CN104977407A - Colloidal gold immunochromatography test paper strip for detecting tetracycline drugs, and preparation method thereof - Google Patents
Colloidal gold immunochromatography test paper strip for detecting tetracycline drugs, and preparation method thereof Download PDFInfo
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Abstract
The present invention discloses a colloidal gold immunochromatography test paper strip for detecting tetracycline drugs, and a preparation method thereof, wherein the colloidal gold immunochromatography test paper strip comprises a sample pad, a conjugation pad, a nitrocellulose coating membrane, a water absorption pad and a PVC bottom plate, the sample pad, the conjugation pad, the nitrocellulose coating membrane and the water absorption pad are sequentially overlapped on the PVC bottom plate, a tetracycline monoclonal antibody-colloidal gold marker is coated on the conjugation pad, a detection line and a quality control line are arranged on the nitrocellulose coating membrane, tetracycline drug antigen is coated in the detection line, and goat anti-mouse IgG is coated in the quality control line. The present invention further discloses a preparation method of the colloidal gold immunochromatography test paper strip, wherein the method comprises colloidal gold preparing, gold-labeled antibody preparing and purification, and test paper strip assembly. The colloidal gold immunochromatography test paper strip of the present invention has characteristics of strong specificity, high sensitivity, easy operation, and rapid and accurate detection, and is suitable for on-site use.
Description
Technical field
The present invention relates to the technical field that aquatic products detect, particularly relate to a kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof.
Background technology
China be an ocean with freshwater resources than more rich country, aquatic products also just naturally become indispensable food in China resident daily life.Due to the development of China's aquaculture, the aquatic products foreign trade turnover of China also increases year by year.But recent years, the average annual export growth rate of China's aquatic products but shows as fluctuated.In 2002, the animal derived food of import prohibition China especially of European Union, make China be subject to the economic loss of nearly 100,000,000 yuans, trace it to its cause, mainly medicament residue exceeded standard.
In recent years, along with the development of each side technology, increasing medicine is applied to aquaculture, and the healthy of consumer in serious threat, also limit the export trade of China in aquatic products etc., establish and improve and improve the detection means of related fields and method just seems especially important.Be widely used in the various kinds of drug in aquaculture, antibiotic use amount has just exceeded the half of total amount.In China, the use amount of tetracycline medication accounts for 57% of the total use amount of microbiotic.Meanwhile, the use of some developed country's tetracycline medications is equally commonplace.The such as U.S., has half to be exactly tetracycline medication in the use total amount of all veterinary drugs.Tetracycline medication can be commonly used biocidal efficacies, the low price self mainly due to it with broad-spectrum high efficacy, and can either be used for animal in prevention and therapy aquaculture process suffered from the disease, again can as the feed addictive promoting cultivated animals growth.But user, when using such medicine, also exists again a series of abuse phenomenons such as a large amount of such medicine of use, long-time such medicine of use, causes serious medicament residue.
Present stage, high performance liquid chromatography, capillary electrophoresis, microbial method, euzymelinked immunosorbent assay (ELISA), electrochemical methods etc. is mainly utilized to carry out quantitative and qualitative analysis detection to tetracycline medication etc. both at home and abroad.It is also relatively high that high performance liquid chromatography has stronger automaticity degree, degree of accuracy, but also also exist that instrument price is high, pretreatment process is complicated, professional requirement be highly unfavorable for widely used shortcoming; The instrument and equipment that capillary electrophoresis is easy and simple to handle, analysis speed is fast but use in experimentation and reagent chemicals more, also higher to environmental requirement, be not suitable for on-the-spot test etc.; Microbial method is on the less demanding of sample itself but there is poor specificity, be subject to other medicines impact shortcoming; Euzymelinked immunosorbent assay (ELISA) is disposable can process multiple sample, high specificity, highly sensitive but higher to the professional requirement of operator itself, is unsuitable for universal use; The detection sensitivity of chemoluminescence method is high, easily realize robotization but there is the shortcoming high to environmental requirement, is unsuitable for on-the-spot test.For the shortcoming of current each class methods, be badly in need of a kind ofly quick and precisely to be applicable to Site Detection and to require to environment and operator itself detection method that lower tetracycline medication is residual.
In view of this, the present inventor studies and devises a kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, and this case produces thus.
Summary of the invention
The object of the present invention is to provide a kind of high specificity, highly sensitive, easy and simple to handle, detect quick, accurate and be applicable to colloidal gold immuno-chromatography test paper strip of the special detection tetracycline medication of onsite application and preparation method thereof, filling up the blank not yet having the colloidal gold immuno-chromatography test paper strip of tetracycline medication in aquatic products at present.
For achieving the above object, the present invention solves the technical scheme of its technical matters and is:
A kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, comprise sample pad, pad, cellulose nitrate coated film, adsorptive pads and PVC base plate, described sample pad, described pad, described cellulose nitrate coated film and described adsorptive pads are overlapped on described PVC base plate successively, and described pad bag is provided with Tetracyclines monoclonal antibody-colloid gold label thing; Described cellulose nitrate coated film is provided with detection line and nature controlling line, and described detection line is coated with tetracycline medication antigen, and described nature controlling line is coated with sheep anti-mouse igg.
A preparation method for the colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, comprises the following steps:
The preparation of step one, collaurum: get 1 250 mL beaker, add 100 mL 0.01% chlorauric acid solutions, in constant temperature blender with magnetic force, serviceability temperature shelves 400 DEG C heat it, after being heated to fluidized state, actual temperature during boiling is 100 DEG C, after keeping boiling 30 s, temperature shelves are become 200 DEG C, rotating speed shelves are adjusted to 200 r/min simultaneously, 500 μ L 4% citric acid three sodium solutions are added fast in beaker, solution in beaker heats 10 min after becoming redness again, i.e. obtained collaurum;
The preparation of step 2, golden labeling antibody and purifying: getting and mixing up pH is that described collaurum 1 mL of 7.9 is in 1.5 mL centrifuge tubes, 9.6 μ g/ml are added in centrifuge tube, extension rate is the Tetracyclines monoclonal antibody of 6, leaves standstill 30 min with after vortex instrument vortex 2 min; In centrifuge tube, add 100 μ L 10% BSA solution, with vortex instrument vortex 2 min, leave standstill 30 min, obtained golden labeling antibody, i.e. Tetracyclines monoclonal antibody-colloid gold label thing;
Simultaneously, by obtained golden labeling antibody solution in 4 DEG C of 2500 r/min, centrifugal 20 min, remove lower sediment, supernatant is forwarded in another centrifuge tube, in 4 DEG C of 12000 r/min, centrifugal 30 min, remove supernatant, precipitation collaurum dilute solution is diluted to 1/10 of original volume, saves backup in 4 DEG C;
The assembling of step 3, test strips: (1) by cellulose nitrate coated film, i.e. NC film, and adsorptive pads is affixed on PVC base plate respectively, and adsorptive pads pushes down 1-2 mm above NC film; (2) the PVC base plate pasting NC film and adsorptive pads is cut into the wide test strips of 4 mm on cutting cutter; (3) sample pad and pad are cut into the wide bar of 4 mm, pad are soaked in golden labeling antibody solution simultaneously, and in 37 DEG C of oven dry, obtained gold mark pad; (4) the gold mark pad after drying is cut into the long fritter of 1 cm and is affixed on PVC base plate, and 1-2 mm below NC film pushed down by gold mark pad, pushes down gold mark pad below 1-2 mm simultaneously, prune sample pad according to test strips length by sample pad; (5) the NC film between pad and absorption pad wraps successively by tetracycline medication antigen and sheep anti-mouse igg, respectively as detection line and control line, wherein, the antigen coated concentration of tetracycline medication is 1.0 mg/mL; Sheep anti-mouse igg bag is 0.75 mg/mL by concentration.
As the optimal way of embodiment, in described step 3 (3), also comprise the step of sample pad and gold mark pad being carried out to pre-treatment, wide rectangular of 4 mm is cut into respectively by sample pad and gold-marking binding pad cutting cutter, soak by the PBS solution that the pH that sucrose, 5% BSA and the final volume concentration containing 5% is 0.1% is 0.01 mol/L of 7.9, drained by solution after immersion, under 37 DEG C of conditions, dry 12 h for subsequent use.
As the optimal way of embodiment, to the cleaning of test container used before also comprising step one: first will test container used and soak 24 h in potassium dichromate washing lotion, and carry out several times cleaning with tap water after taking-up, then clean 3 times with ultrapure water; Be carry out silicidation after soaking 1 min in the chloroform of 5% in dichlorosilane content by the container cleaned up; The container crossed through silicidation is positioned under room temperature condition dry, to be dried complete after to clean up with ultrapure water and in baking oven 37 DEG C carry out drying and process, for subsequent use.
Compared with prior art, the present invention has following advantages:
1, the colloidal gold immuno-chromatography test paper strip of tetracycline medication in detection aquatic products of the present invention, has high specificity, highly sensitive, the advantages such as detection time is short;
2, test strip of the present invention is without any need for specific apparatus, equipment, can execute-in-place, and testing cost is low;
3, test strip of the present invention is easy and simple to handle, does not need professional to operate;
4, the present invention mark pad carried out pre-treatment to sample pad and gold, sample pad and gold are marked water-intake capacity and releasability that spacer has excellence, and then substantially increases detectability;
5, the present invention cleans test container used, finally enhances the stability of golden labeling antibody.
figure of description
Fig. 1 is the structural representation of colloidal gold immuno-chromatography test paper strip of the present invention;
Fig. 2 is the sensitivity test result figure of the embodiment of the present invention 3 Performance Detection;
Fig. 3 is the specificity test result figure of the embodiment of the present invention 3 Performance Detection; Wherein, tetracycline, aureomycin, terramycin, penicillin, chloromycetin, streptomysin test is respectively from left to right;
Fig. 4 is the stability test result figure of the embodiment of the present invention 3 Performance Detection; Wherein, No. 1 for preserve at normal temperatures, No. 2 for preserve under 37 DEG C of conditions;
Fig. 5 is the reperformance test result figure of the embodiment of the present invention 3 Performance Detection;
Fig. 6 is the test strips test result figure of the embodiment of the present invention 4 test sample; Wherein, former state, 2 times of dilutions, 4 times of dilutions, 8 times of dilutions, 16 times of dilutions are respectively in figure from left to right.
Embodiment
embodiment 1
As shown in Figure 1, present invention is disclosed a kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, comprise sample pad 1, pad 2, cellulose nitrate coated film 3, adsorptive pads 4 and PVC base plate 5, described sample pad 1, described pad 2, described cellulose nitrate coated film 3 and described adsorptive pads 4 are overlapped on described PVC base plate 5 successively: described pad 2 bag is provided with Tetracyclines monoclonal antibody-colloid gold label thing; Described cellulose nitrate coated film 3 is provided with detection line 31 and nature controlling line 32, and described detection line 31 is coated with tetracycline medication antigen, and described nature controlling line 32 is coated with sheep anti-mouse igg.
embodiment 2
A preparation method for the colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, comprises the following steps:
The preparation of step one, collaurum: get 1 250 mL beaker, add 100 mL 0.01% chlorauric acid solutions, in constant temperature blender with magnetic force, serviceability temperature shelves 400 DEG C heat it, after being heated to fluidized state, actual temperature during boiling is 100 DEG C, after keeping boiling 30 s, temperature shelves are become 200 DEG C, rotating speed shelves are adjusted to 200 r/min simultaneously, 500 μ L 4% citric acid three sodium solutions are added fast in beaker, solution in beaker heats 10 min after becoming redness again, i.e. obtained collaurum;
The preparation of step 2, golden labeling antibody and purifying: getting and mixing up pH is that described collaurum 1 mL of 7.9 is in 1.5 mL centrifuge tubes, 9.6 μ g/ml are added in centrifuge tube, extension rate is the Tetracyclines monoclonal antibody of 6, leaves standstill 30 min with after vortex instrument vortex 2 min; In centrifuge tube, add 100 μ L 10% BSA solution, with vortex instrument vortex 2 min, leave standstill 30 min, obtained golden labeling antibody, i.e. Tetracyclines monoclonal antibody-colloid gold label thing;
Simultaneously, by obtained golden labeling antibody solution in 4 DEG C of 2500 r/min, centrifugal 20 min, remove lower sediment, supernatant is forwarded in another centrifuge tube, in 4 DEG C of 12000 r/min, centrifugal 30 min, remove supernatant, precipitation collaurum dilute solution is diluted to 1/10 of original volume, saves backup in 4 DEG C;
The assembling of step 3, test strips: (1) by cellulose nitrate coated film, i.e. NC film, and adsorptive pads is affixed on PVC base plate respectively, and adsorptive pads pushes down 1-2 mm above NC film; (2) the PVC base plate pasting NC film and adsorptive pads is cut into the wide test strips of 4 mm on cutting cutter; (3) sample pad and pad are cut into the wide bar of 4 mm, pad are soaked in golden labeling antibody solution simultaneously, and in 37 DEG C of oven dry, obtained gold mark pad; (4) the gold mark pad after drying is cut into the long fritter of 1 cm and is affixed on PVC base plate, and 1-2 mm below NC film pushed down by gold mark pad, pushes down gold mark pad below 1-2 mm simultaneously, prune sample pad according to test strips length by sample pad; (5) the NC film between pad and absorption pad wraps successively by tetracycline medication antigen and sheep anti-mouse igg, respectively as detection line and control line, wherein, the antigen coated concentration of tetracycline medication is 1.0 mg/mL; Sheep anti-mouse igg bag is 0.75 mg/mL by concentration.
As the optimal way of embodiment, in described step 3 (3), also comprise the step of sample pad and gold mark pad being carried out to pre-treatment, wide rectangular of 4 mm is cut into respectively by sample pad and gold-marking binding pad cutting cutter, soak by the PBS solution that the pH that sucrose, 5% BSA and the final volume concentration containing 5% is 0.1% is 0.01 mol/L of 7.9, drained by solution after immersion, under 37 DEG C of conditions, dry 12 h for subsequent use.
As the optimal way of embodiment, to the cleaning of test container used before also comprising step one: first will test container used and soak 24 h in potassium dichromate washing lotion, and carry out several times cleaning with tap water after taking-up, then clean 3 times with ultrapure water; Be carry out silicidation after soaking 1 min in the chloroform of 5% in dichlorosilane content by the container cleaned up; The container crossed through silicidation is positioned under room temperature condition dry, to be dried complete after to clean up with ultrapure water and in baking oven 37 DEG C carry out drying and process, for subsequent use.Whether clean container used in this test is, and the quality of the final colloidal gold solution obtained has very large difference.If there is impurity or dust in container, so they just can affect the formation of gold grain to a certain extent, the final homogeneity affecting particle in colloidal gold solution, also can cause the bad stability of the golden labeling antibody obtained in follow-up test, so container needs through above-mentioned a series of process before using simultaneously.
embodiment 3 Performance Detection
3.1 sensitivity test
Be that the standard items of tetracycline, aureomycin, terramycin are formulated as the different concentration gradient such as 100 μ g/kg, 50 μ g/kg, 25 μ g/kg, 15 μ g/kg by the PBS solution of the 0.01mol/L of 7.9 with pH, then detect with the tetracycline medication colloidal gold immuno-chromatography test paper strip assembled, result as shown in Figure 2.
As shown in Figure 2, when the content of tetracycline standard items is 15 μ g/L, pass through visual inspection, there is a light redness at the T line place of test strips, and when the content of tetracycline standard items is 20 μ g/L, test strips shows a line (nature controlling line), so test strips is 20 μ g/L for the Monitoring lower-cut of tetracycline standard items; When the content of aureomycin standard items is 15 μ g/L, pass through visual inspection, also there is a light redness at test strips T line place, darker than carrying out when tetracycline detects, and when the concentration of standard items is 20 μ g/L, test strips shows a line, so test strips is 20 μ g/L for the detection sensitivity of aureomycin; When the content of terramycin standard items is 15,20 μ g/L, by visual inspection, test strips shows two lines, and when the content of terramycin standard items is 25 μ g/L, test strips shows a line, so the obtained test strips of this test is 25 μ g/L for the Determination Limit of terramycin.
3.2 specificity tests
With structure and tetracycline medication similar penicillin, chloromycetin, streptomysin etc., the specificity of test strips is tested.Be that each standard items are diluted to different concentration gradients by the PBS solution of 0.01 mol/L of 7.9 with pH, negative with PBS solution test, test with a collection of test strips, observations, result as shown in Figure 3.
As shown in Figure 3, test strips is greater than 20 μ g/L at tetracycline concentration, chlortetracycline concentration is greater than 15 μ g/L, terramycin concentration all shows positive when being greater than 25 μ g/L.And when analog concentration is less than 200 μ g/L, tetracycline medication colloidal gold immuno-chromatography test paper strip all without obvious cross reaction, proves that the specificity of obtained test strips is better for analogs such as penicillin, chloromycetin, streptomysins.
3.3 stability test
The test strips assembled is positioned over (built-in drying agent) in different polybags, seal and airtightly to preserve respectively under room temperature and 37 DEG C of conditions, after placing 15 d, the test strips standard items preserved under selecting different placement condition and negative sample are tested, the stability of checking tetracycline medication colloidal gold immuno-chromatography test paper strip, result as shown in Figure 4.
Find in test, under normal temperature condition, preserve 15 d, the color not too large change of ELISA test strip line and nature controlling line.As can be seen from Figure 4, the test strips of preserving under 37 DEG C of conditions, during test, detection line is darker than what deposit under normal temperature condition with the color of nature controlling line.The reason occurring this phenomenon is mainly because the antigen-antibody in the test strips of preserving under 37 DEG C of conditions remains higher immunocompetence in certain hour section and make the specific reaction of antigen and antibody more easily carry out because the raising of temperature obtains more energy of activation.
3.4 reperformance test
With the sample of the ELISA test strip different batches of same batch; Detect with the test strips of the different batches sample to same batch, observations, result as shown in Figure 5 again.
As can be seen from Figure 5, in same batch, carry out 8 test strips of replica test, 2 for detect negative sample (comprise be less than ELISA test strip limit positive), test strips all shows two lines, is feminine gender; Article 6, for measuring positive, test strips all only has line colour developing (only showing nature controlling line), is the positive.In proving same batch, the repeatability of test strips is better.
Meanwhile, adopt 3 batches to amount to 12 test strips (each batch 4) and detect concentration known sample, the measurement result of final three batches is consistent, proves that the repeatability of the test strips of different batches is better.
embodiment 4 sample test
Sample preparation: the shrimp tissue samples shredded with balance 10 about g, carries out homogeneous with homogenizer to this tissue samples, and homogeneous completes and obtains about 1 g with electronic balance, puts it in 5 mL centrifuge tubes; The Mclvaine-EDTA solution that 2 mL pH are 4.0 is added wherein, with whirlpool instrument vortex after 3 minutes, when temperature is lower than 15 DEG C, with hydro-extractor centrifugal 15 min under 4000 r/min rotating speeds with liquid-transfering gun; With liquid-transfering gun get 1mL above-mentioned centrifugal after supernatant add in 5 mL centrifuge tubes, add the NaOH solution of 0.1 mL 4 mol/L wherein, with whirlpool instrument vortex 2 min, then 2 mL normal hexanes are added wherein, with whirlpool instrument vortex 2 min, leave standstill wait lower floor's solution to be shifted out after its layering etc. to be detected.
4.1 test strips tests
Choose prawn that market is sold as subjects, take this shrimp tissue of 2 part of 10 about g, label 1,2, two increment product are as parallel experiment, two increment product all process according to foregoing sample treatment, the test strips of getting extract preparation detects, and result as shown in Figure 6.
As can be seen from Figure 6, the positive is shown as after testing through extracting the sample stoste obtained, and the sensitivity of this test strips is mixed timestamp is that the 10 μ g/L(i.e. content of three class medicines is 10 μ g/L), tetracycline medication content 60 more than the μ g/kg in the shrimp that can use in this test of preliminary judgement.For determining the content of tetracycline medication in this sample further, 2 times, 4 times, 8 times, 16 times dilutions being carried out to this sample stoste, as can be seen from Figure 6 there will be obvious two lines testing result now when extension rate is 16 times for negative.To sum up, the tetracycline medication content of this shrimp of preliminary judgement is near 480 ~ 960 μ g/kg.
4.2 LC-MS tests
Process according to the pre-treating method of LC-MS in Tetracyclines detection of veterinary drugs in food method in GB/T 21317-2007 animal derived food, during upper machine, chromatographic column adopts Inertsil C8-3, mobile phase adopts the formic acid solution of chromatographically pure methyl alcohol and 0.2%, flow velocity is 300 μ L/min, column temperature is 30 DEG C, and sample size is 30 μ L.
When this test utilizes LC-MS to test the tetracycline medication in shrimp is residual, sampling amount is 1 g, and extract use amount is 15 mL, and adding scalar is mixed mark 500 μ g/kg, sample and mark-on test in extract all dilute 4 times, be finally settled to 2 mL.Learn after tested, the content in upper machine sample used is 38.53 μ g/kg, and recovery of standard addition is 72.1%, is 641.3 μ g/kg through calculating the content of the tetracycline medication finally drawn in sample, consistent with the result that test strips method records.
All distortion that those of ordinary skill in the art can directly derive from the disclosure of invention or associate, all should think protection scope of the present invention.
Claims (4)
1. colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, comprise sample pad, pad, cellulose nitrate coated film, adsorptive pads and PVC base plate, described sample pad, described pad, described cellulose nitrate coated film and described adsorptive pads are overlapped on described PVC base plate successively, it is characterized in that: described pad bag is provided with Tetracyclines monoclonal antibody-colloid gold label thing; Described cellulose nitrate coated film is provided with detection line and nature controlling line, and described detection line is coated with tetracycline medication antigen, and described nature controlling line is coated with sheep anti-mouse igg.
2. the preparation method of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication and preparation method thereof, is characterized in that: comprise the following steps:
The preparation of step one, collaurum: get 1 250 mL beaker, add 100 mL 0.01% chlorauric acid solutions, in constant temperature blender with magnetic force, serviceability temperature shelves 400 DEG C heat it, after being heated to fluidized state, actual temperature during boiling is 100 DEG C, after keeping boiling 30 s, temperature shelves are become 200 DEG C, rotating speed shelves are adjusted to 200 r/min simultaneously, 500 μ L 4% citric acid three sodium solutions are added fast in beaker, solution in beaker heats 10 min after becoming redness again, i.e. obtained collaurum;
The preparation of step 2, golden labeling antibody and purifying: getting and mixing up pH is that described collaurum 1 mL of 7.9 is in 1.5 mL centrifuge tubes, 9.6 μ g/ml are added in centrifuge tube, extension rate is the Tetracyclines monoclonal antibody of 6, leaves standstill 30 min with after vortex instrument vortex 2 min; In centrifuge tube, add 100 μ L 10% BSA solution, with vortex instrument vortex 2 min, leave standstill 30 min, obtained golden labeling antibody, i.e. Tetracyclines monoclonal antibody-colloid gold label thing;
Simultaneously, by obtained golden labeling antibody solution in 4 DEG C of 2500 r/min, centrifugal 20 min, remove lower sediment, supernatant is forwarded in another centrifuge tube, in 4 DEG C of 12000 r/min, centrifugal 30 min, remove supernatant, precipitation collaurum dilute solution is diluted to 1/10 of original volume, saves backup in 4 DEG C;
The assembling of step 3, test strips: (1) by cellulose nitrate coated film, i.e. NC film, and adsorptive pads is affixed on PVC base plate respectively, and adsorptive pads pushes down 1-2 mm above NC film; (2) the PVC base plate pasting NC film and adsorptive pads is cut into the wide test strips of 4 mm on cutting cutter; (3) sample pad and pad are cut into the wide bar of 4 mm, pad are soaked in golden labeling antibody solution simultaneously, and in 37 DEG C of oven dry, obtained gold mark pad; (4) the gold mark pad after drying is cut into the long fritter of 1 cm and is affixed on PVC base plate, and 1-2 mm below NC film pushed down by gold mark pad, pushes down gold mark pad below 1-2 mm simultaneously, prune sample pad according to test strips length by sample pad; (5) the NC film between pad and absorption pad wraps successively by tetracycline medication antigen and sheep anti-mouse igg, respectively as detection line and control line, wherein, the antigen coated concentration of tetracycline medication is 1.0 mg/mL; Sheep anti-mouse igg bag is 0.75 mg/mL by concentration.
3. the preparation method of a kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication as claimed in claim 2 and preparation method thereof, it is characterized in that: in described step 3 (3), also comprise the step of sample pad and gold mark pad being carried out to pre-treatment, wide rectangular of 4 mm is cut into respectively by sample pad and gold-marking binding pad cutting cutter, soak by the PBS solution that the pH that sucrose, 5% BSA and the final volume concentration containing 5% is 0.1% is 0.01 mol/L of 7.9, drained by solution after immersion, under 37 DEG C of conditions, dry 12 h for subsequent use.
4. the preparation method of a kind of colloidal gold immuno-chromatography test paper strip detecting tetracycline medication as claimed in claim 2 and preparation method thereof, it is characterized in that: to the cleaning of test container used before also comprising step one: first will test container used and soak 24 h in potassium dichromate washing lotion, carry out several times cleaning with tap water after taking-up, then clean 3 times with ultrapure water; Be carry out silicidation after soaking 1 min in the chloroform of 5% in dichlorosilane content by the container cleaned up; The container crossed through silicidation is positioned under room temperature condition dry, to be dried complete after to clean up with ultrapure water and in baking oven 37 DEG C carry out drying and process, for subsequent use.
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| CN106645679A (en) * | 2017-01-17 | 2017-05-10 | 吉林大学 | Method for detecting tetracycline (TET) based on colloidal gold (AuNPs) chromatography test strip of aptamer |
| CN106771268A (en) * | 2017-01-06 | 2017-05-31 | 何伟 | A kind of people's abo blood group reverse type colloidal gold strip and its preparation method and application |
| CN107176627A (en) * | 2016-03-11 | 2017-09-19 | 上海多佳水处理科技有限公司 | Traceable phosphate-free water treatment agent |
| CN110887964A (en) * | 2019-11-07 | 2020-03-17 | 西北农林科技大学 | Sensitive probe, method for detecting tetracycline and application |
| CN112305216A (en) * | 2020-10-26 | 2021-02-02 | 杭州电子科技大学 | Carbon nano-immune label chromatography test strip and method for detecting antibiotics in animal hair |
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| CN112305216A (en) * | 2020-10-26 | 2021-02-02 | 杭州电子科技大学 | Carbon nano-immune label chromatography test strip and method for detecting antibiotics in animal hair |
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