CN103376316A - Test strip for detecting streptomycin and method - Google Patents
Test strip for detecting streptomycin and method Download PDFInfo
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- CN103376316A CN103376316A CN2012101276467A CN201210127646A CN103376316A CN 103376316 A CN103376316 A CN 103376316A CN 2012101276467 A CN2012101276467 A CN 2012101276467A CN 201210127646 A CN201210127646 A CN 201210127646A CN 103376316 A CN103376316 A CN 103376316A
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Abstract
The invention discloses a test strip for detecting streptomycin and dihydrostreptomycin and a method. The test strip comprises test paper and a micropore reagent, wherein a streptomycin specific antibody marked by streptomycin is lyophilized in the micropore reagent. The streptomycin specific antibody can be a streptomycin monoclonal antibody or a streptomycin polyclonal antibody. The test paper is formed by sequentially connecting a sample absorption cushion, a reaction film, a water absorption cushion, a protective film and a bottom plate. The reaction film comprises a detection area and a quality control area. The detection area is wrapped by a streptomycin hapten-carrier protein conjugate, and the quality control area is wrapped by an antiantibody. The method for detecting streptomycin by the test strip is simple, quick, intuitive, accurate, wide in application range, low in cost and easy to popularize and use.
Description
Technical field
The present invention relates to a kind of test strips and method that detects streptomysin and dihydrostreptomycin, be specifically related to a kind of colloidal gold strip for detection of the sample streptomycin.
Background technology
Streptomysin (Streptomycin) is a kind of important aminoglycoside antibiotics, be widely used in the treatment of Animal diseases, streptomysin is to all kinds of inflammation of chicken, sheep, ox, pig, such as infectious coryza of chicken, pig acute bronchitis, pig dysentery characterized by white mucous stool, cow endometritis etc. good curative effect is arranged, therefore often use as feed addictive.If incorrect use can cause Determination of Streptomycin Residues to exceed standard.Studies have shown that streptomysin can cause irreversible loss to kidney and auditory nerve, long-term use also can make bacterium produce drug resistance.In order to ensure the safety of animal derived food, the regulation streptomysin is 200 μ g/L in milk in No. 235 file of the Ministry of Agriculture " animal food herbal medicine maximum residue limit(MRL) ", is 600 μ g/kg in muscle, fat, liver, is 1000 μ g/kg in kidney.
The at present detection of Determination of Streptomycin Residues mainly contains instrumental analysis and immunological method.Wherein instrumental method is mainly used high performance liquid chromatography (HPLC), vapor-phase chromatography (GC), that these methods have is highly sensitive, the result accurately, the advantage such as good reproducibility, false positive be few, but still there is certain shortcoming, complicated such as sample pretreatment process, the instrumentation degree is high and expensive, analysis speed is slow, is not suitable for carrying out extensive Site Detection etc.Colloidal gold immuno-chromatography test paper strip has been widely used in during medicament residue detects, have easy and simple to handle, quick, without any need for advantages such as equipment, become at present one of immunology detection developing direction.
Summary of the invention
An object of the present invention is to provide a kind of test strips that detects streptomysin and dihydrostreptomycin.
Test strips provided by the present invention comprises test paper, micropore reagent, and micropore reagent has the micropore plug, and test paper comprises reaction film, absorption of sample pad, adsorptive pads, diaphragm, base plate; Freeze-drying has streptomysin specific antibody-colloid gold label thing in the described micropore reagent; The streptomysin specific antibody can be streptomysin monoclonal antibody or streptomysin polyclonal antibody; Comprise detection zone and Quality Control district on the reaction film; When the streptomysin specific antibody was the streptomysin monoclonal antibody, detection zone was coated with streptomysin hapten-carrier protein conjugate, and the Quality Control district is coated with the sheep anti mouse antiantibody; When the streptomysin specific antibody was the streptomysin polyclonal antibody, detection zone was coated with streptomysin hapten-carrier protein conjugate, and the Quality Control district is coated with goat-anti rabbit antiantibody.
Described streptomysin hapten-carrier protein conjugate is to be obtained by streptomysin haptens and carrier protein couplet, and described carrier protein can be bovine serum albumin(BSA), oralbumin, thyroprotein, hemocyanin, human serum albumins.
Streptomysin specific antibody in described streptomysin specific antibody-colloid gold label thing is to prepare as immunogene with streptomysin hapten-carrier protein conjugate.
Described diaphragm sticks on the absorption of sample pad, is the test side, and the above has the MAX mark line.
Described detection zone is positioned at an end of the diaphragm that is bordering on the MAX mark, and described Quality Control district is positioned at the end away from the diaphragm that the MAX mark is arranged.
Described base plate can be the material that PVC base plate or other hard do not absorb water; Described absorption of sample pad can be suction strainer paper or filter paper for oil; Described adsorptive pads is thieving paper; Described reaction film can be nitrocellulose filter or cellulose acetate membrane; Described diaphragm is PE material diaphragm.
Another object of the present invention provides a kind of method for preparing above-mentioned test strips, and it comprises step:
1) the preparation freeze-drying has the micropore reagent of streptomysin specific antibody-colloid gold label thing;
2) preparation has the reaction film in the Quality Control district of the detection zone of coated streptomysin hapten-carrier protein conjugate and coated antiantibody;
3) with 2) reaction film and absorption of sample pad, adsorptive pads, diaphragm, the base plate that prepare be assembled into test paper;
4) with 1) and 3) freeze-drying for preparing has micropore reagent and the test paper of streptomysin specific antibody-colloid gold label thing to be assembled into test strips.
Specifically, step comprises:
1) with streptomysin and reacting ethylenediamine, preparation streptomysin haptens;
2) with streptomysin haptens and carrier protein couplet, preparation streptomysin hapten-carrier protein conjugate;
3) with streptomysin hapten-carrier protein conjugate immune mouse, mouse boosting cell and murine myeloma cell are passed through to merge, screen, obtain secreting the hybridoma cell strain of streptomysin monoclonal antibody or with streptomysin hapten-carrier protein immunization rabbit, obtain the streptomysin polyclonal antibody;
4) extract mouse IgG or rabbit igg immune health goat, obtain sheep anti-mouse igg antibody or goat-anti rabbit antiantibody;
5) with trisodium citrate and gold chloride reaction preparation collaurum;
6) the streptomysin specific antibody with preparation joins in the collaurum of preparation, obtains streptomysin specific antibody-colloid gold label thing;
7) with after streptomysin specific antibody-colloid gold label thing freeze-drying is in micropore reagent, micropore reagent is added the micropore plug;
8) bovine serum albumin(BSA) (final concentration of bovine serum albumin(BSA) in damping fluid is 0.5% volumn concentration), pH are 7.2 with containing, the 0.1mol/L phosphate buffer soaks 2h, 37 ℃ of lower oven dry 2h with the absorption of sample pad;
9) pasting in order absorption of sample pad, reaction film, adsorptive pads and diaphragm on the base plate;
10) micropore reagent, the test paper for preparing is assembled into test strips, preserved 12 months under 2～8 ℃ of conditions.
Another object of the present invention provides a kind of residual method of above-mentioned ELISA test strip sample streptomycin of using, and it comprises step:
(1) sample-pretreating method;
(2) detect with test strips;
(2) analyzing and testing result.
When using the ELISA test strip sample among the present invention, sample solution to be checked is dripped in micropore reagent, behind the mixing (milk sample needs incubated at room 5min), it is downward to indicate MAX mark line end, spread to reaction film after the golden labeling antibody combination in micropore of micropore reagent after insertion is hatched, sample liquid to be checked; If the content of sample liquid streptomycin to be checked is high, then the streptomysin in the sample liquid to be checked can combine with golden labeling antibody in the diffusion process, and then the antigen-combining site of streptomysin on the complete closed gold labeling antibody, stop golden labeling antibody streptomysin hapten-carrier protein conjugate on reaction film to be combined, detection zone does not develop the color, antiantibody then can be combined with golden labeling antibody, the colour developing of Quality Control district; If the low or nothing of the content of sample liquid streptomycin to be checked, then the antigen binding site on the golden labeling antibody can not be closed, and then golden labeling antibody can be combined by streptomysin hapten-carrier albumen coupling the detection zone colour developing on reaction film, antiantibody also can be combined with golden labeling antibody simultaneously, the colour developing of Quality Control district.If the Quality Control district does not develop the color, then test paper lost efficacy.As shown in Figure 4.
Positive: (C) demonstrates band when the Quality Control district, and detection zone (T) does not develop the color, and is judged to the positive, with "+" expression.
Negative: as when Quality Control district (C) and detection zone (T) all demonstrate band, to be judged to feminine gender, to represent with "-".
Invalid: (C) do not show band when the Quality Control district, and test paper lost efficacy.
That test strips of the present invention has is highly sensitive, high specificity, cost is low, simple to operate, detection time short, be fit to various units uses, stores advantage simple, long shelf-life.Wherein, adopt the streptomysin monoclonal antibody of high specific, guaranteed the reliability of testing result; In micropore reagent, in testing process, golden labeling antibody is fully contacted with sample liquid to be checked golden labeling antibody freeze-drying, fully reaction, thus reduce error, increase the reaction sensitivity of whole system.With the method for ELISA test strip streptomysin of the present invention, easy, quick, directly perceived, accurate, applied widely, cost is low, easily promote the use of.
Description of drawings
Fig. 1 is the test paper cross-sectional view.
Fig. 2 is the test paper vertical view.
Fig. 3 is micropore reagent figure.
Fig. 4 is as a result process decision chart of detection paper.
Embodiment
Employed experimental technique is conventional method if no special instructions among the following embodiment.
One, test paper (Fig. 1)
Described test paper is comprised of base plate, absorption of sample pad, reaction film, adsorptive pads, diaphragm;
Described absorption of sample pad 1, reaction film 2, adsorptive pads 3 and diaphragm 7 stick on the base plate 6 successively in order, the end of absorption of sample pad links to each other with reaction film, the end of reaction film links to each other with adsorptive pads, the top of absorption of sample pad aligns with the top of base plate, and the end of adsorptive pads aligns with the end of base plate;
The absorption of sample pad end of described test paper is pasted with diaphragm, and diaphragm 7 covers the test side on the absorption of sample pad, is printed on MAX printed words (Fig. 2) at the test side diaphragm;
Detection zone 4 and Quality Control district 5 are arranged on the described reaction film, all be the ribbon vertical with the appearance of described test paper, detection zone is positioned at an end of the diaphragm that is bordering on the MAX mark, and the Quality Control district is positioned at the end away from the diaphragm that the MAX mark is arranged.Detection zone is coated with streptomysin hapten-carrier protein conjugate (conjugate of streptomysin haptens-oralbumin), and the Quality Control district is coated with the sheep anti mouse antiantibody;
Described base plate is the PVC base plate; Described absorption of sample pad is suction strainer paper; Described adsorptive pads is thieving paper; Described reaction film is nitrocellulose filter; Described diaphragm is PE material diaphragm.
Two, micropore reagent (Fig. 3)
Freeze-drying has streptomysin monoclonal antibody-colloid gold label thing on the described micropore reagent, freeze-drying amount 0.20～0.25 μ g/mL; Described micropore reagent has micropore plug 9.
Above-mentioned test paper, micropore reagent set are dressed up test strips, in 2～8 ℃ of environment, preserve the term of validity 12 months.
The preparation method of test strips described in embodiment 2 embodiment 1
One, the preparation of test strips
The preparation method of test strips mainly may further comprise the steps:
1) the preparation freeze-drying has the micropore reagent of streptomysin monoclonal antibody-colloid gold label thing;
2) preparation has the reaction film in the Quality Control district of the detection zone of coated streptomysin hapten-carrier protein conjugate and coated sheep anti mouse antiantibody;
3) with 2) reaction film and absorption of sample pad, adsorptive pads, diaphragm, the base plate that prepare be assembled into test paper;
4) with 1) and 3) freeze-drying for preparing has micropore reagent and the test paper of streptomysin monoclonal antibody-colloid gold label thing to be assembled into test strips.
Following substep is described in detail:
(1) preparation of each parts
1. the synthetic and evaluation of streptomysin hapten-carrier protein conjugate
(1) the haptenic preparation of streptomysin
0.58g streptomysin and the mixed liquor of 0.07g ethylenediamine in 50ml methyl alcohol at room temperature reacted 5-10 hour, steaming desolventizes, and quantitatively obtains the streptomysin haptens.
(2) immunogenic preparation
Get streptomysin haptens 20mg and obtain solution I with the water-soluble solution of 2ml; Get 3% glutaraldehyde 0.5ml and dropwise join in the solution I, stirring at room reaction 24h obtains solution II; Get bovine serum albumin(BSA) (BSA) 50mg with the water-soluble solution III that solves of 4ml; Solution II is slowly added in the solution III, and the stirring at room reaction is spent the night; Add NaBH
4The 30mg reduction; PBS with 0.02M dialysed three days, changed dislysate every day three times, got the streptomysin immunogene.
(3) preparation of coating antigen
Get oralbumin (OVA) 50mg and obtain solution I with the water-soluble solution of 4ml; Get each 50mg of carbodiimides (EDC) and N-hydroxy-succinamide (NHS) and add in the solution I with the water-soluble solution of 2ml is fully rear, stirring at room reaction 30min obtains solution II; Get streptomysin haptens 25mg and obtain solution III with the water-soluble solution of 3ml; Solution II is slowly joined in the solution III, and stirring at room reaction 24h gets the streptomysin coating antigen.
(4) evaluation of streptomysin hapten-carrier protein conjugate
Carrier protein, streptomysin haptens, streptomysin hapten-carrier protein conjugate are made into the solution of 0.5mg/mL with the PBS of pH7.4, return to zero with 0.01mol/L pH7.4PBS, in the interscan of wavelength 200～800nm scope, obtain the absorption curve of carrier protein, streptomysin haptens, streptomysin hapten-carrier protein conjugate with ultraviolet spectrophotometer.Different absorption curves appears in the three, shows the success of streptomysin haptens and carrier protein couplet.
2. the preparation of streptomysin monoclonal antibody
(1) animal immune
The immunogene that step 1 obtains is injected in the Balb/c Mice Body, and immunizing dose is 150 μ g/, makes it produce antiserum.
(2) Fusion of Cells and cloning
Get immune Balb/c mouse boosting cell, merge in 8: 1 (quantitative proportion) ratios and SP2/0 myeloma cell, screening obtains the streptomysin monoclonal hybridoma strain of stably excreting streptomysin monoclonal antibody.
(3) cell cryopreservation and recovery
Hybridoma is made 5 * 10 with cryopreserving liquid
6The 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) preparation and purification of monoclonal antibody
Increment cultivation: hybridoma is placed cell culture medium, under 37 ℃ of conditions, cultivate, with sad-saturated ammonium sulfate method the nutrient solution that obtains is carried out purifying, obtain monoclonal antibody ,-20 ℃ of preservations.
Described cell culture medium is to add calf serum and sodium bicarbonate in the RPMI1640 nutrient culture media, making the final concentration of calf serum in cell culture medium is 20% (quality percentage composition), and making the final concentration of sodium bicarbonate in cell culture medium is 0.2% (quality percentage composition); The pH of described cell culture medium is 7.4.
3. the 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.
4. the preparation of streptomysin monoclonal antibody-colloid gold label thing
(1) preparation of collaurum
With two ionized waters that boil off 1% gold chloride is diluted to 0.01% (quality percentage composition), get 100ml and place conical flask, be heated to boiling with the thermostatic electromagnetic stirrer, under continuous high temperature, the lasting stirring, add 2.5ml 1% trisodium citrate, continuing at the uniform velocity to be heated with stirring to solution is bright and stops when red, return to original volume with deionized water after being cooled to room temperature, 4 ℃ of preservations.The collaurum outward appearance for preparing is pure, bright, nothing precipitates and floating thing.
(2) preparation of streptomysin monoclonal antibody-colloid gold label thing
Under magnetic agitation, transfer the pH value to 7.2 of collaurum with 0.2mol/L sal tartari, in colloidal gold solution, add above-mentioned streptomysin monoclonal antibody by the standard that adds 10～50 μ g antibody in every milliliter of colloidal gold solution, continue to stir and evenly mix 10min, adding 10% bovine serum albumin(BSA) (BSA), to make its final concentration in colloidal gold solution be 1% (volumn concentration), leaves standstill 10min.12000rpm, 4 ℃ of centrifugal 40min abandon supernatant, precipitation is with redissolving the damping fluid washed twice, it is resuspended to be with volume that the redissolution damping fluid of initial collaurum volume 1/10 will precipitate, put 4 ℃ for subsequent use.
Redissolution damping fluid: the 0.02mol/L phosphate buffer that contains bovine serum albumin(BSA) (BSA) 0.2%～0.5% (volumn concentration), Tween-80 0.05%～0.2% (quality percentage composition), pH7.2.
With streptomysin monoclonal antibody-colloid gold label thing freeze-drying to micropore reagent
In micropore reagent microwell plate, add 100 μ l streptomysin monoclonal antibody-colloid gold label things, put into freeze drier, be under-50 ℃ of conditions at condenser temperature, behind the pre-freeze 3h, vacuum drying 15h again, can take out, obtain the micropore reagent that freeze-drying has streptomysin monoclonal antibody-colloid gold label thing, sealing is preserved.Streptomysin monoclonal antibody-colloid gold label thing freeze-drying amount 0.20～0.25 μ g/mL.
6. the preparation of absorption of sample pad
The absorption of sample pad placed contain bovine serum albumin(BSA) (bovine serum albumin(BSA) is 0.5% (volumn concentration) at the final concentration of damping fluid), pH7.2,0.1mol/L phosphate buffer and soak 2h, 37 ℃ of baking 2h are for subsequent use.
7. the preparation of reaction film
Coated process: with phosphate buffer streptomysin haptens-oralbumin conjugate is diluted to 10mg/mL, with Isoflow point film instrument it is coated in detection zone (T) on the nitrocellulose filter, package amount is 1.0 μ g/cm
2Phosphate buffer with 0.01mol/L, pH7.4 is diluted to 200 μ g/mL with sheep anti-mouse igg antibody, with Isoflow point film instrument it is coated in Quality Control district (C) on the nitrocellulose filter, and package amount is 1.0 μ g/cm
2Coated good reaction film is placed dry 2h under 37 ℃ of conditions, for subsequent use.
(2) assembling of each parts
1. the assembling of test paper
Stick in order successively described absorption of sample pad, reaction film, adsorptive pads, diaphragm on the described base plate; The end of absorption of sample pad links to each other with the top of reaction film, and the end of reaction film links to each other with the top of adsorptive pads, and the top of absorption of sample pad aligns with the top of base plate, and the end of adsorptive pads aligns with the end of base plate; Bonding protective film on the test paper absorption of sample pad that assembles is printed on the MAX mark line on the diaphragm.
2. the assembling of test strips
Test paper and micropore reagent set that above-mentioned steps 1 obtains are dressed up test strips, in 2～8 ℃ environment, store the term of validity 12 months.
The detection of embodiment 3 sample streptomycins
1. sample pre-treatments
Need when detecting honey sample first honey sample to be carried out pre-treatment:
Accurately take by weighing 0.5g honey in the 4ml centrifuge tube, add 1.5ml 0.2mol/L phosphate buffer, whirling motion 1min, mixing is to be checked.(note: 1. carry out weighing after pair honey without crystallization need stir; 2. if any crystallized honey, place that not to be higher than 60 ℃ of water-baths warm, stir after waiting honey all to melt, carry out weighing after being cooled to room temperature.)
2. use the ELISA test strip sample
Detect milk sample
From original packing, take out requisite number purpose micropore reagent and test paper, and carry out mark; Draw milk sample to be checked, pipette 200 μ l in micropore with micropipettor, slowly suction and reagent mixing fully and in the micropore, room temperature (20 ℃-25 ℃) is hatched 5min; To be printed on " MAX " line end and insert down in the micropore, and make it fully to immerse in the solution; After room temperature (20 ℃-25 ℃) is hatched 5min, take out test paper, result of determination.
Detect honey sample
From original packing, take out requisite number purpose micropore reagent and test paper, and carry out mark; Draw honey extract to be checked, pipette 100 μ l in micropore with micropipettor, slowly reagent mixing in suction and abundant and the micropore; To be printed on " MAX " line end and insert down in the micropore, and make it fully to immerse in the solution; After room temperature (20C-25 ℃) is hatched 5～10min, take out test paper, according to the synoptic diagram result of determination.
3. Analysis of test results
Positive: (C) demonstrates band when the Quality Control district, and detection zone (T) does not develop the color, and is judged to the positive, with "+" expression, such as Fig. 4 (a); Negative: as when Quality Control district (C) and detection zone (T) all demonstrate band, to be judged to feminine gender, to represent with "-", such as Fig. 4 (b); Invalid: (C) do not show band when the Quality Control district, and test paper lost efficacy, such as Fig. 4 (c) with (d).
Determining of embodiment 4 Lateral Flow Strip parameters
1. detectability test
Add respectively streptomysin in the blank honey sample, dihydrostreptomycin standard items (available from Sigma company) to final concentration is 0,10,20,40 μ g/kg, carrying out honey sample with test strips detects, the result is: when streptomysin, when dihydrostreptomycin standard items concentration is 0,10 μ g/kg, demonstrate macroscopic two red lines on the test paper, be negative; When streptomysin, when dihydrostreptomycin standard items concentration is 20,40 μ g/kg, test paper Quality Control district colour developing, but detection zone do not develop the color, and is positive, and shows that this test strips is to honey sample streptomysin, dihydrostreptomycin detectability 20 μ g/kg.
Add respectively streptomysin in the blank milk sample, dihydrostreptomycin standard items to final concentration is 0,40,80,160 μ g/L, carrying out milk sample with test strips detects, the result is: when streptomysin, when dihydrostreptomycin standard items concentration is 0,40 μ g/L, test paper demonstrates macroscopic two red lines, is negative; When streptomysin, when dihydrostreptomycin standard items concentration is 80,160 μ g/L, test paper Quality Control district colour developing, but detection zone do not develop the color, and is positive, and shows that this test strips is to milk sample streptomysin, dihydrostreptomycin detectability 80 μ g/L.
2. false positive rate, false negative rate test
Get known content of streptomycin greater than 20 parts of the honey positive of 20 μ g/kg and content of streptomycin less than 20 parts of the honey negative samples of 20 μ g/kg, and known content of streptomycin greater than the milk positive of 80 μ g/L and 20 parts of content of streptomycin less than 20 parts of the milk negative samples of 80 μ g/L, test strips with 3 batches of productions detects respectively, the results are shown in Table 1, table 2.
Table 1 detects the positive result
The result shows: during with the positive honey sample of the ELISA test strip of 3 batches of productions, the result is entirely positive, and the positive sample coincidence rate is 100% as can be known, and false negative rate is 0.When detecting 20 parts of negative honey samples, the result is entirely negative, and negative match-rate is 100% as can be known, and false positive rate is 0.
During with the positive milk sample of the ELISA test strip of 3 batches of productions, the result is entirely positive, and the positive sample coincidence rate is 100% as can be known, and false negative rate is 0.When detecting 20 parts of negative milk samples, the result is entirely negative, and negative match-rate is 100% as can be known, and false positive rate is 0.Detection streptomysin test strips of the present invention can be to honey and the residual fast detecting of carrying out of milk sample streptomycin.
3. specific test
Specificity cross reacting rate commonly used represents, refers to the ability of the antigenic determinant generation combination that antibody is different from structure.The phosphate buffer of the normal other drug (melamine, sulfamido, fluoroquinolones, chloromycetin, macrolides, tetracycline medication) of examining in the sample with pH7.2,0.2mol/L diluted, detect with the test strips described in the embodiment 1.The result shows that during with these ELISA test strip 500 μ g/L melamines, sulfamido, fluoroquinolones, chloromycetin, macrolides, tetracycline medication, test strips Quality Control district and detection zone all develop the color, and illustrate that this test strips is to these medicine no cross reactions.
Claims (9)
1. test strips that detects streptomysin and dihydrostreptomycin; it is characterized in that comprising test paper and micropore reagent; described test paper comprises reaction film, absorption of sample pad, adsorptive pads, diaphragm, base plate; detection zone and Quality Control district are arranged on the described reaction film; detection zone is coated with streptomysin hapten-carrier protein conjugate; the Quality Control district is coated with antiantibody, and freeze-drying has streptomysin specific antibody-colloid gold label thing in the described micropore reagent.
2. test strips according to claim 1 is characterized in that described test paper is sticked on the base plate successively by absorption of sample pad, reaction film, adsorptive pads, diaphragm to form, and has the micropore plug on the described micropore reagent.
3. test paper according to claim 2 is characterized in that described diaphragm sticks on the absorption of sample pad, is the test side, and the above has the MAX mark line.
4. test strips according to claim 1 is characterized in that described detection zone is positioned at an end of the diaphragm that is bordering on the MAX mark, and described Quality Control district is positioned at the end away from the diaphragm that the MAX mark is arranged.
5. test strips according to claim 1, it is characterized in that described streptomysin hapten-carrier protein conjugate is obtained by streptomysin haptens and carrier protein couplet, described carrier protein can be bovine serum albumin(BSA), oralbumin, thyroprotein, hemocyanin, human serum albumins.
6. according to claim 1 or 5 each described test strips, it is characterized in that described streptomysin haptens structural formula is as follows:
7. test strips according to claim 1, it is characterized in that the streptomysin specific antibody in described streptomysin specific antibody-colloid gold label thing is to prepare as immunogene with streptomysin hapten-carrier protein conjugate, described streptomysin specific antibody can be streptomysin monoclonal antibody or streptomysin polyclonal antibody.
8. method for preparing each described test strips of claim 1-7, it comprises step:
1) the preparation freeze-drying has the micropore reagent of streptomysin specific antibody-colloid gold label thing;
2) preparation has the reaction film in the Quality Control district of the detection zone of coated streptomysin hapten-carrier protein conjugate and coated antiantibody;
3) with 2) reaction film and absorption of sample pad, adsorptive pads, diaphragm, the base plate that prepare be assembled into test paper;
4) with 1) and 3) freeze-drying for preparing has micropore reagent and the test paper of streptomysin specific antibody-colloid gold label thing to be assembled into test strips.
9. method that the test sample streptomycin is residual, it comprises step:
1) sample pre-treatments;
2) detect with each described test strips of claim 1-7;
3) analyzing and testing result.
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| CN104678096A (en) * | 2013-11-27 | 2015-06-03 | 中国农业大学 | Streptomycin and tetracycline two-in-one gold label micropore test paper strip |
| CN106771140A (en) * | 2016-11-22 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | A kind of detection kit of food streptomycin |
| CN106771157A (en) * | 2016-11-30 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | A kind of fortimicin detection method and detection card |
| CN106771125A (en) * | 2016-11-25 | 2017-05-31 | 南方医科大学 | A kind of time resolution immunity detection reagent of real-time monitoring anthracycline chemotherapy medicine blood concentration |
| CN109884296A (en) * | 2019-02-26 | 2019-06-14 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of kit of the quick detection African swine fever using the direct labeled primer of nanogold |
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| CN104678096A (en) * | 2013-11-27 | 2015-06-03 | 中国农业大学 | Streptomycin and tetracycline two-in-one gold label micropore test paper strip |
| CN106771140A (en) * | 2016-11-22 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | A kind of detection kit of food streptomycin |
| CN106771125A (en) * | 2016-11-25 | 2017-05-31 | 南方医科大学 | A kind of time resolution immunity detection reagent of real-time monitoring anthracycline chemotherapy medicine blood concentration |
| CN106771157A (en) * | 2016-11-30 | 2017-05-31 | 百奥森（江苏）食品安全科技有限公司 | A kind of fortimicin detection method and detection card |
| CN109884296A (en) * | 2019-02-26 | 2019-06-14 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of kit of the quick detection African swine fever using the direct labeled primer of nanogold |
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| CN103376316B (en) | 2016-07-27 |
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