CN117192095A - Fluorescent pad treatment fluid for drug detection fluorescent immunochromatography test paper, and preparation method and application thereof - Google Patents

Abstract

The invention discloses a fluorescent pad treatment liquid for drug detection fluorescent immunochromatography test paper, and a preparation method and application thereof. The treatment fluid comprises a dispersing agent, a surfactant and a buffer; the dispersing agent comprises any one or a combination of at least two of tween-20, PEG or PVP; the surfactant comprises any one or a combination of at least two of surfactants S6, S9 or S17; the buffer comprises any one or a combination of at least two of tris, sodium dihydrogen phosphate, sodium bisulphite or sodium borate. The invention utilizes multicomponent synergy, can effectively improve the physical and chemical properties of the fluorescent pad, so that fluorescent particles are rapidly and uniformly released after a sample is added, and the release is complete without any residue; the background is clean and white after detection, which is helpful for distinguishing and judging the negative and the positive; the accuracy, sensitivity and stability of the detection result of the drug detection fluorescent immunochromatography test paper are improved.

Description

Fluorescent pad treatment fluid for drug detection fluorescent immunochromatography test paper, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of drug detection, relates to a fluorescent pad treatment liquid for a drug detection fluorescent immunochromatographic test paper, a preparation method and application thereof, and particularly relates to a fluorescent pad treatment liquid for a hair poisoning detection fluorescent immunochromatographic test paper, a preparation method and application thereof.

Background

The traditional rapid detection technology for drug-taking personnel mainly adopts a urine colloidal gold method for only about 24-48 hours. Compared with biological detection materials such as urine, blood, saliva and the like, the hair detection material has the advantages of stable property, convenient material taking, easy storage, long detection time limit, wide application range, less pollution opportunity and the like. Because the content of urine poisoning products and metabolites thereof is very high, the traditional colloidal gold immunoassay technology can be achieved. However, hair intoxicants and their metabolites are very low in content, and therefore, techniques with higher detection sensitivity and better tamper resistance are required for hair detection. Fluorescent Immunochromatography (FICA) is a novel membrane detection technique based on antigen-antibody reaction and chromatographic phenomena. Compared with the colloidal gold technology, the fluorescent immunochromatography technology has the characteristics of quantitative analysis, high sensitivity and the like, and simultaneously has higher operation requirements on the preparation of test strips.

The fluorescent pad (or combination pad) is an important component of the fluorescent immunochromatographic test paper, and mainly has the function of adsorbing fluorescent particles marked with corresponding proteins (such as drug antibodies), and a polyester film is commonly used as a material. The fluorescent pad determines the activity (including sensitivity and specificity), release effect and long-term stability of the fluorescent particles labeled with the corresponding proteins, and has a great influence on the effect of the fluorescent immunochromatographic test paper. The current preparation method of the fluorescent bonding pad is mainly to directly spray or soak the marked concentrated fluorescent particles on a polyester film, or to spray or soak the marked concentrated fluorescent particles on the polyester film after the polyester film is soaked by conventional treatment liquid. The fluorescent labeling binding pad prepared by the method is slow and incomplete in release in the detection process, is easy to cause low detection sensitivity, poor specificity and batch-to-batch instability, and is easy to cause false negative results and false positive results.

In conclusion, development of the fluorescent pad treatment liquid capable of effectively improving detection accuracy, sensitivity and stability has important significance in the field of drug fluorescence detection.

Disclosure of Invention

Aiming at the defects and actual demands of the prior art, the invention provides a fluorescent pad treatment liquid for a drug detection fluorescent immunochromatographic test paper, a preparation method and application thereof, in particular to a fluorescent pad treatment liquid for a hair poisoning product detection fluorescent immunochromatographic test paper, a preparation method and application thereof, so as to improve the accuracy, stability and sensitivity of a drug detection fluorescent immunochromatographic test paper detection result.

In order to achieve the above purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides a fluorescent pad treatment fluid for a drug detection fluorescent immunochromatographic test strip, the treatment fluid comprising a dispersing agent, a surfactant and a buffer; the dispersing agent comprises any one or a combination of at least two of tween-20, PEG or PVP; the surfactant comprises any one or a combination of at least two of surfactants S6, S9 or S17; the buffering agent comprises any one or a combination of at least two of tris (hydroxymethyl) aminomethane, sodium dihydrogen phosphate, sodium bisulphite and sodium borate.

In the invention, a brand new fluorescent pad treatment liquid for drug detection fluorescent immunochromatography test paper is designed, contains a dispersing agent, a surfactant and a buffer agent, and can effectively improve the physical and chemical properties of the fluorescent pad by utilizing multi-component cooperation, so that fluorescent particles are rapidly and uniformly released after a sample is added, and the release is complete without any residue; the background is clean and white after detection, which is helpful for distinguishing and judging the negative and the positive; the correct detection rate of the product is improved, and the accuracy and the sensitivity of the detection result of the drug detection fluorescent immunochromatography test paper are improved.

Preferably, the mass percentage of the surfactant in the treatment liquid is 0.1% -10%, including but not limited to 0.2%, 0.3%, 0.5%, 1%, 2%, 3%, 5%, 8%, 9%, 9.5%, 9.6%, 9.8% or 9.9%.

Preferably, the mass percentage of the dispersing agent in the treatment fluid is 0.01% -0.5%, including but not limited to 0.02%, 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.45%, 0.48% or 0.49%.

Preferably, the buffer in the treatment fluid is 0.25% -3% by mass, including but not limited to 0.26%, 0.27%, 0.28%, 0.5%, 1%, 1.5%, 2%, 2.5%, 2.8% or 2.9%.

Preferably, the treatment liquid further contains a preservative and/or water.

Preferably, the preservative comprises any one or a combination of at least two of Proclin300, BIT-10.

Preferably, the mass percent of the preservative in the treatment fluid is 0.02% -5%, including but not limited to 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, 4.6%, 48% or 4.9%.

Preferably, the fluorescent pad treatment liquid for the drug detection fluorescent immunochromatographic test paper comprises, by mass, 100% of a buffer agent, 0.25% -3% of a dispersing agent, 0.01% -0.5% of a surfactant, 0.1% -10% of a preservative, and the balance of water.

Preferably, the buffer comprises 0.05 to 2 mass percent of tris and 0.2 to 1.0 mass percent of sodium dihydrogen phosphate.

In a second aspect, the present invention provides a method for preparing the fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test paper according to the first aspect, the method comprising:

and mixing the buffering agent, the dispersing agent and the surfactant to obtain the treatment liquid.

In a third aspect, the present invention provides an application of the fluorescent pad treatment solution for detecting drugs according to the first aspect in preparing products for detecting drugs.

In a fourth aspect, the present invention provides a method of preparing a drug fluorescence detection test strip, the method comprising:

contacting the fluorescent pad treatment liquid for drug detection fluorescent immunochromatographic test paper according to the first aspect with a fluorescent pad, and coating an antibody marked by a fluorescent micro-marker on the fluorescent pad; drawing the antigen solutions of drugs on the detection films respectively to form detection lines corresponding to different drugs; and sequentially adhering the sample pad, the fluorescent pad, the detection film and the absorption pad on the surface of the substrate from one end to the other end of the substrate to obtain the drug fluorescent detection test strip.

According to the invention, the fluorescent pad treatment liquid is coated on the polyester film to obtain the polyester film containing the fluorescent pad treatment liquid, and then the antibody marked by the fluorescent micro-marker is uniformly sprayed on the polyester film to obtain the fluorescent pad, so that the physical or chemical properties of the fluorescent pad can be effectively improved by the fluorescent pad treatment liquid, fluorescent particles are rapidly and uniformly released after a sample is added, and the fluorescent pad treatment liquid is completely released without any residue; the background is clean and white after detection, which is helpful for distinguishing and judging the negative and the positive; the correct detection rate of the product is improved.

Preferably, the fluorescent label comprises any one or a combination of at least two of fluorescent microspheres, morphine, anilides, ketamine, tetrahydrocannabinic acid, cannabinoids or cocaine antibodies.

Preferably, the drug comprises any one or a combination of at least two of morphine, phenylpropanamine, ketamine, tetrahydrocannabinic acid, cannabinoids or cocaine.

In the invention, the method for preparing the drug fluorescence detection test strip specifically comprises the following steps:

and (1) preparing a fluorescent pad:

spraying the fluorescent pad treatment liquid on a blank polyester film, and drying at 40-50 ℃ for 12-24 hours;

diluting the fluorescent microsphere marked monoclonal antibody and marked rabbit IgG according to the proportion of 20% -60% and 5% -20% respectively by adopting Tris buffer solution, adding 0.1-0.5 g/mL of sucrose and 0.01-0.1 g/mL of trehalose, spraying the mixture on a polyester film treated by the treatment solution with the spraying amount of 0.5-1.5 mu L/cm, and then drying the polyester film at the temperature of 30-40 ℃ for 12-24 hours;

(II) sample pad treatment: spraying the sample pad treatment liquid on glass fiber, and drying at 40-50 ℃ for 12-24 h;

and (III) preparing a coating film: diluting the drug-antigen hemoglobin conjugate and the goat anti-rabbit IgG to 0.1-1.0 mg/mL by adopting a coating buffer solution, adding coomassie brilliant blue into the goat anti-rabbit IgG, preparing the concentration of the coomassie brilliant blue to 0.1-1 mg/mL, coating the goat anti-rabbit IgG diluent and the drug-antigen diluent on a quality control line and a test line on an NC film respectively according to the coating amount of 0.5-1.5 mu L/cm after the preparation, and then drying the NC film at 40-50 ℃ for 20-30 h;

(IV) assembling: the sample pad, the fluorescent pad, the NC film and the water absorbing paper are sequentially adhered on the PVC bottom plate, the water absorbing paper is adhered on the upper part of the nitrocellulose film to generate partial overlapping of 1-2 mm, the lower end of the nitrocellulose film is arranged below the fluorescent pad, the partial overlapping of 1-2 mm exists between the nitrocellulose film and the fluorescent pad, the other end of the fluorescent pad is overlapped with the sample pad by 1-2 mm, and the other end of the sample pad is aligned with the PVC lining plate.

In a fifth aspect, the present invention provides a fluorescent pad treatment solution for a drug detection fluorescent immunochromatographic test strip according to the first aspect and an application of the method for preparing a drug fluorescence detection test strip according to the fourth aspect in drug detection.

Compared with the prior art, the invention has the following beneficial effects:

the invention designs a brand new fluorescent pad treatment liquid for drug detection fluorescent immunochromatography test paper, which contains a dispersing agent, a surfactant and a buffer agent, and can effectively improve the physical and chemical properties of a fluorescent pad by utilizing multi-component cooperation, so that fluorescent particles are rapidly and uniformly released after a sample is added, and the release is complete without any residue; the background is clean and white after detection, which is helpful for distinguishing and judging the negative and the positive; the correct detection rate of the product is improved, and the accuracy, sensitivity and stability of the detection result of the drug detection fluorescent immunochromatography test paper are improved.

Drawings

FIG. 1 is a schematic diagram of the detection of a positive sample in the present invention;

FIG. 2 is a schematic diagram of the detection of a negative sample according to the present invention;

FIG. 3 is a schematic diagram of the structure of the test paper of the present invention.

Detailed Description

The technical means adopted by the invention and the effects thereof are further described below with reference to the examples and the attached drawings. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting thereof.

The specific techniques or conditions are not identified in the examples and are described in the literature in this field or are carried out in accordance with the product specifications. The reagents or equipment used were conventional products available for purchase through regular channels, with no manufacturer noted.

The invention adopts the technical principle that: after the sample to be detected (such as a hair sample) is processed, the sample to be detected is dissolved in a certain amount of lysate, fully and uniformly mixed and reacted for 1-2 min, then the sample is dripped at a sample pad of a sample adding area, an antigen to be detected in a positive sample is combined with a fluorescent microsphere marked antibody in a fluorescent pad and is subjected to forward chromatography through capillary action, when the sample reaches an NC film of a detection area, the antigen fixed on the detection line does not have the fluorescent microsphere marked antibody combined with the antigen, namely, a detection line does not have fluorescent reaction, and a strong positive result is shown as shown in figure 1. The antigen of the object to be detected does not exist in the negative sample, and the fluorescent marked antibody in the fluorescent pad is chromatographed on the NC film along with capillary action and combined with the antigen protein conjugate coated on the detection area, so that a strong fluorescent reaction is shown as shown in figure 2. It can be seen that the amount of the fluorescent microsphere labeled antibody bound on the line is inversely proportional to the amount of the analyte in the sample, the fluorescent label bound on the quality control line is irrelevant to the amount of the analyte in the sample, and other fluorescent labels continue to be chromatographed to reach the absorption region. After chromatography is finished, the fluorescence intensity of a reading detection line and a quality control line of a fluorescence reader is adopted, the T/C value is calculated, and the content of an object to be detected in the sample can be calculated through a standard curve built in the instrument.

Example 1

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.1% of surfactant (S6), 0.01% of dispersing agent (tween-20), 0.05% of Tris, 0.5% of sodium dihydrogen phosphate, 0.02% of preservative (Proclin 300) and the balance of water.

Example 2

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: surfactant (S6) 0.23%, dispersing agent (tween-20) 0.12%, tris 0.1%, sodium dihydrogen phosphate 0.3%, preservative (Proclin 300) 2% and the balance water.

Example 3

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.5% of surfactant (S6), 0.2% of dispersing agent (tween-20), 0.5% of Tris, 0.2% of sodium dihydrogen phosphate, 1.5% of preservative (Proclin 300) and the balance of water.

Example 4

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 1.8% of surfactant (S6), 0.27% of dispersing agent (tween-20), 0.8% of Tris, 0.56% of sodium dihydrogen phosphate, 4% of preservative (Proclin 300) and the balance of water.

Example 5

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 10% of surfactant (S6), 0.5% of dispersing agent (tween-20), 2% of Tris, 1% of sodium dihydrogen phosphate, 5% of preservative (Proclin 300) and the balance of water.

Example 6

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.1% of surfactant (S6), 0.01% of dispersing agent (tween-20), 0.05% of Tris, 0.5% of sodium borate, 0.02% of preservative (Proclin 300) and the balance of water.

Example 7

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.1% of surfactant (S6), 0.01% of dispersing agent (PEG), 0.05% of Tris, 0.5% of sodium dihydrogen phosphate, 0.02% of preservative (Proclin 300) and the balance of water

Example 8

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.1% of surfactant (S9), 0.01% of dispersing agent (tween-20), 0.05% of Tris, 0.5% of sodium dihydrogen phosphate, 0.02% of preservative (Proclin 300) and the balance of water.

Example 9

The embodiment provides a treatment fluid of a fluorescence immunochromatographic test paper fluorescence pad for drug detection, which comprises the following components in percentage by mass as 100 percent: 0.05% of surfactant (S6), 0.05% of surfactant (S17), 0.01% of dispersing agent (tween-20), 0.05% of Tris, 0.5% of sodium dihydrogen phosphate, 0.02% of preservative (Proclin 300) and the balance of water.

Comparative example 1

This comparative example provides a treatment solution of a fluorescent pad of a fluorescent immunochromatographic test paper for drug detection, which differs from example 1 only in that: the surfactant is not contained (S6), and the mass percentages thereof are distributed to the masses of Tris and sodium dihydrogen phosphate in proportion.

Comparative example 2

This comparative example provides a treatment solution of a fluorescent pad of a fluorescent immunochromatographic test paper for drug detection, which differs from example 1 only in that: the dispersion agent (tween-20) was not contained, and its mass percentage was proportionally distributed to the mass of Tris and sodium dihydrogen phosphate.

Preparation example 1

Preparation of a fluorescence detection kit for detecting drugs in hair.

(1) Preparation of fluorescent-labeled proteins

Preparing 0.05M 2-morpholinoethanesulfonic acid (MES) solution, adjusting the pH, adding 10% fluorescent microspheres, rotating at 50rpm for 60min, taking down and sequentially adding N-hydroxysuccinimide (NHS) and carbodiimide (EDC) solutions, continuously rotating for 60min, centrifuging at 15000rpm for 15min after rotation is finished, removing supernatant, adding 0.5mg antibody for rotating reaction for 2.5h after the MES buffer solution is fully dissolved, adding 10% blocking agent after the reaction is finished, rotating for 3h, centrifuging at 15000rpm for 15min, and adding 1mL Tris buffer solution for re-dissolving after the supernatant is removed;

(2) Sample pad treatment

Fully dissolving and uniformly stirring the prepared sample pad treatment liquid, spraying the solution on glass fibers, drying the glass fibers in a drying oven at 37 ℃ for 24 hours, taking out, sealing, drying and storing;

(3) Fluorescent pad preparation

The fluorescent pad treatment solutions prepared in each example and comparative example are fully dissolved and uniformly stirred, then sprayed on blank polyester fiber films with proper sizes respectively, and dried in an oven with the temperature of 40-50 ℃ for 12-24 hours, taken out, sealed, dried and stored.

The fluorescent microsphere marked monoclonal antibody and rabbit IgG are diluted with Tris buffer solution according to the proportion of 40% and 10%, and then 0.5g/mL of sucrose and 0.2g/mL of trehalose are added. After being fully and uniformly dissolved, the preparation method comprises the steps of spraying the solution on a treated polyester fiber film with a spraying amount of 1.0 mu L/cm, drying the polyester fiber film in a baking oven at 37 ℃ for 24 hours, and sealing and storing the polyester fiber film in a dark place;

(4) Preparation of coating film

Diluting the drug-antigen hemoglobin conjugate and goat anti-rabbit IgG to 0.5mg/mL and 0.8mg/mL by adopting PBS (phosphate buffer solution), adding coomassie brilliant blue into the goat anti-rabbit IgG, preparing the concentration of coomassie brilliant blue to 1.0mg/mL, coating the goat anti-rabbit IgG diluent and the drug-antigen diluent on a quality control line and a test line on an NC film respectively according to the coating amount of 1.0 mu L/cm after the preparation, and then placing the mixture in a 45 ℃ oven for drying for 24 hours and then sealing and preserving at normal temperature;

(5) Hair lysate preparation

Preparing a lysate, wherein the formula is as follows: 0.5% PVP-10000,6% sodium sulfite, 0.8% casein, 1.2% keratinase and 5% Proclin300, and the treatment liquid is fully dissolved, uniformly stirred, canned, sealed, dried and stored.

The fittings such as the sample pad, the fluorescent pad, the NC film, the water absorbing paper and the like are sequentially adhered on the PVC bottom plate, as shown in fig. 3, the water absorbing pad is adhered on the upper part of the nitrocellulose film, the partial overlapping of 2mm is generated, the lower end of the nitrocellulose film is arranged below the fluorescent pad, the partial overlapping of 2mm exists between the nitrocellulose film and the nitrocellulose film, the other end of the fluorescent pad is overlapped with the sample pad by 2mm, and the other end of the sample pad is aligned with the PVC lining plate. Cutting into 4.0mm strips after assembly, and placing into hair reagent card for pressing.

Preparation example 2

Preparation of morphine hair detection reagent

Mixing morphine antibody (Qingdao Handson, HDS-23026) and rabbit IgG fluorescent mark according to the proportion of 40% and 10%, adding trehalose and sucrose after mixing, spraying on the treated polyester fiber film with the spraying amount of 1.0 mu L/cm after full concussion and dissolution, drying in a drying oven at 37 ℃ for 24 hours, sealing and storing at normal temperature after drying, and when the flowing direction of the sample is, the sample reaches a detection area and then reaches a quality control line, as shown in figure 1, the reagent line composition is as follows: the detection line is a shaking pill detection line, morphine antigen-bovine serum albumin conjugate is coated, the concentration is 0.3mg/mL, the dividing amount is 1.0 mu L/cm, the quality control line is coated with 0.8mg/mL goat anti-rabbit IgG, and the dividing amount is 1.0 mu L/cm. And (5) after coating, putting the plate film into a drying oven at 45 ℃ for drying for 24 hours, and sealing and storing at normal temperature after drying.

The morphine hair detection agent is produced as follows:

coating morphine antigen and rabbit IgG fluorescent marker on the treated polyester fiber film, spot-coating morphine antigen and quality control line antibody on detection zone and quality control zone of nitrocellulose film by spot-coating machine, fully drying to make nitrocellulose film firmly adsorb raw material, compounding the above-mentioned polyester fiber film, glass fiber and nitrocellulose film on PVC plastic sheet, placing the compounded plastic sheet on cutting machine, cutting into single test paper, placing single test paper into matched plastic box, placing aluminium foil bag, pyrolysis liquor, instruction book and ID card into packaging bag, sealing, and making sample inspection of the product to be inspected, its sensitivity, specificity and stability are passed, and its specific reagent structure diagram is shown in figure 3.

Preparation example 3

Preparation of methamphetamine hair detection reagent

Mixing methamphetamine antibody (Qingdao Handson, HDS-23024) and rabbit IgG according to the proportion of 40% and 10% after fluorescent labeling, adding trehalose and sucrose after mixing, sufficiently shaking and dissolving, spraying on a polyester fiber film after treatment in a spraying amount of 1.0 mu L/cm, drying in a drying oven at 37 ℃ for 24 hours, sealing and storing at normal temperature after drying, and enabling a sample to reach a detection area and then a quality control line according to the flowing direction of the sample, wherein the reagent line comprises the following components as shown in figure 1: the detection line is a methamphetamine detection line, the methamphetamine antigen-bovine serum albumin conjugate is coated, the concentration is 0.5mg/mL, the dividing amount is 1.0 mu L/cm, the quality control line is coated with 0.8mg/mL of goat anti-rabbit IgG, and the dividing amount is 1.0 mu L/cm. And (5) after coating, putting the plate film into a drying oven at 45 ℃ for drying for 24 hours, and sealing and storing at normal temperature after drying.

The production steps of the methamphetamine hair detection reagent are as follows:

coating a polyester fiber film with a methamphetamine antibody and a rabbit IgG fluorescent marker, spot-coating the methamphetamine antigen and the quality control line antibody on a detection zone and a quality control zone of a nitrocellulose film by a spot-coating machine, sufficiently drying to ensure that the nitrocellulose film firmly adsorbs raw materials, compositing the polyester fiber film, glass fiber and the nitrocellulose film on a PVC plastic sheet, placing the composited plastic sheet on a cutting machine, cutting into single test paper, placing the single test paper into a plastic box matched with the single test paper, placing an aluminum foil bag, a lysate, a specification and an ID card into a packaging bag, sealing, waiting to be detected, and performing spot-checking on the waiting to be detected product to ensure that the sensitivity, the specificity and the stability of the waiting to be detected product are qualified, wherein a specific reagent structure diagram is shown in figure 3.

Test case

(1) The fluorescent pads treated by the treatment solutions in the examples 1-9 and the comparative examples 1-2 are respectively prepared into morphine hair detection kit and methamphetamine hair detection kit, 10 human negative hair lysates are respectively detected, the test is repeated for 5 times, the supernatant of the negative hair lysates is sucked by a dropper, 3 drops of the supernatant are dripped into a sample-adding hole of a reagent plate, a reagent card is reacted for 3 minutes outside the machine after the sample is added, the reagent card is immediately inserted into an instrument after the reaction is finished, a test button is clicked, and the system automatically reads the card and gives a test result to record the negative accuracy.

(2) The fluorescent pads treated by the treatment liquid in each example and the comparative example are respectively prepared into morphine hair detection kit and methamphetamine hair detection kit, 5 positive hair (positive sample is confirmed by LCMS) lysate is respectively detected, the test is repeated for 10 times, the supernatant of the positive hair lysate is sucked by a dropper, 3 drops of the supernatant are dripped into a sample-adding hole of a reagent plate, a reagent card is shown in a figure III, the reagent card is reacted for 3 minutes outside the machine after the sample is added, the reagent card is immediately inserted into an instrument after the reaction is finished, a test button is clicked, and the system automatically reads the card and gives a test result to record the positive accuracy.

(3) The fluorescent pads treated by the treatment solutions in the examples and the comparative examples are respectively prepared into morphine hair detection kits and methamphetamine hair detection kits, each concentration point of 2ng/mL of morphine reference and 2ng/mL of methamphetamine reference prepared by using negative hair lysate is respectively detected and tested repeatedly for 10 times, the fluorescent value reacted by the instrument is recorded, the CV value of the T/C is calculated according to the ratio T/C of the fluorescent value of the detection line to the fluorescent value of the quality control line.

The morphine hair detection kit negative sample results are shown in table 1, the methamphetamine hair detection kit negative sample results are shown in table 2, the LCMS confirms the positive sample concentration results are shown in table 3, the morphine hair detection kit positive sample results are shown in table 4, the methamphetamine hair detection kit positive sample results are shown in table 5, the morphine hair detection kit detection reference results are shown in table 6, and the methamphetamine hair detection kit detection reference results are shown in table 7.

TABLE 1

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TABLE 2

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TABLE 3 Table 3

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TABLE 4 Table 4

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TABLE 5

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TABLE 6

TABLE 7

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The results show that:

(1) The test paper prepared by using the treatment liquid of the examples 1-9 has no false positive result in the detection of a negative sample, which shows that the treatment liquid of the fluorescent pad promotes the release of fluorescent particles in the fluorescent pad, so that the fluorescent particles are completely released, the test paper prepared by using the treatment liquid of the comparative examples 1-2 is lack of 1 component (surfactant S6 or diffusant tween-20) compared with the test paper of the example 1, partial false positive condition can occur in the detection of the negative sample, which shows that the surfactant S6 and diffusant tween-20 have synergistic effect in promoting the strip running effect of a drug sample in hair, and shows that the treatment liquid of the fluorescent pad designed by the invention can promote the release of the fluorescent particles, so that the background of the test paper is white and clean after the detection, and the false positive result is reduced.

(2) The test paper prepared by the treatment liquid in the embodiment 1-9 has a positive detection rate of more than 95% in the detection of positive samples, which shows that the treatment liquid for the fluorescent pad promotes the release of fluorescent particles in the fluorescent pad to ensure that the fluorescent particles are completely released, compared with the test paper prepared by the treatment liquid in the comparative embodiment 1-2, 1 component (surfactant S6 or dispersing agent tween-20) is absent, partial false negative condition can occur in the detection of positive and negative samples, the positive detection rate is less than 80%, which shows that the surfactant S6 and the dispersing agent tween-20 have a synergistic effect in promoting the strip running effect of drug samples in hair, and shows that the treatment liquid for the fluorescent pad designed by the invention can promote the release of fluorescent particles and improve the positive detection rate.

(3) The CV value of the fluorescent value T/C of the standard sample of 2ng/mL is less than 10% by using the test paper prepared by the treatment liquid of the embodiment 1-9, which shows that the treatment liquid of the hair drug fluorescent pad can reduce the reagent difference, the fluorescent pad treatment liquid of the invention promotes the uniform release of fluorescent particles, improves the specific combination of drug antigen and antibody, has strong anti-interference capability, compared with the test paper prepared by the treatment liquid of the embodiment 1-2, 1 component (surfactant S6 or dispersing agent tween-20) is absent, the CV value of the fluorescent value T/C of the standard sample of 2ng/mL is more than 20% by lacking the surfactant S6 or dispersing agent tween-20, which shows that the fluorescent pad treatment liquid of the invention has a synergistic effect on promoting the specific combination of hair drug, and the fluorescent pad treatment liquid designed by the invention can promote the uniform release of fluorescent particles, improve the specific combination of drug antigen and antibody and reduce other non-specific interference.

(4) Example 6 shows that the effect of sodium dihydrogen phosphate is better than that of sodium tetraborate, as compared with example 1, the buffer composition is different, and the comprehensive detection rate result and CV value of fluorescence value T/C of 2ng/mL standard substance are shown. Example 7 shows that the diffuser composition is different from example 1, and the integrated detection rate result and CV value of fluorescence value T/C of 2ng/mL standard show that the tween-20 result is superior to PEG. Example 8 is different from example 1 in the surfactant composition, and the experimental results show that the S6 results are superior to S9. Example 9 shows that the synergistic effect of the two surfactants S6 and S17 is better than the S6 effect as compared to example 1.

In conclusion, the fluorescent pad treatment liquid composed of the surfactant, the dispersing agent and the buffer agent can be creatively found to be capable of rapidly detecting drugs aiming at hair, improving the specificity and stability of detection reagents, accelerating the reaction time of drug molecules, being more suitable for rapid screening in public places, being safer in components and not causing environmental pollution.

The applicant states that the detailed method of the present invention is illustrated by the above examples, but the present invention is not limited to the detailed method described above, i.e. it does not mean that the present invention must be practiced in dependence upon the detailed method described above. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.

Claims (10)

1. The fluorescent pad treatment fluid for the drug detection fluorescent immunochromatographic test paper is characterized by comprising a dispersing agent, a surfactant and a buffer;

the dispersing agent comprises any one or a combination of at least two of tween-20, PEG or PVP;

the surfactant comprises any one or a combination of at least two of surfactants S6, S9 or S17;

the buffer comprises any one or a combination of at least two of tris, sodium dihydrogen phosphate, sodium bisulphite or sodium borate.

2. The fluorescent pad treatment fluid for drug detection fluorescent immunochromatographic test paper according to claim 1, wherein the mass percentage of the surfactant in the treatment fluid is 0.1% -10%;

preferably, the mass percentage of the dispersing agent in the treatment fluid is 0.01% -0.5%;

preferably, the mass percentage of the buffer in the treatment liquid is 0.25% -3%.

3. The fluorescent pad treatment fluid for drug detection fluorescent immunochromatographic test paper according to claim 1 or 2, characterized in that the treatment fluid further contains a preservative and/or water;

preferably, the preservative comprises any one or a combination of at least two of Proclin300, BIT-10;

preferably, the mass percentage of the preservative in the treatment fluid is 0.02% -5%.

4. The fluorescent pad treatment fluid for drug detection fluorescent immunochromatographic test paper according to any one of claims 1 to 3, which is characterized in that the treatment fluid contains 0.25% to 3% of buffer, 0.01% to 0.5% of dispersing agent, 0.1% to 10% of surfactant and 0.02% to 5% of preservative by mass percentage based on 100%, and the balance being water;

preferably, the buffer comprises 0.05 to 2 mass percent of tris and 0.2 to 1.0 mass percent of sodium dihydrogen phosphate.

5. The method for preparing the fluorescent pad treatment fluid for drug detection fluorescent immunochromatographic test paper according to any one of claims 1 to 4, which is characterized by comprising the following steps:

and mixing the buffering agent, the dispersing agent and the surfactant to obtain the treatment liquid.

6. The use of the fluorescent pad treatment fluid for drug detection fluorescent immunochromatographic test strips according to any one of claims 1 to 4 for the preparation of products for drug detection.

7. A method of preparing a drug fluorescence detection test strip, the method comprising:

contacting a fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test paper according to any one of claims 1 to 4 with a fluorescent pad, and coating an antibody labeled with a fluorescent marker on the fluorescent pad; drawing the antigen solutions of drugs on the detection films respectively to form detection lines corresponding to different drugs;

and sequentially adhering the sample pad, the fluorescent pad, the detection film and the absorption pad on the surface of the substrate from one end to the other end of the substrate to obtain the drug fluorescent detection test strip.

8. The method of preparing a drug fluorescence detection test strip of claim 7, wherein the fluorescent label comprises any one or a combination of at least two of fluorescent microspheres, morphine, phenylpropanams, ketamine, tetrahydrocannabinic acid, cannabinoid or cocaine antibodies.

9. The method of preparing a drug fluorescence detection test strip of claim 7 or 8, wherein the drug comprises any one or a combination of at least two of morphine, anilines, ketamine, tetrahydrocannabinic acid, a cannabinoid or cocaine.

10. Use of the fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test paper according to any one of claims 1 to 4 and the method for preparing the drug fluorescence detection test paper according to any one of claims 7 to 9 in drug detection.