Luminescence immunoassay method, device and electronic device based on portable device

Abstract

The present invention relates to the field of chemiluminescence immunoassay, and discloses a luminescence immunoassay method based on a portable device, including: placing an antibody-polymer microsphere composite material, a luminescence reaction reagent, and a buffer cleaning solution in a predetermined cavity of an original microfluidic paper chip to obtain a target microfluidic paper chip, adding a sample to be tested to a card reader device to obtain a card reader device to be tested, inserting the target microfluidic paper chip into the card reader device to be tested to obtain a paper chip-card reader device, pressing the luminescence reaction reagent and the buffer cleaning solution to the antibody-polymer microsphere composite material to obtain a microsphere antibody complex, applying an electric field to the microsphere antibody complex, and querying the antigen content of the sample to be tested according to the luminescence amount. The present invention also provides a luminescence immunoassay device, an electronic device, and a computer-readable storage medium based on a portable device. The present invention can solve the problems of high price, complex operation, difficult maintenance, and difficult management of consumables existing in the mainstream products of "electrochemical luminescence immunoassay method".

Description

Technical Field

[0001] The present invention relates to the field of chemiluminescence immunoassay, and particularly to a luminescence immunoassay method, device, electronic device and computer-readable storage medium based on a portable device. Background Art

[0002] With the increasing harm of viruses to humans, chemiluminescence immunoassay technology has developed rapidly. There are three common directions for chemiluminescence immunoassay technology, namely: "direct chemiluminescence method", "enzymatic chemiluminescence method" and "electrochemiluminescence immunoassay method". Among them, the "electrochemiluminescence immunoassay method" has the characteristics of high sensitivity, strong specificity, fast detection speed, wide linear dynamic range, etc. Therefore, the "electrochemiluminescence immunoassay method" has begun to be widely used in medical, food safety detection and other aspects.

[0003] However, the mainstream products of the "electrochemiluminescence immunoassay method" are all large-scale, high-throughput, fully automated devices, which have problems such as high price, complex operation, difficult maintenance and difficult management of consumables. Summary of the Invention

[0004] The present invention provides a luminescence immunoassay method, device and computer-readable storage medium based on a portable device, and its main purpose is to solve the problems of high price, complex operation, difficult maintenance and difficult management of consumables existing in the mainstream products of the "electrochemiluminescence immunoassay method".

[0005] To achieve the above object, a luminescence immunoassay method based on a portable device provided by the present invention includes:

[0006] Obtain an antibody-polymer microsphere composite material, a luminescence reaction reagent and a buffer cleaning solution, and place the antibody-polymer microsphere composite material, the luminescence reaction reagent and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively to obtain a target microfluidic paper chip;

[0007] Obtain a sample to be tested, add the sample to be tested into a pre-constructed card reader device to obtain a to-be-read card reader device, and insert the target microfluidic paper chip into the to-be-read card reader device to obtain a paper chip-card reader device;

[0008] Press the luminescence reaction reagent and the buffer cleaning solution to the cavity where the antibody-polymer microsphere composite material is located for reaction to obtain a microsphere antibody complex;

[0009] Apply an electric field to the microsphere antibody complex by using the paper chip-card reader device, detect the luminescence amount of the microsphere antibody complex, and query the antigen content of the sample to be tested according to the luminescence amount.

[0010] Optionally, the process for obtaining the antibody-polymer microbead composite material includes:

[0011] Obtain polymer microbeads, antibody I, avidin, and biotin;

[0012] Bind the avidin and biotin to the polymer microbeads to obtain polymer microbeads;

[0013] Utilize the binding sites of the polymer microbeads to link antibody I to obtain the antibody-polymer microbead composite material.

[0014] Optionally, the process for obtaining the luminescence reaction reagent includes:

[0015] Obtain a luminescent molecule and antibody II;

[0016] Link the luminescent molecule and antibody II to obtain the antibody-luminescence composite molecule;

[0017] Obtain a reaction reagent, and mix the antibody-luminescence composite molecule and the reaction reagent to obtain the luminescence reaction reagent.

[0018] Optionally, the process for constructing the original microfluidic paper chip includes:

[0019] Obtain a microfluidic paper chip cassette;

[0020] According to the process of electrochemiluminescence immunoassay, design the position layout of a preset reagent storage cavity, a buffer cleaning solution storage cavity, a paper chip area, a microsphere antibody composite cavity, an electrode area, a waste liquid collection cavity, and a sampling point on the microfluidic paper chip cassette;

[0021] Add a pre-constructed paper chip to the paper chip area, and add a pre-constructed water-absorbing polymer to the waste liquid collection cavity to obtain the original microfluidic paper chip.

[0022] Optionally, placing the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning solution into the predetermined cavities of the pre-constructed original microfluidic paper chip respectively to obtain the target microfluidic paper chip includes:

[0023] Fix the antibody-polymer microbead composite material on a pre-constructed polymer glass slide to obtain a bead-loaded polymer glass slide;

[0024] Place the luminescence reaction reagent and the buffer cleaning solution into a pre-constructed flexible film respectively to obtain a buffer cleaning film bag and a predetermined number of luminescence reagent film bags;

[0025] Fix the bead-carrying polymer glass slide in the microsphere antibody composite cavity of the pre-constructed original microfluidic paper chip, and place the luminescent reagent thin film bag and the buffer cleaning thin film bag in the reagent storage cavity and the buffer cleaning solution storage cavity of the original microfluidic paper chip respectively to obtain the target microfluidic paper chip.

[0026] Optionally, the step of pressing the luminescent reaction reagent and the buffer cleaning solution into the cavity where the antibody-polymer microsphere composite material is located for reaction to obtain the microsphere antibody complex includes:

[0027] Receive a test instruction, and according to the test instruction, start the piezoelectric ceramic in the paper chip card reader device to obtain a serialized start of the piezoelectric ceramic;

[0028] Use the serialized start of the piezoelectric ceramic to press out part of the reagent in the buffer cleaning thin film bag to obtain the first cleaning solution to be seeped;

[0029] Utilize the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the first cleaning solution to be seeped to the microsphere antibody composite cavity of the target microfluidic paper chip to clean the microsphere antibody composite cavity and obtain the antigen-antibody composite molecule to be detected;

[0030] Use the serialized start of the piezoelectric ceramic to press out the reagent in the luminescent reagent thin film bag to obtain the luminescent reagent to be seeped;

[0031] Utilize the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the luminescent reagent to be seeped to the microsphere antibody composite cavity to react with the antigen-antibody composite molecule to be detected to obtain the microsphere antibody complex to be cleaned;

[0032] Use the serialized start of the piezoelectric ceramic to press out the reagent in the buffer cleaning thin film bag to obtain the second cleaning solution to be seeped;

[0033] Utilize the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the second cleaning solution to be seeped to the microsphere antibody composite cavity to clean the microsphere antibody composite cavity and obtain the microsphere antibody complex.

[0034] Optionally, the step of querying the antigen content of the sample to be detected according to the luminescence amount includes:

[0035] Obtain a pre-constructed luminescence amount - antigen content comparison table;

[0036] According to the luminescence amount, query the antigen content of the sample to be detected in the luminescence amount - antigen content comparison table.

[0037] To solve the above problems, the present invention further provides a luminescent immunoassay device based on a portable device, the device comprising:

[0038] A target microfluidic paper chip manufacturing module, configured to obtain an antibody-polymer microsphere composite material, a luminescence reaction reagent, and a buffer cleaning solution, and place the antibody-polymer microsphere composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively, to obtain a target microfluidic paper chip;

[0039] A paper chip-reader device acquisition module, configured to obtain a sample to be tested, add the sample to be tested to a pre-constructed reader device to obtain a to-be-read reader device, and insert the target microfluidic paper chip into the to-be-read reader device to obtain a paper chip-reader device;

[0040] A microsphere antibody complex acquisition module, configured to press the luminescence reaction reagent and the buffer cleaning solution to the cavity where the antibody-polymer microsphere composite material is located for reaction to obtain a microsphere antibody complex;

[0041] An optoelectronic detection module, configured to apply an electric field to the microsphere antibody complex by using the paper chip-reader device, detect the luminescence amount of the microsphere antibody complex, and query the antigen content of the sample to be tested according to the luminescence amount.

[0042] To solve the above problems, the present invention further provides an electronic device, the electronic device comprising:

[0043] A memory, storing at least one instruction; and

[0044] A processor, executing the instruction stored in the memory to implement the above-mentioned luminescent immunoassay method based on a portable device.

[0045] To solve the above problems, the present invention further provides a computer-readable storage medium, in which at least one instruction is stored, and the at least one instruction is executed by a processor in an electronic device to implement the above-mentioned luminescent immunoassay method based on a portable device.

[0046] Compared with the mainstream products of the "electrochemical immunoassay" described in the background art, which have problems such as high price, complex operation, difficult maintenance, and difficult management of consumables, the embodiments of the present invention obtain the target microfluidic paper chip by placing the antibody-polymer microsphere composite material, the chemiluminescence reaction reagent, and the buffer cleaning solution in the predetermined cavities of the original microfluidic paper chip respectively, thus having two important elements for detecting the antigen content in the sample to be detected, namely: the antibody-polymer microsphere composite material and the chemiluminescence reaction reagent. Then, the sample to be detected is obtained by using the card reader device, and the target microfluidic paper chip is inserted into the card reader device to obtain the paper chip-card reader device. Various reagents are pressed into the cavity where the antibody-polymer microsphere composite material is located through the paper chip-card reader device for reaction to obtain the microsphere antibody complex. Finally, an electric field is applied to the microsphere antibody complex by using the paper chip-card reader device, so that the microsphere antibody complex cavity generates a chemiluminescence effect, and the antigen content in the sample to be detected is judged according to the chemiluminescence amount detected by the photodetector, realizing the process of chemiluminescence immunoassay. Therefore, the chemiluminescence immunoassay method, device, electronic device, and computer-readable storage medium based on a portable device proposed by the present invention can solve the problems of high price, complex operation, difficult maintenance, and difficult management of consumables existing in the mainstream products of the "electrochemical immunoassay". BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 FIG. is a schematic flowchart of a chemiluminescence immunoassay method based on a portable device provided by an embodiment of the present invention;

[0048] Figure 2 is Figure 1 a detailed implementation flowchart of one of the steps;

[0049] Figure 3 is Figure 1 a detailed implementation flowchart of another step;

[0050] Figure 4 FIG. is a functional module diagram of a chemiluminescence immunoassay device based on a portable device provided by an embodiment of the present invention;

[0051] Figure 5 FIG. is a structural schematic diagram of an electronic device for implementing the chemiluminescence immunoassay method based on a portable device provided by an embodiment of the present invention.

[0052] The realization, functional characteristics, and advantages of the object of the present invention will be further described with reference to the embodiments and the accompanying drawings. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0053] It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0054] An embodiment of the present application provides a chemiluminescent immunoassay method based on a portable device. The execution subject of the chemiluminescent immunoassay method based on the portable device includes, but is not limited to, at least one of electronic devices such as a server, a terminal, etc. that can be configured to execute the method provided in the embodiment of the present application. In other words, the chemiluminescent immunoassay method based on the portable device can be executed by software or hardware installed on a terminal device or a server device, and the software can be a blockchain platform. The server includes, but is not limited to: a single server, a server cluster, a cloud server, or a cloud server cluster, etc.

[0055] Referring to Figure 1 As shown, it is a schematic flow chart of the chemiluminescent immunoassay method based on a portable device provided by an embodiment of the present invention. In this embodiment, the chemiluminescent immunoassay method based on the portable device includes:

[0056] S1. Obtain an antibody-polymer microsphere composite material, a chemiluminescent reaction reagent, and a buffer cleaning solution, and place the antibody-polymer microsphere composite material, the chemiluminescent reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively to obtain a target microfluidic paper chip.

[0057] It can be understood that the antibody-polymer microsphere composite material refers to a composite material obtained by connecting polymer microspheres, antibody 1, avidin, and biotin. The basic structure of the polymer microspheres is uniformly dispersed microspheres prepared from polymer Fe3O4, and the surface can be connected with antibody 1 through chemical bonds to enable it to have the ability to carry out immune reactions. In addition, dendritic branching can be carried out on the polymer microspheres through biotin and avidin, and then multiple luminescent functional groups can be connected, thereby increasing the density of the luminescent groups and simultaneously reducing the detection limit.

[0058] It can be explained that the chemiluminescent reaction reagent refers to a reagent composed of an antibody-luminescent composite molecule and a reaction reagent, where the reaction reagent refers to a solvent that assists in carrying out chemiluminescent immunoassay reactions, including: a luminescent substrate, an alkaline solution, and other various reagents required for electrochemiluminescent immunoassay reactions. The antibody-luminescent composite molecule refers to a composite molecule obtained by combining antibody 2 with a luminescent molecule. The antibody-luminescent composite molecule can carry out chemiluminescent immunoassay reactions using an antigen, and then display the content of the antigen.

[0059] It should be understood that the original microfluidic paper chip refers to an original paper chip box obtained by carrying out functional zoning for chemiluminescent immunoassay reactions on a pre-constructed microfluidic paper chip box body, and adding a paper chip and a water-absorbing polymer to the zoned paper chip area and the waste liquid collection cavity respectively. The functional zoning of the original microfluidic paper chip can be a reagent storage cavity, a buffer cleaning solution storage cavity, a paper chip area, a microsphere antibody composite cavity, an electrode area, a waste liquid collection cavity, and a sampling point.

[0060] Interpretive, the microfluidic paper chip cartridge can be formed by digging grooves in materials such as plastic and glass and covering them with a cover plate. The paper chip is essentially a chromatographic sheet with capillary seepage function composed of materials such as filter paper, non-woven fabric, nitrocellulose, and electrospun fibers, mainly using capillary seepage to make various reaction reagents flow. The water-absorbing polymer can also be replaced by any material that can continuously absorb liquid, such as sponge and gel. Its main function is to make various reaction reagents leave the paper chip and promote capillary seepage.

[0061] In the embodiment of the present invention, the process for obtaining the antibody-polymer microsphere composite includes:

[0062] Obtaining polymer microspheres, antibody I, avidin, and biotin;

[0063] Binding the avidin and biotin to the polymer microspheres to obtain polymer microspheres;

[0064] Using the binding sites of the polymer microspheres to connect antibody I to obtain the antibody-polymer microsphere composite.

[0065] Specifically, refer to Figure 2 As described, the process for obtaining the luminescent reaction reagent includes:

[0066] S11. Obtaining a luminescent molecule and antibody II;

[0067] S12. Connecting the luminescent molecule and antibody II to obtain the antibody-luminescent composite molecule;

[0068] S13. Obtaining a reaction reagent, mixing the antibody-luminescent composite molecule and the reaction reagent to obtain the luminescent reaction reagent.

[0069] Optionally, a luminescent immunoassay can be completed using the sandwich immunoassay method. The sandwich immunoassay method refers to using antibody I in the antibody-polymer microsphere composite to capture the antigen, and then using antibody II in the antibody-luminescent composite molecule to react with the captured antigen, thereby connecting the antibody-polymer microsphere composite, the antibody-luminescent composite molecule, and the antigen together to facilitate subsequent steps.

[0070] Specifically, as Figure 3 shown, the construction process of the original microfluidic paper chip includes:

[0071] S14. Obtaining a microfluidic paper chip cartridge;

[0072] S15. According to the process of electrochemiluminescence immunoassay, on the microfluidic paper chip cassette body, design the position layout of preset reagent storage cavities, buffer cleaning solution storage cavities, paper chip area, microsphere antibody composite cavity, electrode area, waste liquid collection cavity and sampling point;

[0073] S16. Add the pre-constructed paper chip to the paper chip area, and add the pre-constructed water-absorbing polymer to the waste liquid collection cavity to obtain the original microfluidic paper chip.

[0074] It is understandable that the electrode area is located below the microsphere antibody composite cavity, which is convenient for applying an electric field to the microsphere antibody composite cavity subsequently.

[0075] Specifically, placing the antibody-polymer microsphere composite material, luminescence reaction reagent and buffer cleaning solution in the preset cavities of the pre-constructed original microfluidic paper chip respectively to obtain the target microfluidic paper chip includes:

[0076] Fix the antibody-polymer microsphere composite material on the pre-constructed polymer glass slide to obtain a bead-carrying polymer glass slide;

[0077] Place the luminescence reaction reagent and the buffer cleaning solution in the pre-developed flexible film respectively to obtain a buffer cleaning film bag and a predetermined number of luminescence reagent film bags;

[0078] Fix the bead-carrying polymer glass slide in the microsphere antibody composite cavity of the pre-constructed original microfluidic paper chip, and place the luminescence reagent film bag and the buffer cleaning film bag in the reagent storage cavity and the buffer cleaning solution storage cavity of the original microfluidic paper chip respectively to obtain the target microfluidic paper chip.

[0079] It is understandable that the flexible film refers to a pre-developed film bag that can store various reagents for a long time. The film bag has a pressure-sensitive liquid valve, and when the film bag is subjected to a certain intensity of pressure, the pressure-sensitive liquid valve will be automatically opened, enabling the reagent therein to flow out through the pressure-sensitive liquid valve.

[0080] S2. Obtain a test sample, add the test sample to the pre-constructed card reader device to obtain a card reader device to be tested, and insert the target microfluidic paper chip into the card reader device to be tested to obtain a paper chip-card reader device.

[0081] It is understandable that after inserting the target microfluidic paper chip into the card reader device, the sampling point of the target microfluidic paper chip will coincide with the sample adding point of the card reader device, so that the pre-added test sample will react with the antibody-polymer microsphere composite material in the target microfluidic paper chip to obtain a complex of antibody and antigen and a test sample solution.

[0082] It should be understood that the paper chip-reader device contains a card slot, a constant temperature system, a piezoelectric ceramic and its controller, an electrode control system, a photoelectric sensor, a power control system, and a user operation interface system.

[0083] In an embodiment of the present invention, the obtaining of the sample to be tested and adding the sample to be tested to the sample addition point of a pre-constructed card reader device to obtain a card reader device to be tested includes:

[0084] Obtaining the sample to be tested by using a pre-constructed syringe;

[0085] Injecting the sample to be tested into the card reader device from the sample addition point through the syringe to obtain the card reader device to be tested.

[0086] It can be understood that there is a sealing structure at the sample addition point of the card reader device, which can ensure the tightness of the card reader device during the insertion and extraction of the syringe.

[0087] In an embodiment of the present invention, the inserting of the target microfluidic paper chip into the card reader device to be tested to obtain a paper chip-card reader device includes:

[0088] Opening the lockable cover plate of the card reader device to be tested to obtain the interface of the target microfluidic paper chip;

[0089] Inserting the target microfluidic paper chip into the interface and locking the lockable cover plate to obtain the paper chip-card reader device.

[0090] It should be understood that there is a light-shielding cover plate at the interface where the target microfluidic paper chip is inserted into the card reader device to be tested. By locking the light-shielding cover plate, it can ensure that the inside of the card reader device to be tested is completely dark and there is no light leakage gap at the interface, thereby ensuring the smooth progress of the chemiluminescence immunoassay reaction.

[0091] S3. Pressing the chemiluminescent reaction reagent and the buffer cleaning liquid into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex.

[0092] In an embodiment of the present invention, the pressing of the chemiluminescent reaction reagent and the buffer cleaning liquid into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex includes:

[0093] Receiving a test instruction, and starting the piezoelectric ceramic in the paper chip card reader device according to the test instruction to obtain a serialized start of the piezoelectric ceramic;

[0094] Using the serialized start of the piezoelectric ceramic to press out part of the reagent in the buffer cleaning film bag to obtain a first liquid for infiltration and cleaning;

[0095] By using the capillary seepage action of the paper chip and the water absorption action of the water-absorbing polymer, the first cleaning liquid to be seeped is drained to the microsphere antibody composite cavity of the target microfluidic paper chip to clean the microsphere antibody composite cavity, and a test antigen-antibody composite molecule is obtained.

[0096] By using the serialized activation of the piezoelectric ceramic, the reagent in the reagent film bag of the luminescent reagent is pressed out to obtain the luminescent reagent to be seeped.

[0097] By using the capillary seepage action of the paper chip and the water absorption action of the water-absorbing polymer, the luminescent reagent to be seeped is drained to the microsphere antibody composite cavity to react with the test antigen-antibody composite molecule, and a microbead antibody complex to be cleaned is obtained.

[0098] By using the serialized activation of the piezoelectric ceramic, the reagent in the buffer cleaning film bag is pressed out to obtain the second cleaning liquid to be seeped.

[0099] By using the capillary seepage action of the paper chip and the water absorption action of the water-absorbing polymer, the second cleaning liquid to be seeped is drained to the microsphere antibody composite cavity to clean the microsphere antibody composite cavity, and the microbead antibody complex is obtained.

[0100] It can be understood that when receiving the test instruction input by the user on the paper chip-reader device, the piezoelectric ceramics in the paper chip-reader device will be sequentially activated. There is a reagent film bag of the luminescent reagent or a buffer cleaning film bag under each piezoelectric ceramic. The reagents in the reagent film bag of the luminescent reagent and the buffer cleaning film bag are pressed into the microsphere antibody composite cavity in a certain order by the activated piezoelectric ceramics, and the immunoassay, cleaning, and luminescence reaction are gradually completed.

[0101] It can be explained that when receiving the test instruction, it is first necessary to activate the piezoelectric ceramic above the buffer cleaning film bag to complete the cleaning of the microsphere antibody composite cavity, remove the solvent in the test sample, and facilitate subsequent reactions. Then, activate the piezoelectric ceramic above the reagent film bag of the luminescent reagent, and the antibody-luminescence composite molecule flows into the microsphere antibody composite cavity along with the luminescence reaction reagent, and the antibody II in the antibody-luminescence composite molecule undergoes an immunoassay with the antigen that may be contained in the antibody-polymer microsphere composite material, so that the antigen that may be contained in the test sample is connected to the antibody-luminescence composite molecule and the antibody-polymer microsphere composite material.

[0102] It can be understood that the reagents in the reagent film bag of the luminescent reagent and the buffer cleaning film bag can be sequentially reacted through the pressure-sensitive liquid valve and the paper chip and flow through the microsphere antibody composite cavity according to the preset reagent addition sequence of the luminescent immunoassay, and finally, the water-absorbing polymer in the waste liquid collection cavity is used to collect the waste liquid.

[0103] In the embodiment of the present invention, before the luminescent reaction reagent and the buffer cleaning fluid are fluidized into the cavity where the antibody-polymer microbead composite material is located for reaction, the method further includes:

[0104] Starting a thermostat in the paper chip-card reader device, and using the thermostat to adjust the temperature in the paper chip-card reader device to a predetermined temperature;

[0105] The predetermined temperature is maintained by the constant temperature device to obtain a constant temperature condition.

[0106] It can be understood that the predetermined temperature refers to a temperature preset by the user that is suitable for the luminescent immunoreaction.

[0107] S4. Apply an electric field to the microbead-antibody complex using the paper chip-card reader device, and detect the luminescence of the microbead-antibody complex, and query the antigen content of the sample to be tested based on the luminescence.

[0108] It can be explained that the photodetector can be various optical signal detection devices such as silicon-based sensors, photomultiplier tubes, single photon counters, etc. When the paper chip-card reader device energizes the electrodes, if the sample to be tested contains antigens, the microbead antibody complex will emit photons under the action of the electric field, which can be detected by the photodetector. The higher the antigen content in the sample to be tested, the more photons will be emitted.

[0109] In the embodiment of the present invention, querying the antigen content of the sample to be tested according to the luminescence amount includes:

[0110] Obtain a pre-constructed luminescence-antigen content comparison table;

[0111] According to the luminescence amount, the antigen content of the sample to be tested is queried in the luminescence amount-antigen content comparison table.

[0112] It is understandable that when the antigen content in the sample to be tested is higher, the luminescence amount is greater, and the two are in direct proportion. Therefore, the luminescence amount-antigen content comparison table can be established based on the corresponding relationship between the luminescence amount and the antigen content.

[0113] In the embodiment of the present invention, the process of constructing the luminescence amount-antigen content comparison table includes:

[0114] Statistics of historical antigen content and corresponding historical luminescence data;

[0115] Taking the historical antigen content as an independent variable and the historical luminescence data as a dependent variable, a calculation formula of antigen content-luminescence is constructed;

[0116] The unit luminescence amount of the unit antigen is calculated using the antigen content-luminescence amount calculation formula;

[0117] According to the unit luminescence of the unit antigen, the luminescence amount-antigen content comparison table is constructed.

[0118] It is understandable that the historical antigen content and historical luminescence data are data related to the past antigen content and the corresponding luminescence. The unit luminescence of the unit antigen refers to the luminescence corresponding to each unit amount of the antigen.

[0119] Compared with the background technology, the mainstream products of "electrochemical immunoluminescence method" have the problems of high price, complex operation, difficult maintenance and difficult management of consumables. In the embodiment of the present invention, the antibody-polymer microbead composite material, the luminescent reaction reagent and the buffer cleaning solution are respectively placed in the predetermined cavity of the original microfluidic paper chip to obtain the target microfluidic paper chip, which has two important elements for detecting the antigen content in the sample to be tested, namely: the antibody-polymer microbead composite material and the luminescent reaction reagent, and then the sample to be tested is obtained by using the card reader device, and the target microfluidic paper chip is inserted into the card reader device to obtain the paper chip-card reader device, and various reagents are pressed into the cavity where the antibody-polymer microbead composite material is located by the paper chip-card reader device to react to obtain the microbead antibody complex, and finally the paper chip-card reader device is used to apply an electric field to the microbead antibody complex, so that the microbead antibody composite cavity produces a luminescent effect, and the antigen content in the sample to be tested is judged according to the luminescence detected by the photoelectric detector, thereby realizing the process of luminescent immunoassay. Therefore, the luminescent immunoassay method, apparatus, electronic device and computer-readable storage medium based on portable devices proposed in the present invention can solve the problems of mainstream products of "electrochemical immunoluminescence method" such as high price, complex operation, difficult maintenance and difficult management of consumables.

[0120] like Figure 4 FIG. 1 is a functional module diagram of a luminescent immunoassay device based on a portable device provided in one embodiment of the present invention.

[0121] The portable device-based luminescent immunoassay device 100 of the present invention can be installed in an electronic device. According to the functions to be implemented, the portable device-based luminescent immunoassay device 100 can include a target microfluidic paper chip manufacturing module 101, a paper chip-card reader device acquisition module 102, a microbead antibody complex acquisition module 103 and a photoelectric detection module 104. The module of the present invention can also be referred to as a unit, which refers to a series of computer program segments that can be executed by an electronic device processor and can complete fixed functions, which are stored in the memory of the electronic device.

[0122] The target microfluidic paper chip manufacturing module 101 is configured to obtain an antibody-polymer microbead composite material, a luminescence reaction reagent, and a buffer cleaning solution, and place the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively, so as to obtain a target microfluidic paper chip;

[0123] The paper chip-reader device acquisition module 102 is configured to obtain a sample to be tested, add the sample to be tested to a pre-constructed reader device to obtain a to-be-read reader device, and insert the target microfluidic paper chip into the to-be-read reader device to obtain a paper chip-reader device;

[0124] The microbead antibody complex acquisition module 103 is configured to press the luminescence reaction reagent and the buffer cleaning solution into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex;

[0125] The photoelectric detection module 104 is configured to apply an electric field to the microbead antibody complex by using the paper chip-reader device, detect the luminescence amount of the microbead antibody complex, and query the antigen content of the sample to be tested according to the luminescence amount.

[0126] Specifically, each module in the luminescence immunoassay device 100 based on a portable device in the embodiment of the present invention adopts the same technical means as those in the Figures 1 to 4 luminescence immunoassay method based on a portable device described above, and can produce the same technical effects, which will not be elaborated herein.

[0127] As Figure 5 shown, it is a schematic structural diagram of an electronic device for implementing the luminescence immunoassay method based on a portable device provided by an embodiment of the present invention.

[0128] The electronic device 1 may include a processor 10, a memory 11, a bus 12, and a communication interface 13, and may further include a computer program stored in the memory 11 and executable on the processor 10, such as a luminescence immunoassay program based on a portable device.

[0129] Among them, the memory 11 at least includes one type of readable storage medium, and the readable storage medium includes flash memory, mobile hard disk, multimedia card, card-type memory (such as SD or DX memory, etc.), magnetic memory, magnetic disk, optical disc, etc. The memory 11 can be an internal storage unit of the electronic device 1 in some embodiments, such as the mobile hard disk of the electronic device 1. The memory 11 can also be an external storage device of the electronic device 1 in some other embodiments, such as a plug-in mobile hard disk, a Smart Media Card (SMC), a Secure Digital (SD) card, a Flash Card, etc. equipped on the electronic device 1. Further, the memory 11 can also include both the internal storage unit and the external storage device of the electronic device 1. The memory 11 can be used not only to store application software installed in the electronic device 1 and various types of data, such as the code of the chemiluminescence immunoassay program based on a portable device, etc., but also to temporarily store data that has been output or will be output.

[0130] The processor 10 can be composed of integrated circuits in some embodiments. For example, it can be composed of a single packaged integrated circuit, or can also be composed of multiple integrated circuits with the same or different functions packaged, including the combination of one or more central processing units (CPU), microprocessors, digital processing chips, graphics processors, and various control chips, etc. The processor 10 is the control core (Control Unit) of the electronic device, connecting various components of the entire electronic device through various interfaces and lines, and by running or executing programs or modules stored in the memory 11 (such as the chemiluminescence immunoassay program based on a portable device, etc.), and calling the data stored in the memory 11, to execute various functions of the electronic device 1 and process data.

[0131] The bus can be a peripheral component interconnect (PCI) bus or an extended industry standard architecture (EISA) bus, etc. This bus can be divided into an address bus, a data bus, a control bus, etc. The bus is set to realize the connection and communication between the memory 11 and at least one processor 10, etc.

[0132] Figure 5 Only the electronic device with components is shown. Those skilled in the art can understand that Figure 5The structures shown do not constitute a limitation on the electronic device 1, and it may include fewer or more components than shown, or combine certain components, or have different component arrangements.

[0133] For example, although not shown, the electronic device 1 may further include a power source (such as a battery) for supplying power to each component. Preferably, the power source can be logically connected to the at least one processor 10 through a power management device, so as to implement functions such as charge management, discharge management, and power consumption management through the power management device. The power source may also include any components such as one or more DC or AC power sources, a recharge device, a power failure detection circuit, a power converter or inverter, a power status indicator, etc. The electronic device 1 may also include various sensors, a Bluetooth module, a Wi-Fi module, etc., which will not be elaborated here.

[0134] Furthermore, the electronic device 1 may further include a network interface. Optionally, the network interface may include a wired interface and / or a wireless interface (such as a WI-FI interface, a Bluetooth interface, etc.), which is generally used to establish a communication connection between the electronic device 1 and other electronic devices.

[0135] Optionally, the electronic device 1 may further include a user interface. The user interface may be a display, an input unit (such as a keyboard), and optionally, the user interface may also be a standard wired interface or a wireless interface. Optionally, in some embodiments, the display may be an LED display, a liquid crystal display, a touch liquid crystal display, and an OLED (Organic Light-Emitting Diode) toucher, etc. Among them, the display may also be appropriately referred to as a display screen or a display unit, which is used to display the information processed in the electronic device 1 and to display a visual user interface.

[0136] It should be understood that the above embodiments are only for illustration purposes and are not limited by this structure in the scope of the patent application.

[0137] The luminescence immunoassay program based on the portable device stored in the memory 11 of the electronic device 1 is a combination of multiple instructions. When running in the processor 10, it can:

[0138] Obtain an antibody-polymer microsphere composite material, a luminescence reaction reagent, and a buffer cleaning solution, and place the antibody-polymer microsphere composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively to obtain a target microfluidic paper chip;

[0139] Obtain a sample to be tested, add the sample to be tested to a pre-constructed card-reading device to obtain a card-reading device to be processed, and insert the target microfluidic paper chip into the card-reading device to be processed to obtain a paper chip-card-reading device;

[0140] Press the luminescence reaction reagent and the buffer cleaning liquid into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex;

[0141] Apply an electric field to the microbead antibody complex by using the paper chip-card-reading device, detect the luminescence amount of the microbead antibody complex, and query the antigen content of the sample to be tested according to the luminescence amount.

[0142] Specifically, for the specific implementation method of the above instructions by the processor 10, reference can be made to Figures 1 to 4 the description of the relevant steps in the corresponding embodiment, which will not be elaborated here.

[0143] Further, if the module / unit integrated in the electronic device 1 is implemented in the form of a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. The computer-readable storage medium can be volatile or non-volatile. For example, the computer-readable medium can include: any entity or device capable of carrying the computer program code, a recording medium, a USB flash drive, a mobile hard disk, a magnetic disk, an optical disc, a computer memory, a read-only memory (ROM, Read-Only Memory).

[0144] The present invention also provides a computer-readable storage medium, where the readable storage medium stores a computer program, and when the computer program is executed by a processor of an electronic device, it can implement:

[0145] Obtain an antibody-polymer microbead composite material, a luminescence reaction reagent, and a buffer cleaning liquid, and place the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning liquid in predetermined cavities of a pre-constructed original microfluidic paper chip respectively to obtain a target microfluidic paper chip;

[0146] Obtain a sample to be tested, add the sample to be tested to a pre-constructed card-reading device to obtain a card-reading device to be processed, and insert the target microfluidic paper chip into the card-reading device to be processed to obtain a paper chip-card-reading device;

[0147] Press the luminescence reaction reagent and the buffer cleaning liquid into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex;

[0148] Apply an electric field to the microbead antibody complex by using the paper chip-card-reading device, detect the luminescence amount of the microbead antibody complex, and query the antigen content of the sample to be tested according to the luminescence amount.

[0149] In several embodiments provided by the present invention, it should be understood that the disclosed devices, apparatuses, and methods can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative. For example, the division of the modules is only a logical function division, and there can be other division methods in actual implementation.

[0150] The modules described as separate components may or may not be physically separated, and the components shown as modules may or may not be physical units, that is, they can be located in one place or distributed to multiple network units. Some or all of the modules can be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0151] In addition, in each embodiment of the present invention, the functional modules can be integrated in a processing unit, or each unit can exist physically alone, or two or more units can be integrated in one unit. The above-mentioned integrated unit can be implemented in the form of hardware or in the form of a combination of hardware and software functional modules.

[0152] For those skilled in the art, it is obvious that the present invention is not limited to the details of the above-mentioned exemplary embodiments, and without departing from the spirit or basic characteristics of the present invention, the present invention can be implemented in other specific forms.

[0153] Therefore, from any point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be included in the present invention. Any associated drawing marks in the claims should not be regarded as limiting the claimed rights.

[0154] The blockchain referred to in the present invention is a new application mode of computer technologies such as distributed data storage, peer-to-peer transmission, consensus mechanism, and encryption algorithms. Blockchain, essentially a decentralized database, is a string of data blocks generated by using cryptographic methods. Each data block contains information about a batch of network transactions, which is used to verify the validity (anti-counterfeiting) of the information and generate the next block. The blockchain can include a blockchain underlying platform, a platform product service layer, and an application service layer, etc.

[0155] In addition, obviously, the word "including" does not exclude other units or steps, and the singular does not exclude the plural. The multiple units or apparatuses stated in the system claims can also be implemented by one unit or apparatus through software or hardware. Words such as "second" are used to denote names and do not represent any specific order.

[0156] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A chemiluminescent immunoassay method based on a portable device, characterized in that The method includes: Obtaining an antibody-polymer microbead composite material, a luminescence reaction reagent, and a buffer cleaning solution, and respectively placing the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip to obtain a target microfluidic paper chip; Obtaining a sample to be measured, adding the sample to be measured to a pre-constructed card reader device to obtain a to-be-read card reader device, and inserting the target microfluidic paper chip into the to-be-read card reader device to obtain a paper chip-card reader device; Pressing the luminescence reaction reagent and the buffer cleaning solution into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex; Applying an electric field to the microbead antibody complex by using the paper chip-card reader device, detecting the luminescence amount of the microbead antibody complex, and querying the antigen content of the sample to be measured according to the luminescence amount; The process of obtaining the antibody-polymer microbead composite material includes: Obtaining polymer microbeads, antibody I, avidin, and biotin; Binding the avidin and biotin to the polymer microbeads to obtain polymer microbeads; Using the binding sites of the polymer microbeads to connect antibody I to obtain the antibody-polymer microbead composite material; The process of obtaining the luminescence reaction reagent includes: Obtaining a luminescent molecule and antibody II; Connecting the luminescent molecule and antibody II to obtain an antibody-luminescence composite molecule; Obtaining a reaction reagent, and mixing the antibody-luminescence composite molecule and the reaction reagent to obtain the luminescence reaction reagent; The process of constructing the original microfluidic paper chip includes: Obtaining a microfluidic paper chip box body; According to the process of electrochemiluminescence immunoassay, designing the position layout of a preset reagent storage cavity, a buffer cleaning solution storage cavity, a paper chip area, a microsphere antibody complex cavity, an electrode area, a waste liquid collection cavity, and a sampling point on the microfluidic paper chip box body; Adding a pre-constructed paper chip to the paper chip area, and adding a pre-constructed water-absorbing polymer to the waste liquid collection cavity to obtain the original microfluidic paper chip; The step of respectively placing the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of the pre-constructed original microfluidic paper chip to obtain a target microfluidic paper chip includes: Fixing the antibody-polymer microbead composite material on a pre-constructed polymer glass slide to obtain a bead-carrying polymer glass slide; Respectively placing the luminescence reaction reagent and the buffer cleaning solution in a pre-constructed flexible film to obtain a buffer cleaning film bag and a predetermined number of luminescence reagent film bags; Fixing the bead-carrying polymer glass slide in the microsphere antibody complex cavity of the pre-constructed original microfluidic paper chip, and respectively placing the luminescence reagent film bag and the buffer cleaning film bag in the reagent storage cavity and the buffer cleaning solution storage cavity of the original microfluidic paper chip to obtain the target microfluidic paper chip; The step of pressing the luminescence reaction reagent and the buffer cleaning solution into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex includes: Receive a test instruction, and according to the test instruction, start the piezoelectric ceramics in the paper chip card reader device to obtain a serialized start of the piezoelectric ceramics; Use the serialized start of the piezoelectric ceramics to press out part of the reagent in the buffer cleaning film bag to obtain a first cleaning solution to be seeped; Use the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the first cleaning solution to be seeped to the microsphere antibody composite cavity of the target microfluidic paper chip to clean the microsphere antibody composite cavity and obtain a to-be-detected antigen-antibody composite molecule; Use the serialized start of the piezoelectric ceramics to press out the reagent in the luminescent reagent film bag to obtain a luminescent reagent to be seeped; Use the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the luminescent reagent to be seeped to the microsphere antibody composite cavity to react with the to-be-detected antigen-antibody composite molecule to obtain a microbead antibody complex to be cleaned; Use the serialized start of the piezoelectric ceramics to press out the reagent in the buffer cleaning film bag to obtain a second cleaning solution to be seeped; Use the capillary seepage effect of the paper chip and the water absorption effect of the water-absorbing polymer to drain the second cleaning solution to be seeped to the microsphere antibody composite cavity to clean the microsphere antibody composite cavity and obtain the microbead antibody complex; The querying of the antigen content of the to-be-detected sample according to the luminescence amount includes: Obtain a pre-constructed luminescence amount-antigen content comparison table; According to the luminescence amount, query the antigen content of the to-be-detected sample in the luminescence amount-antigen content comparison table.

2. A chemiluminescent immunoassay device based on a portable device, characterized in that, The device includes: A target microfluidic paper chip manufacturing module, configured to obtain an antibody-polymer microbead composite material, a luminescence reaction reagent, and a buffer cleaning solution, and place the antibody-polymer microbead composite material, the luminescence reaction reagent, and the buffer cleaning solution in predetermined cavities of a pre-constructed original microfluidic paper chip respectively to obtain a target microfluidic paper chip; A paper chip-card reader device acquisition module, configured to obtain a to-be-detected sample, add the to-be-detected sample to a pre-constructed card reader device to obtain a to-be-read card device, and insert the target microfluidic paper chip into the to-be-read card device to obtain a paper chip-card reader device; A microbead antibody complex acquisition module, configured to press the luminescence reaction reagent and the buffer cleaning solution into the cavity where the antibody-polymer microbead composite material is located for reaction to obtain a microbead antibody complex; An optoelectronic detection module, configured to apply an electric field to the microbead antibody complex by using the paper chip-card reader device, detect the luminescence amount of the microbead antibody complex, and query the antigen content of the to-be-detected sample according to the luminescence amount.

3. An electronic device, characterized in that, The electronic device includes: At least one processor; and, A memory communicatively connected to the at least one processor; wherein, The memory stores instructions executable by the at least one processor, and the instructions are executed by the at least one processor so that the at least one processor can execute the luminescence immunoassay method based on a portable device as claimed in claim 1.

4. A computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by a processor, it implements the luminescence immunoassay method based on a portable device as described in claim 1.

Images