A fluorescent pad processing solution for drug detection fluorescent immuno-chromatographic test paper, and a preparation method and application thereof

Abstract

The application discloses a fluorescent pad processing liquid for a drug detection fluorescent immunochromatography test paper and a preparation method and application thereof. The processing liquid comprises a diffusion agent, a surfactant and a buffer; the diffusion 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; and the buffer comprises any one or a combination of at least two of Tris, sodium dihydrogen phosphate, sodium bisulfite or sodium borate. The application utilizes multi-component synergy, can effectively improve the physical and chemical properties of the fluorescent pad, makes the fluorescent particles rapidly and uniformly release after sample addition, and is completely released without any residue; the background is clean and white after detection, which is helpful for distinguishing and judging the negative and positive; and the application increases the accuracy, sensitivity and stability of the drug detection fluorescent immunochromatography test paper detection result.

Description

Technical Field

[0001] This invention belongs to the field of drug detection technology, and relates to a fluorescent pad treatment solution for a fluorescent immunochromatographic test strip for drug detection, its preparation method and application, and particularly to a fluorescent pad treatment solution for a fluorescent immunochromatographic test strip for drug detection in hair, its preparation method and application. Background Technology

[0002] Traditional rapid drug testing techniques, primarily using colloidal gold methods on urine, have an detection time of only about 24–48 hours. Compared to biological samples such as urine, blood, and saliva, hair samples offer advantages such as stability, ease of collection, preservation, longer detection time, wider applicability, and lower risk of contamination. While traditional colloidal gold immunochromatography (FICA) is sufficient to detect drugs and their metabolites in very high concentrations in urine, the concentrations in hair are extremely low. Therefore, techniques with higher sensitivity and better resistance to interference are needed for hair testing. FICA is a novel membrane detection technique based on antigen-antibody reactions and chromatographic phenomena. Compared to colloidal gold technology, FICA offers quantitative analysis and high sensitivity, but also requires more sophisticated procedures in test strip preparation.

[0003] The fluorescent pad (or conjugation pad) is a crucial component of fluorescent immunochromatographic test strips. Its main function is to adsorb fluorescent particles labeled with corresponding proteins (such as drug antibodies). Polyester membranes are commonly used materials. The fluorescent pad determines the activity (including sensitivity and specificity), release efficiency, and long-term stability of the fluorescent particles labeled with the corresponding proteins, significantly impacting the effectiveness of the immunochromatographic test strip. Currently, the preparation methods for fluorescent conjugation pads mostly involve directly spraying or immersing pre-labeled, concentrated fluorescent particles onto a polyester membrane, or treating the polyester membrane with a conventional treatment solution before spraying or immersing the pre-labeled, concentrated fluorescent particles onto the polyester membrane. Fluorescently labeled conjugation pads prepared using these methods release slowly and incompletely during detection, easily leading to low sensitivity, poor specificity, and batch-to-batch instability, resulting in false negative and false positive results.

[0004] In conclusion, developing fluorescent pad treatment solutions that effectively improve the accuracy, sensitivity, and stability of drug fluorescence detection is of great significance to the field of drug fluorescence detection. Summary of the Invention

[0005] To address the shortcomings of existing technologies and practical needs, this invention provides a fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test strips, its preparation method, and its application. In particular, it relates to a fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test strips in hair, its preparation method, and its application, aiming to improve the accuracy, stability, and sensitivity of drug detection fluorescent immunochromatographic test strip results.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] In a first aspect, the present invention provides a fluorescent pad treatment solution for a fluorescent immunochromatographic test strip for drug detection, the treatment solution comprising a dispersant, a surfactant, and a buffer; the dispersant comprising any one or a combination of at least two of Tween-20, PEG, or PVP; the surfactant comprising any one or a combination of at least two of surfactants S6, S9, or S17; and the buffer comprising any one or a combination of at least two of tris(hydroxymethyl)aminomethane, sodium dihydrogen phosphate, sodium bisulfite, and sodium borate.

[0008] This invention presents a novel fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test strips. This solution contains a dispersant, a surfactant, and a buffer. Utilizing the synergistic effect of these multiple components, the physical and chemical properties of the fluorescent pad are effectively improved. This results in the rapid, uniform, and complete release of fluorescent particles after sample addition, leaving no residue. The background after detection is clean and white, aiding in the differentiation between negative and positive results. This improves the product's accurate detection rate and increases the accuracy and sensitivity of the drug detection fluorescent immunochromatographic test strip results.

[0009] Preferably, the mass percentage of surfactant in the treatment solution is 0.1% to 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%.

[0010] Preferably, the mass percentage of the dispersant in the treatment liquid is 0.01% to 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%.

[0011] Preferably, the mass percentage of the buffer in the treatment solution is 0.25% to 3%, including but not limited to 0.26%, 0.27%, 0.28%, 0.5%, 1%, 1.5%, 2%, 2.5%, 2.8%, or 2.9%.

[0012] Preferably, the treatment solution also contains a preservative and / or water.

[0013] Preferably, the preservative includes any one or a combination of at least two of Proclin 300 and BIT-10.

[0014] Preferably, the preservative in the treatment solution is 0.02% to 5% by mass, 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%.

[0015] Preferably, the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection contains, by weight percentage, 0.25%–3% buffer, 0.01%–0.5% dispersant, 0.1%–10% surfactant, and 0.02%–5% preservative, with the balance being water.

[0016] Preferably, the buffer comprises tris(hydroxymethyl)aminomethane and sodium dihydrogen phosphate, wherein the mass percentage of tris(hydroxymethyl)aminomethane is 0.05% to 2% and the mass percentage of sodium dihydrogen phosphate is 0.2% to 1.0%.

[0017] In a second aspect, the present invention provides a method for preparing the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection as described in the first aspect, the preparation method comprising:

[0018] The buffer, dispersant, and surfactant are mixed to obtain the treatment solution.

[0019] Thirdly, the present invention provides the use of the fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test strips as described in the first aspect in the preparation of products for drug detection.

[0020] Fourthly, the present invention provides a method for preparing a fluorescent drug detection test strip, the method comprising:

[0021] The fluorescent pad treatment solution of the fluorescent immunochromatographic test strip for drug detection described in the first aspect is used to contact the fluorescent pad, and the antibody labeled with the fluorescent microlabel is coated on the fluorescent pad; the antigen solution of the drug is streaked on the detection membrane to form detection lines corresponding to different drugs; the sample pad, fluorescent pad, detection membrane and absorbent pad are sequentially adhered to the surface of the substrate from one end to the other to obtain the drug fluorescent test strip.

[0022] In this invention, a fluorescent pad treatment solution is coated onto a polyester membrane to obtain a polyester membrane containing the fluorescent pad treatment solution. Then, an antibody labeled with a fluorescent microlabel is uniformly sprayed onto the polyester membrane to obtain a fluorescent pad. The fluorescent pad treatment solution can effectively improve the physical or chemical properties of the fluorescent pad, so that the fluorescent particles are released rapidly and uniformly after the sample is added, and the release is complete without any residue. The background after detection is clean and white, which helps to distinguish between negative and positive results and improves the correct detection rate of the product.

[0023] Preferably, the fluorescent label comprises any one or a combination of at least two of the following: fluorescent microspheres, morphine, amphetamines, ketamine, tetrahydrocannabinoids, cannabinoids, or cocaine antibodies.

[0024] Preferably, the drug includes any one or a combination of at least two of morphine, amphetamines, ketamine, tetrahydrocannabinol, cannabinoids, or cocaine.

[0025] In this invention, the method for preparing the drug fluorescent test strip specifically includes:

[0026] (I) Preparation of fluorescent pads:

[0027] The fluorescent pad treatment solution was sprayed onto a blank polyester film and dried at 40–50°C for 12–24 hours.

[0028] The fluorescent microsphere-labeled monoclonal antibody and labeled rabbit IgG were diluted with Tris buffer at a ratio of 20%–60% and 5%–20%, respectively. Then, 0.1–0.5 g / mL of sucrose and 0.01–0.1 g / mL of trehalose were added, and the mixture was sprayed onto the polyester film treated with the treatment solution at a spraying rate of 0.5–1.5 μL / cm. The film was then dried at 30–40 °C for 12–24 h.

[0029] (II) Sample pad treatment: Spray the sample pad treatment solution onto the glass fiber and dry it at 40-50℃ for 12-24 hours;

[0030] (III) Preparation of coating membrane: The drug-antigen hemoglobin conjugate and goat anti-rabbit IgG were diluted to 0.1-1.0 mg / mL using coating buffer. Coomassie brilliant blue was added to the goat anti-rabbit IgG to a concentration of 0.1-1 mg / mL. After preparation, the above dilutions of goat anti-rabbit IgG and drug-antigen were coated onto the control line and test line of the NC membrane at a coating amount of 0.5-1.5 μL / cm, respectively. The membrane was then dried at 40-50℃ for 20-30 h.

[0031] (iv) Assembly: On the PVC base plate, the sample pad, fluorescent pad, NC membrane and absorbent paper are attached in sequence. The absorbent paper is attached to the upper part of the nitrocellulose membrane, creating a 1-2 mm overlap. The lower end of the nitrocellulose membrane is placed below the fluorescent pad, with a 1-2 mm overlap between the two. The other end of the fluorescent pad overlaps the sample pad by 1-2 mm, and the other end of the sample pad is aligned with the PVC backing plate.

[0032] Fifthly, the present invention provides the application of the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection described in the first aspect and the method for preparing the fluorescent test strip for drug detection described in the fourth aspect in drug detection.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] This invention presents a novel fluorescent pad treatment solution for drug detection fluorescent immunochromatographic test strips. Containing a dispersant, surfactant, and buffer, this multi-component synergistic effect effectively improves the physical and chemical properties of the fluorescent pad, resulting in rapid, uniform, and complete release of fluorescent particles after sample addition, leaving no residue. The resulting background is clean and white, aiding in the differentiation between negative and positive results. This improves the product's accurate detection rate and enhances the accuracy, sensitivity, and stability of the drug detection fluorescent immunochromatographic test strip results. Attached Figure Description

[0035] Figure 1 This is a schematic diagram illustrating the principle of positive sample detection in this invention;

[0036] Figure 2 This is a schematic diagram illustrating the principle of negative sample detection in this invention;

[0037] Figure 3 This is a schematic diagram of the test strip structure of the present invention. Detailed Implementation

[0038] To further illustrate the technical means and effects of this invention, the following description, in conjunction with embodiments and accompanying drawings, provides a further explanation of the invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it.

[0039] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0040] The technical principle of this invention is as follows: After processing the sample to be tested (such as a hair sample), it is dissolved in a certain amount of lysis buffer. After thorough mixing and reaction for 1-2 minutes, it is dropped onto the sample pad in the sample application area. The analyte antigen in the positive sample binds to the fluorescent microsphere-labeled antibody in the fluorescent pad and is chromatographyd forward through capillary action. When it reaches the NC membrane in the detection area, the antigen fixed on the detection line does not have any fluorescent microsphere-labeled antibody bound to it, that is, the detection line shows no fluorescence reaction, presenting a strong positive result. Figure 1 As shown. In negative samples, the analyte antigen is absent. The fluorescently labeled antibody in the fluorescent pad is chromatographically deposited onto the NC membrane via capillary action, binding to the antigen-protein conjugate coated in the detection area, exhibiting a strong fluorescent reaction, as shown. Figure 2As shown, the amount of fluorescently labeled antibody bound to the detection line is inversely proportional to the amount of analyte in the sample, while the amount of fluorescent label bound to the control line is independent of the amount of analyte in the sample. Other fluorescent labels continue chromatography to reach the absorption region. After chromatography, the fluorescence intensity of the detection line and control line is read using a fluorescence reader, and the T / C value is calculated. The content of the analyte in the sample can then be calculated using the instrument's built-in standard curve.

[0041] Example 1

[0042] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.1% surfactant (S6), 0.01% dispersant (Tween-20), 0.05% Tris, 0.5% sodium dihydrogen phosphate, 0.02% preservative (Proclin 300), and the balance being water.

[0043] Example 2

[0044] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.23% surfactant (S6), 0.12% dispersant (Tween-20), 0.1% Tris, 0.3% sodium dihydrogen phosphate, 2% preservative (Proclin 300), and the balance being water.

[0045] Example 3

[0046] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.5% surfactant (S6), 0.2% dispersant (Tween-20), 0.5% Tris, 0.2% sodium dihydrogen phosphate, 1.5% preservative (Proclin 300), and the balance being water.

[0047] Example 4

[0048] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 1.8% surfactant (S6), 0.27% dispersant (Tween-20), 0.8% Tris, 0.56% sodium dihydrogen phosphate, 4% preservative (Proclin 300), and the balance being water.

[0049] Example 5

[0050] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 10% surfactant (S6), 0.5% dispersant (Tween-20), 2% Tris, 1% sodium dihydrogen phosphate, 5% preservative (Proclin 300), and the balance being water.

[0051] Example 6

[0052] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.1% surfactant (S6), 0.01% dispersant (Tween-20), 0.05% Tris, 0.5% sodium borate, 0.02% preservative (Proclin 300), and the balance being water.

[0053] Example 7

[0054] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage (100%): surfactant (S6) 0.1%, dispersant (PEG) 0.01%, Tris 0.05%, sodium dihydrogen phosphate 0.5%, preservative (Proclin 300) 0.02%, and the balance being water.

[0055] Example 8

[0056] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.1% surfactant (S9), 0.01% dispersant (Tween-20), 0.05% Tris, 0.5% sodium dihydrogen phosphate, 0.02% preservative (Proclin 300), and the balance being water.

[0057] Example 9

[0058] This embodiment provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which comprises the following components by weight percentage: 0.05% surfactant (S6), 0.05% surfactant (S17), 0.01% dispersant (Tween-20), 0.05% Tris, 0.5% sodium dihydrogen phosphate, 0.02% preservative (Proclin 300), and the balance being water.

[0059] Comparative Example 1

[0060] This comparative example provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which differs from Example 1 only in that it does not contain surfactant (S6), and its mass percentage is proportionally allocated to the mass of Tris and sodium dihydrogen phosphate.

[0061] Comparative Example 2

[0062] This comparative example provides a treatment solution for the fluorescent pad of a fluorescent immunochromatographic test strip for drug detection, which differs from Example 1 only in that it does not contain a dispersant (Tween-20), and its mass percentage is proportionally allocated to the mass of Tris and sodium dihydrogen phosphate.

[0063] Preparation Example 1

[0064] Preparation of a fluorescent detection kit for detecting drugs in hair.

[0065] (1) Preparation of fluorescently labeled proteins

[0066] Prepare a 0.05M 2-morpholinoethanesulfonic acid (MES) solution, adjust the pH, add 10% fluorescent microspheres, rotate at 50 rpm for 60 min, remove the solution and add N-hydroxysuccinimide (NHS) and carbodiimide (EDC) solutions sequentially, continue rotating for 60 min, centrifuge at 15000 rpm for 15 min after rotation, remove the supernatant, add MES buffer solution to fully dissolve, add 0.5 mg antibody and rotate for 2.5 h, after the reaction is complete, add 10% blocking agent, rotate to block for 3 h, centrifuge at 15000 rpm for 15 min, remove the supernatant and reconstitute with 1 mL Tris buffer;

[0067] (2) Sample pad treatment

[0068] After thoroughly dissolving and stirring the prepared sample pad treatment solution, spray it onto the glass fiber and place it in an oven at 37°C for 24 hours to dry. Then, remove it, seal it, and store it in a dry place.

[0069] (3) Preparation of fluorescent pads

[0070] After thoroughly dissolving and stirring the fluorescent pad treatment solutions prepared in each embodiment and comparative example, they were sprayed onto blank polyester fiber films of appropriate size, placed in an oven at 40-50°C and dried for 12-24 hours, then removed, sealed, dried and stored.

[0071] Fluorescent microsphere-labeled monoclonal antibodies and rabbit IgG were diluted with Tris buffer at ratios of 40% and 10%, respectively, and then 0.5 g / mL of sucrose and 0.2 g / mL of trehalose were added. After thorough dissolution, the mixture was sprayed onto the treated polyester fiber membrane at a rate of 1.0 μL / cm, and then dried in a 37°C oven for 24 h before being stored in a sealed container away from light.

[0072] (4) Preparation of coating membrane

[0073] The drug-antigen hemoglobin conjugate and goat anti-rabbit IgG were diluted with PBS to 0.5 mg / mL and 0.8 mg / mL, respectively. Coomassie Brilliant Blue was added to the goat anti-rabbit IgG to prepare a concentration of 1.0 mg / mL. After preparation, the above goat anti-rabbit IgG dilution and drug-antigen dilution were coated onto the control line and test line of the NC membrane at a coating amount of 1.0 μL / cm, respectively. The membrane was then dried in a 45℃ oven for 24 h and then stored at room temperature in a sealed container.

[0074] (5) Preparation of hair lysis solution

[0075] Prepare the lysis buffer with the following formula: 0.5% PVP-10000, 6% sodium sulfite, 0.8% casein, 1.2% keratinase, and 5% Proclin 300. After thoroughly dissolving and stirring the solution, bottle it, seal it, and store it in a dry place.

[0076] Sample pads, fluorescent pads, NC film, absorbent paper, and other accessories are sequentially glued onto the PVC base plate. Figure 3 As shown, the absorbent pad is attached to the upper part of the nitrocellulose membrane, creating a 2mm overlap. The lower end of the nitrocellulose membrane is placed below the fluorescent pad, with a 2mm overlap between them. The other end of the fluorescent pad overlaps the sample pad by 2mm, and the other end of the sample pad is aligned with the PVC backing. After assembly, the sample pad is cut into 4.0mm reagent strips, placed in the hair reagent card, and pressed firmly for later use.

[0077] Preparation Example 2

[0078] Preparation of morphine hair test reagent

[0079] After morphine antibody (Qingdao Handerson, HDS-23026) and rabbit IgG fluorescent labeling were completed, they were mixed at a ratio of 40% and 10%. Trehalose and sucrose were added after mixing, and the mixture was thoroughly shaken to dissolve. The solution was then sprayed onto the treated polyester fiber membrane at a rate of 1.0 μL / cm and dried in an oven at 37℃ for 24 hours. After drying, the membrane was sealed and stored at room temperature. Following the sample flow direction, the sample first reaches the detection zone and then the control line. Figure 1 As shown, the reagent lines are composed as follows: the test line is an ecstasy test line, coated with morphine antigen-bovine serum albumin conjugate at a concentration of 0.3 mg / mL, with a stroke volume of 1.0 μL / cm; the control line is coated with 0.8 mg / mL goat anti-rabbit IgG, with a stroke volume of 1.0 μL / cm. After coating, the plates are dried in an oven at 45℃ for 24 hours, and then sealed and stored at room temperature.

[0080] The production steps of morphine hair follicle test reagent are as follows:

[0081] Morphine antibody and rabbit IgG fluorescent marker were coated onto a treated polyester fiber membrane. Morphine antigen and control line antibody were spotted onto the detection and control zones of the nitrocellulose membrane using a dotting machine. After thorough drying, the nitrocellulose membrane was allowed to firmly adsorb the raw materials. The polyester fiber membrane, glass fiber, and nitrocellulose membrane were then laminated onto a PVC plastic sheet. The laminated plastic sheet was placed on a cutting machine and cut into single-use test strips. The single-use test strips were placed into a matching plastic box. An aluminum foil bag, lysis buffer, instruction manual, and ID card were placed into the packaging bag, sealed, and ready for testing. Samples were randomly selected for testing to ensure their sensitivity, specificity, and stability were met before shipment. The specific reagent structure diagram is shown below. Figure 3 As shown.

[0082] Preparation Example 3

[0083] Preparation of Methamphetamine Hair Detection Reagent

[0084] Methamphetamine antibody (Qingdao Handerson, HDS-23024) and rabbit IgG fluorescently labeled were mixed at a ratio of 40% and 10%, respectively. Trehalose and sucrose were added, and the mixture was thoroughly shaken to dissolve. The solution was then sprayed onto the treated polyester fiber membrane at a rate of 1.0 μL / cm and dried in a 37℃ oven for 24 hours. After drying, the membrane was sealed and stored at room temperature. Following the sample flow direction, the sample first reaches the detection zone and then the control line. Figure 1 As shown, the reagent lines are composed as follows: the detection line is a methamphetamine detection line coated with methamphetamine antigen-bovine serum albumin conjugate at a concentration of 0.5 mg / mL, with a stroke volume of 1.0 μL / cm; the control line is coated with 0.8 mg / mL goat anti-rabbit IgG, with a stroke volume of 1.0 μL / cm. After coating, the plates are dried in an oven at 45°C for 24 hours, and then sealed and stored at room temperature.

[0085] The production steps of the methamphetamine hair follicle test kit are as follows:

[0086] Methamphetamine antibody and rabbit IgG fluorescent marker were coated onto a polyester fiber membrane. Methamphetamine antigen and control line antibody were spotted onto the detection and control zones of the nitrocellulose membrane using a spotting machine. The membrane was thoroughly dried to ensure strong adsorption of the raw materials. The polyester fiber membrane, glass fiber, and nitrocellulose membrane were then laminated onto a PVC plastic sheet. The laminated plastic sheet was placed on a cutting machine and cut into single-use test strips. Each test strip was placed into a matching plastic box. An aluminum foil bag, lysis buffer, instruction manual, and ID card were placed inside the packaging bag, sealed, and ready for testing. Samples were randomly selected for testing to ensure sensitivity, specificity, and stability; those that passed were released from the factory. A detailed reagent structure diagram is shown below. Figure 3 As shown.

[0087] Test case

[0088] (1) The fluorescent pads treated with the solutions in Examples 1-9 and Comparative Examples 1-2 were used to prepare morphine hair follicle detection kits and methamphetamine hair follicle detection kits, respectively. Ten samples of negative hair lysis buffer were tested, and the tests were repeated five times. The supernatant of the negative hair lysis buffer was aspirated with a dropper and added 3 drops to the sample wells of the reagent plate. The reagent card was placed as shown in the image. Figure 3 As shown, after adding the sample, react outside the instrument for 3 minutes. After the reaction is complete, immediately insert the reagent card into the instrument and click the "Test" button. The system will automatically read the card and give the test result, recording the negative accuracy rate.

[0089] (2) The fluorescent pads treated with the above-mentioned solutions in the examples and comparative examples were used to prepare morphine hair detection kits and methamphetamine hair detection kits, respectively. Five samples of positive hair lysates (concentration of positive samples confirmed by LCMS) were tested, and the tests were repeated 10 times. The supernatant of the positive hair lysate was aspirated with a dropper and added 3 drops to the sample wells of the reagent plate. The reagent card was then... Figure 3 As shown, after adding the sample, react outside the instrument for 3 minutes. After the reaction is complete, immediately insert the reagent card into the instrument and click the "Test" button. The system will automatically read the card and give the test result, and record the positive accuracy rate.

[0090] (3) The fluorescent pads treated with the treatment solutions in the above examples and comparative examples were prepared into morphine hair test kits and methamphetamine hair test kits, respectively. The morphine reference standard of 2 ng / mL and methamphetamine reference standard of 2 ng / mL prepared with negative hair lysis buffer were tested repeatedly 10 times at each concentration point. The fluorescence value of the instrument was recorded. The CV value of T / C was calculated by the ratio of the fluorescence value of the detection line to the fluorescence value of the quality control line.

[0091] Table 1 shows the results of negative samples from the morphine hair follicle test kit; Table 2 shows the results of negative samples from the methamphetamine hair follicle test kit; Table 3 shows the concentration results of positive samples confirmed by LCMS; Table 4 shows the results of positive samples from the morphine hair follicle test kit; Table 5 shows the results of positive samples from the methamphetamine hair follicle test kit; Table 6 shows the results of the morphine hair follicle test kit reference material; and Table 7 shows the results of the methamphetamine hair follicle test kit reference material.

[0092] Table 1

[0093]

[0094]

[0095]

[0096]

[0097]

[0098] Table 2

[0099]

[0100]

[0101]

[0102]

[0103] Table 3

[0104]

[0105]

[0106] Table 4

[0107]

[0108]

[0109]

[0110] Table 5

[0111]

[0112]

[0113]

[0114] Table 6

[0115]

[0116] Table 7

[0117]

[0118]

[0119] The results show that:

[0120] (1) The test strips prepared using the treatment solutions of Examples 1-9 showed no false positive results in the detection of negative samples, indicating that the fluorescent pad treatment solution of the present invention promotes the release of fluorescent particles in the fluorescent pad and ensures complete release of fluorescent particles. The test strips prepared using the treatment solutions of Comparative Examples 1-2 lack one component (surfactant S6 or dispersant Tween-20) compared with Example 1, and some false positives occurred in the detection of negative samples. This indicates that surfactant S6 and dispersant Tween-20 have a synergistic effect in promoting the running effect of drug samples in hair. This shows that the fluorescent pad treatment solution designed in the present invention can promote the release of fluorescent particles, making the background white and clean after detection, and reducing false positive results.

[0121] (2) The test strips prepared using the treatment solutions of Examples 1-9 showed a positive detection rate of >95% in the detection of positive samples, indicating that the fluorescent pad treatment solution of the present invention promotes the release of fluorescent particles in the fluorescent pad and ensures complete release of fluorescent particles. The test strips prepared using the treatment solutions of Comparative Examples 1-2 lack one component (surfactant S6 or dispersant Tween-20) compared with Example 1, and some false negatives occurred in the detection of positive and negative samples, with a positive detection rate of <80%. This indicates that surfactant S6 and dispersant Tween-20 have a synergistic effect in promoting the running of drug samples in hair, and that the fluorescent pad treatment solution designed in the present invention can promote the release of fluorescent particles and improve the positive detection rate.

[0122] (3) The CV value of the test strips prepared using the treatment solutions of Examples 1-9 for detecting the fluorescence value T / C of 2 ng / mL standard was <10%, indicating that the hair drug fluorescent pad treatment solution of the present invention can reduce the variability of reagents, indicating that the fluorescent pad treatment solution of the present invention promotes the uniform release of fluorescent particles, improves the specific binding of drug antigen and antibody, and has strong anti-interference ability. The test strips prepared using the treatment solutions of Comparative Examples 1-2 lack one component (surfactant S6 or dispersant Tween-20) compared with Example 1. The CV value of the fluorescence value T / C of 2 ng / mL standard without surfactant (S6) or dispersant Tween-20 was >20%, indicating that surfactant S6 or dispersant Tween-20 has a synergistic effect in promoting the specific binding of drugs in hair, indicating that the fluorescent pad treatment solution designed in the present invention can promote the uniform release of fluorescent particles, improve the specific binding of drug antigen and antibody, and reduce other non-specific interference.

[0123] (4) Compared with Example 1, Example 6 had a different buffer composition. The overall detection rate results and the CV value of the fluorescence value (T / C) of the 2 ng / mL standard showed that sodium dihydrogen phosphate was more effective than sodium tetraborate. Compared with Example 1, Example 7 had a different dispersant composition. The overall detection rate results and the CV value of the fluorescence value (T / C) of the 2 ng / mL standard showed that Tween-20 was more effective than PEG. Compared with Example 1, Example 8 had a different surfactant composition. The experimental results showed that S6 was more effective than S9. Compared with Example 1, Example 9 had a different surfactant composition. The results showed that the synergistic effect of surfactants S6 and S17 was better than that of S6.

[0124] In summary, this invention creatively discovers that a fluorescent pad treatment solution composed of surfactants, dispersants, and buffers can rapidly detect drugs in hair, improve the specificity and stability of the detection reagent, accelerate the reaction time of drug molecules, and is more suitable for rapid screening in public places. In addition, the components used are relatively safe and will not cause environmental pollution.

[0125] The applicant declares that the detailed method of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above detailed method, that is, it does not mean that the present invention must rely on the above detailed method to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A fluorescent pad treatment solution for a fluorescent immuno-chromatographic test strip for the detection of a drug in a hair sample, characterized in that, The treatment fluid includes a dispersant, a surfactant, and a buffer. The dispersant is Tween-20; The surfactant is S6; The buffer is a combination of tris(hydroxymethyl)aminomethane and sodium dihydrogen phosphate; The treatment solution contains surfactant S6 at a mass percentage of 0.1% to 10%, dispersant Tween-20 at a mass percentage of 0.01% to 0.5%, and buffer at a total mass percentage of 0.25% to 3%, with the mass ratio of tris(hydroxymethyl)aminomethane to sodium dihydrogen phosphate being (0.05% to 2%): (0.2% to 1.0%).

2. The fluorescent pad treatment solution for a fluorescent immuno-chromatographic test strip for drug detection according to claim 1, characterized by, The treatment solution also contains preservatives and / or water.

3. The fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection according to claim 2, characterized in that, The preservatives include any one or a combination of at least two of Proclin 300 and BIT-10.

4. The fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection according to claim 2, characterized in that, The preservative in the treatment solution is 0.02% to 5% by mass.

5. The fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection according to claim 1, characterized in that, The treatment solution contains, by mass percentage, 0.25%~3% buffer, 0.01%~0.5% dispersant, 0.1%~10% surfactant and 0.02%~5% preservative, with the balance being water.

6. The method for preparing the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection according to any one of claims 1-5, characterized in that, The preparation method includes: The buffer, dispersant, and surfactant are mixed to obtain the treatment solution.

7. The use of the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection as described in any one of claims 1-5 in the preparation of products for drug detection.

8. A method for preparing a fluorescent drug detection test strip, characterized in that, The method includes: The fluorescent pad treatment solution of the fluorescent immunochromatographic test strip for drug detection according to any one of claims 1-5 is used to contact the fluorescent pad, and the antibody labeled with the fluorescent marker is coated on the fluorescent pad; the antigen solution of the drug is streaked on the detection membrane to form detection lines corresponding to different drugs; The sample pad, fluorescent pad, detection membrane, and absorbent pad are sequentially adhered to the surface of the substrate from one end to the other to obtain the drug fluorescent test strip.

9. The method for preparing a fluorescent drug detection test strip according to claim 8, characterized in that, The fluorescent markers include fluorescent microspheres.

10. The method for preparing a fluorescent drug detection strip according to claim 8 or 9, characterized in that, The drugs include any one or a combination of at least two of the following: morphine, amphetamines, ketamine, tetrahydrocannabinol, cannabinoids, or cocaine.

11. The application of the fluorescent pad treatment solution for the fluorescent immunochromatographic test strip for drug detection according to any one of claims 1-5 or the method for preparing the fluorescent test strip for drug detection according to any one of claims 8-10 in drug detection.

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