CN111239399B - Test strip and method for detecting profenofos - Google Patents
Test strip and method for detecting profenofos Download PDFInfo
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Abstract
The invention discloses a test strip and a method for detecting profenofos. The test strip comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate, wherein the reaction membrane is provided with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody, and the conjugate release pad is sprayed with profenofos monoclonal antibody-colloidal gold marker. The invention also provides a method for detecting profenofos in vegetable and fruit samples by using the test strip. The test strip provided by the invention has the advantages of simplicity in operation, high sensitivity, high detection speed, low cost, suitability for large-batch sample screening and the like, and can meet the requirements of the food supervision departments in China for on-site monitoring and detection.
Description
Technical Field
The invention relates to a test strip and a method for detecting profenofos, in particular to a colloidal gold test strip for detecting profenofos, which is particularly suitable for detecting profenofos residues in vegetables and fruits.
Background
Profenofos (profenofos) is a moderately toxic non-systemic broad-spectrum organophosphorus insecticide with contact and stomach toxicity effects and has good control effect on pests on crops. However, as the usage amount increases, the pollution generated by the vegetable used is not negligible. Although a small amount of profenofos residue does not cause immediate and direct toxicity to human bodies, long-term eating of such contaminated vegetables and fruits can cause diseases such as cancers, infertility, endocrine disorders and the like, and can also harm nerve centers, so that the people can die due to cramps. The national standard GB 2763-2019 (maximum residual quantity of pesticides in food safety national Standard food) prescribes the residual quantity of profenofos in different foods.
The currently reported methods for detecting profenofos are mainly the instrument methods such as gas chromatography, gas chromatography-mass spectrometry, liquid chromatography-tandem mass spectrometry and the like. The methods are operated under laboratory conditions, the pretreatment of the samples is tedious and time-consuming, expensive instruments and equipment are needed to be equipped, 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 method is difficult to meet the requirements of rapid detection of a large number of samples and on-site samples. Therefore, the development of the colloidal gold test strip which is simple and rapid and is suitable for profenofos residue in vegetables and fruits can meet the requirements of on-site screening and monitoring of a large number of samples and can better meet the requirements of food supervision departments in China and the like for detection work.
Disclosure of Invention
The invention aims to provide a colloidal gold test strip capable of detecting profenofos residues in vegetables and fruits, and provides a detection method which is efficient, accurate, simple and convenient and suitable for on-site monitoring and screening of a large number of samples.
The invention provides a test strip for detecting profenofos, which comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate; the reaction membrane is provided with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody; the conjugate release pad is sprayed with profenofos monoclonal antibody-colloidal gold labels.
The profenofos monoclonal antibody is prepared by taking profenofos hapten-carrier protein conjugate as an immunogen.
The profenofos hapten-carrier protein conjugate is obtained by coupling profenofos hapten with carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroxine or human serum albumin, the profenofos hapten is obtained by condensation reaction of trichlorfon serving as an initial raw material with butanethiol, ethanol and 5-bromo-3-chloro-2-hydroxybenzoic acid in sequence, and the molecular structural formula is as follows:
the sample absorbing pad, the conjugate releasing pad, the reaction membrane and the water absorbing pad are sequentially stuck on the bottom plate, and 1/3-1/2 of the conjugate releasing pad is covered under the sample absorbing pad.
The bottom plate can be a PVC bottom plate or other hard non-water-absorbing materials; the sample absorption pad can be suction filter paper or oil filter paper; the conjugate release pad may be a glass wool or polyester material; the water absorbing pad is water absorbing paper; the reaction membrane may be a nitrocellulose membrane or a cellulose acetate membrane.
Another object of the present invention is to provide a method for preparing the above test strip, comprising the steps of:
1) Preparing a conjugate release pad sprayed with profenofos monoclonal antibody-colloidal gold labels;
2) Preparing a reaction membrane with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;
3) And (3) assembling the conjugate release pad, the reaction membrane, the sample absorption pad, the water absorption pad and the bottom plate which are prepared in the steps 1) and 2) into the test strip.
Specifically, the method comprises the following steps:
1) The method comprises the steps of taking phosphorus oxychloride as an initial raw material, and carrying out condensation reaction with butanethiol, ethanol and 5-bromo-3-chloro-2-hydroxybenzoic acid to prepare profenofos hapten;
2) Coupling profenofos hapten with carrier protein to prepare profenofos hapten-carrier protein conjugate;
3) Immunizing a mouse by using profenofos hapten-carrier protein conjugate, and obtaining a hybridoma cell strain secreting profenofos monoclonal antibody by fusing and screening spleen cells of the mouse and myeloma cells of the mouse;
4) Extracting mouse IgG to immunize healthy goats to obtain goat anti-mouse anti-antibody;
5) Respectively coating profenofos hapten-carrier protein conjugate and 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 the reaction of trisodium citrate and chloroauric acid;
7) Adding the prepared profenofos monoclonal antibody into the prepared colloidal gold to obtain a profenofos monoclonal antibody-colloidal gold marker;
8) Spraying a profenofos monoclonal antibody-colloidal gold marker on a conjugate release pad, drying at 37 ℃ for 1h, taking out, and storing in a dry environment for later use;
9) Soaking the sample absorption pad in phosphate buffer solution containing 0.5% bovine serum albumin and having pH of 7.2 and 0.1mol/L for 2h, and drying at 37 ℃ for 2h;
10 A sample absorbing pad, a conjugate releasing pad, a reaction membrane, and a water absorbing pad are sequentially stuck on the bottom plate, and 1/3 area of the conjugate releasing pad from the initial end is covered by the sample absorbing pad. Finally cutting into small strips with the width of 3mm, adding a plastic box, vacuum packaging, and preserving for 12 months at the temperature of 4-30 ℃.
The invention also provides a method for detecting profenofos residues in vegetables and fruits by using the test strip, which comprises the following steps:
(1) Sample pretreatment;
(2) Detecting by using a test strip;
(3) And analyzing the detection result.
The profenofos rapid detection test strip adopts a highly specific antibody antigen reaction and immunochromatographic analysis technology, a profenofos monoclonal antibody-colloidal gold marker is fixed on a conjugate release pad, and profenofos in a sample is combined with the profenofos monoclonal antibody-colloidal gold marker on the conjugate release pad in the flowing process to form a drug-antibody-colloidal gold marker. And the drugs in the sample compete with the profenofos hapten-carrier protein conjugate on the reaction membrane detection line for combining with the profenofos monoclonal antibody-colloidal gold marker, and whether profenofos residues are contained in the sample liquid to be detected is judged according to the red stripe depth of the detection line.
During detection, a sample is dripped into a test strip card hole after treatment, when the concentration of profenofos in the sample is lower than the detection limit or zero, a monoclonal antibody-colloidal gold marker is combined with profenofos 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 or consistent with that of the C line; if the concentration of profenofos in the sample is equal to or higher than the detection limit, the monoclonal antibody-colloidal gold-labeled substance will bind to profenofos entirely, so that no red band or less coloration than the C-line will occur at the T-line because of competition reactions without binding to profenofos hapten-carrier protein conjugate. As shown in fig. 3.
Negative: and when the quality control line (C) shows red stripes, the detection line (T) also shows red stripes, and the color of the (T) line is close to or deeper than that of the (C), the judgment is negative.
Positive: and judging positive when the quality control line (C) shows red stripes and the detection line (T) does not develop color or the color of the detection line (T) is lighter than that of the detection line (C).
Invalidation: when the quality control line (C) does not display red stripes, whether the detection line (T) displays red stripes or not, the test strip is judged to be invalid.
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 shelf life. The method for detecting profenofos residue by using the test strip is simple, convenient, quick, visual, accurate, wide in application range, low in cost and easy to popularize and use.
Drawings
FIG. 1 is a diagram showing the synthesis of profenofos hapten.
Fig. 2 is a schematic diagram of a cross-sectional structure of the test strip.
Fig. 3 is a test strip detection result judgment chart.
Detailed Description
The invention is further illustrated below in conjunction with specific examples. It is to be understood that these examples are for illustration of the invention only and are not intended to limit the scope of the invention.
Example 1 preparation of test strip for detecting profenofos
The preparation method of the test strip mainly comprises the following steps:
1) Preparing a conjugate release pad sprayed with profenofos monoclonal antibody-colloidal gold labels;
2) Preparing a reaction membrane with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;
3) And (3) assembling the conjugate release pad, the reaction membrane, the sample absorption pad, the water absorption pad and the bottom plate which are prepared in the steps 1) and 2) into the test strip.
The following is a stepwise detailed description:
1. synthesis of profenofos hapten (synthetic route see figure 1)
1) Dissolving 1.67g of phosphorus trichloride in 100mL of carbon tetrachloride, adding ice bath, cooling to 0-5 ℃, adding 1.8g of butanethiol, adding 0.78g of sodium bicarbonate, continuously stirring for 2 hours, stopping the reaction, adding 80mL of water at 0-5 ℃, transferring to a separating funnel, rapidly oscillating, standing for layering, removing a water phase, concentrating an organic phase, and evaporating to obtain an intermediate 1;
2) Adding 50mL of absolute ethyl alcohol with the temperature of 0-5 ℃ into the intermediate 1 for dissolution, adding 1.38g of absolute potassium carbonate, continuously stirring for 1h at the temperature, stopping the reaction, adding 120mL of water with the temperature of 0-5 ℃ and 150mL of chloroform, sufficiently oscillating, standing in a separating funnel for layering, removing a water phase, concentrating an organic phase, and evaporating to dryness to obtain an intermediate 2;
3) And (3) adding 80mL of acetonitrile into the intermediate 2 for dissolution, adding 2.58g of 5-bromo-3-chloro-2-hydroxybenzoic acid and 1.23g of KOH, stirring at room temperature for 3 hours, stopping the reaction, removing the acetonitrile by rotary evaporation, adding 70mL of water, adding 100mL of ethyl acetate, vibrating for extraction, standing, removing the water phase, drying the organic phase by anhydrous sodium sulfate, evaporating to dryness, loading on a silica gel column, eluting and separating by using a mixed solvent with the volume ratio of dichloromethane to methanol being 10:1, and thus obtaining the profenofos hapten.
2. Preparation of immunogens
17mg of profenofos hapten is taken, 1mL of dioxane is added for dissolution, 13.3mg of N-hydroxysuccinimide (NHS) and 16.2mg of carbodiimide (EDC) are added for reaction at room temperature for 3 hours, and hapten solution A is obtained; taking 50mg of Bovine Serum Albumin (BSA), and adding 0.05mol/L PB buffer solution for dissolution to obtain solution B; and (3) dropwise adding the solution A into the solution B, reacting for 12 hours at 4 ℃, dialyzing and purifying for 3 days by using 0.02mol/L PBS, changing the solution 3 times a day to obtain the profenofos hapten-BSA conjugate, namely the immunogen, and sub-packaging and preserving at-20 ℃.
3. Preparation of coating Material
Taking 9.3mg of profenofos hapten, adding 1mL of N, N-Dimethylformamide (DMF) for dissolution, adding 8.3mg of NHS and 10.1mg of EDC, and reacting for 3 hours at room temperature to obtain hapten solution A; taking 50mg of Ovalbumin (OVA), and adding 0.05mol/L PB buffer solution for dissolution to obtain solution B; and (3) dropwise adding the solution A into the solution B, reacting for 12 hours at 4 ℃, dialyzing and purifying for 3 days by using 0.02mol/L PBS, changing the solution 3 times a day to obtain the profenofos hapten-OVA conjugate, namely the profenofos hapten-OVA conjugate, and subpackaging and preserving at-20 ℃.
4. Preparation of profenofos monoclonal antibody
(1) Immunization of animals
The immunogen obtained in the step 2 is injected into Balb/c mice, and the immune dose is 150 mug/mouse, so that antisera are generated.
(2) Cell fusion and cloning
Spleen cells of an immunized Balb/c mouse are fused with SP2/0 myeloma cells according to the proportion of 8:1 (quantitative proportion), cell supernatant is measured by adopting indirect competition ELISA, and positive holes are screened. Cloning the positive hole by limiting dilution method until obtaining hybridoma cell strain for stably secreting monoclonal antibody.
(3) Cell cryopreservation and resuscitation
The hybridoma cells were prepared into 1X 10 by using a frozen stock solution 6 Cell suspensions of individual/mL were stored in liquid nitrogen for long periods. And (3) taking out the frozen storage tube during recovery, immediately putting into a 37 ℃ water bath for medium-speed thawing, centrifuging to remove frozen storage liquid, and transferring into a culture flask for culture.
(4) Preparation and purification of monoclonal antibodies
Incremental culture method: the hybridoma cells are placed in a cell culture medium, cultured at 37 ℃, and the obtained culture solution is purified by an octanoic acid-saturated ammonium sulfate method to obtain monoclonal antibodies, and the monoclonal antibodies are preserved at-20 ℃.
The cell culture medium is prepared by adding calf serum and sodium bicarbonate into RPMI1640 culture medium, wherein the final concentration of the calf serum in the cell culture medium is 20% (mass fraction), and the final concentration of the sodium bicarbonate in the cell culture medium is 0.2% (mass fraction); the pH of the cell culture medium was 7.4.
5. Preparation of goat anti-mouse antibody
Sheep is used as immune animals, and a murine antibody is used as immunogen to immunize pathogen-free sheep, so that the goat anti-mouse antibody is obtained.
6. Preparation of profenofos monoclonal antibody-colloidal gold marker
(1) Preparation of colloidal gold
Diluting 1% chloroauric acid into 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 high temperature and continuous stirring, stopping stirring and heating until the solution is transparent red, cooling to room temperature, recovering to original volume with deionized water, and preserving at 4deg.C. The prepared colloidal gold has pure appearance, is transparent, has no sediment or floating matters, and has a wine red color when observed in sunlight.
(2) Preparation of profenofos monoclonal antibody-colloidal gold marker
Under the magnetic stirring, adjusting the pH value of the colloidal gold to 7.2 by using 0.2mol/L potassium carbonate solution, adding the profenofos monoclonal antibody into the colloidal gold solution according to the standard of adding 20-50 mu g of antibody into each milliliter of the colloidal gold solution, and continuously stirring and uniformly mixing for 30min; after 10min of standing, 10% BSA was added to give a final concentration of 1% in the colloidal gold solution, and the mixture was left for 10min. Centrifuging at 12000r/min and 4deg.C for 40min, discarding supernatant, washing the precipitate twice with redissolving buffer, re-suspending the precipitate with redissolving buffer with volume 1/10 of that of initial colloidal gold, and standing at 4deg.C for use.
Reconstitution buffer: 0.1 to 0.3 percent of BSA, 0.05 to 0.2 percent of Tween-80 and 0.02mol/L of phosphate buffer solution with pH value of 7.2.
7. Preparation of conjugate release pads
The conjugate release pad was soaked in phosphate buffer containing 0.5% BSA, pH 7.2, 0.5mol/L, soaked uniformly for 1h, and baked at 37℃for 3 h. And uniformly spraying the prepared profenofos monoclonal antibody-colloidal gold marker on a conjugate release pad by using an isolow film spraying instrument, spraying 0.01mL profenofos monoclonal antibody-colloidal gold marker on each 1cm conjugate release pad, placing the conjugate release pad in a 37 ℃ environment (humidity is less than 20%) for 60min, taking out the conjugate release pad, and placing the conjugate release pad in a dry environment (humidity is less than 20%) for storage for later use.
8. Preparation of sample absorbent pad
The sample absorption pad is placed in phosphate buffer solution containing 0.5% bovine serum albumin with pH of 7.2 and 0.1mol/L for soaking for 2 hours, and is dried at 37 ℃ for 2 hours for standby.
9. Preparation of reaction film
The profenofos hapten-ovalbumin conjugate is coated on a reaction membrane to form a detection line, and the goat anti-mouse anti-antibody is coated on the reaction membrane to form a quality control line.
The coating process comprises the following steps: diluting profenofos hapten-ovalbumin conjugate to 1mg/mL by using a phosphate buffer solution, and coating the profenofos hapten-ovalbumin conjugate on a detection line (T line) on a nitrocellulose membrane by using an isolow spot membrane tester, wherein the coating amount is 1.0 mu L/cm; the goat anti-mouse antibody was diluted to 200. Mu.g/mL with 0.01mol/L phosphate buffer at pH7.4, and coated on a quality control line (C line) on a nitrocellulose membrane in an isolow spot film apparatus in an amount of 1.0. Mu.L/cm. And (5) drying the coated reaction film for 2 hours at 37 ℃ for standby.
10. Assembly of test strips
According to the cross-section structure of the test strip shown in figure 2, a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3) and a water absorption pad (4) are sequentially stuck on a PVC bottom plate (7); the 1/3 area of the initial end of the conjugate release pad is covered by the sample absorption pad, the tail end of the conjugate release pad is connected with the initial end of the reaction membrane, the tail end of the reaction membrane 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 bottom plate, and the tail end of the water absorption pad is aligned with the tail end of the PVC bottom plate; the reaction film is provided with a detection line (5) and a quality control line (6), and the detection line (T line) and the quality control line (C line) are strip-shaped strips which are perpendicular to the length of the test strip; the detection line is positioned at one side close to the tail end of the conjugate release pad; the quality control line is positioned at one side far away from the tail end of the conjugate release pad; cutting the test paper strip into small strips with the width of 3mm by a machine, putting the test paper strip into a special plastic card, and storing the test paper strip in an environment of 4-30 ℃ for 12 months.
Example 2 detection of profenofos in vegetables and fruits
1. Sample pretreatment
Cutting the sample into fragments smaller than 1cm or small pieces after removing mud before detection; weighing (1.00+/-0.05) g of the sheared sample into a 10mL polystyrene centrifuge tube, adding 6mL of phosphate buffer solution, covering a cover, manually oscillating for 30s, standing for 1min, and taking supernatant as a sample liquid to be detected.
2. Detection by test strips
Sucking 80 mu L of sample liquid to be detected by a micropipette and vertically dripping the 80 mu L of sample liquid into a sample adding hole; the flow of the liquid was started to time, the reaction was continued for 10 minutes, and the result was judged.
3. Analyzing the detection result
Negative (-). The color development of the T line is deeper than or consistent with that of the C line, which indicates that the profenofos concentration in the sample is lower than the detection limit, as shown in figures 3a and 3b.
Positive (+): the T line is lighter than the C line or the T line is not developed, which indicates that the profenofos concentration in the sample is equal to or higher than the detection limit, as shown in figures 3C and 3d.
Invalidation: the absence of line C indicates an incorrect procedure or that the test strip has failed due to deterioration, as shown in fig. 3e, 3f.
Example 3 sample detection example
1. Limit of detection test
Taking blank cauliflower, cabbage, radish leaf, common cabbage and orange samples, respectively adding profenofos to the samples to reach final concentrations of 0.05mg/kg, 0.1mg/kg and 0.2mg/kg, taking a test strip for detection, and repeatedly measuring each sample for three times.
When the test paper strip is used for detecting samples of cauliflower, cabbage, radish leaf, common cabbage and orange, when the concentration of the profenofos in the samples is 0.05mg/kg, the test paper strip shows that the color development of the T line is deeper than or consistent with that of the C line, and the color development of the T line is negative; when the adding concentration of profenofos is 0.1mg/kg and 0.2mg/kg, the test strip shows that the color development of T line is lighter than that of C line or the color development of T line is not positive, which indicates that the detection limit of the test strip to profenofos in vegetables and fruits is 0.1mg/kg.
2. False positive rate and false negative rate test
Taking 20 parts of blank cauliflower, cabbage mustard, radish leaves, common cabbage and orange samples, adding profenofos to the samples to obtain a final concentration of 0.1mg/kg, and respectively detecting the samples by using 3 batch test strips, so as to calculate the negative-positive rate.
The results show that: when positive samples are detected by using test strips produced in 3 batches, the results are positive, the positive coincidence rate is 100%, and the false negative rate is 0; when a negative sample was detected, the result was all negative, and it was found that the negative coincidence rate was 100% and the false positive rate was 0. The test strip for detecting profenofos can be used for rapidly detecting profenofos residues in vegetable and fruit samples.
3. Specificity test
When the test strip is used for detecting 1000 mug/kg of parathion, omethoate, diazinon, phoxim, quetiapine, triazophos, methamidophos, aqueous amine thiophosphor, dimethoate, methyl parathion, phenthoate, malathion and other organophosphorus pesticides, the test strip shows that the color development of T line is deeper than that of C line or consistent with that of C line, and is negative, thus indicating that the test strip has no cross reaction to the drugs.
Claims (5)
1. The test strip for detecting profenofos comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate, wherein the reaction membrane is provided with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody, and the conjugate release pad is sprayed with profenofos monoclonal antibody-colloidal gold marker; the profenofos monoclonal antibody is prepared by taking profenofos hapten-carrier protein conjugate as immunogen; the profenofos hapten-carrier protein conjugate is obtained by coupling profenofos hapten with carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroxine or human serum albumin;
the method is characterized in that the method for synthesizing profenofos hapten comprises the following steps: 1) Dissolving 1.67g of phosphorus trichloride in 100mL of carbon tetrachloride, adding ice bath, cooling to 0-5 ℃, adding 1.8g of butanethiol, adding 0.78g of sodium bicarbonate, continuously stirring for 2 hours, stopping the reaction, adding 80mL of water at 0-5 ℃, transferring to a separating funnel, rapidly oscillating, standing for layering, removing a water phase, concentrating an organic phase, and evaporating to obtain an intermediate 1;
2) Adding 50mL of absolute ethyl alcohol with the temperature of 0-5 ℃ into the intermediate 1 for dissolution, adding 1.38g of absolute potassium carbonate, continuously stirring for 1h at the temperature of 0-5 ℃, stopping the reaction, adding 120mL of water with the temperature of 0-5 ℃ and 150mL of chloroform with the temperature of 0-5 ℃, sufficiently vibrating, standing in a separating funnel for layering, removing a water phase, concentrating an organic phase, and evaporating to dryness to obtain an intermediate 2;
3) Adding 80mL of acetonitrile into the intermediate 2 for dissolution, adding 2.58g of 5-bromo-3-chloro-2-hydroxybenzoic acid and 1.23g of KOH, stirring for 3 hours at room temperature, stopping the reaction, removing the acetonitrile by rotary evaporation, adding 70mL of water, adding 100mL of ethyl acetate, vibrating for extraction, standing, removing the water phase, drying the organic phase by anhydrous sodium sulfate, evaporating to dryness, loading on a silica gel column, eluting and separating by using a mixed solvent with the volume ratio of dichloromethane to methanol being 10:1, and obtaining profenofos hapten, wherein the molecular structural formula is as follows:
2. the test strip of claim 1, wherein the sample absorbing pad, the conjugate releasing pad, the reaction membrane, and the absorbent pad are sequentially attached to the base plate.
3. The test strip of any one of claims 1-2, wherein the conjugate release pad is 1/3 to 1/2 coated under the sample absorbent pad.
4. A method of preparing the test strip of any one of claims 1-3, comprising the steps of:
1) Preparing a conjugate release pad sprayed with profenofos monoclonal antibody-colloidal gold labels;
2) Preparing a reaction membrane with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;
3) And (3) assembling the conjugate release pad, the reaction membrane, the sample absorption pad, the water absorption pad and the bottom plate which are prepared in the steps 1) and 2) into the test strip.
5. A method for detecting profenofos residue in a vegetable or fruit sample comprising the steps of:
1) Sample pretreatment;
2) Detecting with the test strip of any one of claims 1-3;
3) And analyzing the detection result.
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| CN112067814B (en) * | 2020-08-31 | 2023-07-11 | 北京勤邦科技股份有限公司 | Test strip and method for detecting difenoconazole |
| CN112114147B (en) * | 2020-09-01 | 2023-01-20 | 北京望尔生物技术有限公司 | Test strip and method for detecting pyraclostrobin |
| CN112595844B (en) * | 2020-11-17 | 2023-07-07 | 北京勤邦科技股份有限公司 | Test strip and method for detecting fenpyrazamine |
| CN113156127B (en) * | 2021-04-01 | 2022-08-19 | 北京勤邦生物技术有限公司 | Test strip and method for detecting chlorpyrifos |
| CN116643049B (en) * | 2023-07-27 | 2023-09-29 | 云南省农业科学院质量标准与检测技术研究所 | Profenofos pesticide colloidal gold marker based on modified nano gold material and application thereof |
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