobuterol hydrochloride sees embodiment one.
The detection step of qualitative detection Clenbuterol fluorescent micro-ball immune chromatography test paper strip is following:
Qualitative detection: the fresh pig urine sample is recovered room temperature, directly adds sample aperture, after ten minutes; Test card is put into simple and easy fluorescence detector detection window, and fluorescent microsphere is launched the fluorescence of particular color under LED lamp source excitation; After the monochromatic filter filtration, selection can only see through the optical filter of bluish-green coloured light, observes from watch window; Can observe two blue-green fluorescence bands clearly at detection zone and Quality Control district respectively, explain to detect in the sample not comprise Clenbuterol.
Embodiment three: detect the preparation of the fluorescent micro-ball immune chromatography test paper strip of residual furazolidone in the sample
One, the preparation of chromatograph test strip
1. the preparation of nitrocellulose filter;
(1) preparation furazolidone artificial antigen;
Adopt diazonium method coupling furazolidone-OVA (chicken egg white), as envelope antigen.
(2) preparation in detection zone and Quality Control district: the concentration of regulating furazolidone-OVA conjugate with the PBS (wherein comprising 5% sucrose and 0.05%Tween-20) of 0.01M pH7.2 is 1.0mg/mL; The solution of gained sprays on film as detection zone; The concentration of regulating anti-mouse antibody with the PBS (wherein comprising 5% sucrose and 0.05%Tween-20) of 0.01M pH7.2 is 1.0mg/mL, and the solution of gained sprays on film as the Quality Control district, and the spray film amount in two districts is 0.74 μ l/cm; The two zone position 5mm of being separated by; Quality Control offset thieving paper one end 2mm after 37 ℃ of oven dry are handled and spent the night, preserves subsequent use down in the environment of drying at room temperature.
2. the preparation of fluorescent microsphere pad;
The preparation of the anti-furazolidone monoclonal antibody of fluorescent microsphere mark: get 1mg and be enclosed with fluorescent microsphere that phenanthroline joins ruthenium dye at the centrifugal 10～15min of 1000 * g, centrifugal back collecting precipitation uses the borate buffer solution adjusting microballoon concentration of 0.01M pH4.8 to be OD
450=0.2.Add the 50mg/mL EDC of 90 μ l and the 5mg/mLNHS of 150 μ l then respectively, with the borate buffer solution dissolving of pH 4.8; The vibration mixing, behind incubated at room 10～30min, the centrifugal 5～15min of 1000 * g; Deposition is regulated microballoon concentration OD with the borate buffer solution dissolving of pH 4.8
450Be 0.2-1.0, add the anti-furazolidone monoclonal antibody of 1～10 μ g in the 1mL fluorescent microsphere, after fully mixing; Behind the stirring at room reaction 3hr, ultrapure water centrifuge washing 3 times, after deposition is precipitated to initial volume with PBS (wherein comprising 5% sucrose and the 0.05%Tween-20) redissolution of 0.01M pH7.2; With BIODOT Dispensing System; According to the amount of 4 μ l/cm be sprayed into glass fibre membrane (on 30 * 0.8cm), 25 ℃ of vacuum drying 1～2hr, it is subsequent use to be put in dry environment.
3. assembling test strips: overlap joint ground is pasted successively on the PVC plastic bottom board: (1) filter paper and sample pad, sample pad are a kind of glass fibre membrane of handling through 0.01～5%TWEEN-20; (2) be dispersed with the fluorescent microsphere pad of the anti-furazolidone monoclonal antibody of fluorescent microsphere mark; (3) have anti-furazolidone artificial antigen as detection zone with the nitrocellulose filter of anti-mouse antibody as the Quality Control district arranged; (4) thieving paper.After assembling, be put in the ready test paper barrel, the aluminium foil bag of packing into behind the adding drying agent, seals preservation, under the environment of drying at room temperature, can preserve at least 1 year.
Two, residual furazolidone in the qualitative and detection by quantitative sample
1. qualitative detection: get 1mL milk sample, after 5min is boiled in water-bath, centrifugal under 500 * g; Removal upper strata fat is drawn 70 μ l supernatants, adds on the sample pad and detects; After ten minutes, test card is put into simple and easy fluorescence detector detection window, fluorescent microsphere is under LED lamp source excitation; The fluorescence of emission particular color, after the monochromatic filter filtration, selection can only see through the optical filter of red light; Observe from watch window, on simple and easy fluorescence detector, observe detection zone and do not have the fluorescence band, a red fluorescence band clearly can be seen by the Quality Control district.Explain to detect in the sample and comprise furazolidone.
2. detection by quantitative:
(1) on the sample pad of test strips, add sample, behind the reaction 10min, test strip is put into the detection detection window;
(2) fluorescent microsphere sends strong fluorescence under LED lamp source excitation;
(3) emitted fluorescence is assembled pipe through photoelectricity after the CCD scanning system converges, and sends into photomultiplier; Light signal is enhanced; Through signal conversion element, through after the software processes, the power of fluorescence shows on the display of data output with the height of numerical value.
(4) in the testing process, prepare the sample of known series concentration, measure the numerical value of its corresponding fluorescence intensity, thereby set up a typical curve according to this series of values and corresponding concentration with the furazolidone standard items.Get 1mL milk sample, centrifugal under 500 * g after 5min is boiled in water-bath, remove upper strata fat, draw 70 μ l supernatants, detect.The numerical value that detects the data output of sample is 0.85, and looking into canonical plotting, to draw the content that detects furazolidone in the sample be 0.2ng/ml.
Embodiment four: detect the preparation of the fluorescent micro-ball immune chromatography test paper strip of hydrochloric acid Ractopamine residual in the sample and clenobuterol hydrochloride
One, the preparation of chromatograph test strip
1. the preparation of nitrocellulose filter;
(1) preparation Clenbuterol (Clen) and Ractopamine (Rac) artificial antigen;
The Clen-BSA conjugate adopts the preparation of diazonium method, and the Rac-BSA conjugate adopts the preparation of EDC method
(2) preparation of detection zone: use the concentration of PBS adjusting Clenbuterol-BSA conjugate of 0.01M pH7.2 to be 0.5mg/mL; The concentration of regulating Ractopamine-BSA conjugate is 0.8mg/mL; Then gained solution is sprayed on respectively and forms two detection zones on the film; The two zone position 5mm of being separated by after 37 ℃ of oven dry are handled and spent the night, preserves subsequent use down in the environment of drying at room temperature.
2. the preparation of fluorescent microsphere pad;
The preparation of the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark: (method for making is as implementing row 1)
The preparation of the anti-Ractopamine monoclonal antibody of fluorescent microsphere mark:
Get 1mg and be enclosed with fluorescent microsphere that phenanthroline joins ruthenium dye at the centrifugal 10～15min of 1000 * g, centrifugal back collecting precipitation uses the borate buffer solution adjusting microballoon concentration of 0.01M pH4.8 to be OD
450=0.2.Add the 20mg/mL EDC of 20 μ l and the 10mg/mLNHS of 100ul then respectively, with the borate buffer solution dissolving of pH 4.8; The vibration mixing, behind incubated at room 10～30min, centrifugal 5～15min under 1000 * g; Deposition is regulated microballoon concentration OD with the borate buffer solution dissolving of pH 4.8
450Be 0.2-1.0, add the anti-Ractopamine monoclonal antibody of 1～10 μ g in the 1mL fluorescent microsphere, fully mix; Behind the stirring at room reaction 3hr, ultrapure water centrifuge washing 3 times, deposition is precipitated to initial volume with 7.2PBS (wherein comprising 5% sucrose and the 0.05%Tween-20) redissolution of 0.01M; Use BIODOT Dispensing System then; According to the amount of 4 μ l/cm be sprayed into glass fibre membrane (on 30 * 0.8cm), 25 ℃ of vacuum drying 1～2hr, it is subsequent use to be put in dry environment.
3. assembling test strips: overlap joint ground is pasted successively on the PVC base plate: (1) filter paper and sample pad, sample pad are a kind of glass fibre membrane of handling through 0.01～5%Tween-20; (2) be dispersed with the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark and the fluorescent microsphere pad of anti-Ractopamine monoclonal antibody; (3) anti-Ractopamine artificial antigen and the anti-clenbuterol artificial antigen nitrocellulose filter as two detection zones is arranged; (4) thieving paper.Be put into after assembling in the ready test paper barrel, the aluminium foil bag of packing into behind the adding drying agent, seals preservation, under the environment of drying at room temperature, can preserve at least 1 year.
Two, residual Ractopamine and clenobuterol hydrochloride in the qualitative and detection by quantitative sample
1. qualitative detection: get 1g fresh pork appearance, add the PBS of 1mL0.01～0.1 MpH7.2,5min is boiled in water-bath; After returning to room temperature, get 70 μ l supernatants and be added to sample pad, after ten minutes; Test card is put into simple and easy fluorescence detector detection window, and fluorescent microsphere is launched the fluorescence spectrum of particular color under LED lamp source excitation; After filtering through the dichromatism monochromatic filter respectively, observe, a red fluorescence band occurring with respect to the Ractopamine detection zone from watch window; Do not having any band with respect to the Clenbuterol detection zone, explaining and contain Clenizole Hydrochloride.
2. detection by quantitative:
After the step of above-mentioned qualitative detection, carry out following detection by quantitative step:
(1) emitted fluorescence is assembled pipe through photoelectricity after the CCD scanning system converges, and sends into photomultiplier; Light signal is enhanced; Through signal conversion element, through after the software processes, the power of fluorescence shows on the display of data output with the height of numerical value.
(2) in the testing process, prepare the sample of known series concentration, measure the numerical value of its corresponding fluorescence intensity, thereby set up a typical curve according to this series of values and corresponding concentration with the Clenbuterol standard items.Get 1g fresh pork appearance; Add the PBS of 1mL0.01～0.1 MpH7.2,5min is boiled in water-bath, return to room temperature after; Add 70 μ l supernatant samples; Detect, at last according to the numerical value 0.17 of the data output that detects sample, looking into canonical plotting, to draw the content that detects furazolidone in the sample be 4.4ng/mL.
Embodiment five: detect the preparation of the fluorescent micro-ball immune chromatography test paper strip of furazolidone and clenobuterol hydrochloride
One, the preparation of chromatograph test strip
1. the preparation of nitrocellulose filter;
(1) preparation Clenbuterol and furazolidone artificial antigen;
Embodiment one, two is seen in the preparation of Clenbuterol and furazolidone artificial antigen.
(2) preparation of detection zone: use the concentration of PBS adjusting furazolidone-OVA conjugate of 0.01M pH7.2 to be 1.0mg/mL; The concentration of regulating Clenbuterol-BSA conjugate is 0.5mg/mL; Then gained solution is sprayed on respectively and forms two detection zones on the film; The two zone position 5mm of being separated by after 37 ℃ of oven dry are handled and spent the night, preserves subsequent use down in the environment of drying at room temperature.
2. the preparation of fluorescent microsphere pad;
The anti-clenbuterol monoclonal antibody of preparation fluorescent microsphere mark: (method for making such as embodiment 1)
The anti-furazolidone monoclonal antibody of preparation fluorescent microsphere mark: (method for making such as embodiment 2)
3. assembling test strips: overlap joint ground is pasted successively on the PVC base plate: (1) filter paper and sample pad, sample pad are a kind of glass fibre membrane of handling through 0.01～5%TWEEN-20; (2) be dispersed with the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark and the fluorescent microsphere pad of anti-furazolidone monoclonal antibody; (3) anti-furazolidone artificial antigen and the anti-clenbuterol artificial antigen nitrocellulose filter as two detection zones is arranged; (4) thieving paper.Be put into after assembling in the ready test paper barrel, the aluminium foil bag of packing into behind the adding drying agent, seals preservation, under the environment of drying at room temperature, can preserve at least 1 year.
Two, residual clenobuterol hydrochloride and furazolidone in the qualitative and detection by quantitative sample
1. qualitative detection: get 1mL milk sample, after 5min was boiled in water-bath, 500 * g was centrifugal, removed upper strata fat; Draw 70 μ l supernatants, detect,, test card is put into simple and easy fluorescence detector detection window adding liquid sample on the sample pad after ten minutes; Fluorescent microsphere is under LED lamp source excitation, and the fluorescence spectrum of emission particular color is after the monochromatic filter filtration; Observe from watch window, any band does not appear in detection zone, explains that furazolidone and clenobuterol hydrochloride all contain.
2. detection by quantitative:
(1) in advance the sample pad of sample adding test paper, behind the reaction 10min, test strip is put into detection window;
(2) fluorescent microsphere sends strong fluorescence under LED lamp source excitation;
(3) emitted fluorescence is assembled pipe through photoelectricity after the CCD scanning system converges, and sends into photomultiplier; Light signal is enhanced; Through signal conversion element, through after the software processes, the power of fluorescence shows on the display of data output with the height of numerical value.
(4) in the experimentation, prepare the sample of known series concentration respectively with furazolidone standard items and Clenbuterol standard items, measure the numerical value of its corresponding fluorescence intensity respectively, thereby set up a typical curve respectively according to this series of values and corresponding concentration.Get 1mL milk sample; After 5min was boiled in water-bath, 500 * g was centrifugal, removed upper strata fat; Draw 70 μ l supernatants; Detect, at last the numerical value according to the data output that detects sample is respectively 0.33 and 0.92, looks into canonical plotting and draws that the content of furazolidone and Clenbuterol is respectively 2ng/mL and 0.1ng/mL in the detection sample.
The joint inspection method that embodiment 4 adopts detects the material similar not of the same race in a kind of sample.Can also realize that in the same sample detection of inhomogeneity material like embodiment 5, realizes joint inspection to hormone medicine and nitrofurans.Similarly, can be extrapolated to three inspections and many inspections, be applicable to the detection of antibiotics, sulfamido, furans, anti parasitic class and hormone medicine.
Claims (2)
1. preparation method who detects the fluorescent micro-ball immune chromatography test paper strip of residual animal medicine; Said test strips is made up of base plate, filter paper, sample pad, fluorescent microsphere pad, nitrocellulose filter and thieving paper; Filter paper, sample pad, fluorescent microsphere pad, nitrocellulose filter and thieving paper are pasted in overlap joint ground successively on said base plate; Be coated with the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark on the described fluorescent microsphere pad, be coated with on the described nitrocellulose filter Clenbuterol artificial antigen as detection zone be coated with anti-mouse antibody as the Quality Control district;
The preparation method of said test strips comprises following step:
(1) preparation of nitrocellulose filter:
1. prepare Clenbuterol-BSA conjugate artificial antigen:
Adopt diazonium method coupling Clenbuterol-BSA;
2. the preparation in detection zone and Quality Control district:
Clenbuterol-BSA conjugate and anti-mouse antibody encapsulate on the nitrocellulose filter: use the concentration of phosphate buffer adjusting Clenbuterol-BSA conjugate of 0.01M pH7.2 to be 0.5mg/ml; The solution of gained sprays on film as detection zone; Use the concentration of the phosphate buffer adjusting anti-mouse antibody of 0.01M pH7.2 to be 0.5mg/mL, the solution of gained sprays on film as the Quality Control district, and the spray film amount in two districts is 0.74 μ l/cm; The two zone position 5mm of being separated by; Quality Control offset thieving paper one end 2mm after 37 ℃ of oven dry are handled and spent the night, preserves subsequent use down in the environment of drying at room temperature;
Phosphate buffer described in the step (1) is the phosphate buffer that comprises 5% sucrose and 0.05% Tween-20;
(2) preparation of fluorescent microsphere pad:
The preparation of the anti-clenbuterol monoclonal antibody of fluorescent microsphere mark: get the centrifugal 10～15min of fluorescent microsphere 1000 * g that 1mg is enclosed with cumarin, centrifugal back collecting precipitation uses the borate buffer solution adjusting microballoon concentration of 0.01M pH4.8 to be OD
450=0.2, the 20mg/mL that adds 80 μ l respectively then is to ethyl-N, N-dimethyl propyl carbodiimide; With 2～20mg/mL nitrogen HOSu NHS of 130 μ l, with the borate buffer solution dissolving of pH 4.8, behind the vibration mixing; Behind incubated at room 10～30min; Centrifugal 5～the 15min of 1000 * g, deposition is regulated microballoon concentration OD with the borate buffer solution dissolving of pH 4.8
450Be 0.2-1.0, add the anti-clenbuterol monoclonal antibody of 1 μ g in the 1mL fluorescent microsphere, after fully mixing; Stirring at room reaction 3 hours, ultrapure water centrifuge washing 3 times is after deposition is precipitated to initial volume with the phosphate buffer redissolution of 0.01M pH7.2; Use the BIODOT operating platform; Amount according to 8 μ l/cm is sprayed on the glass fibre membrane of 30 * 0.8cm, 25 ℃ of vacuum drying 1～2 hour, and it is subsequent use to be put in dry environment;
Phosphate buffer described in the step (2) is the phosphate buffer that comprises 5% sucrose and 0.05% Tween-20;
(3) assembling test strips: overlap joint ground is pasted successively on the PVC of 5.5 * 30cm base plate: 1. filter paper and sample pad; Sample pad is the glass fibre membrane of a kind of treated 1 * 30cm; The sample pad disposal route: with the phosphate buffer of the 0.01M pH7.4 that contains 5% sucrose and 0.01～5% Tween-20 soak moistening after; Placed 37 ℃ of oven dryings 2 hours, and be positioned over dry environment and preserve subsequent use; 2. be dispersed with the fluorescent microsphere pad of 0.8 * 30cm of fluorescent microsphere mark anti-clenbuterol monoclonal antibody; 3. have the Clenbuterol artificial antigen as detection zone with the nitrocellulose filter of anti-mouse antibody as the 2.5 * 30cm in Quality Control district arranged; 4. the thieving paper of 1.2 * 30cm; Put into ready test paper barrel after assembling, the aluminium foil bag of packing into behind the adding drying agent, seals preservation, preserves subsequent use in the environment of drying at room temperature.
2. fluorescent micro-ball immune chromatography test paper strip according to the detection residual animal medicine of the described preparation method of claim 1 preparation.
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| CN2007101661660A CN101435823B (en) | 2007-11-12 | 2007-11-12 | Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof |
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| CN2007101661660A CN101435823B (en) | 2007-11-12 | 2007-11-12 | Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof |
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| CN2007101661660A Active CN101435823B (en) | 2007-11-12 | 2007-11-12 | Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5753517A (en) * | 1996-03-29 | 1998-05-19 | University Of British Columbia | Quantitative immunochromatographic assays |
| CN1321247A (en) * | 1998-10-02 | 2001-11-07 | 基因诊断有限公司 | Assay using porosity-reduction to inhibit migration |
| CN1645146A (en) * | 2005-02-03 | 2005-07-27 | 厦门大学 | Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip |
| CN1847851A (en) * | 2006-04-03 | 2006-10-18 | 河南省农业科学院生物技术研究所 | Test peper strip for fast detection of Rct opamine residue |
-
2007
- 2007-11-12 CN CN2007101661660A patent/CN101435823B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5753517A (en) * | 1996-03-29 | 1998-05-19 | University Of British Columbia | Quantitative immunochromatographic assays |
| CN1321247A (en) * | 1998-10-02 | 2001-11-07 | 基因诊断有限公司 | Assay using porosity-reduction to inhibit migration |
| CN1645146A (en) * | 2005-02-03 | 2005-07-27 | 厦门大学 | Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip |
| CN1847851A (en) * | 2006-04-03 | 2006-10-18 | 河南省农业科学院生物技术研究所 | Test peper strip for fast detection of Rct opamine residue |
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