CN110441517B - Test strip and method for detecting 6-benzyladenine - Google Patents
Test strip and method for detecting 6-benzyladenine Download PDFInfo
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Abstract
The invention discloses a test strip and a method for detecting 6-benzyladenine. The test strip comprises test paper and a microporous reagent, wherein the test paper comprises a reaction membrane, a sample absorption pad, a water absorption pad and a bottom plate, a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody are arranged on the reaction membrane, and a 6-benzyladenine monoclonal antibody-colloidal gold marker is freeze-dried on the microporous reagent. The method for detecting 6-benzyladenine by using the test strip is simple, convenient, rapid, visual, accurate, convenient to carry, wide in application range, low in cost and easy to popularize and use.
Description
Technical Field
The invention relates to a test strip and a method for detecting 6-benzyladenine, in particular to a colloidal gold test strip for detecting 6-benzyladenine, which is particularly suitable for detecting 6-benzyladenine residues in bean sprouts.
Background
6-benzyladenine (6-BA) is an artificially synthesized cytokinin, which is a white or off-white crystal, is poorly soluble in water, is slightly soluble in ethanol, and is stable in acid and alkali. Has the function of inhibiting the decomposition of chlorophyll, nucleic acid and protein in plant leaves, is mainly used for the cultivation of grains and fruit trees and gardening, and can be applied to all growth stages of crops. 6-BA is currently widely used as a growth regulator for rootless bean sprouts. Excessive intake of 6-BA in human body can stimulate skin mucosa, cause injury of esophagus and gastric mucosa, and cause nausea, vomiting and the like. The original national food and drug administration, ministry of agriculture, national health and family planning committee of China stipulate in the bulletin (No. 11 in 2015) about the prohibition of using 6-benzyladenine and other substances in the bean sprout production process: the producers cannot use 6-benzyladenine, 4-sodium chlorophenoxyacetate, gibberellin, etc. in the production process of bean sprouts, and the bean sprouts owners cannot run bean sprouts containing 6-benzyladenine, 4-sodium chlorophenoxyacetate, gibberellin, etc.
The reported methods for detecting 6-benzyladenine mainly comprise liquid chromatography, liquid chromatography tandem mass spectrometry and other instrument methods. The methods are operated under laboratory conditions, the sample pretreatment is complicated and time-consuming, expensive instruments and equipment are required, the detection cost is high, the time consumption is long, the operation is complex, the method has great limitation in the practical application process, and the requirement for rapidly detecting a large number of samples and field samples is difficult to meet. Therefore, the colloidal gold test strip which is simple and rapid and is suitable for 6-benzyladenine residue in bean sprouts is developed, a large number of samples can be screened and monitored on site, and detection work of food supervision departments and the like in China can be better met.
Disclosure of Invention
The invention aims to provide a colloidal gold test strip capable of detecting 6-benzyladenine residues in bean sprouts, and provides a detection method which is efficient, accurate, simple and convenient and is suitable for field monitoring and large-scale sample screening.
The test strip for detecting 6-benzyladenine provided by the invention comprises test paper and a micropore reagent; the test paper comprises a bottom plate, a sample absorption pad, a reaction membrane and a water absorption pad which are sequentially connected, wherein the reaction membrane is provided with a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody; the micropore reagent is freeze-dried with a 6-benzyladenine monoclonal antibody-colloidal gold marker which is provided with a micropore plug.
The 6-benzyladenine monoclonal antibody is prepared by taking a 6-benzyladenine hapten-carrier protein conjugate as an immunogen.
The 6-benzyladenine hapten-carrier protein conjugate is obtained by coupling 6-benzyladenine hapten and carrier protein, the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroxine or human serum albumin, the 6-benzyladenine hapten is obtained by reacting amino-6-benzyladenine and 4- (3-oxopropyl) -benzoic acid, and the molecular structural formula is as follows:
the bottom plate can be a PVC bottom plate or other hard non-absorbent materials; the sample absorption pad can be suction filter paper or oil filter paper; the water absorption pad is absorbent paper; the reaction membrane can be a nitrocellulose membrane or a cellulose acetate membrane.
Another object of the present invention is to provide a method for preparing the test strip, which comprises the steps of:
1) Preparing a microporous reagent freeze-dried with a 6-benzyladenine monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the prepared reaction membrane in the step 2) with a sample absorption pad, a water absorption pad and a base plate to form test paper;
4) And (2) assembling the microporous reagent prepared in the steps 1) and 3) and freeze-dried with the 6-benzyladenine monoclonal antibody-colloidal gold marker and test paper into the test paper strip.
Specifically, the steps include:
1) Reacting amino-6-benzyladenine with 4- (3-oxopropyl) -benzoic acid to prepare a 6-benzyladenine hapten;
2) Coupling the 6-benzyladenine hapten with carrier protein to prepare a 6-benzyladenine hapten-carrier protein conjugate;
3) Immunizing a mouse by using the 6-benzyladenine hapten-carrier protein conjugate, and fusing and screening spleen cells and myeloma cells of the mouse to obtain a hybridoma cell strain secreting the 6-benzyladenine monoclonal antibody;
4) Extracting mouse IgG to immunize a healthy goat to obtain a goat anti-mouse anti-antibody;
5) Respectively coating the 6-benzyladenine hapten-carrier protein conjugate and the goat anti-mouse anti-antibody on a detection line (T) and a quality control line (C) of a reaction membrane;
6) Preparing colloidal gold by reacting trisodium citrate with chloroauric acid;
7) Adding the prepared 6-benzyladenine monoclonal antibody into the prepared colloidal gold to obtain a 6-benzyladenine monoclonal antibody-colloidal gold marker;
8) Freeze-drying the 6-benzyladenine monoclonal antibody-colloidal gold marker in a micropore reagent, and adding a micropore plug into the micropore reagent;
9) Soaking the sample absorption pad in 0.5% bovine serum albumin-containing phosphate buffer solution with pH of 7.2 and 0.1mol/L for 2h, and drying at 37 deg.C for 2h;
10 A sample absorbing pad, a reaction film and a water absorbing pad are sequentially adhered on the bottom plate, and finally the mixture is cut into small strips with the width of 3mm, added with a plastic box and vacuum-packaged;
11 Prepared micropore reagent and test paper are assembled into test paper strips, and the test paper strips are stored for 12 months at the temperature of 2-8 ℃.
The invention also aims to provide a method for detecting 6-benzyladenine residues in bean sprouts by using the test strip, which comprises the following steps:
(1) Pretreating a sample;
(2) Detecting with a test strip;
(3) And analyzing the detection result.
The 6-benzyladenine rapid detection test strip adopts a highly specific antibody antigen reaction and immunochromatography analysis technology, a 6-benzyladenine monoclonal antibody-colloidal gold marker is lyophilized in a microporous reagent, and 6-benzyladenine in a sample is fully reacted with the monoclonal antibody-colloidal gold marker to form a drug-antibody-colloidal gold marker; and then the reagent is dropped into a test strip card hole, the medicine in the sample competes with the 6-benzyladenine hapten-carrier protein conjugate on the reaction membrane detection line to combine with the 6-benzyladenine monoclonal antibody-colloidal gold marker in the flowing process, and whether the sample liquid to be detected contains 6-benzyladenine residue is judged according to the depth of a red strip of the detection line.
During detection, a sample is treated and then dropped into a microporous reagent, the microporous reagent is added into a test strip card hole after full reaction, when the concentration of 6-benzyladenine in the sample is lower than the detection limit or zero, a monoclonal antibody-colloidal gold marker can be combined with a 6-benzyladenine hapten-carrier protein conjugate fixed on a reaction membrane in the chromatography process, a red strip appears on a detection line (T) and a quality control line (C), and the color development of the T line is deeper than that of the C line or is consistent with that of the C line; if the concentration of 6-benzyladenine in the sample is equal to or higher than the detection limit, the monoclonal antibody-colloidal gold label will bind to the 6-benzyladenine in its entirety, and thus no red band or lighter color than the C line will appear at the T line because the competitive reaction will not bind to the 6-benzyladenine hapten-carrier protein conjugate. As shown in fig. 4.
Negative: when the quality control line (C) shows a red strip, the detection line (T) also shows a red strip, and the color of the line (T) is close to or deeper than that of the line (C), the line (C) is judged to be negative.
Positive: when the quality control line (C) shows a red band, and the detection line (T) does not show color or the color of the line (T) is lighter than that of the line (C), the test result is judged to be positive.
And (4) invalidation: when the quality control line (C) does not show a red strip, the test strip is judged to be invalid whether the detection line (T) shows a red strip or not.
The test strip has the advantages of high sensitivity, strong specificity, low cost, simple operation, short detection time, suitability for various units, simple storage and long quality guarantee period. Wherein, the 6-benzyladenine monoclonal antibody with high specificity is adopted, so that the reliability of the detection result is ensured; the gold-labeled antibody is freeze-dried in the microporous reagent, and in the detection process, the gold-labeled antibody can be fully contacted with a sample solution to be detected to fully react, so that the error is reduced, and the reaction sensitivity of the whole system is increased. The method for detecting 6-benzyladenine by using the test strip is simple, convenient, rapid, visual, accurate, wide in application range, low in cost and easy to popularize and use.
Drawings
FIG. 1 is a diagram of the synthesis of 6-benzyladenine hapten.
FIG. 2 is a schematic cross-sectional view of the test strip.
FIG. 3 is a diagram of a microwell reagent.
FIG. 4 is a diagram showing the test result of the test strip.
Detailed Description
The invention is further illustrated below with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Example 1 preparation of test strip for detecting 6-benzyladenine
The preparation method of the test strip mainly comprises the following steps:
1) Preparing a microporous reagent freeze-dried with a 6-benzyladenine monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the prepared reaction membrane in the step 2) with a sample absorption pad, a water absorption pad and a base plate to form test paper;
4) And (2) assembling the microporous reagent prepared in the steps 1) and 3) and freeze-dried with the 6-benzyladenine monoclonal antibody-colloidal gold marker and test paper into the test paper strip.
The following steps are detailed:
1. synthesis of 6-benzyladenine hapten (synthetic route see figure 1)
Taking 2.4g of amino-6-benzyladenine, adding 70mL of ethanol for dissolving, adding 0.45g of NaOH, fully stirring, adding 1.85g of 4- (3-oxopropyl) -benzoic acid, heating at 60 ℃ for reaction for 4h, cooling to room temperature, adding 0.38g of sodium borohydride, continuing stirring for 2h, stopping the reaction, adding 120mL of water, adding 6mol/L of hydrochloric acid for adjusting the pH value to 6, adding 80mL of ethyl acetate, extracting, separating a water phase, evaporating an organic phase by rotation, and evaporating to dryness to obtain a red oily substance, and recrystallizing by 165mL of acetonitrile/n-hexane (1, V/V) to obtain 1.9g of a 6-benzyladenine hapten product with the yield of 47.26%.
2. Preparation of immunogens
Taking 15mg of 6-benzyladenine hapten product, adding 1mL of N, N-Dimethylformamide (DMF), dissolving and clarifying, adding 13mg of carbodiimide (EDC) and 9.4mg of N-hydroxysuccinimide (NHS), fully and uniformly mixing, and reacting for 4h to obtain hapten activating solution A; taking 60mg of Bovine Serum Albumin (BSA), and adding 0.05mol/L PB for dissolving to obtain solution B; dripping the A solution into the B solution, reacting at 4 deg.C for 24h, dialyzing with 0.02mol/L PBS for purifying for three days, changing solution 3 times per day to obtain 6-benzyl adenine hapten-BSA conjugate as immunogen, packaging, and storing at-20 deg.C.
3. Preparation of coating antigen
Taking 9.6mg of a 6-benzyladenine hapten product, adding 1mL of DMF, dissolving and clarifying, adding 7.7mg of EDC and 6.4mg of NHS, fully and uniformly mixing, and reacting for 4h to obtain a hapten activating solution A; dissolving Ovalbumin (OVA) 50mg with PB 0.05mol/L to obtain solution B; dripping the A solution into the B solution, reacting at 4 deg.C for 24h, dialyzing and purifying with 0.02mol/L PBS for three days, changing the solution 3 times per day to obtain 6-benzyladenine hapten-OVA conjugate, i.e. coating antigen, packaging, and storing at-20 deg.C.
4. Preparation of 6-benzyladenine monoclonal antibody
(1) Animal immunization
Injecting the immunogen obtained in the step 2 into Balb/c mice at an immune dose of 150 mug/mouse to generate antiserum.
(2) Cell fusion and cloning
Taking immune Balb/c mouse spleen cells, fusing the immune Balb/c mouse spleen cells with SP2/0 myeloma cells according to the proportion of 8 (quantitative ratio), measuring cell supernatant by adopting indirect competition ELISA, and screening positive holes. Cloning the positive hole by using a limiting dilution method until obtaining a hybridoma cell strain which stably secretes the monoclonal antibody.
(3) Cell cryopreservation and recovery
Preparing hybridoma cells into 1 × 10 frozen stock solution 6 Cell suspension per mL, stored for long periods in liquid nitrogen. Taken out at the time of resuscitationAnd (4) freezing and storing the tube, immediately putting the tube into a water bath at 37 ℃ for quick melting, centrifuging to remove frozen stock solution, and transferring the tube into a culture bottle for culture.
(4) Preparation and purification of monoclonal antibodies
An incremental culture method: placing the hybridoma cell in cell culture medium, culturing at 37 deg.C, purifying the obtained culture solution by octanoic acid-saturated ammonium sulfate method to obtain monoclonal antibody, and storing at-20 deg.C.
The cell culture medium is prepared by adding calf serum and sodium bicarbonate into RPMI1640 culture medium to make the final concentration of calf serum in the cell culture medium 20% (mass fraction) and the final concentration of sodium bicarbonate in the cell culture medium 0.2% (mass fraction); the pH of the cell culture medium was 7.4.
5. Preparation of goat anti-mouse anti-antibody
The sheep is taken as an immune animal, and the pathogen-free sheep is immunized by taking the murine antibody as an immunogen to obtain the goat anti-mouse antibody.
6. Preparation of 6-benzyladenine monoclonal antibody-colloidal gold marker
(1) Preparation of colloidal gold
Diluting 1% chloroauric acid to 0.01% (mass fraction) with double distilled deionized water, placing 100mL into a conical flask, heating to boil with a constant temperature electromagnetic stirrer, adding 2.5mL 1% trisodium citrate under continuous stirring at high temperature, stirring at constant speed, heating until the solution is bright red, cooling to room temperature, recovering the volume with deionized water, and storing at 4 deg.C. The prepared colloidal gold has pure appearance, is transparent and bright, has no sediment or floating objects, and is wine red when observed in sunlight.
(2) Preparation of 6-benzyladenine monoclonal antibody-colloidal gold marker
Under magnetic stirring, adjusting the pH value of the colloidal gold to 7.2 by using 0.2mol/L potassium carbonate solution, adding the 6-benzyladenine monoclonal antibody into the colloidal gold solution according to the standard that 20-50 mu g of antibody is added into each milliliter of the colloidal gold solution, and continuously stirring and uniformly mixing for 30min; after standing for 10min, 10% BSA was added so that the final concentration in the colloidal gold solution became 1%, and the mixture was allowed to stand for 10min. Centrifuging at 12000r/min at 4 deg.C for 40min, discarding supernatant, washing the precipitate with redissolving buffer twice, resuspending the precipitate with redissolving buffer with volume 1/10 of the original volume of colloidal gold, and standing at 4 deg.C for use.
Redissolving buffer solution: 0.1 to 0.3 percent of BSA, 0.05 to 0.2 percent of Tween-80 and 0.02mol/L phosphate buffer solution with pH of 7.2.
7. Preparation of microporous reagents
Adding 100 mu L of 6-benzyladenine monoclonal antibody-colloidal gold marker into a micropore reagent micropore plate, putting the micropore reagent micropore plate into a freeze dryer, pre-freezing for 3h at the temperature of a cold trap under the condition of minus 50 ℃, then carrying out vacuum drying for 15h, taking out the reagent, obtaining the micropore reagent freeze-dried with the 6-benzyladenine monoclonal antibody-colloidal gold marker, and sealing and storing.
8. Preparation of sample absorbent pad
The sample absorption pad is placed in a phosphate buffer solution containing 0.5 percent of bovine serum albumin, with the pH value of 7.2 and 0.1mol/L for soaking for 2h, and is dried for 2h at 37 ℃ for standby.
9. Preparation of the reaction film
Coating the 6-benzyladenine hapten-ovalbumin conjugate on a reaction membrane to form a detection line, and coating the goat anti-mouse anti-antibody on the reaction membrane to form a quality control line.
Coating process: diluting the 6-benzyladenine hapten-ovalbumin conjugate to 1mg/mL by using a phosphate buffer solution, and coating the conjugate on a detection line (T line) on a nitrocellulose membrane by using an Isoflow point membrane instrument, wherein the coating amount is 1.0 mu L/cm; the goat anti-mouse anti-antibody was diluted to 200. Mu.g/mL with 0.01mol/L, pH7.4 phosphate buffer, and coated on a quality control line (C line) on a nitrocellulose membrane in an amount of 1.0. Mu.L/cm using an Isoflow dot-membrane apparatus. And (3) drying the coated reaction membrane at 37 ℃ for 2 hours for later use.
10. Assembly of test strips
According to the section structure of the test paper shown in the attached figure 2, a sample absorption pad (1), a reaction membrane (2) and a water absorption pad (3) are sequentially adhered to a PVC bottom plate (6); the tail end of the sample absorption pad is connected with the initial end of the reaction film, the tail end of the reaction film is connected with the initial end of the water absorption pad, the initial end of the sample absorption pad is aligned with the initial end of the PVC base plate, and the tail end of the water absorption pad is aligned with the tail end of the PVC base plate; the reaction membrane is provided with a detection line (4) and a quality control line (5), and the detection line (T line) and the quality control line (C line) are strip-shaped bands vertical to the long phase of the test paper; the detection line is positioned at one side close to the tail end of the sample absorption pad; the quality control line is positioned on one side far away from the tail end of the sample absorption pad; the test paper was cut into small strips of 3mm in width by a machine and mounted in a specially made plastic card.
According to the microwell reagent diagram of FIG. 3, a microwell reagent 7 has a plug 8 thereon.
The test paper and the micropore reagent are assembled into the test paper strip, and the test paper strip is stored in an environment with the temperature of 2-8 ℃ and has the validity period of 12 months.
Example 2 detection of 6-benzyladenine in Bean sprouts
1. Pretreatment of samples
Homogenizing the bean sprout sample with homogenizer (do not cut and grind into powder); weighing 2.0g +/-0.05 g of sample into a 10mL polystyrene centrifuge tube, adding 3mL phosphate buffer solution, and whirling for 1min by using a vortex mixer to mix uniformly; centrifuging at 4000r/min for 5min at room temperature (20-25 ℃), and taking supernatant as sample liquid to be detected.
2. Detection with test strips
Sucking 100 mu L of sample liquid to be detected into the micropores by using a micropipettor, slowly sucking, and fully and uniformly mixing with the reagent in the micropores; after incubation for 3min at room temperature (20-25 ℃), sucking 90 mu L of the mixed solution and vertically dripping the mixed solution into a sample adding hole; the liquid flow was started, the reaction was carried out for 8min, and the results were judged.
3. Analyzing the results of the detection
Negative (-): the color development of the T line is darker than that of the C line or consistent with that of the C line, which indicates that the concentration of 6-benzyladenine in the sample is lower than the detection limit, as shown in FIGS. 4a and 4b.
Positive (+): the T line is lighter or not developed than the C line, indicating that the concentration of 6-benzyladenine in the sample is equal to or higher than the detection limit, as shown in FIGS. 4C and 4d.
And (4) invalidation: the absence of line C indicates an incorrect procedure or the test strip has deteriorated and failed, as shown in FIGS. 4e and 4f.
Example 3 sample testing example
1. Limit of detection test
Taking blank bean sprout samples, adding 6-benzyladenine into the blank bean sprout samples to the final concentrations of 25 mug/kg, 50 mug/kg and 100 mug/kg respectively, taking test strips for detection, and repeatedly measuring each sample for three times.
When the bean sprout sample is detected by using the test strip, when no 6-benzyladenine exists in the bean sprout sample and the addition concentration of the 6-benzyladenine is 25 mu g/kg, the color development of the T line is darker than that of the C line or is consistent with that of the C line on the test strip, and the test strip is negative; when the addition concentration of the 6-benzyladenine is 50 mug/kg and 100 mug/kg, the test strip shows that the color development of the T line is lighter than that of the C line or the T line is not developed, and the test strip is positive, which shows that the detection limit of the test strip on the 6-benzyladenine in the bean sprouts is 50 mug/kg.
2. Test for false positive and false negative rates
Taking 20 parts of blank bean sprout samples and positive bean sprout samples added with 6-benzyladenine to the final concentration of 50 mug/kg respectively, detecting by using test strips produced in 3 batches respectively, and calculating the negative and positive rates. The results are shown in Table 1.
Table 1 bean sprout sample test results
The results show that: when 3 test strips produced in batches are used for detecting positive bean sprout samples, the results are all positive, and the positive coincidence rate is 100 percent and the false negative rate is 0; when 20 negative bean sprout samples are detected, the results are all negative, the negative coincidence rate is 100 percent, and the false positive rate is 0. The test strip for detecting 6-benzyladenine can be used for rapidly detecting the 6-benzyladenine residue in the bean sprout sample.
3. Specificity test
When the test strip is used for detecting 1000 mu g/kg of 4-chlorophenoxyacetic acid, 2,4, 5-trichlorophenoxyacetic acid, gibberellin, naphthylacetic acid, indoleacetic acid, indolebutyric acid, paclobutrazol, thidiazuron and other plant growth regulators, the test strip shows that the T-line color development is deeper than or consistent with the C-line color development and is negative, which indicates that the test strip has no cross reaction to the medicines.
Claims (4)
1. A test strip for detecting 6-benzyladenine comprises a test strip and a microporous reagent, wherein the test strip comprises a reaction membrane, a sample absorption pad, a water absorption pad and a bottom plate, the reaction membrane is provided with a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody, and the microporous reagent is freeze-dried with a 6-benzyladenine monoclonal antibody-colloidal gold marker; the 6-benzyladenine monoclonal antibody is prepared by taking a 6-benzyladenine hapten-carrier protein conjugate as an immunogen; the 6-benzyladenine hapten-carrier protein conjugate is obtained by coupling 6-benzyladenine hapten and carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroxine or human serum albumin, and the synthesis method of the 6-benzyladenine hapten is as follows: taking 2.4g of amino-6-benzyladenine, adding 70mL of ethanol for dissolution, adding 0.45g of NaOH, fully stirring, adding 1.85g of 4- (3-oxopropyl) -benzoic acid, heating at 60 ℃ for reaction for 4h, cooling to room temperature, adding 0.38g of sodium borohydride, continuing stirring for 2h, stopping the reaction, adding 120mL of water, adding 6mol/L of hydrochloric acid for adjusting the pH value to 6, adding 80mL of ethyl acetate, extracting, separating out a water phase, carrying out rotary evaporation and evaporation on an organic phase to dryness to obtain a red oily substance, recrystallizing acetonitrile/n-hexane 165mL with a volume ratio of 1 to obtain a 6-benzyladenine hapten, wherein the molecular structural formula of the 6-benzyladenine hapten is as follows:
2. the test strip of claim 1, wherein the test strip is formed by sequentially adhering a sample absorption pad, a reaction membrane and a water absorption pad on a bottom plate, and the microporous reagent is provided with a microporous plug.
3. A method of making the test strip of any one of claims 1-2, comprising the steps of:
1) Preparing a microporous reagent freeze-dried with a 6-benzyladenine monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a 6-benzyladenine hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the prepared reaction membrane in the step 2) with a sample absorption pad, a water absorption pad and a base plate to form test paper;
4) And (2) assembling the microporous reagent prepared in the steps 1) and 3) and freeze-dried with the 6-benzyladenine monoclonal antibody-colloidal gold marker and test paper into the test paper strip.
4. A method for detecting 6-benzyladenine residue in a bean sprout sample comprises the following steps:
1) Pretreating a sample;
2) Performing a test using the test strip of any one of claims 1-2;
3) And analyzing the detection result.
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| CN201910643893.4A CN110441517B (en) | 2019-07-17 | 2019-07-17 | Test strip and method for detecting 6-benzyladenine |
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| CN110441517B true CN110441517B (en) | 2023-01-20 |
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