Progesterone immunochromatography detection test strip as well as preparation method and application thereof

Abstract

The invention discloses a progesterone immunochromatography detection test strip as well as a preparation method and application thereof. The progesterone immunochromatography detection test strip comprises a sample pad, a combination pad, a chromatography membrane and a water absorption pad which are sequentially overlapped on a bottom plate, wherein the combination pad is coated with a fluorescent marker; a detection line and a quality control line are arranged on the chromatography membrane; the detection line is coated with a progesterone molecularly imprinted polymer. A progesterone molecularly imprinted polymer replaces a conventional progesterone antigen to serve as a detection line, the combination pad is coated with a fluorescent marker, progesterone in a sample flows through the combination pad to be combined with the fluorescent marker and flows to the detection line of the chromatographic membrane, and a compound formed by the progesterone and the fluorescent marker can be specifically recognized by the molecularly imprinted polymer; qualitative and quantitative detection of progesterone is realized, and high-precision and high-sensitivity detection of a sample can be realized in cooperation with a fluorescence detector. The progesterone molecularly imprinted polymer is used for replacing the conventional progesterone antigen, so that the defects of long preparation period, easiness in inactivation, high cost and the like of the traditional biological antibody are overcome.

Description

Technical Field

[0001] The invention relates to a progesterone immunochromatographic test strip, a preparation method and application thereof, and belongs to the technical field of immunochromatographic detection. Background Art

[0002] Progesterone (Prog for short, also known as progesterone, progesterone, progesterone, progestin, progestogen, progestin or progesterone in Chinese) is a steroid involved in the female menstrual cycle, pregnancy and the embryo of humans or animals. It is the most important progestogen. The value of progesterone can be measured to determine whether the ovary has ovulated, and the function of the corpus luteum can also be evaluated by measuring progesterone. Progesterone plays an important role in the reproductive activities of women or female mammals, such as maintaining pregnancy, initiating labor, and breast development. Therefore, the detection of progesterone is particularly important for pregnant women or animals. Progesterone is generally detected by drawing peripheral venous blood. Clinical detection methods include instrumental methods: chemiluminescence instrument, liquid chromatography-mass spectrometry and high-performance chromatography. These instruments are costly and complex to operate. They have high requirements for detection personnel and environment and require supporting reagents.

[0003] Immunochromatography is a detection and analysis method that combines immunotechnology and chromatography technology. It is widely used in on-site detection because of its fast detection speed, strong specificity, and the fact that it does not require professional instruments to determine the results or professional personnel to operate. At present, small molecule antigens such as progesterone on the market can only be detected by competitive method because they only have a single determinant cluster and steric hindrance. The detection principle of the competitive method is that after the sample containing progesterone molecules is dripped onto the sample pad, it is chromatographed to the binding pad and combined with the colloidal gold-antibody complex to form a colloidal gold marker and move toward the T line together. If the sample contains progesterone molecules, the progesterone molecules in the sample compete with the progesterone molecules coated on the T line for binding to the antibodies on the colloidal gold-antibody complex. Since the colloidal gold-antibody complex cannot bind to the progesterone on the T line, no line is generated at the T line position; if there are no progesterone molecules in the sample, the colloidal gold-antibody complex will bind to the progesterone molecules on the T line, forming a visual line at the T line. However, in the competitive method, the sensitivity and specificity of the test are greatly affected by the progesterone molecules and antibodies, and the accuracy is relatively insufficient. Progesterone is a small molecule substance, and the preparation of antigens and antibodies is relatively difficult. In addition, the storage conditions are harsh, and animals need to be sacrificed. Summary of the invention

[0004] The first object of the present invention is to provide a progesterone immunochromatographic test strip to solve the problems of narrow linear range, large sensitivity fluctuation, insufficient accuracy and inability to quantitatively detect of existing progesterone immunochromatographic test strips; using progesterone molecular imprinting polymer instead of conventional progesterone molecules as the detection line avoids the problems of difficult progesterone preparation and harsh storage conditions.

[0005] The second object of the present invention is to provide a method for preparing a progesterone immunochromatographic test strip to solve the above problems.

[0006] The third object of the present invention is to provide a progesterone immunochromatographic test strip for use in progesterone detection to solve the above problems.

[0007] In order to achieve the above object, the technical solution adopted by the progesterone immunochromatographic test strip of the present invention is:

[0008] A progesterone immunochromatographic test strip comprises a sample pad, a conjugation pad, a chromatographic membrane and a water-absorbing pad which are overlapped in sequence on a bottom plate, wherein the conjugation pad is coated with a fluorescent marker; the chromatographic membrane is provided with a detection line and a quality control line; and the detection line is coated with a progesterone molecular imprinting polymer.

[0009] The progesterone molecular imprinted polymer of the present invention replaces conventional progesterone molecules as the detection line, and the conjugate pad is coated with a fluorescent marker. The progesterone in the sample flows through the conjugate pad and combines with the fluorescent marker containing the progesterone monoclonal antibody, and continues to be chromatographed to the detection line of the nitrocellulose membrane. The complex formed by the combination of progesterone and the fluorescent marker can be specifically recognized by the molecular imprinted polymer, thereby realizing the qualitative and quantitative detection of progesterone. In combination with a corresponding fluorescence detector, the sample to be tested can be detected with high precision and high sensitivity.

[0010] Specifically, the chromatography membrane is a nitrocellulose membrane; and the bottom plate is a PVC bottom plate.

[0011] Preferably, the fluorescent marker is a conjugate of time-resolved fluorescent microspheres and progesterone monoclonal antibodies; the concentration of the progesterone monoclonal antibodies is 10-50 μg / mL.

[0012] The present invention uses time-resolved fluorescent microspheres Eu, which have a large Stokes shift (>150nm) and a fluorescence lifetime 5 to 6 orders of magnitude higher than that of background substances, and can effectively eliminate the interference of various nonspecific fluorescence in the sample to be tested, and significantly improve the sensitivity of progesterone detection.

[0013] Specifically, the quality control line is coated with a specific antibody that binds to the progesterone monoclonal antibody. For example, if the progesterone monoclonal antibody in the fluorescent marker is a mouse IgG antibody, the specific antibody is an anti-mouse IgG antibody such as a goat anti-mouse IgG antibody or a rabbit anti-mouse IgG antibody.

[0014] Preferably, the progesterone molecular imprinted polymer is prepared by copolymerizing and removing the template molecule using progesterone molecule as a template, methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinking agent, and benzophenone as an initiator.

[0015] More preferably, the progesterone: methacrylic acid: ethylene glycol dimethacrylate: benzophenone = 20-25 mg: 24-26 μL: 110-120 μL: 5-6 mg.

[0016] Molecularly imprinted polymers (MIPs) are artificially synthesized polymer materials that have selective recognition ability for template molecules by simulating the principle of antigen-antibody interaction in nature. Whether they can effectively bind to template molecules is closely related to the selection of template molecules, functional monomers, crosslinking agents, initiators and solvents, and the proportion of each component. The present invention screens and optimizes materials such as functional monomers, crosslinking agents, initiators and solvents, and proves through experiments that the prepared progesterone molecular immunoimprinted polymer can effectively bind to natural progesterone molecules, and is applied to the progesterone immunochromatographic test strip detection line, and can specifically identify the complex formed by the combination of progesterone and fluorescent markers, thereby realizing highly sensitive and accurate qualitative and quantitative detection of progesterone.

[0017] Preferably, the copolymerization condition is to react under ultraviolet light source for 25-35 minutes.

[0018] The present invention targets the template molecule progesterone, screens out the reaction components, optimizes the polymerization reaction conditions through a large number of experiments, and conducts the polymerization reaction under the above conditions. The recognition holes formed inside the obtained progesterone molecular imprinting polymer are consistent with the progesterone molecule in terms of recognition site and spatial size. Experiments have proved that the polymer can specifically recognize the complex formed by the combination of progesterone and a fluorescent marker on the detection line of the progesterone immunochromatographic test strip.

[0019] Preferably, the copolymerization is carried out in an organic solvent, with 0.125-0.130 mL of organic solvent corresponding to 1 mg of progesterone molecule.

[0020] The organic solvent can fully dissolve the components of the copolymerization reaction and provide homogeneous and stable reaction conditions. Further preferably, the organic solvent is acetonitrile.

[0021] In order to achieve the above object, the technical scheme adopted by the method for preparing the progesterone immunochromatographic test strip of the present invention is:

[0022] A method for preparing a progesterone immunochromatographic test strip comprises the following steps:

[0023] 1) Add the fluorescent marker to the conjugate pad and dry it;

[0024] 2) coating the progesterone molecular imprinting polymer solution on the detection line of the chromatographic membrane, coating the specific antibody solution that binds to the progesterone monoclonal antibody in the fluorescent marker on the quality control line of the chromatographic membrane, and drying;

[0025] 3) Assembling the sample pad, the dried conjugate pad, the dried chromatography membrane, the absorbent pad and the substrate into a fluorescent immunochromatographic test strip.

[0026] The preparation method of the fluorescent immunochromatographic test strip of the present invention has a simple process. Since the progesterone molecular imprinting polymer is used as the detection line instead of the conventional progesterone molecule, and the conjugate pad is coated with a fluorescent marker, the progesterone in the sample flows through the conjugate pad and combines with the fluorescent marker containing the progesterone monoclonal antibody, and continues to be chromatographed to the detection line of the nitrocellulose membrane. The complex formed by the combination of the progesterone and the fluorescent marker can be specifically recognized by the molecular imprinting polymer, thereby realizing the qualitative and quantitative detection of the progesterone. In combination with a corresponding fluorescence detector, the sample to be tested can be detected with high precision and high sensitivity.

[0027] Specifically, the progesterone immunochromatographic test strip is prepared by the following steps:

[0028] (1) Preparation of fluorescent markers and conjugate pads: Activating fluorescent microspheres using DEC and NHS methods, coupling the activated fluorescent microspheres with progesterone monoclonal antibodies, and coating the fluorescent markers obtained by the reaction on the conjugate pads;

[0029] (2) Preparation of progesterone molecular imprinted polymer: progesterone and methacrylic acid are dissolved in acetonitrile for prepolymerization reaction, ethylene glycol dimethacrylate and benzophenone are added to the prepolymerization system for polymerization reaction under nitrogen environment to obtain a polymer, and the template molecule progesterone is removed to obtain the progesterone molecular imprinted polymer;

[0030] (3) Preparation of chromatographic membrane: coating the progesterone molecular imprinting polymer prepared in step (2) on the chromatographic membrane detection line, and coating the goat anti-mouse IgG on the quality control line;

[0031] (4) Assembly of progesterone immunochromatographic test strip: The sample pad is treated with a sample pad treatment solution, and after drying, the sample pad, the binding pad, the chromatographic membrane and the absorbent pad are sequentially attached to the bottom plate to assemble the test strip.

[0032] Preferably, the concentration of the progesterone molecularly imprinted polymer in the progesterone molecularly imprinted polymer solution is 15-25 mg / mL; and the concentration of the specific antibody in the specific antibody solution is 0.5-1 mg / mL.

[0033] More preferably, the coating speed is 0.5-0.7 μL / mm.

[0034] In order to achieve the above object, the technical solution adopted by the application of the progesterone immunochromatographic test strip of the present invention in progesterone detection is:

[0035] The invention discloses an application of a progesterone immunochromatographic test strip in progesterone detection.

[0036] The progesterone immunochromatographic test strip of the present invention has high detection sensitivity and good accuracy, has a wide detection range, is linearly correlated within the range of 0-100 ng / mL, and can realize qualitative and quantitative detection of progesterone.

[0037] Preferably, the detection is a quantitative detection.

[0038] More preferably, the sample to be tested is dropped onto the sample pad, and the reaction is carried out for 10 to 15 minutes under light-proof conditions before fluorescence detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is a schematic diagram of the structure of the progesterone immunochromatographic test strip in Example 1 of the present invention;

[0040] Figure 2 The standard curve of progesterone detection of the progesterone immunochromatographic test strip in Example 3 of the present invention;

[0041] Figure 3 This is the correlation curve of the progesterone immunochromatographic test strip in the range of 0-20 ng / mL for progesterone detection in Experimental Example 1 of the present invention;

[0042] Among them, 1-sample pad; 2-binding pad; 3-nitrocellulose membrane; 4-detection line; 5-quality control line; 6-absorbent pad; 7-PVC bottom plate. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical solutions and advantages of the present application clearer, the technical solutions of the present application will be clearly and completely described below in conjunction with the specific embodiments of the present application and the corresponding drawings. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments, and are not intended to limit the scope of the present invention. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0044] Example 1 A progesterone immunochromatographic test strip

[0045] The progesterone immunochromatographic test strip of the present embodiment is a fluorescent immunochromatographic test strip for qualitative and quantitative detection of progesterone. Figure 1As shown, it comprises a sample pad 1, a conjugate pad 2, a nitrocellulose membrane 3 and a water-absorbing pad 6 which are overlapped in sequence on a PVC bottom plate 7; the conjugate pad is coated with a fluorescent marker, and the fluorescent marker is a conjugate of time-resolved (Eu) fluorescent microspheres and progesterone monoclonal antibodies; a detection line (T line) 4 and a quality control line (C line) 5 are provided on the chromatographic membrane; the detection line is coated with a progesterone molecular imprinting polymer; and the quality control line is coated with goat anti-mouse IgG.

[0046] When the progesterone immunochromatographic test strip of this embodiment detects progesterone in a sample to be tested, after the chromatography begins, the progesterone antigen in the sample flows through the binding pad and combines with the fluorescent marker containing the progesterone monoclonal antibody, and continues to flow to the T line of the nitrocellulose membrane through the chromatography. The complex formed by the combination of progesterone and the fluorescent marker can be specifically recognized by the molecular imprinting polymer, so that the fluorescence intensity of the T line increases with the increase of the progesterone concentration, and the excess complex flows to the C line and combines with the goat anti-mouse IgG to show fluorescence. Finally, the fluorescence signal values ​​of the T line and the C line are read using a fluorescent immunoassay analyzer, thereby realizing the qualitative and quantitative detection of progesterone.

[0047] Example 2 Preparation method of progesterone immunochromatographic test strip

[0048] The preparation method of the fluorescent immunochromatographic test strip of this embodiment comprises the following steps:

[0049] 1. Preparation of fluorescent microspheres and progesterone monoclonal antibody coupling complex:

[0050] (1) Activation of fluorescent microspheres

[0051] 200 μL of time-resolved fluorescent microspheres (300 nm in diameter) were taken and added to 800 μL of activation buffer (50 mmol / LMES, pH 6.0). The suspended fluorescent microspheres were fully mixed with a vortex mixer, and centrifuged at 4°C, 10,000 rpm for 15 min. The supernatant was carefully aspirated with a micropipette, and the step was repeated once.

[0052] Weigh NHS and EDC separately in two centrifuge tubes, and use activation buffer to dissolve the activator at a concentration of 1%. Add 10 μL NHS to the time-resolved fluorescent microspheres, mix thoroughly and quickly with a vortex mixer, then pipette 10 μL EDC into it, mix quickly, stir magnetically at room temperature for 30 minutes, centrifuge at 4°C, 10,000 rpm for 30 minutes, and carefully remove the supernatant;

[0053] Add 1 mL of coupling buffer (10 mmol / L PBS, pH 7.4), resuspend the fluorescent microspheres by ultrasound to disperse them evenly (50 W, working for 2 s, interval of 2 s, lasting for 3 min; the subsequent ultrasound conditions were the same), centrifuge at 4 ° C, 10000 rpm for 30 min, carefully remove the supernatant, and repeat this step once.

[0054] (2) Conjugation of fluorescent microspheres with progesterone monoclonal antibodies

[0055] Add 1 mL of coupling buffer to the activated fluorescent microspheres, resuspend and disperse by ultrasound, add progesterone monoclonal antibody (Cat. No.: AT01PROGAb) to a final concentration of 20 μg / mL, mix thoroughly with a vortex mixer, and perform coupling reaction for 2 hours at room temperature. Centrifuge at 4°C, 10,000 rpm for 30 minutes, carefully remove the supernatant, add 1 mL of coupling buffer to resuspend, centrifuge under the same conditions to remove the supernatant, and repeat once.

[0056] Add 1 mL of blocking solution (PBS solution containing 5% BSA) and block for 1 hour under rotation at room temperature. Collect the precipitate by centrifugation, add 1 mL of preservation solution (PBS buffer containing 1% BSA, 5% trehalose, 10% sucrose, 0.05% Proclin and 0.5% PVP, pH 7.4), and resuspend by ultrasonication. Repeat twice to obtain the fluorescent marker.

[0057] 2. Preparation of progesterone molecularly imprinted polymer:

[0058] 23.6 mg of template molecule progesterone and 25.6 μL of functional monomer methacrylic acid were taken into a round-bottom flask, and fully dissolved in 3 mL of acetonitrile. The prepolymerization reaction was carried out for 3 hours. 114 μL of cross-linking agent ethylene glycol dimethacrylate and 5.4 mg of initiator benzophenone were added to the prepolymerization system. Ultrasonic oscillation was used to mix the polymerization solution evenly. Nitrogen was passed into the reaction solution for 10 minutes. The polymerization time was 30 minutes under ultraviolet light to obtain a polymer. After the reaction was completed, phosphate buffer (PBS, pH 7.4) was passed to rinse for 10 minutes, and then an elution solution (PBS (pH 7.4): acetic acid = 9:1, volume ratio) was passed to elute for about 33 minutes, and then PBS (pH 7.4) was passed to rinse for about 75 minutes. The template molecule progesterone was removed to obtain a molecularly imprinted polymer.

[0059] 3. The formation of chromatography test strips:

[0060] The XYZ three-dimensional spraying platform of BioDot Company was used to spray fluorescent microspheres and progesterone monoclonal antibody complexes on the binding pad at a speed of 0.5 μL / mm, and dried in a blast drying oven at 37°C for use; the sample pad was treated with sample pad treatment solution (10% sucrose, 0.1% BSA, 0.1% Tween 20 dissolved in 10mM PBS buffer) for 10 minutes, and dried at 37°C for use; a membrane scribing instrument was used to scribble sheep anti-mouse IgG secondary antibody solution (the concentration of sheep anti-mouse IgG secondary antibody was 1 mg / mL) at the quality control line (C line) position of the nitrocellulose membrane at a speed of 0.5 μL / mm, and the molecular imprinting polymer solution (the concentration of molecular imprinting polymer was 20 mg / mL) was scribed at the detection line (T line) position, the two lines were 5 mm apart, and dried in a blast drying oven, and then the sample pad, fluorescent pad, nitrocellulose membrane and absorbent paper were sequentially pasted on the PVC bottom plate, each pad and NC membrane covered 1 mm, and the test strip ( Figure 1 ).

[0061] Example 3 Application of progesterone immunochromatographic test strips in progesterone detection

[0062] This embodiment provides the application of the progesterone immunochromatographic test strip in the detection of progesterone, and the specific implementation operation is as follows:

[0063] The progesterone molecule was diluted with 10mM PB+0.5% TWEEN-20 buffer to prepare progesterone standards with progesterone concentrations of 0, 1, 5, 10, 30, 50, 80, and 100 ng / mL. 80 μL of the progesterone standard was added to the sample pad of the progesterone immunochromatographic test strip in Example 1, and the fluorescence detection was performed after reacting for 15 minutes under light-proof conditions. The fluorescence signal values ​​of the T line and the C line were read using a fluorescent immunoassay analyzer, and the value of T / C was calculated. The results are shown in Table 1.

[0064] Table 1 Test results of progesterone standards with different concentrations in the range of 0-100 ng / mL

[0065]

[0066]

[0067] From the data in Table 1, it can be seen that within the range of 0-100 ng / mL, as the progesterone concentration increases, the T line signal value increases, the C line signal value changes very little, and the T / C value increases. It can be concluded that the detection range of the test strip is 0-100 ng / mL, and within this range, the antigen concentration is positively correlated with the T / C value, which can achieve quantitative detection of the sample. According to the T / C values ​​at different concentrations, the corresponding standard curve can be established, such as Figure 2As shown, the concentration of progesterone in the sample is calculated according to the strength of the T-line fluorescence signal. The present invention establishes a specific recognition progesterone immunochromatographic test strip that is linearly correlated in the range of 0-100 ng / mL.

[0068] (Y=0.0265X+0.1951, R2=0.9745).

[0069] Experimental Example 1 Accuracy Test of Progesterone Immunochromatographic Test Strips

[0070] The accuracy of the progesterone immunochromatographic test strip was tested in the range of 0-20 ng / mL to determine the minimum detection limit of the test strip. The results are shown in Table 2. With a 2-unit concentration gradient, there is a linear correlation in the range of 0-20 ng / mL. The linear relationship is as shown in Figure 3 As shown, the minimum detection limit of this test strip is 2ng / mL, indicating that the detection sensitivity of the test strip in this application is relatively high.

[0071] Table 2 Test results of progesterone standards at different concentrations in the range of 0-20 ng / mL

[0072]

[0073] Experimental Example 2 Stability Test of Progesterone Immunochromatographic Test Strips

[0074] The average of 10 T / C values ​​of 2 concentration samples (20 ng / mL and 70 ng / mL progesterone standard) was taken as the mean, and the coefficient of variation CV was calculated 10 times. The experimental results are shown in Table 3. The experimental results show that the coefficient of variation of the test strips is less than 11%.

[0075] Table 3 Stability test of 10 batches of progesterone immunochromatographic test strips

[0076] Sample concentration Mean Standard Deviation CV 20 0.985945 0.10609 0.107602 70 2.297693 0.237898 0.103538

[0077] Experimental Example 3 Practical Application of Progesterone Immunochromatographic Test Strips

[0078] Take 9 progesterone test strips, add progesterone standards with concentrations of 20, 50, and 70 ng / mL to the sample addition area (each concentration is repeated 3 times), and after chromatography for 10 minutes, the instrument reads the C and T line signals, and calculates the progesterone antigen concentration according to the standard curve in Example 3. The results are shown in Table 4. It can be seen from the table that the progesterone immunochromatographic test strip of the present invention can effectively detect progesterone in actual samples, and the accuracy is within 11%. Compared with commercially available test strips, when the progesterone molecule content in the sample to be tested is small, the result of the test strip detection of the present application is closer to the actual content of the sample.

[0079] Table 4 Progesterone immunochromatographic test strips of the present invention and commercially available test strips

[0080]

[0081] In summary, the progesterone immunochromatographic test strip provided by the present invention has a wide detection range and can realize rapid and accurate qualitative and quantitative detection of progesterone in a sample to be tested, which is of great significance for clinical testing and has broad market application prospects.

[0082] Finally, it is noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A progesterone immunochromatographic test strip, comprising a sample pad, a conjugate pad, a chromatographic membrane and a water-absorbing pad sequentially overlapped on a bottom plate, characterized in that: The conjugate pad is coated with a fluorescent marker; the chromatographic membrane is provided with a detection line and a quality control line; the detection line is coated with a progesterone molecular imprinting polymer.

2. The progesterone immunochromatographic test strip according to claim 1, characterized in that: The fluorescent marker is a conjugate of time-resolved fluorescent microspheres and progesterone monoclonal antibodies; the concentration of the progesterone monoclonal antibodies is 10-50 μg / mL.

3. The progesterone immunochromatographic test strip according to claim 1, characterized in that: The progesterone molecular imprinting polymer is prepared by copolymerizing and removing the template molecule, using progesterone molecule as a template, methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinking agent, and benzophenone as an initiator.

4. The progesterone immunochromatographic test strip according to claim 3, characterized in that: The progesterone: methacrylic acid: ethylene glycol dimethacrylate: benzophenone = 20-25 mg: 24-26 μL: 110-120 μL: 5-6 mg.

5. A method for preparing the progesterone immunochromatographic test strip according to any one of claims 1 to 4, characterized in that: The following steps are involved: 1) Add the fluorescent marker to the conjugate pad and dry it; 2) coating the progesterone molecular imprinting polymer solution on the detection line of the chromatographic membrane, coating the specific antibody solution that binds to the progesterone monoclonal antibody in the fluorescent marker on the quality control line of the chromatographic membrane, and drying; 3) Assemble the sample pad, the dried conjugate pad, the dried chromatography membrane, the absorbent pad and the bottom plate into a fluorescent immunochromatographic test strip.

6. The method for preparing the progesterone immunochromatographic test strip according to claim 5, characterized in that: The concentration of the progesterone molecular imprinted polymer in the progesterone molecular imprinted polymer solution is 15-25 mg / mL; the concentration of the specific antibody in the specific antibody solution is 0.5-1 mg / mL.

7. The method for preparing the progesterone immunochromatographic test strip according to claim 6, characterized in that: The coating speed is 0.5-0.7 μL / mm.

8. Use of the progesterone immunochromatographic test strip according to any one of claims 1 to 4 in progesterone detection.

9. Use of the progesterone immunochromatographic test strip according to claim 8 in progesterone detection, characterized in that: The detection is a quantitative detection.

10. Use of the progesterone immunochromatographic test strip according to claim 8 in progesterone detection, characterized in that: Add the sample to be tested onto the sample pad, allow to react for 10 to 15 minutes in a dark environment, and then perform fluorescence detection.

Images