Fluorescence immunochromatography instant detection system based on mobile terminal and application of fluorescence immunochromatography instant detection system

Abstract

The invention discloses a fluorescence immunochromatography instant detection system based on a mobile terminal and application thereof, the instant detection system comprises: a carrier, which is internally provided with an excitation light source assembly and a test strip loading channel; the test strip loading channel comprises a pulling and inserting opening, a limiting groove and light blocking groove covers, the pulling and inserting opening is formed in the side edge of the carrier and located on one side of the limiting groove, the limiting groove is used for loading the test strip after the test strip is inserted into the pulling and inserting opening, and the light blocking groove covers are arranged on the two sides of the limiting groove respectively and used for blocking a sample pad, a combination pad and absorbent paper of the test strip. Only the nitrocellulose membrane of the test strip is exposed; the carrier cover covers the carrier to form a cassette; a light hole is formed in the carrying cover, and the light hole, the nitrocellulose membrane and exciting light of the exciting light source assembly are located on the same optical path; and the mobile terminal is loaded on the loading cover. The system is simple in structure, low in manufacturing cost, convenient to operate and very suitable for being used in mobile medical centers or basic laboratories.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biochemical experimental instruments, and more particularly to a fluorescence immunoassay instant detection system based on a mobile terminal and application thereof. BACKGROUND

[0002] The immunoassay test strip is a widely used instant detection technology, which has the advantages of simple operation and portability, and is most widely used in the medical field. For example, the immunoassay test strip for triple detection of influenza A, influenza B and new coronavirus antigens disclosed in Chinese patent CN119438583A. The fluorescence immunoassay test strip is a kind of immunoassay test strip using fluorescent microspheres labeled antigen or antibody as detection probe, based on indirect method or double antigen sandwich detection of target. It can realize quantitative detection of signal through dry fluorescence immunoassay analyzer. Compared with the traditional colloidal gold immunoassay test strip, the fluorescence immunoassay test strip has lower detection background, higher sensitivity, more accurate quantification, wider detection range and more stable results, which is superior to the limitation of colloidal gold qualitative or semi-quantitative. For example, the time-resolved fluorescence immunoassay test strip for combined detection of IL-6 and PCT disclosed in Chinese patent CN109975557A. However, the dry immunoassay fluorescence analyzer is expensive, bulky and needs to be operated by computer, which is poor in portability. For example, the immunofluorescence instrument disclosed in Chinese design patent CN306930374S. In order to further meet the portability, some people have disclosed a handheld dry fluorescence immunoassay analyzer (Chinese design patent CN309343065S). Although the handheld dry fluorescence immunoassay analyzer brings portability, it also brings higher price, which cannot be popularized and popularized.

[0003] In recent years, smart phones have been used to build portable test strip detection platforms due to their small size, smart digitalization, instant information processing and one per capita, such as the test strip result rapid reading device based on smart phone digital image processing disclosed in CN111830252A. This patent uses photo recognition of the entire test strip, which does not greatly avoid the non-reactive area (such as sample pad, conjugate pad and absorbent paper), which is easy to cause interference of environmental signal. Because the sample pad, conjugate pad and absorbent paper of the existing test strip are non-removable fluorescent materials, they will emit interfering fluorescence under ultraviolet excitation light, thereby affecting image processing. In addition, the sample pad, conjugate pad, nitrocellulose membrane (reaction area) and absorbent paper are not flat at the joint parts, which will also interfere with fluorescence analysis. SUMMARY

[0004] Therefore, it is necessary to provide a fluorescence immunoassay instant detection system based on a mobile terminal and application thereof in view of the above technical problems.

[0005] In order to solve the above technical problems, the first aspect of the present application proposes a mobile terminal-based fluorescence immunoassay rapid detection system, which adopts the technical solutions as follows: The mobile terminal-based fluorescence immunoassay rapid detection system comprises: A carrier, which is internally provided with an excitation light source assembly and a test strip loading channel; the test strip loading channel comprises a plug-in port, a limiting groove, and a light-blocking groove cover, the plug-in port is arranged on the side of the carrier and located on one side of the limiting groove, the limiting groove is used for loading the test strip after the test strip is inserted into the plug-in port, and the light-blocking groove cover is arranged on both sides of the limiting groove to shield the sample pad, the conjugate pad, and the absorbent paper of the test strip, and only the nitrocellulose membrane of the test strip is exposed; A carrier cover, which forms a dark box after being arranged on the carrier; the carrier cover is provided with a light transmission hole, which is on the same optical path as the excitation light of the excitation light source assembly and the nitrocellulose membrane; A mobile terminal, which is loaded on the carrier cover, the camera module of the mobile terminal corresponds to the light transmission hole to take a picture of the exposed nitrocellulose membrane area; the mobile terminal is internally provided with a fluorescence intensity analysis application program to convert the received T-line and C-line fluorescence signals into T / C values.

[0006] Further, the excitation light source assembly comprises a lamp panel capable of emitting excitation light, an excitation light source switch, an adjustable resistor for adjusting the intensity of excitation light, and a first battery, which are electrically connected with the lamp panel.

[0007] Further, the lamp panel comprises a circuit board, lamp beads arranged on the front surface of the circuit board, and a heat sink located on the back surface.

[0008] Further, the test strip uses aggregation-induced luminescence nanoparticles (AIENP) with a wavelength of 312 nm for excitation and a wavelength of 465 nm for emission as a detection probe to label a monoclonal antibody, and the fluorescence signal thereof can be recognized by the mobile terminal.

[0009] Further, the carrier and the carrier cover are made of black polylactic acid to reduce the interference of background fluorescence signals.

[0010] Further, a heat dissipation assembly is further included to maintain the temperature stability inside the dark box, thereby ensuring the stability of the excitation light intensity; the heat dissipation assembly comprises a heat dissipation fan, a fan switch electrically connected with the heat dissipation fan, and a second battery.

[0011] Further, the heat dissipation fan is arranged on the top of the carrier.

[0012] Further, a filter is arranged at the light transmission hole.

[0013] The second aspect of the application provides the application of any of the above mobile terminal based fluorescent immunoassay chromatographic instant detection system in immune detection analysis, and the application comprises the following steps: (1) Turn on the light source switch and the cooling fan switch in advance to ensure stable light intensity; (2) Take out the fluorescent immunoassay chromatographic test strip and place it horizontally on the operation table; (3) Dilute the standard sample with the sample buffer to a series of gradient concentrations, and drop them on the sample pad of the test strip respectively for chromatographic reaction; (4) Insert the test strip after reaction into the test strip loading channel along the horizontal direction of the insertion port, and take a photo with the camera of the intelligent terminal; (5) Open the fluorescence intensity analysis application program, test the T line and C line fluorescence intensity of each concentration standard sample, output the T / C value, and fit the standard curve of the concentration and the T / C value; (6) After the sample and the serum standard sample are processed in the same way, the mobile terminal based fluorescent immunoassay chromatographic instant detection system is used to test the T / C value, and the specific concentration value is calculated by substituting the standard curve, so that quantitative analysis is realized.

[0014] Compared with the prior art, the application has the following beneficial effects: The mobile terminal based fluorescent immunoassay chromatographic instant detection system provided by the application has a newly added light blocking groove cover on the carrier, which is arranged on both sides of the limiting groove and can accurately shield the sample pad, the combination pad and the absorbent paper of the test strip, and only expose the nitrocellulose membrane as the core reaction area. The addition of the light blocking groove cover effectively eliminates the fluorescence interference and focusing deviation problems of the non-reaction area, greatly improves the stability and accuracy of the fluorescence analysis, and solves the technical problems that the sample pad, the combination pad and the absorbent paper of the existing fluorescent immunoassay chromatographic test strip are all non-fluorescent materials, which will produce interfering fluorescence under ultraviolet excitation light, and the image processing accuracy is affected. At the same time, it also solves the technical problem that the flatness difference exists in the connection parts of the sample pad, the combination pad, the nitrocellulose membrane (reaction area) and the absorbent paper, which causes the focusing of the photo to be not on the same level and affects the fluorescence analysis effect.

[0015] The mobile terminal-based fluorescent immunochromatographic instant detection system of the application eliminates the dependence of traditional fluorescent immunochromatographic test strips on dry-type immunofluorescence analyzers, and truly realizes on-site instant, rapid and intelligent analysis and detection results; the application solves the problems of the traditional dry-type immunofluorescence analyzer, such as being heavy and not portable, and truly realizes the exquisite and portable fluorescent immunochromatographic test strip analysis system based on 3D printing technology and a smart phone, greatly reduces the cost of the instrument, and solves the problem of high price of the traditional dry-type immunofluorescence analyzer; compared with the traditional immunological detection method, the detection time is greatly shortened, and the mobile terminal-based fluorescent immunochromatographic instant detection system is suitable for remote areas, primary laboratories, families and on-site detection. BRIEF DESCRIPTION OF DRAWINGS

[0016] In order to more clearly illustrate the scheme in the application or the prior art, the drawings needed to be used in the following embodiment or prior art description will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0017] Figure 1 It is a structural exploded view of the mobile terminal-based fluorescent immunochromatographic instant detection system of the application. Figure 2 It is a structural view of the mobile terminal-based fluorescent immunochromatographic instant detection system of the application. Figure 3 It is an optical detection path diagram of the mobile terminal-based fluorescent immunochromatographic instant detection system of the application. Figure 4 It is the influence of the mobile terminal-based fluorescent immunochromatographic instant detection system of the application on the detection results with or without a light blocking groove cover. Figure 5 A is a structural view of the fluorescent test strip in the application, Figure 5 B is a negative detection result schematic diagram of the fluorescent test strip, Figure 5 C is a positive detection result schematic diagram of the fluorescent test strip. Figure 6 A is an AIENP transmission electron microscope (TEM) diagram, Figure 6 B is an excitation and emission spectrum scanning diagram of AIENP, Figure 6 C is a SEM diagram of T line of a negative fluorescent test strip, Figure 6 D is a SEM diagram of T line of a positive fluorescent test strip. Figure 7 A is a mobile phone page screenshot of the mobile terminal-based fluorescent immunochromatographic instant detection system of the application for detecting a series of gradient concentrations (0.04-2 ng / mL) of SARS-CoV-2 N protein standard, Figure 7B is the signal trend chart of the detection of the novel coronavirus N protein by the series gradient concentration (0.04-2 ng / mL) of the novel coronavirus N protein standard; Figure 7 C is the standard curve chart of the detection of the novel coronavirus N protein by the mobile terminal-based fluorescent immunochromatography instant detection system. DETAILED DESCRIPTION

[0018] In order to make the personnel in the technical field better understand the application scheme, the technical solutions in the embodiments of the application will be clearly and completely described below in combination with the drawings in the embodiments of the application. Obviously, the described embodiments are only a part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by the person skilled in the art without creative labor should belong to the protection scope of the application.

[0019] In order to make the personnel in the technical field better understand the application scheme, the technical solutions in the embodiments of the application will be clearly and completely described below in combination with the drawings in the embodiments of the application.

[0020] Example 1: Mobile terminal-based fluorescent immunochromatography instant detection system As shown in FIGS. Figure 1 , 2 and 3, the mobile terminal-based fluorescent immunochromatography instant detection system comprises: The carrier 1 is internally provided with an excitation light source assembly and a test strip loading channel. The test strip loading channel comprises a plug-in port 14, a limiting groove and a light blocking groove cover 12, 13. The plug-in port 14 is arranged at the side of the carrier and located at one side of the limiting groove. The limiting groove is used for loading the test strip 15 after the test strip is inserted into the plug-in port 14. The light blocking groove cover 12, 13 is arranged at both sides of the limiting groove, respectively, to shield the sample pad 22, the conjugate pad 23 and the absorbent paper 27 of the test strip, and only expose the nitrocellulose membrane 24 of the test strip. The carrier cover 16 is arranged above the carrier 1 to form a dark box. The carrier cover 16 is provided with a light transmission hole, which is on the same optical path as the excitation light of the excitation light source assembly and the nitrocellulose membrane 24. The excitation light is inclined at an angle of 45 degrees to the test strip nitrocellulose membrane 24 area (i.e. the excitation light beam is inclined at an angle of 45 degrees to the nitrocellulose membrane 24). The smart phone camera 20, the optical filter 17 and the nitrocellulose membrane 24 are on the same reflection optical path.

[0021] The mobile terminal, such as a smart phone 19 or a tablet, is loaded on the carrier cover 16. The camera module of the mobile terminal corresponds to the light transmission hole. The mobile terminal is internally provided with a fluorescence intensity analysis application program. The ratio of the fluorescence signals of the T line and the C line is converted into a T / C value.

[0022] The excitation light source assembly includes a lamp board capable of emitting an excitation light source, an excitation light source switch 7, an adjustable resistor 6 for adjusting the intensity of the excitation light, a first battery 4 (12V), and a DC female connector 5, all electrically connected to the lamp board. The lamp board includes a circuit board, LED beads 2 on the front of the circuit board, and a heat sink 3 on the back. The DC female connector 5 serves as the charging port for the first battery 4, and the two are integrated. The positive terminal of the first battery 4 is connected to the LED beads 2, the LED beads 2 are connected to the adjustable resistor 6, the adjustable resistor 6 is connected to the excitation light source switch 7, and the excitation light source switch 7 is connected to the negative terminal of the first battery 4, forming a closed-loop circuit.

[0023] The mobile terminal-based real-time fluorescence immunochromatographic assay system also includes a heat dissipation component to maintain a stable internal temperature of the dark chamber, thereby ensuring stable excitation light intensity. The heat dissipation component includes a cooling fan 8 located on top of the carrier 1, a fan switch 11 electrically connected to the cooling fan 8, and a second battery 9 (3.7V). It also includes a Type-C charging port 10 connected to the second battery 9. The Type-C charging port 10 serves as the charging port for the second battery 9, and the two are integrated. The positive terminal of the second battery 9 is connected to the cooling fan 8, the cooling fan 8 is connected to the fan switch 11, and the fan switch 11 is connected to the negative terminal of the second battery 9, forming a closed-loop circuit.

[0024] The carrier 1 and the cover 16 are made of black polylactic acid, making the entire interior of the dark box black and reducing interference from background fluorescence signals. The cover 16 can be fitted onto the carrier 1. The smartphone 19 is mounted on the cover 16, which is shaped like a phone case. The size of its camera 20 is completely embedded in the camera frame 18 on the cover 16. The light-transmitting hole is opened on the camera frame 18, and a filter 17 is provided at the light-transmitting hole.

[0025] The test strip 15 consists of a PVC base plate 21, a sample pad 22, a conjugate pad 23, a nitrocellulose membrane 24, and absorbent paper 27. The conjugate pad 23 is coated with an AIENP-labeled monoclonal antibody. The monoclonal antibody is labeled with AIENP, which is excited by 312 nm light and emits light at 465 nm, and its fluorescence signal can be recognized by a mobile terminal. The detection T line 25 and control C line 26 on the nitrocellulose membrane 24 are coated with monoclonal antibody 2 and goat anti-mouse IgG, respectively.

[0026] The test strip 15 is horizontally inserted into the test strip loading channel in the carrier 1 along the insertion port 14. The light-blocking groove covers 12 and 13 are respectively located on both sides of the limiting groove to block the sample pad 22, conjugate pad 23 and absorbent paper 27 of the test strip, leaving only the nitrocellulose membrane 24 of the test strip exposed to be irradiated and photographed by laser for analysis and detection of the fluorescence intensity of the T line 25 and the quality control C line 26.

[0027] The smartphone 19 is a smart mobile device with macro photography capabilities. It has a display screen and runs on Android 10.0 or later, enabling mobile sharing of analysis results. Based on the built-in light intensity analysis application of the smartphone 19, data processing and analysis are performed on the captured images to calculate the grayscale area of ​​the detection T-line and quality control C-line and save the T / C value.

[0028] Furthermore, the test strips were tested using a mobile terminal-based real-time fluorescence immunochromatography system with and without a light-blocking cover. The results showed that the coefficient of variation of the T / C value was lower when the light-blocking cover was present. Figure 4 This indicates that adding a light-blocking groove cover to cover the non-reactive area of ​​the test strip in this invention helps to reduce interference with the detection signal. When the same test strip is equipped with a light-blocking groove cover, the coefficient of variation of the test results is significantly reduced, which greatly improves the stability and accuracy of the fluorescence immunochromatographic detection results. The design of the light-blocking groove cover has important improvement significance.

[0029] Addressing the technical challenge of interference fluorescence generated by non-fluorescent materials in the non-reactive areas (sample pad, conjugate pad, absorbent paper) of existing test strips under UV excitation, this invention adds light-blocking covers to both sides of the carrier limiting groove. This precisely shields the non-reactive areas, exposing only the core reactive area (nitrocellulose membrane), thus eliminating the impact of interfering fluorescence on image processing at its source. Furthermore, it resolves the issue of inconsistent focusing caused by unevenness at the joints of the test strip components. The light-blocking covers ensure that imaging is focused only on the flat nitrocellulose membrane area, guaranteeing focusing stability and improving the imaging quality of fluorescence analysis.

[0030] Example 2: A mobile terminal-based fluorescence immunochromatographic point detection system for detecting the N protein of the novel coronavirus. Synthesis and labeling of AIENP: 10 mg TCPE and 2.5 mg ZrCl4 were dissolved separately in DMF, sonicated until completely dissolved, and then 2 mL of acetic acid was added and stirred to mix. The mixture was then stirred at 85 °C and 500 rpm for 30 min, followed by the addition of 120 μL of deionized water and a further 2 h of reaction. After the reaction, the solution was cooled to room temperature, centrifuged at 10,000 rpm for 15 min to collect the product, and washed with anhydrous ethanol. Finally, the product was resuspended in 4 mL of ethanol and stored at 4 °C. 10 μL of AIENP stock solution was diluted with 990 μL of deionized water, and then labeled with an appropriate amount of mAb1 for 30 min. 100 μL of 10% BSA was added for blocking for 30 min, and the AIENP-mAb1 probe was collected by centrifugation at 10,000 rpm for 15 min and resuspended in 150 μL of reconstitution solution (10% sucrose, 5% trehalose, 1% BSA, 0.5% Tween, 50 mM Tris-HCl, pH 8.0).

[0031] Test strip preparation: Figure 5 A is a schematic diagram of the fluorescent test strip used to detect the N protein of the novel coronavirus in this implementation case. The test strip 15 consists of a PVC base plate 21, a sample pad 22, a conjugate pad 23, a nitrocellulose membrane 24, and absorbent paper 27. The conjugate pad 23 is sprayed with AIENP-labeled monoclonal antibody with a particle size of 250 nm. The detection T line 25 and the control C line 26 on the nitrocellulose membrane 24 are coated with monoclonal antibody 2 and goat anti-mouse IgG, respectively.

[0032] The specific preparation process of the test strip is as follows: Spray AIENP-mAb1 at 5 μL / cm onto the conjugate pad and dry at 56℃ for later use; Coat the T line with 1 mg / mL monoclonal antibody 2 at 1 μL / cm and the C line with 0.5 mg / mL goat anti-mouse IgG, and dry at 56℃ for later use; Sequentially attach the nitrocellulose membrane, sample pad, conjugate pad and absorbent paper to the PVC base plate and cut into 4 mm wide test strips.

[0033] Figure 5 B and Figure 5 C represents the negative and positive results of the fluorescent test strip for detecting the N protein of the novel coronavirus. When the sample contains the N protein, it will be detected by the AIENP-labeled monoclonal antibody on the binding pad 23 and captured by the monoclonal antibody 2 coated on the detection T line 25, showing the T line. Excess AIENP-labeled monoclonal antibody is captured by the goat anti-mouse IgG coated on the quality control C line 26, showing the C line.

[0034] In addition, the AIENP-labeled monoclonal antibodies were characterized by transmission electron microscopy, such as... Figure 6As shown in Figure A, the labeled AIENP particles have a diameter of approximately 250 nm. To maximize the luminescence efficiency of AIENP, an optimal wavelength excitation source and filter were fitted to the mobile terminal-based fluorescence immunochromatographic point detection system. Excitation and emission spectra of AIENP were scanned, and the results showed that the optimal wavelength for AIENP excitation light was 312 nm, and the optimal wavelength for emission light was 465 nm. Figure 6 B), therefore, the excitation light source used in this invention patent is a wavelength of 312 nm, and the filter is a wavelength of 465 nm; furthermore, the T-line 25 region of the nitrocellulose membrane 24 after detecting negative samples was characterized by scanning electron microscopy, showing that there was no AIENP binding in the T-line region (B). Figure 6 C), and AIENP binding was found in the T-line 25 region of the nitrocellulose membrane 24 after the positive sample was detected (C). Figure 6 D).

[0035] Aggregation-induced emission (AIE) fluorescent molecules exhibit a significant luminescence enhancement effect in the aggregated state, primarily due to their unique mechanism of restricted intramolecular motion. Compared to conventional fluorescent microspheres, AIE molecules can effectively utilize the aggregation process, fundamentally overcoming the problem of fluorescence quenching that easily occurs in the aggregated state of traditional fluorescent materials, thus greatly improving fluorescence efficiency. Combining AIE materials with immunochromatography technology can fully leverage their advantages such as high quantum yield, excellent photostability, and strong resistance to photobleaching. Based on this, AIE immunochromatographic test strips are prepared, further transforming the excellent properties of the materials into high sensitivity, high accuracy, and high reliability at the product level. However, traditional AIE fluorescent microspheres involve encapsulating AIE molecules within polystyrene microspheres and then labeling them with antibodies via chemical bonds. This process is not only cumbersome but also time-consuming and labor-intensive. Examples include the AIE fluorescent microspheres used in the fluorescent immunochromatographic test strips disclosed in Chinese invention patent application CN119438577A, and the AIE fluorescent microspheres used in the ELISA based on aggregation-induced emission ratio fluorescent probes disclosed in Chinese invention patent application CN114778825A. In contrast, this invention uses a specific method to prepare and synthesize AIE nanoparticle probes. The synthesized AIENP does not require polystyrene microspheres as a fluorescent molecular carrier, and monoclonal antibody labeling can be completed through electrostatic adsorption. This significantly reduces the time required for monoclonal antibody labeling from several hours to tens of minutes, resulting in higher labeling efficiency.

[0036] The testing steps are as follows: (1) Pre-activate the light source switch and cooling fan switch to ensure stable excitation light intensity; (2) Take out the fluorescence immunochromatographic test strip and place it horizontally on the operating table; (3) Dilute the standard to a series of gradient concentrations using sample buffer, and add them dropwise onto the sample pad of the test strip for chromatographic reaction; Specifically: The novel coronavirus N protein was sequentially diluted to a gradient concentration using sample diluent (2 ng / mL, 1 ng / mL, 0.5 ng / mL, 0.25 ng / mL, 0.125 ng / mL, 0.063 ng / mL, 0.031 ng / mL, 0.016 ng / mL, 0.008 ng / mL, and 0.004 ng / mL); each concentration was tested in triplicate. 80 μL of the standard was added to the sample pad and the test was timed for 15 min. (4) Insert the reacted test strip into the carrier horizontally along the test insertion port, and turn on the smartphone camera; (5) Open the fluorescence intensity analysis application, test the fluorescence intensity of the T line and C line of each concentration standard, output the T / C value, and fit the relevant standard curve of standard concentration and T / C value; Specifically: Open the smartphone results analysis application app, click the Open button to take out the photo of the test strip result, click the Auto button to display the fluorescence intensity scan peaks of the detection T line and the control C line and the fluorescence intensity ratio of the detection T line and the control C line (T / C), click Save to export and save the data; with the N protein concentration as the x-axis and the T / C value as the y-axis, fit a linear equation to achieve quantitative analysis.

[0037] Screenshots of the mobile terminal-based fluorescence immunochromatographic real-time detection system's detection results for a range of concentrations (0.004-2 ng / mL) of novel coronavirus N protein standards are shown below. Figure 7 Plotting concentration on the x-axis and T / C value on the y-axis, the fitted equation is Y = 1.0772 - 1.13977 * 0.07454. x (R) 2 =0.999).

[0038] (6) After the sample and serum standard are processed in the same way, the T / C value is tested using a mobile terminal-based fluorescence immunochromatographic instant detection system. The specific concentration value is calculated by substituting it into the standard curve to achieve quantitative analysis. Specifically, the actual sample detection process is as follows: take 300 μL of sample diluent to elute the nasopharyngeal swab, take 80 μL of nasopharyngeal swab sample eluent and drop it onto the sample pad, time for 15 min, and then use a mobile terminal-based real-time fluorescence immunochromatography detection system to analyze the T / C value. Substitute it into the fitting equation to calculate the N protein concentration in the sample.

[0039] In the description of this invention, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," and "circumferential" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0040] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0041] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection, an electrical connection, or a connection that allows communication between them; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0042] Obviously, the embodiments described above are only some embodiments of this application, not all embodiments. The accompanying drawings show preferred embodiments of this application, but do not limit the patent scope of this application. This application can be implemented in many different forms; rather, the purpose of providing these embodiments is to provide a more thorough and comprehensive understanding of the disclosure of this application. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing specific embodiments, or make equivalent substitutions for some of the technical features. Any equivalent structures made using the content of this application's specification and drawings, directly or indirectly applied to other related technical fields, are similarly within the scope of patent protection of this application.

Claims

1. A mobile terminal-based real-time fluorescence immunochromatographic detection system, characterized in that, It includes: The carrier contains an excitation light source assembly and a test strip loading channel. The test strip loading channel includes an insertion port, a limiting groove, and a light-blocking groove cover. The insertion port is located on the side of the carrier and on one side of the limiting groove. The limiting groove is used to load the test strip after it is inserted into the insertion port. The light-blocking groove cover is located on both sides of the limiting groove to cover the sample pad, conjugate pad, and absorbent paper of the test strip, exposing only the nitrocellulose membrane of the test strip. A cover is placed over the carrier to form a dark box; the cover has a light-transmitting hole, which is on the same optical path as the excitation light of the nitrocellulose membrane and the excitation light source assembly; A mobile terminal is mounted on the cover, and the camera module of the mobile terminal corresponds to the light-transmitting hole to take pictures of the exposed nitrocellulose membrane area; the mobile terminal is equipped with a fluorescence intensity analysis application to convert the received light source signal into a T / C value.

2. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 1, characterized in that, The excitation light source assembly includes a lamp board capable of emitting an excitation light source, an excitation light source switch electrically connected to the lamp board, an adjustable resistor for adjusting the intensity of the excitation light, and a first storage battery.

3. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 2, characterized in that, The lamp panel includes a circuit board, LED beads disposed on the front side of the circuit board, and a heat sink located on the back side.

4. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 1, characterized in that, The test strip uses aggregation-induced emission nanoparticles excited by 312 nm wavelength light and emitted by 465 nm wavelength light as detection probes to label monoclonal antibodies, and its fluorescence signal can be recognized by mobile terminals.

5. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 1, characterized in that, The carrier and cap are formed of black polylactic acid.

6. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 1, characterized in that, It also includes a heat dissipation component to maintain a stable internal temperature of the dark box; the heat dissipation component includes a cooling fan, a fan switch electrically connected to the cooling fan, and a second battery.

7. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 6, characterized in that, The cooling fan is located on the top of the carrier.

8. The mobile terminal-based real-time fluorescence immunochromatographic detection system according to claim 1, characterized in that, A filter is provided at the light-transmitting hole.

9. The application of the mobile terminal-based real-time fluorescence immunochromatographic detection system according to any one of claims 1 to 8 in immunoassay analysis, characterized in that, The application includes the following steps: (1) Pre-activate the light source switch and cooling fan switch to ensure stable excitation light intensity; (2) Take out the fluorescence immunochromatographic test strip and place it horizontally on the operating table; (3) Dilute the standard to a series of gradient concentrations using sample buffer, and add them dropwise onto the sample pad of the test strip for chromatographic reaction; (4) Insert the reacted test strip into the test strip loading channel in the horizontal direction along the insertion port, and turn on the camera of the smart terminal to take a picture; (5) Open the fluorescence intensity analysis application, test the fluorescence intensity of the T line and C line of each concentration standard, output the T / C value, and fit the relevant standard curve of standard concentration and T / C value; (6) After the sample and serum standard are processed in the same way, the T / C value is tested using a mobile terminal-based fluorescence immunochromatographic instant detection system. The specific concentration value is calculated by substituting it into the standard curve to achieve quantitative analysis.

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