CN108931640B - Test strip for detecting organophosphorus pesticide and application thereof - Google Patents
Test strip for detecting organophosphorus pesticide and application thereof Download PDFInfo
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Abstract
The invention discloses a test strip for detecting organophosphorus pesticide 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 an organophosphorus pesticide 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 an organophosphorus pesticide monoclonal antibody-colloidal gold marker. The invention also provides a method for detecting organophosphorus pesticide residues in crops such as vegetables, fruits and the like by using the organophosphorus pesticide 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 organophosphorus pesticide and application thereof, in particular to a colloidal gold test strip for detecting organophosphorus pesticide, which is particularly suitable for detecting organophosphorus pesticide residues in crops such as vegetables, fruits and the like.
Background
The organophosphorus pesticide is organic compound pesticide containing phosphorus element, and is mainly used for preventing and controlling plant diseases, pests and weeds. Organophosphorus pesticides are widely used in agricultural production, so that residues with different degrees are generated in crops, the harm to human bodies is mainly acute toxicity, and a series of neurotoxicity symptoms such as sweating, tremor, confusion and language disorder can be generated after large dose or repeated contact, and severe patients can suffer from respiratory paralysis and even die.
At present, most of organophosphorus pesticides are detected by instrument methods, such as gas chromatography, liquid chromatography and the like. Generally, the detection cost of the instrument method is high, the operation is complex, and the cost and the time consumption are high for detecting a large batch of samples in a basic laboratory, so that the method is not an ideal detection method. The method overcomes the defects of an instrument method and is beneficial to improving the supervision means and the supervision level.
Disclosure of Invention
The invention aims to provide the test strip for detecting the organophosphorus pesticide residue, which has the advantages of high sensitivity, simple operation, low cost and short detection time.
The test strip for detecting organophosphorus pesticide residues 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 an organophosphorus pesticide 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 an organophosphorus pesticide monoclonal antibody-colloidal gold marker.
The organophosphorus pesticide hapten-carrier protein conjugate is obtained by coupling organophosphorus pesticide hapten and carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroid protein and human serum albumin.
The organophosphorus pesticide monoclonal antibody is prepared by taking an organophosphorus pesticide hapten-carrier protein conjugate as an immunogen and is obtained by secreting an organophosphorus pesticide 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 1/3-1/2 of the conjugate release pad is covered under the sample absorption pad.
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 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 an organophosphorus pesticide monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with an organophosphorus pesticide 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 the compound a and hydroxylamine hydrochloride and other substances to obtain an organophosphorus pesticide hapten;
2) Coupling the organophosphorus pesticide hapten with carrier protein to obtain an organophosphorus pesticide hapten-carrier protein conjugate;
3) Immunizing a mouse by using an organophosphorus pesticide hapten-carrier protein conjugate, and fusing and screening splenocytes and myeloma cells of the mouse to obtain an organophosphorus pesticide 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 organophosphorus pesticide monoclonal antibody prepared in the step 3) into the colloidal gold prepared in the step 5) to obtain an organophosphorus pesticide monoclonal antibody-colloidal gold marker;
7) Spraying the monoclonal antibody-colloidal gold marker of the organophosphorus pesticide on a conjugate release pad, drying at 37 ℃ for 1h, taking out, and storing in a dry environment for later use;
8) Coating the organophosphorus pesticide 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 conjugate releasing pad, a reaction film and a water absorbing pad are sequentially adhered on the bottom plate, the sample absorbing pad covers the conjugate releasing 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 4-30 ℃.
The invention also aims to provide a method for detecting organophosphorus pesticide residues in vegetables and crops by using the test strip, which comprises the following steps:
(1) Pretreating a sample;
(2) Detecting by using a test strip;
(3) And analyzing the detection result.
The test paper strip for rapidly detecting the organophosphorus pesticide adopts a highly specific antibody-antigen reaction and competitive inhibition immunochromatography analysis technology, an organophosphorus pesticide monoclonal antibody-colloidal gold marker is fixed on a conjugate release pad, and the organophosphorus pesticide in a sample is combined with the organophosphorus pesticide monoclonal antibody-colloidal gold marker on the conjugate release pad in the flowing process to form a drug-antibody-colloidal gold marker. The method comprises the steps of combining the drug in a sample with an organophosphorus pesticide hapten-carrier protein conjugate on a reaction membrane detection line in a competitive manner to form an organophosphorus pesticide monoclonal antibody-colloidal gold marker, and judging whether the sample liquid to be detected contains organophosphorus pesticide residues or not according to the existence of red strips or the color depth of the red strips of the detection line.
During detection, a sample is treated and then dropped into a test strip hole, when the concentration of the organophosphorus pesticide in the sample is lower than the detection limit or zero, the monoclonal antibody-colloidal gold marker is combined with the organophosphorus pesticide 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 the organophosphorus pesticide in the sample is equal to or higher than the detection limit, the monoclonal antibody-colloidal gold label will bind to all of the organophosphorus pesticide, and thus no red band will appear at the T-line because of the competitive reaction with the organophosphorus pesticide hapten-carrier protein conjugate.
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. The method for detecting organophosphorus pesticide 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 organophosphorus pesticide hapten.
FIG. 2 is a schematic cross-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 test strip for detecting organophosphorus pesticide
The preparation method of the test strip mainly comprises the following steps:
1) Preparing a conjugate release pad sprayed with an organophosphorus pesticide monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with an organophosphorus pesticide 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 organophosphorus pesticide hapten
(1) Taking 1.0g of the compound a, adding 30ml of methanol for dissolution, taking 0.27g of hydroxylamine hydrochloride, adding 1ml of water for dissolution, adding into a methanol solution, adding 0.46g of potassium carbonate, heating and refluxing for 24h, stopping the reaction, and cooling to room temperature to obtain a solution A; taking 0.31g of zinc powder, adding 5ml of water, stirring, adding 0.2ml of glacial acetic acid, stirring for 30min at 90 ℃, adding all the solution A, continuing stirring for 4h, stopping the reaction, adding 100ml of water and 70ml of ethyl acetate multiplied by 3, extracting for three times, combining organic phases, evaporating to dryness, and recrystallizing by using 120ml of ethyl acetate/n-hexane (v/v, 1/10) to obtain a compound b0.81g with the yield of 81%;
(2) Dissolving 0.81g of the compound b in 50ml of hydrobromic acid, carrying out reflux reaction at 100 ℃ for 3h, stopping the reaction, adding 6mol/L sodium hydroxide solution to adjust the pH value to 7, adding 100ml of ethyl acetate multiplied by 3, extracting for three times, combining organic phases, evaporating to dryness, loading on a silica gel column, and eluting with ethyl acetate/petroleum ether (v/v, 1/5) to obtain 0.7g of a compound c, wherein the yield is 92.1%;
(3) Taking 0.7g of the compound c, adding 30mL of methanol for dissolution, adding 0.55g of diethoxy-chlorphos, adding 0.78g of KOH, reacting for 5 hours at 55 ℃, stopping the reaction, adding water, adding 6mol/L hydrochloric acid for adjusting the pH value to 6, adding 120mL multiplied by 3 of dichloromethane, extracting for three times, combining organic phases, evaporating to dryness, and recrystallizing by 60mL of absolute ethyl alcohol/acetonitrile (v/v, 1/1) to obtain 1.3g of the compound d, wherein the yield is 86.67 percent, and the hapten product is obtained.
The hydrogen with chemical shift delta =11 is the absorption peak of the carboxyl hydrogen of the spacer arm, and the absorption peaks of the methylene hydrogen on the spacer arm are 1.84, 2.33 and 1.56 through nuclear magnetic data, and the existence of the peaks proves the success of the spacer arm coupling.
2. Preparation of immunogens
Taking 38mg of a hapten product of the compound d, adding 2ml of DMF (dimethyl formamide) for dissolving, adding 17mg of N-succinimide and 29mg of EDC, and stirring at room temperature for 2 hours to obtain a hapten activating solution A; taking 100mg of bovine serum albumin, adding 0.05mol/L of CB buffer solution for dissolving to obtain B solution, dripping A solution into the B solution, stirring at 4 ℃ for 24h, dialyzing and purifying by 0.02mol/L of PB buffer solution for 3 days, changing the solution 3 times every day to obtain organophosphorus-BSA immunogen, and storing at-20 ℃ for later use.
3. Preparation of coating antigen
Taking 21mg of a hapten product of a compound d, adding 2ml of DMF (dimethyl formamide) for dissolving, adding 13mg of N-succinimide and 20mg of EDC, and stirring at room temperature for 2 hours to obtain a hapten activating solution A; dissolving ovalbumin 100mg in CB buffer solution 0.05mol/L to obtain solution B, dripping A solution into solution B, stirring at 4 deg.C for 24h, dialyzing and purifying with 0.02M PB buffer solution for 3 days, changing solution 3 times per day to obtain organophosphorus-OVA immunogen, and storing at-20 deg.C for use.
4. Preparation of organophosphorus pesticide 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 the 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 organophosphorus pesticide 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 material.
(2) Preparation of organophosphorus pesticide monoclonal antibody-colloidal gold marker
Adjusting the pH value of the colloidal gold to 7.0 by using 0.2mol/L potassium carbonate solution under magnetic stirring, adding the organophosphorus pesticide monoclonal antibody into the colloidal gold solution according to the standard of adding 20-50 mu g of the organophosphorus pesticide monoclonal antibody into each milliliter of the colloidal gold solution, continuously stirring and uniformly mixing for 30min, adding 10% of BSA (bovine serum albumin) so that the final concentration of the BSA in the colloidal gold solution is 1% (volume fraction), 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. And uniformly spraying the prepared organophosphorus pesticide monoclonal antibody-colloidal gold marker on a conjugate release pad by using an Isoflow film spraying instrument, spraying 0.01ml of organophosphorus pesticide monoclonal antibody-colloidal gold marker on every 1cm of conjugate release pad, placing in an environment at 37 ℃ (humidity is less than 20%) for 60min, taking out, and placing in a dry environment (humidity is less than 20%) for storage.
8. Preparation of the reaction film
Coating the organophosphorus pesticide 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 organophosphorus pesticide hapten-ovalbumin conjugate to 10mg/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 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 of the end away from the conjugate release pad; the test paper strip is cut into small strips with the width of 3mm by a machine, and the small strips are arranged in a special plastic card and can be stored for 12 months at the temperature of 4-30 ℃.
EXAMPLE 2 detection of organophosphorus pesticide residue in sample
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 to be detected into the sample adding hole, timing when the liquid flows, reacting for 5-10 min, and judging the result.
2. Analyzing the results of the detection
The results were read 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;
and (4) invalidation: indicating that retesting is required.
Example 3 sample testing example
1. Limit of detection test
Taking samples of blank Chinese cabbage and apple, respectively adding organophosphorus pesticide to the samples until the final concentrations are 0.1, 0.2 and 0.4mg/kg, taking test strips for detection, and repeatedly measuring each sample for three times.
When the test paper strip is used for detecting samples of the Chinese cabbage and the apple, the analyzer shows that the samples are negative when the adding concentration of the organophosphorus pesticide in the samples is 0.1 mg/kg; when the concentration of the organophosphorus pesticide added therein was 0.2, 0.4mg/kg, the analyzer showed positive.
2. Test for false positive and false negative rates
Taking 20 parts of each of the cabbage and apple positive samples with the known organophosphorus pesticide content of 0.2mg/kg and 20 parts of each of the cabbage and apple negative samples with the organophosphorus pesticide content of less than 0.1mg/kg, detecting by using three batches of test strips, and calculating the negative and positive rates of the three batches of test strips. The results are shown in tables 1 and 2.
TABLE 1 cabbage 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 cabbage and apple samples, the results are all positive, the coincidence rate of the positive samples is 100 percent, and the false negative rate is 0; when 20 negative samples of the Chinese cabbage and the apple 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 the organophosphorus pesticide can be used for rapidly detecting the organophosphorus pesticide residue in Chinese cabbage and apples.
3. Specificity test
The organophosphorus pesticide test paper is used for detecting 500mg/kg carbofuran, dichlorvos 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 strip has no cross reaction to 500mg/kg carbofuran, dichlorvos and other medicaments.
Claims (6)
1. A test strip for detecting organophosphorus pesticides 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 an organophosphorus pesticide 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 an organophosphorus pesticide monoclonal antibody-colloidal gold marker, the organophosphorus pesticide hapten-carrier protein conjugate is obtained by coupling organophosphorus pesticide hapten and carrier protein, the carrier protein is bovine serum albumin, hemocyanin, thyroxine and human serum albumin, and the preparation method of the organophosphorus pesticide hapten comprises the following steps:
(1) Taking 1.0g of the compound a, adding 30ml of methanol for dissolution, taking 0.27g of hydroxylamine hydrochloride, adding 1ml of water for dissolution, adding into a methanol solution, adding 0.46g of potassium carbonate, heating and refluxing for 24h, stopping the reaction, and cooling to room temperature to obtain a solution A; taking 0.31g of zinc powder, adding 5ml of water, stirring, adding 0.2ml of glacial acetic acid, stirring for 30min at 90 ℃, adding all the solution A, continuing stirring for 4h, stopping reaction, adding 100ml of water, multiplying 70ml of ethyl acetate by 3, extracting for three times, combining organic phases, evaporating to dryness, and recrystallizing with 120ml of ethyl acetate/n-hexane (v/v, 1/10) to obtain a compound b0.81g, wherein the yield is 81%;
(2) Dissolving 0.81g of the compound b in 50ml of hydrobromic acid, carrying out reflux reaction at 100 ℃ for 3h, stopping the reaction, adding 6mol/L sodium hydroxide solution to adjust the pH value to 7, adding 100ml of ethyl acetate multiplied by 3, extracting for three times, combining organic phases, evaporating to dryness, loading on a silica gel column, and eluting with ethyl acetate/petroleum ether (v/v, 1/5) to obtain 0.7g of a compound c, wherein the yield is 92.1%;
(3) Taking 0.7g of the compound c, adding 30mL of methanol for dissolution, adding 0.55g of diethoxy-chlorphos, adding 0.78g of KOH, reacting at 55 ℃ for 5 hours, stopping the reaction, adding water, adding 6mol/L hydrochloric acid for adjusting the pH value to 6, adding 120mL multiplied by 3 of dichloromethane, extracting for three times, combining organic phases, evaporating to dryness, and recrystallizing by 60mL of absolute ethyl alcohol/acetonitrile (v/v, 1/1) to obtain 1.3g of the compound d, wherein the yield is 86.67 percent, namely a hapten product;
the molecular structural formula of the organophosphorus pesticide hapten is as follows:
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 claim 1, 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 organophosphorus pesticide monoclonal antibody is prepared by using an organophosphorus pesticide hapten-carrier protein conjugate as an immunogen, and the goat anti-mouse anti-antibody is obtained by immunizing a goat with a murine antibody.
5. A method of making the test strip of any one of claims 1-4, comprising the steps of:
1) Preparing a conjugate release pad sprayed with an organophosphorus pesticide monoclonal antibody-colloidal gold marker;
2) Preparing a reaction membrane with a detection line coated with an organophosphorus pesticide 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.
6. A method for detecting organophosphorus pesticide residues in vegetables and fruit crops comprises the following steps:
1) Sample pretreatment;
2) Performing a test using the test strip of any one of claims 1-4;
3) And analyzing the detection result.
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