Test strip for detecting O6-monoacetylmorphine and methamphetamine and detection method

Abstract

The invention belongs to the technical field of drug detection, and particularly relates to a test strip for detecting O6-monoacetylmorphine and methamphetamine and a detection methamphetamine, the test strip comprises a gold-labeled pad, the surface of the gold-labeled pad is sprayed with an O6-monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugate, a mixture of an O6-monoacetylmorphine coating antigen and time-resolved europium fluorescent microsphere-BSA (Bovine Serum Albumin) is sprayed on the NC membrane to serve as a T1 line, a mixture of a methylamphetamine coating antigen and time-resolved europium fluorescent microsphere-BSA is sprayed on the NC membrane to serve as a T2 line, and a mixture of a goat anti-mouse secondary antibody and time-resolved europium fluorescent microsphere-BSA is sprayed on the NC membrane to serve as a C line; a to-be-detected sample is dropped on the surface of the sample pad on the surface of the test strip main body for detection. The O6-monoacetylmorphine and methamphetamine detection efficiency, sensitivity and specificity are high, the operation is simple and convenient, and the method can be applied to combined determination of two drugs in a biological detection material.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug detection, and in particular relates to a method for detecting 6 -Test strips and detection methods for monoacetylmorphine and methamphetamine. Background Art

[0002] At present, the problems faced by rapid drug testing are the complexity of drug application scenarios and the complexity of drug detection types. 6 -MAM and methamphetamine is a challenging technical task. 6 -Monoacetylmorphine and methamphetamine are two of the most common drug ingredients, which are highly addictive. They are different in chemical structure. Methamphetamine is the main ingredient of ice, while monoacetylmorphine is the main ingredient of methamphetamine. 6 -Monoacetylmorphine is a characteristic metabolite of heroin. 6 -Monoacetylmorphine and methamphetamine are of great significance in the fields of abuse monitoring, forensic identification, clinical diagnosis and drug research, focusing precisely on drugs and their metabolites, such as O 6 -On-site instant detection of monoacetylmorphine and methamphetamine not only helps grassroots public security organs and forensic doctors to quickly screen suspected drug types and determine the drug use of suspects, but also helps to strengthen drug control and effectively combat drug crimes.

[0003] Traditional detection methods include gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), but their application is limited to laboratories due to the cumbersome sample pretreatment, high requirements for laboratory technicians, and high costs of instrumental analysis methods. Rapid detection technology does not require complex pretreatment and is low in cost, making it suitable for instant detection of trace drugs and their metabolites in biological samples at the forensic scene.

[0004] Lateral flow chromatographic assay (LFCA) has been widely used in the rapid screening of known drugs, and a considerable number of commercial kits have been widely used for the rapid detection of methamphetamine, ketamine, heroin and other drugs. The existing methods for drug detection have the following problems: the detection results are slow, they are interfered by impurities, and they cannot be accurately quantified, which is not conducive to practical use. Summary of the invention

[0005] In order to solve the problems in the prior art, the present invention provides a detection method. 6 -The test strips and detection methods for monoacetylmorphine and methamphetamine have high efficiency, high sensitivity, high specificity, and simple operation, and can be applied to the purpose of joint determination of two drugs in biological samples.

[0006] The present invention solves the technical problem by adopting the following technical solutions:

[0007] The present invention aims to provide a method for detecting 6 -Test strips for monoacetylmorphine and methamphetamine, including a gold-labeled pad and an NC membrane. The surface of the gold-labeled pad is sprayed with O 6 -Monoacetylmorphine and methamphetamine antibody-3D dendritic gold nanoparticle conjugates, NC membrane sprayed with O 6 -A mixture of monoacetylmorphine-coated antigen and time-resolved europium fluorescent microspheres-BSA was used as the T1 line, a mixture of methamphetamine-coated antigen and time-resolved europium fluorescent microspheres-BSA was used as the T2 line, and a mixture of goat anti-mouse secondary antibody and time-resolved europium fluorescent microspheres-BSA was used as the C line.

[0008] Furthermore, O 6 The preparation method of the monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugate comprises: taking the three-dimensional dendritic gold nanoparticles precipitated in PBS, adding O 6 -monoacetylmorphine and methamphetamine antibodies, vortex mix, stir, then add blocking agent, mix, stir, and block. After blocking, centrifuge to remove the supernatant, add PBS resuspension to the precipitate, vortex mix, and obtain O 6 -Monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugates.

[0009] Furthermore, the centrifugation time is 5 min, the centrifugation temperature is 4° C., and the centrifugation speed is 10000 r / min.

[0010] Furthermore, the preparation method of three-dimensional dendritic gold nanoparticles includes: stirring Tris-HCl, adding gold seeds, HAuCl4, and dopamine hydrochloride and continuing to stir, and centrifuging and washing after stirring to obtain three-dimensional dendritic gold nanoparticle precipitation.

[0011] Furthermore, the method for preparing the gold seeds includes: stirring the colloidal gold solution and adding polyvinyl pyrrolidone.

[0012] Furthermore, the preparation method of the colloidal gold solution includes: adding water and chloroauric acid into a conical flask, sealing the flask mouth with plastic wrap, placing the conical flask on a heated magnetic stirrer, turning on stirring and heating, and after the system is slightly boiled, continuing to stir, adding trisodium citrate, and turning off the heat source and stirring after the color is stable, and cooling to room temperature.

[0013] Further, after adding trisodium citrate, the color of the solution changed from black to purple and then to red. The heat source was turned off after the color was stable for 10 minutes, and the stirring was turned off 10 minutes after the heat source was turned off.

[0014] Furthermore, the test strip also includes a bottom plate, a sample pad and a water absorbent pad, and the edges of the NC film, the water absorbent pad and the gold label pad are all deburred.

[0015] A detection O 6 - The test strip detection method for monoacetylmorphine and methamphetamine is to drop the sample to be tested on the sample pad surface of the test strip body. In the colorimetric mode, the T1 line, T2 line and C line on the NC film surface of the test strip body are all black lines, indicating that the sample to be tested does not contain O 6 -monoacetylmorphine and methamphetamine; if the T1 line and T2 line on the NC membrane surface of the test strip body disappear and the C line is normal, it means that the sample to be tested contains O 6 -monoacetylmorphine and methamphetamine; if the T1 line on the NC membrane surface of the test strip body disappears, the T2 line does not disappear, and the C line is normal, it means that the sample to be tested contains O 6 -monoacetylmorphine; if the T1 line on the NC membrane surface of the test strip body does not disappear, the T2 line disappears, and the C line is normal, it means that the sample to be tested contains methamphetamine; if the C line disappears, it means that the test strip is invalid; similarly, in the fluorescence mode, time-resolved fluorescence is used to reduce the interference of matrix effects, and the corresponding T1 line fluorescence appears, indicating that O 6 -Monoacetylmorphine is detected and T2 line fluorescence appears, indicating the detection of methamphetamine. If the C line fluorescence is not quenched, it means that the test strip is invalid.

[0016] Compared with the prior art, the beneficial technical effects of the present invention are:

[0017] 1. A method for detecting 6 -Monoacetylmorphine and methamphetamine detection method and test paper, using competitive colorimetric / fluorescent lateral flow immunochromatography technology, with three-dimensional dendritic gold nanoparticles as markers, under optimal conditions, without complex pretreatment, can construct a rapid detection of O in biological samples within 15 minutes on site 6 - Lateral flow immunochromatographic test strips for monoacetylmorphine and methamphetamine.

[0018] 2. Compared with the traditional method, the lateral flow immunochromatographic test paper used in this application has the advantages of simple operation and rapid detection, and its colorimetric mode is 6 - The detection limit of monoacetylmorphine is as low as 1.15ng / mL, the detection limit of methamphetamine is as low as 1.20ng / mL, and the detection limit of O 6 - The detection limits for monoacetylmorphine are as low as 0.21 ng / mL and for methamphetamine are as low as 0.47 ng / mL.

[0019] 3. Compared with the colloidal gold test strips commonly used in the market, 6-The detection limits of monoacetylmorphine and methamphetamine in colorimetric mode are increased by 14 times and 6 times respectively, and the detection limits in fluorescence mode are further increased by 5 times and 3 times respectively, which can be used for rapid on-site detection.

[0020] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention, it can be implemented in accordance with the contents of the specification. In order to make the above contents of the present invention and its objectives, features and advantages more obvious and easy to understand, the specific implementation methods of the present invention are listed below. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a flow chart for preparing the three-dimensional dendritic gold nanoparticle material and a material characterization diagram in the present invention.

[0022] Figure 2 It is a schematic diagram of the main structure of the test strip of the present invention.

[0023] Figure 3 It is a test strip result interpretation diagram of the present invention.

[0024] Figure 4 Schematic diagram of the comparison of LOD between the present invention and commercial colloidal gold.

[0025] Figure 5 The figure is a test result diagram for specificity verification of the test strip of the present invention.

[0026] 101 - bottom plate, 102 - sample pad, 103 - gold label pad, 104 - NC membrane, 105 - water absorbent pad. DETAILED DESCRIPTION

[0027] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary illustrations and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0028] In addition, unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0029] refer to Figure 2 As shown, detection O 6- A test strip for monoacetylmorphine and methamphetamine, comprising a test strip body, wherein the test strip body comprises a bottom plate 101, a sample pad 102, a gold label pad 103, an NC film 104 and a water absorbent pad 105, wherein the bottom plate 101, the sample pad 102, the gold label pad 103, the NC film 104 and the water absorbent pad 105 in the test strip body have the same width on both sides, and the edges of the NC film 104, the water absorbent pad 105 and the gold label pad 103 in the test strip body are deburred, and a test paper inkjet instrument, a test paper cutting machine, a heating magnetic stirrer, an electronic balance, an electric constant temperature blast drying oven and a low-temperature high-speed centrifuge are used in combination in the preparation of the test strip body.

[0030] O 6 -The preparation and detection methods of monoacetylmorphine and methamphetamine test strips are as follows:

[0031] S1. Preparation of detection reagents, including chloroauric acid, trisodium citrate, dopamine hydrochloride, polyvinyl pyrrolidone (PVP), PBS buffer solution, bovine serum albumin, O 6 -MAM and methamphetamine antibodies, O 6 -Monoacetylmorphine and methamphetamine coated antigen, O 6 -Monoacetylmorphine and methamphetamine standards, goat anti-mouse secondary antibody, gold label pad, NC membrane, absorbent pad, glass fiber membrane (sample pad), bottom plate, distilled water, time-resolved europium fluorescent microspheres-BSA.

[0032] S2. Preparation of colloidal gold. The colloidal gold is prepared by trisodium citrate reduction method. After the preparation is completed, it is stored in a refrigerator at 4°C for later use;

[0033] Soak a 250 mL conical flask and a magnetic stirrer in aqua regia, rinse with ultrapure water, add 2 / 3 volume of water to the conical flask, boil at 280°C, pour out, and repeat the washing for 6 times;

[0034] Add 85mL of water and 3mL of 1% chloroauric acid to a conical flask in sequence, seal the flask mouth with plastic wrap, place the conical flask on a heated magnetic stirrer, start stirring and heating, and continue stirring after the system boils slightly. Add 12mL of 1% trisodium citrate, and the color of the solution changes from black to purple and then to red. Turn off the heat source after the color is stable for 10 minutes, turn off the stirring 10 minutes after the heat source is turned off, and cool naturally to room temperature.

[0035] S3, preparation of Au@PVP. The preparation of Au@PVP is carried out by stirring and mixing. After the preparation is completed, it is stored in a refrigerator at 4°C for later use;

[0036] Add 20 mL of colloidal gold solution to a conical flask, place the conical flask on a magnetic stirrer, start stirring, add 4 mg of polyvinyl pyrrolidone (MW10000), stir evenly, and store in a 4°C refrigerator for later use.

[0037] S4, preparation of three-dimensional dendritic gold nanoparticles. The three-dimensional dendritic gold nanoparticles are prepared by using a seed growth method. After the preparation is completed, the three-dimensional dendritic gold nanoparticles are stored in a refrigerator at 4° C. for later use;

[0038] Add 80mL Tris-HCl (0.01M) to a conical flask, place the conical flask on a magnetic stirrer, start stirring, and add 100μL gold seeds (Au@PVP), 3mL0.1% HAuCl4, and 4mL dopamine hydrochloride (2mg / mL) in sequence. The color of the solution changes from transparent to yellow and then to black during the preparation. The stirring time is 12h, and the three-dimensional dendritic gold nanoparticles are precipitated by centrifugation and washing.

[0039] S5. Three-dimensional gold nanoparticles and O 6 -Conjugation of monoacetylmorphine and methamphetamine antibodies: 1 mg of three-dimensional dendritic gold nanoparticles was placed in a centrifuge tube containing 1.8 mL of PBS, and 5 μg, i.e., 5 μL of 1 mg / mL O 6 -monoacetylmorphine and methamphetamine antibodies, vortex to mix, shake at room temperature for 1 hour, then add 200 μL of blocking agent (such as bovine serum albumin), mix, shake at room temperature for 1 hour, block, after blocking, centrifuge to remove the supernatant, the centrifugation time is 5 minutes, the centrifugation temperature is 4°C, the centrifugation speed is 10000r / min, finally, the two precipitates are mixed and redissolved in 1mL of resuspension solution (PBS), vortex to mix, and O 6 -Monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugates.

[0040] S6, antibody labeling, the O obtained in step S5 6 -Monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugate (1 mg / mL) was evenly sprayed onto the gold label pad using a film sprayer. 25 μL of labeled antibody was sprayed per 1 cm of the gold label pad, which was placed in an oven at 37°C and dried in a dry environment for later use.

[0041] S7. Preparation of lateral flow immunochromatographic test paper: Cut the sample pad into 17.5 mm × 300 mm pieces, soak it in the treatment solution for 2 h, dry it in an oven at 37 °C for later use, and then use a film sprayer to spray O 6-Monoacetylmorphine coated antigen (0.05 mg / mL) and time-resolved europium fluorescent microsphere-BSA (0.08 mg / mL) mixture, methamphetamine coated antigen (0.2 mg / mL) and time-resolved europium fluorescent microsphere-BSA (0.08 mg / mL) mixture and 6 mg / mL goat anti-mouse secondary antibody and time-resolved europium fluorescent microsphere-BSA (0.08 mg / mL) mixture were sprayed on the NC membrane as T1 line, T2 line and C line in turn, with an interval of 3 mm between each line. The sprayed NC membrane was placed in a 37°C oven to dry for use. Finally, the sample pad, gold label pad, NC membrane and absorbent pad were attached to the PVC base plate in turn, leaving a 2 mm gap between the sample pad and the NC membrane and assembled with the gold label pad. The assembled base plate was cut into 4 mm wide test strips on a strip cutter and stored in a dry and cool condition for use.

[0042] S8, O 6 - For the detection of monoacetylmorphine and methamphetamine, add 100 μL of the processed biological sample to the surface of the sample pad on the main surface of the test strip, and then make a qualitative judgment.

[0043] Qualitative judgment: In the colorimetric mode, if the T1 line, T2 line and C line on the NC film surface of the test strip body are all black, it means that the sample to be tested is O 6 - Negative for monoacetylmorphine and methamphetamine, i.e., no O 6 -monoacetylmorphine and methamphetamine; if the T1 line and T2 line on the NC membrane surface of the test strip body disappear and the C line is normal, it means that the sample to be tested is O 6 - positive for monoacetylmorphine and methamphetamine, i.e. containing O 6 -monoacetylmorphine and methamphetamine and the content is greater than LOD; if the T1 line on the NC film surface of the test strip body disappears, the T2 line does not disappear, and the C line is normal, it means that the sample to be tested is O 6 - Monoacetylmorphine positive, i.e. containing O 6 -monoacetylmorphine and the content is greater than LOD; if the T1 line on the NC membrane surface of the test strip body does not disappear, the T2 line disappears, and the C line is normal, it means that the sample to be tested is positive for methamphetamine, that is, it contains methamphetamine and the content is greater than LOD; if the C line disappears, it means that the test strip is invalid. Similarly, in the fluorescence mode, time-resolved fluorescence is used to reduce the interference of matrix effects, and the corresponding T1 line fluorescence appears, representing O 6 -Monoacetylmorphine is detected, and the T2 line fluorescence appears, indicating the detection of methamphetamine. If the C line fluorescence is not quenched, it means that the test strip is invalid.

[0044] Quantitative determination: The color intensity of the test strip is analyzed by a colorimeter, and a series of different concentrations of target (O 6-Monoacetylmorphine: 1.25ng / mL, 2.5ng / mL, 5ng / mL, 10ng / mL, 20ng / mL; methamphetamine: 1.25ng / mL, 2.5ng / mL, 5ng / mL, 10ng / mL, 20ng / mL) test strip color intensity values ​​were used to establish a standard curve and obtain the value of 6 - Colorimetric LOD of monoacetylmorphine and methamphetamine; The fluorescence intensity of the test strip was analyzed by immunofluorescence instrument, and the fluorescence intensity of the test strip was analyzed by adding a series of different concentrations of target (O 6 -Monoacetylmorphine: 0.625ng / mL, 1.25ng / mL, 2.5ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 40ng / mL; methamphetamine: 0.625ng / mL, 1.25ng / mL, 2.5ng / mL, 5ng / mL, 10ng / mL, 20ng / mL) test strip fluorescence intensity values ​​were used to establish a standard curve and obtain the value of 0 6 - Fluorescence LOD of monoacetylmorphine and methamphetamine. The quantification of the analyte can be performed based on the established standard curve.

[0045] The present invention 6 -The specific reaction structure principle in the detection process of monoacetylmorphine and methamphetamine is as follows:

[0046] O 6 -Monoacetylmorphine and methamphetamine antibodies were conjugated with three-dimensional dendritic gold nanoparticles to prepare gold-labeled conjugates, and T line fixed O 6 - monoacetylmorphine coated primary, C line fixed sheep anti-mouse secondary antibody, established competitive lateral flow immunochromatographic test strip, drug users' biological samples contain O 6 - monoacetylmorphine and methamphetamine, the test subject's biological sample processing liquid is dripped onto the sample pad 102, and the test liquid flows forward under the capillary action, passes through the gold label pad 103, and the O in the sample 6 -O in the conjugates of monoacetylmorphine and methamphetamine with gold label pad 103 6 - monoacetylmorphine and methamphetamine antibodies bind, which in turn affects the binding of the antibody to the coating source on the T line, so at this time, the T1 and T2 lines do not appear or the color is weakened; the antibody-three-dimensional dendritic gold nanoparticle conjugate continues to migrate forward and binds to the goat anti-mouse secondary antibody fixed on the C line, showing a black line. When a normal subject who has not taken drugs drops a biological sample on the sample pad 102, because the test solution that is chromatographed forward does not contain O 6-monoacetylmorphine and methamphetamine, so it did not bind to the antibody in the gold-labeled conjugate. When it flowed through the T1 and T2 lines, it bound to the coating source fixed on the T line, and the T1 and T2 lines appeared black at this time; the antibody-three-dimensional dendritic gold nanoparticle conjugate continued to migrate forward and bound to the goat anti-mouse secondary antibody fixed on the C line, showing a black line. Therefore, in the colorimetric mode, for normal subjects, three black lines are displayed on the test paper; for suspected heroin users, T1 is weakened or disappeared, and T2 and C lines are normal. For suspected methamphetamine users, T2 line is weakened or disappeared, and T1 and C lines are normal. For suspected methamphetamine and heroin users, T1 and T2 are weakened or disappeared, and C line is normal. Similarly, in the fluorescence mode, time-resolved fluorescence is used to reduce the interference of matrix effects. The corresponding T line fluorescence appears, representing the detection of the corresponding substance, which can be quantified based on the change in fluorescence intensity.

[0047] refer to Figure 4 As shown, by comparing with the LOD of commercial colloidal gold, it can be seen that 6 -The detection limits of monoacetylmorphine and methamphetamine in colorimetric mode are increased by 14 times and 6 times respectively, and the detection limits in fluorescence mode are further increased by 5 times and 3 times respectively, which can be used for rapid on-site detection.

[0048] The test strips were specifically verified using common drugs, namely ketamine, norketamine, cocaine, tetrahydrocannabinol, MDA, methcathinone, fentanyl, methadone, benzoylecgonine, O 6 -Monoacetylmorphine, methamphetamine, and mixed samples of the above drugs were tested, and the negative control was methanol + phosphate buffer solution.

[0049] The results are as follows Figure 5 As shown in the figure, in the colorimetric mode: only when the target substance exists in the sample, the corresponding T line will disappear, while for the other drugs, the intensity of their T lines is almost the same as that of the blank sample; in the fluorescence mode: only when the target substance exists in the sample, the corresponding T line fluorescence will appear, while for the other drugs, the fluorescence intensity of their T lines is almost the same as that of the blank sample; indicating that the established O 6 - Rapid test strips for monoacetylmorphine and methamphetamine 6 -Monoacetylmorphine and methamphetamine have good selectivity.

[0050] The present invention discloses a colorimetric / fluorescence dual signal system based on a competitive mode to simultaneously detect O 6 -monoacetylmorphine and methamphetamine method, drop the sample to be tested on the sample pad of the test strip, and observe the color development of the T line and the C line after 15 minutes. The present invention is the first to use dendritic gold nanoparticles with the optimal particle size to detect O 6-Monoacetylmorphine and methamphetamine were detected by using the excellent fluorescence quenching effect of dendritic gold nanoparticles, which greatly improved the sensitivity and specificity of immunochromatography, making the linear range of detection reach 1.25-20ng / mL. Compared with the commonly used colloidal gold test strips on the market, the sensitivity and specificity have been significantly improved. The main body of the test strip includes a base plate, a sample pad, a gold label pad, an NC membrane and a water absorbent pad. It uses lateral flow immunochromatography technology and uses dendritic gold as a marker. Under optimal conditions, without complex pretreatment, a method can be constructed that can quickly detect O in biological samples within 15 minutes on site. 6 - Lateral flow immunochromatography technology for monoacetylmorphine and methamphetamine. Compared with the traditional method, the lateral flow immunochromatography test paper used in this application has the advantages of simple operation and rapid detection, and its colorimetric mode is 6 - The detection limit of monoacetylmorphine is as low as 1.15ng / mL, the detection limit of methamphetamine is as low as 1.20ng / mL, and the detection limit of O 6 -The detection limit of monoacetylmorphine is as low as 0.21ng / mL, and the detection limit of methamphetamine is as low as 0.47ng / mL, which can be used for rapid detection on site.

[0051] The serial numbers of the above embodiments of the present invention are only for description and do not represent the advantages or disadvantages of the embodiments.

[0052] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation modes, which are merely illustrative rather than restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are within the protection of the present invention.

Claims

1. A method for detecting 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that It includes a gold-labeled pad and an NC film. The surface of the gold-labeled pad is sprayed with O 6 -Monoacetylmorphine and methamphetamine antibody-3D dendritic gold nanoparticle conjugates, NC membrane sprayed with O 6 -A mixture of monoacetylmorphine-coated antigen and time-resolved europium fluorescent microspheres-BSA was used as the T1 line, a mixture of methamphetamine-coated antigen and time-resolved europium fluorescent microspheres-BSA was used as the T2 line, and a mixture of goat anti-mouse secondary antibody and time-resolved europium fluorescent microspheres-BSA was used as the C line.

2. A method for detecting O as claimed in claim 1 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: O 6 The preparation method of the monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugate comprises: taking the three-dimensional dendritic gold nanoparticles precipitated in PBS, adding O 6 -monoacetylmorphine and methamphetamine antibodies, vortex mix, stir, then add blocking agent, mix, stir, and block. After blocking, centrifuge to remove the supernatant, add PBS resuspension to the precipitate, vortex mix, and obtain O 6 -Monoacetylmorphine and methamphetamine antibody-three-dimensional dendritic gold nanoparticle conjugates.

3. A method for detecting O as claimed in claim 2 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: The centrifugation time was 5 min, the centrifugation temperature was 4°C, and the centrifugation speed was 10000 r / min.

4. A method for detecting O as claimed in claim 2 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: The preparation method of three-dimensional dendritic gold nanoparticles comprises: stirring Tris-HCl, adding gold seeds, HAuCl4 and dopamine hydrochloride and continuing stirring, and centrifugally washing after stirring to obtain three-dimensional dendritic gold nanoparticle precipitation.

5. A method for detecting O as claimed in claim 4 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: The method for preparing the gold seeds comprises: stirring the colloidal gold solution and adding polyvinyl pyrrolidone.

6. A method for detecting O as claimed in claim 5 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: The preparation method of the colloidal gold solution comprises: adding water and chloroauric acid into a conical flask, sealing the flask mouth with plastic wrap, placing the conical flask on a heated magnetic stirrer, starting stirring and heating, and continuing stirring after the system slightly boils, adding trisodium citrate, and turning off the heat source and stirring after the color is stable, and cooling to room temperature.

7. A method for detecting O as claimed in claim 6. 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: After adding trisodium citrate, the color of the solution changes from black to purple and then to red. The heat source is turned off after the color is stable for 10 minutes, and the stirring is turned off 10 minutes after the heat source is turned off.

8. A method for detecting O as claimed in claim 1. 6 -Test strips for monoacetylmorphine and methamphetamine, characterized in that: The test strip also includes a bottom plate, a sample pad and a water absorbent pad. The edges of the NC membrane, the water absorbent pad and the gold standard pad are all deburred.

9. A method of detecting a molecule according to any one of claims 1 to 8. 6 - A method for detecting monoacetylmorphine and methamphetamine with a test strip, characterized in that: The sample to be tested is dropped onto the sample pad surface of the test strip body. In the colorimetric mode, the T1 line, T2 line and C line on the NC film surface of the test strip body are all black, indicating that the sample to be tested does not contain O 6 -monoacetylmorphine and methamphetamine; if the T1 line and T2 line on the NC membrane surface of the test strip body disappear and the C line is normal, it means that the sample to be tested contains O 6 -monoacetylmorphine and methamphetamine; if the T1 line on the NC membrane surface of the test strip body disappears, the T2 line does not disappear, and the C line is normal, it means that the sample to be tested contains O 6 -monoacetylmorphine; if the T1 line on the NC membrane surface of the test strip body does not disappear, the T2 line disappears, and the C line is normal, it means that the sample to be tested contains methamphetamine; if the C line disappears, it means that the test strip is invalid; similarly, in the fluorescence mode, time-resolved fluorescence is used to reduce the interference of matrix effects, and the corresponding T1 line fluorescence appears, indicating that O 6 -Monoacetylmorphine is detected and T2 line fluorescence appears, indicating the detection of methamphetamine. If the C line fluorescence is not quenched, it means that the test strip is invalid.

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