CN111735951B - Test strip for detecting fenpropathrin and application thereof - Google Patents
Test strip for detecting fenpropathrin and application thereof Download PDFInfo
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- CN111735951B CN111735951B CN202010493885.9A CN202010493885A CN111735951B CN 111735951 B CN111735951 B CN 111735951B CN 202010493885 A CN202010493885 A CN 202010493885A CN 111735951 B CN111735951 B CN 111735951B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54306—Solid-phase reaction mechanisms
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2430/00—Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
- G01N2430/10—Insecticides
- G01N2430/12—Pyrethroids
Abstract
The invention discloses a test strip for detecting fenpropathrin and application thereof. The test strip comprises a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3), a water absorption pad (4) and a bottom plate (7), wherein the reaction membrane is provided with a detection line (5) coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line (6) coated with a goat anti-mouse anti-antibody, and the conjugate release pad (2) is sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker. The invention also provides a method for detecting fenpropathrin residues in vegetables and fruits such as cucumbers, apples and the like by applying the fenpropathrin test strip. The test strip provided by the invention has the characteristics of simple operation, high sensitivity, high detection speed, low cost and the like, and is suitable for screening and on-site monitoring of a large number of samples.
Description
Technical Field
The invention relates to a test strip for detecting fenpropathrin and application thereof, in particular to a colloidal gold test strip for detecting fenpropathrin, which is particularly suitable for detecting fenpropathrin residue in vegetables and fruits such as cucumbers, apples and the like.
Background
The pyrethroid pesticide plays an extremely important role in modern agricultural production, and has the characteristics of stable property, wide insecticidal spectrum, high insecticidal activity, short residual period and the like, wherein fenpropathrin is one of more applications. Fenpropathrin is produced and used in large scale for a long time, which seriously harms the ecological environment safety and the quality of agricultural products.
The fenpropathrin is detected by GC and HPLC methods at home and abroad. Compared with the two methods, the immunochemical detection method has the advantages of rapidness, specificity, sensitivity, accuracy, batch monitoring, simple sample treatment, automatic operation and the like for the detection of the medicine. The invention discloses a preparation method of an artificial fenpropathrin antigen, which is applied to a rapid detection test strip, has the advantages of short time, simple operation and low cost, and is suitable for detecting samples in bulk by a basic unit.
Disclosure of Invention
The invention aims to provide the fenpropathrin residue detection test strip which is high in sensitivity, simple to operate, low in cost and short in detection time.
The test strip for detecting fenpropathrin residue provided by the invention comprises a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3), a water absorption pad (4) and a bottom plate (7); the reaction membrane is provided with a detection line (5) coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line (6) coated with a goat anti-mouse anti-antibody, and the conjugate release pad (2) is sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker.
The fenpropathrin hapten-carrier protein conjugate is obtained by coupling fenpropathrin hapten and carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroid protein and human serum albumin.
The fenpropathrin monoclonal antibody is prepared by taking a fenpropathrin hapten-carrier protein conjugate as an immunogen and is obtained by secreting a fenpropathrin monoclonal antibody hybridoma cell strain; the goat anti-mouse anti-antibody is obtained by immunizing a goat with a murine antibody.
The sample absorption pad (1), the conjugate release pad (2), the reaction membrane (3) and the water absorption pad (4) are sequentially adhered to the bottom plate (7), and the conjugate release pad 1/3~1/2 is covered under the sample absorption pad.
The bottom plate can be a PVC bottom plate or other hard non-water-absorbing materials; the sample absorbing pad can be suction filter paper or oil filter paper; the conjugate release pad may be a glass wool or polyester material; 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 conjugate release pad sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a base plate to form the test strip.
Specifically, the steps include:
1) Hapten preparation: carrying out a series of chemical reactions on fenpropathrin alcohol, HCl and other substances to obtain fenpropathrin hapten;
2) Coupling fenpropathrin hapten with carrier protein to obtain a fenpropathrin hapten-carrier protein conjugate;
3) Immunizing a mouse by using a fenpropathrin hapten-carrier protein conjugate, and fusing and screening spleen cells and myeloma cells of the mouse to obtain a fenpropathrin monoclonal hybridoma cell strain;
4) Extracting mouse IgG to immunize healthy goats to obtain goat anti-mouse anti-antibodies;
5) Preparing colloidal gold by reacting trisodium citrate with chloroauric acid;
6) Adding the fenpropathrin monoclonal antibody prepared in the step 3) into the colloidal gold prepared in the step 5) to obtain a fenpropathrin monoclonal antibody-colloidal gold marker;
7) Spraying a fenpropathrin 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;
8) Coating the fenpropathrin hapten-carrier protein 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;
9) Soaking the sample absorption pad in 0.5% bovine serum albumin (volume fraction), pH7.2, 0.1mol/L phosphate buffer solution for 2h, and drying at 37 deg.C for 2h;
10 A sample absorbing pad, a binder release pad, a reaction film and a water absorbing pad are sequentially stuck on a bottom plate, the sample absorbing pad covers the binder release pad, and finally the sample absorbing pad is cut into small strips with the width of 3mm, a plastic box is added, the small strips are packaged in vacuum, and the small strips can be stored for 12 months at the temperature of 4 to 30 ℃.
The invention also aims to provide a method for detecting fenpropathrin residue in vegetables and fruits by using the test strip, which comprises the following steps:
(1) Detecting by using a test strip;
(2) And analyzing the detection result.
The rapid fenpropathrin detection test strip adopts a highly specific antibody-antigen reaction and competitive inhibition immunochromatography analysis technology, a fenpropathrin monoclonal antibody-colloidal gold marker is fixed on a conjugate release pad, and fenpropathrin in a sample is combined with the fenpropathrin monoclonal antibody-colloidal gold marker on the conjugate release pad in the flowing process to form a drug-antibody-colloidal gold marker. The drug in the sample and the fenpropathrin hapten-carrier protein conjugate on the reaction film detection line compete to combine with the fenpropathrin monoclonal antibody-colloidal gold marker, and whether the sample liquid to be detected contains fenpropathrin residue is judged according to whether the red strip of the detection line has zero or light color.
During detection, a sample is treated and then dripped into a test strip hole, when the concentration of the fenpropathrin in the sample is lower than the detection limit or zero, the monoclonal antibody-colloidal gold marker is combined with a fenpropathrin hapten-carrier protein conjugate fixed on a reaction membrane in the chromatography process, and a red strip appears in a detection line (T) and a quality control line (C); if the concentration of fenpropathrin in the sample is equal to or above the limit of detection, the monoclonal antibody-colloidal gold label will bind to the fenpropathrin in its entirety, so that no red band will appear at the T-line because the competition reaction will not bind to the fenpropathrin hapten-carrier protein conjugate.
The test strip provided by the invention has the advantages of high sensitivity, strong specificity, low cost, simplicity in operation, short detection time, suitability for various units, simplicity in storage and long quality guarantee period. The method for detecting fenpropathrin residue 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 synthetic diagram of fenpropathrin hapten.
FIG. 2 is a schematic sectional view of a 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 fenpropathrin test paper strip
The preparation method of the test strip mainly comprises the following steps:
1) Preparing a conjugate release pad sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a PVC base plate to form the test strip.
The following steps are detailed:
1. preparation of fenpropathrin hapten
3.61g of vinyl fenpropathrin is taken and added with 30ml of acetonitrile for dissolution, 1ml of 1mol/L HCl is added for full stirring, 3ml of potassium permanganate aqueous solution containing 1.58g of the mixture is added, the mixture reacts for 3 hours at 50 ℃, the reaction is stopped, 200ml of water is added, 100ml of ethyl acetate is added for extraction for three times, organic phases are combined, anhydrous sodium sulfate is dried and evaporated to dryness to obtain red oily matter, the red oily matter is applied to a silica gel column, and the mixture is eluted by petroleum ether/ethyl acetate (v/v, 5/1) and separated to obtain 3.4g of carboxy fenpropathrin hapten product with the yield of 89.7 percent.
2. Preparation of immunogens
Taking 14mg of a carboxyfenpropathrin hapten product, adding 1ml of DMF for dissolving, adding 12mg of HOBT and 14mg of EDC, and reacting at room temperature for 3 hours to obtain a hapten solution A; taking 50mg of Bovine Serum Albumin (BSA), adding 0.05M CB buffer solution for dissolving to obtain B solution, dripping A solution into the B solution, reacting for 12h at 4 ℃, dialyzing and purifying for 3 days by 0.02M PBS, and changing solution three times every day to obtain a fenpropathrin-BSA conjugate which is an immunogen and storing at-20 ℃ for later use.
3. Preparation of coating antigen
Taking 8.4mg of carboxyfenpropathrin hapten product, adding 1ml of DMSO for dissolving, adding NHS6.8mg and EDC7.7mg, and reacting at room temperature for 3h to obtain a hapten solution A; taking 50mg of egg serum albumin (OVA), adding 0.05M CB buffer solution for dissolving to obtain B solution, dripping A solution into the B solution, reacting for 12h at 4 ℃, dialyzing and purifying for 3 days by 0.02M PBS, and changing solution three times every day to obtain the fenpropathrin-OVA conjugate which is the coating antigen and is stored at-20 ℃ for later use.
4. Preparation of fenpropathrin 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 8:1 (quantitative ratio), measuring cell supernatants by adopting an indirect competitive ELISA method, 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
Making hybridoma cell into 1 × 10 with frozen stock solution 6 Cell suspension per ml, preserved for long periods in liquid nitrogen. Taking out the frozen tube during recovery, immediately putting the tube into a water bath at 37 ℃ for fast melting, centrifuging to remove frozen liquid, 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 fenpropathrin 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 in 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 and heating at constant speed until the solution is bright red, cooling to room temperature, recovering to original volume with deionized water, and storing at 4 deg.C. The prepared colloidal gold has pure appearance, transparency and no precipitate or floating substances.
(2) Preparation of fenpropathrin monoclonal antibody-colloidal gold marker
Under magnetic stirring, adjusting the pH value of the colloidal gold to 7.0 by using 0.2mol/L potassium carbonate solution, adding fenpropathrin monoclonal antibody into the colloidal gold solution according to the standard of adding 20-50 mu g of fenpropathrin into each milliliter of the colloidal gold solution, continuously stirring and uniformly mixing for 30min, adding 10 percent BSA (percent by volume) to ensure that the final concentration of the BSA in the colloidal gold solution is 1 percent, and standing 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: 0.02mol/L phosphate buffer solution containing casein 0.02% -0.1% (mass fraction), tween-80 0.05% -0.2% (mass fraction) and pH7.2.
7. Preparation of conjugate Release pad
The conjugate release pad was soaked in 0.5mol/L phosphate buffer containing bovine serum albumin (concentration of bovine serum albumin in buffer is 0.5%), pH7.2, and the solution was soaked uniformly for 1h and baked at 37 ℃ for 3 h. Uniformly spraying the prepared fenpropathrin monoclonal antibody-colloidal gold marker on a conjugate release pad by using an Isoflow film spraying instrument, spraying 0.01ml of the fenpropathrin monoclonal antibody-colloidal gold marker on each 1cm of the conjugate release pad, placing the conjugate release pad in an environment at 37 ℃ (the humidity is less than 20%) for 60min, taking out the conjugate release pad, and placing the conjugate release pad in a dry environment (the humidity is less than 20%) for storage.
8. Preparation of the reaction film
Coating the fenpropathrin 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 fenpropathrin hapten-ovalbumin conjugate to 10mg/ml by using a phosphate buffer solution, and coating the fenpropathrin hapten-ovalbumin conjugate on a detection line (T line) on a nitrocellulose membrane by using an Isoflow point membrane instrument, wherein the coating amount is 0.8 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 for 2 hours at 37 ℃ for later use.
9. Preparation of sample absorbent pad
The sample absorption pad is soaked in phosphate buffer solution containing 0.5 percent of bovine serum albumin (volume fraction), pH7.2 and 0.1mol/L for 2h, and is baked for 2h at 37 ℃ for standby.
10. Assembly of test strips
Sequentially adhering a sample absorption pad, a conjugate release pad, a reaction membrane and a water absorption pad on a PVC bottom plate; the binder release pad is covered by the sample absorption pad in a 1/3 area from the starting end, the tail end of the binder release pad is connected with the starting end of the reaction film, the tail end of the reaction film is connected with the starting end of the water absorption pad, the starting end of the sample absorption pad is aligned with the starting 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 and a quality control line, and the detection line (T line) and the quality control line (C line) are strip-shaped strips which are vertical to the long phase of the test strip; the detection line is located on the side near the end of the conjugate release pad; the quality control line is positioned on the side away from the end of the conjugate release pad; cutting the test paper into small strips with the width of 3mm by a machine, packaging the small strips in a special plastic card, and storing the small strips for 12 months at the temperature of 4-30 ℃.
EXAMPLE 2 detection of fenpropathrin residue in samples
1. Detection with test strip
And (3) sucking the sample solution to be detected by a suction pipe, vertically and dropwise adding the sample solution into the sample adding hole, timing when the liquid flows, reacting for 5-10min, and judging the result.
2. Analyzing the results of the detection
Reading the results by a colloidal gold analyzer (abbreviated as "analyzer"):
negative (-): indicating that the concentration of the substance to be detected in the sample is lower than the detection limit;
positive (+): indicating that the concentration of the substance to be detected in the sample is equal to or higher than the detection limit;
no effect: indicating that retesting is required.
Example 3 sample testing example
1. Limit of detection test
Taking blank cucumber and apple samples, respectively adding fenpropathrin to the samples until the final concentrations are 25, 50 and 100 mu g/kg, taking test strips for detection, and repeatedly measuring each sample for three times.
When the test paper is used for detecting cucumber and apple samples, the analyzer shows that the samples are negative when the fenpropathrin addition concentration is 25 mug/kg; when the fenpropathrin is added at a concentration of 50, 100 mug/kg, the analyzer shows positive.
2. Test for false positive and false negative rates
20 parts of each of cucumber and apple positive samples with the known fenpropathrin content of 50 mug/kg and 20 parts of each of cucumber and apple negative samples with the known fenpropathrin content of less than 25 mug/kg are taken, three batches of test strips are used for detection, and the negative and positive rates of the samples are calculated. The results are shown in tables 1 and 2.
TABLE 1 cucumber test sample results
TABLE 2 apple test sample results
The results show that: when 3 test strips produced in batches are used for detecting positive cucumber and apple samples, the results are all positive, and the coincidence rate of the positive samples is 100 percent, and the false negative rate is 0; when 20 negative cucumber and apple samples are detected, the results are all negative, and the negative coincidence rate is 100 percent and the false positive rate is 0. The test strip for detecting fenpropathrin can be used for rapidly detecting fenpropathrin residues in cucumbers and apples.
3. Specificity test
Fenpropathrin test paper is used for detecting 500 mu g/kg of deltamethrin, cyhalothrin and other medicaments. The result shows that the quality control line and the detection line of the test strip are both colored and negative. The test paper has no cross reaction to 500 mu g/kg of deltamethrin, cyhalothrin and other medicaments.
Claims (7)
1. The test strip for detecting fenpropathrin comprises a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3), a water absorption pad (4) and a bottom plate (7), and is characterized in that the reaction membrane is provided with a detection line (5) coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line (6) coated with a goat anti-mouse anti-antibody, the conjugate release pad (2) is sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker, and the preparation method of the fenpropathrin hapten is as follows:
3.61g of vinyl fenpropathrin is taken, 30ml of acetonitrile is added for dissolution, 1ml of 1mol/L HCl is added, the mixture is fully stirred, 3ml of potassium permanganate aqueous solution containing 1.58g of the mixture is added, the mixture reacts for 3 hours at 50 ℃, the reaction is stopped, 200ml of water is added, 100ml of ethyl acetate is added, multiplied by 3 is extracted for three times, organic phases are combined, anhydrous sodium sulfate is dried and evaporated to dryness, red oily matter is obtained, the red oily matter is applied to a silica gel column, petroleum ether/ethyl acetate (v/v, 5/1) is eluted and separated, 3.4g of carboxyl fenpropathrin hapten product is obtained, and the yield is 89.7%.
2. The test strip of claim 1, wherein the sample absorbing pad (1), the conjugate releasing pad (2), the reaction membrane (3) and the absorbent pad (4) are sequentially adhered to the bottom plate (7).
3. The strip of any one of claims 1-2, wherein the conjugate release pad 1/3~1/2 is covered under a sample absorbing pad.
4. The test strip of claim 1, wherein the fenpropathrin hapten-carrier protein conjugate is obtained by coupling fenpropathrin hapten and carrier protein, and the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroxine, and human serum albumin.
5. The test strip of claim 1, wherein the fenpropathrin monoclonal antibody is prepared by taking a fenpropathrin hapten-carrier protein conjugate as an immunogen, and the goat anti-mouse anti-antibody is obtained by immunizing a goat with a mouse antibody.
6. A method of making the test strip of any one of claims 1-5, comprising the steps of:
1) Preparing a conjugate release pad sprayed with a fenpropathrin monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with a fenpropathrin hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a base plate to form the test strip.
7. A method for detecting fenpropathrin residues in cucumbers, apples, other vegetables and fruits comprises the following steps:
1) Detecting with the test strip of any one of claims 1-5;
2) And analyzing the detection result.
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