CN215297225U - Novel electrochemical immunodetection system for coronavirus 2019-nCoV - Google Patents
Novel electrochemical immunodetection system for coronavirus 2019-nCoV Download PDFInfo
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- CN215297225U CN215297225U CN202120004857.6U CN202120004857U CN215297225U CN 215297225 U CN215297225 U CN 215297225U CN 202120004857 U CN202120004857 U CN 202120004857U CN 215297225 U CN215297225 U CN 215297225U
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Abstract
The utility model discloses a novel coronavirus 2019-nCoV electrochemical immunodetection system, which comprises an electrochemical immunosensor test paper, a bottom plate and an electrode layer, wherein the electrode layer comprises a counter electrode and a working electrode; the detection instrument is matched with the electrochemical immunosensor test paper for use, and at least comprises a detection signal input module, a test paper insertion module, a signal conversion module and a data processing module, wherein the detection signal input module is used for applying a detection signal to the test paper and outputting a feedback signal, the test paper insertion module is used for realizing connection of the detection instrument and the test paper, the signal conversion module is used for receiving the feedback signal and performing analog-to-digital conversion on the feedback signal, and the data processing module is used for comparing the feedback signal obtained in the detection signal input module with a standard curve of a database and calculating to obtain a result. The utility model discloses utilize antibody and nucleocapsid protein to combine, arouse the change of electric signal on the electrochemistry immunosensor, detect the signal of telecommunication through the electrochemistry method, realize quick, simple, cheap, the novel coronavirus 2019-nCoV's of quantitative determination electrochemistry immunodetection system, detecting instrument size is little, easy operation.
Description
Technical Field
The utility model belongs to the technical field of electrochemistry immunodetection, especially, relate to a novel coronavirus 2019-nCoV's electrochemistry immunodetection system.
Background
The novel coronavirus (2019-nCoV) is newly discovered in the end of 2019, has high infectivity and harmfulness, can be transmitted in modes of droplet, contact with patients and the like, and infected persons can present pneumonia symptoms such as cold and the like, and is an acute respiratory infectious disease. Most of the conventional methods for detecting 2019-nCoV currently are nucleic acid detection and colloidal gold test paper detection, although the nucleic acid detection is the gold standard for clinically determining 2019-nCoV, special instruments are needed, the requirements on equipment, fields and operators are extremely high, the operation is complicated, the time consumption is long, and the infection risk of the detectors is high; although the colloidal gold test paper is simple to operate, the colloidal gold immunochromatographic assay can only carry out rough qualitative test and cannot complete more accurate quantitative test. The electrochemical method has the advantages of sensitivity, high efficiency, low price, easy miniaturization and the like, and is particularly suitable for being made into a portable rapid and accurate diagnosis product, the existing research on 2019-nCoV detection by the electrochemical immunization technology is to utilize a double-antibody sandwich immunization method, a substance with the property of horseradish peroxidase is marked on a sample pad, the method for making the test paper is more complicated, the making cost is higher, and a small-sized instrument matched with detection is not provided; although the 2019-nCoV detection is performed by using a blood glucose instrument, a plurality of substances are labeled by probes, are mixed with a sample to prepare a mixed solution, and the mixed solution is incubated for 0.5 to 3 hours and then can be dropped into a test paper for subsequent detection.
The market lacks a quantitative detection system which is efficient, cheap and miniaturized.
SUMMERY OF THE UTILITY MODEL
In order to overcome the defects of the prior art, the utility model provides a novel coronavirus 2019-nCoV electrochemical immunodetection system which is portable, easy to operate, high in detection speed, high in detection result accuracy and low in use cost.
The utility model provides a technical scheme that its technical problem adopted is: an electrochemical immunodetection system of a novel coronavirus 2019-nCoV, comprising:
the electrochemical immunosensor test paper comprises a bottom plate and an electrode layer at least comprising a counter electrode and a working electrode;
the detecting instrument is matched with the electrochemical immunosensor test paper for use, and at least comprises,
the detection signal input module is used for applying a detection signal to the electrochemical immunosensor test paper and outputting a feedback signal;
the test paper inserting module is used for realizing the connection between a detection instrument and the electrochemical immunosensor test paper;
the signal conversion module is used for receiving the feedback signal of the detection signal input module and performing analog-to-digital conversion on the feedback signal;
and the data processing module is used for comparing and calculating the feedback signal obtained in the detection signal input module with a standard curve of a database to obtain a detection result.
Furthermore, the electrochemical immunosensor test paper also comprises an insulating layer with a sampling hole, wherein a colloidal gold solution layer, a 2019-nCoV antibody layer and a sealing layer are coated in the sampling hole.
Further, the detection signal input module is a square wave pulse input module and/or an alternating current input module.
Further, the alternating current input module applies variable frequency alternating current between a working electrode and a counter electrode of the electrochemical immunosensor test paper; the square wave pulse input module applies a square wave pulse signal between a working electrode and a counter electrode of the electrochemical immunosensor test paper.
Furthermore, the size of the variable frequency alternating current is-0.005V, and the amplitude is 0.005V; the square wave pulse signal is initially-0.4V, the voltage increment is 0.004V, the pulse amplitude is 0.025V, and the frequency is 15 Hz.
Further, the detection instrument further comprises a detection unit,
the temperature input module detects the current ambient temperature and outputs the current ambient temperature into a feedback signal;
and the data processing module compares the feedback signal obtained by the detection signal input module and the feedback signal obtained by the temperature input module with a standard curve of a database for calculation to obtain a detection result.
Furthermore, the electrode layer also comprises a starting electrode; the electrochemical immunosensor test paper is provided with an identification unit which is a resistance element arranged between the working electrode and the starting electrode; or the starting electrode is connected with the working electrode, or the starting electrode is connected with the counter electrode in different connection modes.
Furthermore, the reagent cup for containing the detection solution comprises a reagent cup main body and a reagent cup sealing film packaged on the reagent cup main body.
Furthermore, the device also comprises a reagent tube for containing the detection solution, and the reagent tube comprises a tube cover, a tube body, a sealing film arranged at the top opening of the tube body, and a tube cover sealing piece.
Furthermore, an L-shaped groove is formed in the outer wall of the pipe body, a protruding block capable of being clamped with the L-shaped groove is formed in the inner surface of the pipe cover, and the pipe cover sealing element is arranged on the upper portion of the protruding block.
The utility model discloses utilize finished 2019-nCoV antibody and electrochemistry immunity technique to produce the test paper that can quantitative detection 2019-nCoV nucleocapsid protein (N albumen), use with the detection device cooperation that has the electrochemistry performance simultaneously, reach the purpose of quantitative detection 2019-nCoV nucleocapsid protein (N albumen). The utility model discloses the 2019-nCoV test paper preparation method who produces is simple, and the cost of manufacture is lower, and the test paper of preparation and detecting instrument size are less, portable easy operation, and the testing result is rapid.
The utility model has the advantages that: the method has the advantages that the change of the electric signal on the electrochemical immunosensor is caused by the combination of the antibody fixed on the electrochemical immunosensor and the nucleocapsid protein, the electric signal is detected by an electrochemical method, the electrochemical immunodetection system for quickly, simply, cheaply and quantitatively detecting the novel coronavirus 2019-nCoV is realized, the size of a used detection instrument is small, only the size of a glucometer is large, and the operation of the instrument is simple; because the detection method adopts a single antigen-antibody combination method instead of a double-antibody sandwich method, two antibodies do not need to be used and the antibodies do not need to be modified, the test paper is simpler to manufacture and has lower manufacturing cost.
Drawings
Fig. 1 is a block diagram of a detector module of the novel coronavirus 2019-nCoV detection system of the present invention.
FIG. 2 is a schematic diagram of the reagent cup of the present invention.
FIG. 3 is a structural diagram of the reagent tube of the present invention.
Fig. 4 is a cross-sectional view of the pipe cap and the pipe body of the present invention.
Fig. 5 is a structural diagram of the electrochemical immunosensor strip structure in which the detection signal is an ac signal in the first and third embodiments of the present invention.
Fig. 6 is a nyquist plot of the ac response of the electrochemical immunosensor to varying concentrations 2019-nCoV in accordance with a first embodiment of the present invention.
Fig. 7 is a structural diagram of the electrochemical immunosensor strip according to the second embodiment of the present invention, wherein the detection signal is a square-wave pulse signal.
Fig. 8 is a square wave pulse voltammetry response graph of the electrochemical immunosensor according to the second embodiment of the present invention for different concentrations of 2019-nCoV.
Fig. 9 is a structural diagram of the electrochemical immunosensor strip structure in which the detection signal is an ac signal in the third embodiment of the present invention.
Fig. 10 is a structural diagram of the test strip structure of the electrochemical immunosensor in accordance with the third embodiment of the present invention, wherein the detection signal is a square-wave pulse signal.
Fig. 11 is a structural diagram of the electrochemical immunosensor strip according to the third embodiment of the present invention, wherein the detection signal is a square-wave pulse signal.
Detailed Description
In order to make the technical solution of the present invention better understood, the following figures in the embodiments of the present invention are combined to clearly and completely describe the technical solution in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts shall belong to the protection scope of the present invention.
Example one
A novel electrochemical immunodetection system for coronavirus 2019-nCoV comprises electrochemical immunosensor test paper and a detection instrument matched with the electrochemical immunosensor test paper.
The detection instrument comprises
1) A detection signal input module: a detection signal may be applied to the electrochemical immunosensor strip and a feedback signal may be output. The detection signal can be a square wave pulse signal input or an alternating current signal input, and in the embodiment, the detection signal is an alternating current signal input;
2) a temperature input module: a high-precision thermistor is arranged in a detection instrument, a zero-temperature-drift reference constant voltage of 2.048V is generated through a circuit, and the constant voltage can detect the current ambient temperature and convert the ambient temperature into a feedback signal through a sampling circuit formed by the thermistor and other elements
3) The test paper insertion module is used for realizing an interface with the connecting function of a detection instrument and the test paper of the electrochemical immunosensor, judging whether the test paper of the electrochemical immunosensor is effective or not, outputting a feedback signal and prompting to dip into a detection solution; the instrument applies a direct current voltage to the working electrode 23 and the starting electrode 22 corresponding to the electrochemical immunosensor test paper, when the electrochemical immunosensor test paper is inserted, the working electrode 23 and the starting electrode 22 are communicated with the instrument, so that the instrument automatically finishes starting, and when the electrochemical immunosensor test paper electrode is damaged, the effective identification of the test paper cannot be finished;
4) the signal conversion module: the temperature input module is used for receiving the feedback signals output by the detection signal input module and the temperature input module and carrying out analog-to-digital conversion on the feedback signals;
5) a data processing module: comparing the temperature data and the detection signal data obtained by conversion with a standard curve of a database for calculation to obtain a final detection result of the 2019-nCoV electrochemical immunosensor test paper, and displaying the detection result as negative or positive and the concentration of the negative or positive on a display module;
6) a data transmission module: according to the selection of the user, data can be uploaded to the cloud server and the equipment terminal in a wired or wireless mode, so that the user can conveniently check and analyze the big data; wired modes include USB connection and the like, and wireless modes include Bluetooth, WiFi and the like;
7) a human-computer interaction module: the device comprises a display screen, a display screen and a control module, wherein the display screen can display information such as real-time, detection results and the like, and can be used for manually touching or pressing keys to select corresponding operations;
8) a power supply module: the power is supplied to the whole instrument and can be charged through a battery or a USB;
9) a switch module: the power supply of the instrument is controlled to be switched on and off, and the instrument can be started to check data or perform other operations when the test paper is not used;
10) the low-voltage detection module: detect power module electric quantity and send the warning when low electric quantity, remind to charge or change the battery to avoid detecting that data has the mistake.
Of course, the above-mentioned detecting instrument can also be a blood sugar instrument with an additional detecting signal input module, and realizes the detection of blood sugar and new crown.
Utilize the utility model discloses a detecting system still can realize the quantitative determination to analytes such as glycated hemoglobin.
The electrochemical immunosensor test strip includes a base plate 1, an electrode layer including at least a counter electrode 21 and a working electrode 23, and an insulating layer 3 having a well 32. The inner part of the sampling hole is coated with a colloidal gold solution layer, a 2019-nCoV antibody layer and a sealing layer.
In order to contain the detection solution, a reagent cup 4 and a reagent tube 5 are also designed.
As shown in FIG. 2, a detection solution, i.e., potassium ferricyanide/potassium ferricyanide probe solution, is filled in a reagent cup 4, the reagent cup 4 comprises a reagent cup sealing film 41 and a reagent cup main body 42, when the electrochemical immunosensor test paper needs to be immersed in the detection solution, the reagent cup sealing film 41 is torn, and the electrochemical immunosensor test paper is immersed in the reagent cup 4 for detection.
As shown in fig. 3 and 4, a reagent tube 5 for use with an electrochemical immunoassay system of a novel coronavirus 2019-nCoV, which is convenient for transportation and use, wherein the reagent tube 5 is divided into a tube cap 51, a tube body 52, a tube body sealing film 53 and a tube cap sealing element 512, the tube cap sealing element 512 is a tube cap leather ring 512, the top of the tube cap 51 is provided with an opening, the length and the width of the opening are matched with the width and the thickness of a test paper to realize the attachment of the test paper and the tube cap 51, the material of the opening at the top of the tube cap 51 is preferably an elastic material, the opening can be tightly attached to the test paper without affecting the performance of the test paper, the tube body 52 stores a certain amount of solution, the upper part of the port of the tube body 52 is provided with a tube mouth sealing film 53, the outer surface of the tube body 52 is provided with an L-shaped groove 521, the inner surface of the tube cap 51 is provided with a bump 511, the tube cap leather ring 512 is placed on the upper part of the bump 511, the L-shaped groove 521 and the bump 511 can be clamped and rotated at a certain angle on a vertical horizontal plane, the tight fit of the tube cap 51 and the test tube body 52 is realized, the tube cap leather ring 512 is matched with the L-shaped groove 521 and the bump 511, so that the reagent tube is sealed, the solution is not easy to leak, and the number of the L-shaped groove 521 and the bump 511 can be one to more, and the matching can be realized.
In other embodiments, the reagent tube may be tightly sealed by providing the tube body 52 with external threads and providing the tube cap 51 with internal threads, which are matched with the threads.
In order to realize the detection test paper matched with the detection instrument for use, two types of electrochemical immunosensor test paper are designed.
In this embodiment, the test paper for electrochemical immunosensor includes a substrate 1, an electrode layer 2, and an insulating layer 3, wherein the electrode layer 2 is provided with a counter electrode 21, a start electrode 22, a working electrode 23, and a resistor 24, and the insulating layer 3 is provided with a sample hole 32, a lowest detection line 33, a highest detection line 34, and a blank sample hole 31, as shown in fig. 5.
The electrochemical immunosensor test paper is prepared by the following steps:
preparation of colloidal gold
Preparing colloidal gold by chemical reduction method, adding 250mL of 0.1% HAuCI into 500mL of clean beaker4And (3) heating the solution to boiling under vigorous stirring, then adding 3.75mL of 1% sodium citrate into the solution, continuing to heat and stir for 15min, stopping heating, and continuing to stir for 15min to prepare a colloidal gold solution. The solution was stored in a refrigerator at 4 ℃ until use.
Firstly, gold is used as printing ink to be printed on an electrochemical immunosensor test paper base plate 1 to form a screen printing electrode layer 2, a layer of insulating layer 3 is covered on the screen printing electrode layer 2 and used for protecting electrodes, small holes are formed in the insulating layer 3, a sample adding hole 32 is used as a liquid dropping area and used for dropping antibody solution, and a blank sample adding hole 31 is used for exposing a counter electrode 21. Then 8 mul of colloidal gold solution is dripped on the sample adding hole 32 of the electrode, 2.2 mul of 2019-nCoV finished antibody is dripped on the electrode after drying, and blocking treatment is carried out by 1 percent BSA solution after overnight incubation at 2 ℃ for blocking non-specific binding sites. After blocking, excess BSA solution was washed off, and after drying it in the air, it was stored in a refrigerator at 4 ℃ for further use. In order to ensure the accuracy of the detection, a lowest detection line 33 and a highest detection line 34 may be further provided on the test strip to observe whether the test strip is inserted into the solution to a proper depth.
In this embodiment, the detection signal of the detection signal input module is an ac signal input module, which can generate a small-amplitude sine wave potential with a certain frequency to be applied to the detection paper and obtain a feedback signal; an alternating current signal is applied between the working electrode 23 and the counter electrode 21, and a variable frequency alternating current with-0.005V and the amplitude of 0.005V is applied between the two electrodes, so that the impedance response signal of the test paper between 1Hz and 100000Hz is detected.
An electrochemical immunodetection method of a novel coronavirus 2019-nCoV comprises the following steps:
s1, firstly, dripping a sample to be detected in a spotting area of the electrochemical immunosensor test paper, washing off the sample to be detected with PBS after the sample to be detected and the electrochemical immunosensor test paper fully react, and airing;
s2, inserting the reacted test paper into a test paper inserting module, wherein a direct current signal is always applied to the position, corresponding to the test paper working electrode 23 and the starting electrode 22, of the instrument, when the test paper is not inserted, the working electrode and the starting electrode 22 are in an open circuit state, after the test paper is inserted, the working electrode 23 is communicated with the starting electrode 22, and when the signal value is detected for 3 times continuously, the instrument can be started automatically; a signal input module of the instrument inputs a corresponding detection signal and prompts that the test paper is immersed in a detection solution;
s3, tearing the sealing film, immersing the test paper into a proper depth of the detection solution according to the lowest line 33 and the highest line 34 of the test paper of the electrochemical immunosensor, applying a direct current voltage to the position, corresponding to the working electrode 23 and the counter electrode 21, of the test paper by the instrument, and prompting the start of detection when the instrument detects that the electric signal of the test paper reaches a preset target for 3 times continuously;
s4, inputting a corresponding detection signal by the instrument according to the corresponding test paper, namely applying a variable frequency alternating current with the amplitude of 0.005V and the magnitude of-0.005V between the working electrode 23 and the counter electrode 21 of the test paper, and detecting an impedance response signal of the test paper between 1Hz and 100000 Hz;
s5, the instrument converts the generated response signal through the detection signal input module through the signal conversion module and the data processing module to obtain a specific signal value;
as shown in FIG. 6, which reflects the Nyquist plot of the immunosensor's impedance response to varying concentrations 2019-nCoV, the semi-circle diameter in the left half of the curve in FIG. 6 represents the electron transfer resistance inversely proportional to the electron transfer kinetics, i.e., the electron transfer resistance R of the redox probe at the electrode interfacectAs can be seen from FIG. 6, when 1ng/mL of antigen was added to the sample, the diameter of the semicircle was significantly larger compared to the sample without antigen, indicating that the resistance to charge transfer was significantly increased, indicating that the antigen had successfully bound to the antibody on the sensor and prevented the transfer of electrons to the redox probe, thereby allowing forThe feasibility of detecting 2019-nCoV by the method is proved, and the signal can be used for qualitatively detecting 2019-nCoV and realizing further quantitative operation.
S6, the temperature input module detects the current environment temperature according to the thermistor and converts the environment temperature signal through the signal conversion module and the data processing module to obtain a specific signal value;
s7, comparing the detection signal data and the temperature data obtained by conversion in the steps S5 and S6 with standard curves stored in a database for calculation to obtain a final detection result of the 2019-nCoV test paper, and displaying the result as negative or positive and the concentration of the negative or positive test paper on a display module; the standard curve is detected by using an electrochemical immunosensor to the novel coronavirus 2019-nCoV standard substance with different concentrations at different temperatures based on an alternating-current signal input detection method.
Example two
In this embodiment, the test paper for electrochemical immunosensor includes a substrate 1, an electrode layer 2, and an insulating layer 3, wherein the electrode layer 2 is provided with a counter electrode 21, a start electrode 22, a working electrode 23, and a resistor 24, and the insulating layer 3 is provided with a sample hole 32, a lowest detection line 33, and a highest detection line 34, as shown in fig. 7. It is not provided with a blank well 31.
The manufacturing steps of the electrochemical immunosensor test paper are the same as those of the first embodiment, and are not described again.
In this embodiment, the detection signal of the detection signal input module is a square wave pulse input module, which can be used to generate a square wave pulse signal with fixed frequency and amplitude; a square wave pulse signal is applied between the working electrode 23 and the counter electrode 21, the voltage increment is 0.004V, the pulse amplitude is 0.025V, the frequency is 15Hz, and the current response signal in the voltage range of-0.4V-0.175V is detected.
An electrochemical immunodetection method of a novel coronavirus 2019-nCoV comprises the following steps:
s1, firstly, dripping a sample to be detected in a spotting area of the electrochemical immunosensor test paper, washing off the sample to be detected with PBS after the sample to be detected and the electrochemical immunosensor test paper fully react, and airing;
s2, inserting the reacted test paper into a test paper inserting module, wherein a direct current signal is always applied to the position, corresponding to the test paper working electrode 23 and the starting electrode 22, of the instrument, when the test paper is not inserted, the working electrode and the starting electrode 22 are in an open circuit state, after the test paper is inserted, the working electrode 23 is communicated with the starting electrode 22, and when the signal value is detected for 3 times continuously, the instrument can be started automatically; a signal input module of the instrument inputs a corresponding detection signal and prompts that the test paper is immersed in a detection solution;
s3, tearing the sealing film, immersing the test paper into the detection solution at a proper depth according to the lowest line 33 and the highest line 34 of the test paper, applying a direct current voltage to the position of the test paper corresponding to the working electrode 23 and the counter electrode 21 by the instrument, and prompting the start of detection when the instrument detects that the electric signal of the test paper reaches a preset target for 3 times continuously;
s4, inputting a corresponding detection signal by an instrument according to the corresponding test paper, namely applying a square wave pulse signal with the initial voltage of-0.4V, the voltage increment of 0.004V, the pulse amplitude of 0.025V and the frequency of 15Hz between the working electrode 23 and the counter electrode 21 of the test paper, and detecting a current response signal of the square wave pulse signal within the voltage range of-0.4V-0.175V;
s5, the instrument converts the generated response signal through the detection signal input module through the signal conversion module and the data processing module to obtain a specific signal value;
as shown in fig. 8, it can be seen from fig. 8 that when the sample contains 1ng/mL of antigen, the current value obtained by measuring the potassium ferricyanide solution at the predetermined concentration is lower than that of the sample containing no antigen, indicating that the antigen has been captured by the antibody immobilized on the sensor, and the antigen is a macromolecular protein substance, and thus the transfer of electrons on the surface of the electrode is inhibited by the adhesion to the surface of the sensor, resulting in a reduction in the peak detected at the predetermined concentration of potassium ferricyanide. Therefore, the detection accuracy of the EIS method can be proved, and simultaneously, as another detection means, the 2019-nCoV can be qualitatively and further quantitatively detected by using an SWV (square wave volt-ampere) signal.
S6, the temperature input module detects the current environment temperature according to the thermistor and converts the environment temperature signal through the signal conversion module and the data processing module to obtain a specific signal value;
s7, comparing the detection signal data and the temperature data obtained by conversion in the steps S5 and S6 with standard curves stored in a database for calculation to obtain a final detection result of the 2019-nCoV test paper, and displaying the result as negative or positive and the concentration of the negative or positive test paper on a display module; the standard curve is a standard curve for detecting the novel coronavirus 2019-nCoV standard substance with different concentrations by using an electrochemical immunosensor at different temperatures based on a square wave pulse signal input detection method.
EXAMPLE III
As shown in fig. 9 and 10, the electrode layer 2 of the electrochemical immunosensor test paper is further provided with a resistance element 24 for identifying the type of the test paper by an instrument; the resistive element 24 may be a resistor printed with a different resistance between the working electrode 23 and the start electrode 22, or may be a conductive line printed with ink having a different resistance between the working electrode 23 and the start electrode 22. When the test paper insertion instrument is started, the resistance element 24 is identified, and a signal is fed back to the detection signal input module, so that the detection signal input module selects a detection signal corresponding to the test paper according to different feedback signals.
Of course, as shown in fig. 5 and 11 as an example, the electrochemical immunosensor test strip is connected to the working electrode 23 and the starting electrode 22 through the starting electrode 22 or the starting electrode 22 is connected to the counter electrode 21, a dc signal is applied to the position of the instrument corresponding to the starting electrode 22, the working electrode 23 and the counter electrode 21, after the test strip is inserted, the instrument divides the test strip into two types according to the combination of the response signals corresponding to the starting electrode 22 and the working electrode 23, and the starting electrode 22 and the counter electrode 21, and inputs different detection signals to the different types of test strips.
An electrochemical immunodetection method of a novel coronavirus 2019-nCoV comprises the following steps:
s1, firstly, a sample to be detected in a spotting area of the electrochemical immunosensor test paper is washed away by PBS after the sample to be detected and the electrochemical immunosensor test paper fully react, and the sample is dried;
s2, inserting the reacted test paper into a test paper inserting module, wherein a direct current signal is always applied to the position, corresponding to the test paper working electrode 23 and the starting electrode 22, of the instrument, when the test paper is not inserted, the working electrode and the starting electrode 22 are in an open circuit state, after the test paper is inserted, the working electrode 23 is communicated with the starting electrode 22, and when the signal value is detected for 3 times continuously, the instrument can be started automatically; meanwhile, the test paper is provided with a resistance element 24, and an instrument can detect different signal values according to the difference of the resistance element 24, or the instrument is different according to the combination mode of response signals corresponding to the starting electrode 22 and the working electrode 23, and the starting electrode 22 and the counter electrode 21; according to different detection signals of the instrument, a signal input module of the instrument inputs corresponding detection signals and prompts that the test paper is immersed in a detection solution;
s3, tearing the sealing film, immersing the test paper into the detection solution at a proper depth according to the lowest line 33 and the highest line 34 of the test paper, applying a direct current voltage to the position of the test paper corresponding to the working electrode 23 and the counter electrode 21 by the instrument, and prompting the start of detection when the instrument detects that the electric signal of the test paper reaches a preset target for 3 times continuously;
s4, inputting a corresponding detection signal by the instrument according to the corresponding test paper, wherein the detection signal can be a variable frequency alternating current applied between a working electrode and a counter electrode of the electrochemical immunosensor test paper; or applying a square wave pulse signal between a working electrode and a counter electrode of the electrochemical immunosensor test paper;
s5, the instrument converts the generated response signal through the detection signal input module through the signal conversion module and the data processing module to obtain a specific signal value;
s6, the temperature input module detects the current environment temperature according to the thermistor and converts the environment temperature signal through the signal conversion module and the data processing module to obtain a specific signal value;
s7, comparing the detection signal data and the temperature data obtained by conversion in the steps S5 and S6 with standard curves stored in a database for calculation to obtain a final detection result of the 2019-nCoV test paper, and displaying the result as negative or positive and the concentration of the negative or positive test paper on a display module; the standard curve comprises a standard curve for detecting the novel coronavirus 2019-nCoV standard substance with different concentrations by using an electrochemical immunosensor at different temperatures based on a square wave pulse signal input detection method, and a standard curve for detecting the novel coronavirus 2019-nCoV standard substance with different concentrations by using the electrochemical immunosensor at different temperatures based on an alternating current signal input detection method.
The above detailed description is provided for illustrative purposes, and is not intended to limit the present invention, and any modifications and variations of the present invention are within the spirit and scope of the following claims.
Claims (10)
1. An electrochemical immunodetection system of a novel coronavirus 2019-nCoV, which is characterized by comprising:
the electrochemical immunosensor test paper comprises a bottom plate (1) and an electrode layer at least comprising a counter electrode (21) and a working electrode (23);
the detecting instrument is matched with the electrochemical immunosensor test paper for use, and at least comprises,
the detection signal input module is used for applying a detection signal to the electrochemical immunosensor test paper and outputting a feedback signal;
the test paper inserting module is used for realizing the connection between a detection instrument and the electrochemical immunosensor test paper;
the signal conversion module is used for receiving the feedback signal of the detection signal input module and performing analog-to-digital conversion on the feedback signal;
and the data processing module is used for comparing and calculating the feedback signal obtained in the detection signal input module with a standard curve of a database to obtain a detection result.
2. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: the electrochemical immunosensor test paper further comprises an insulating layer (3) with a sampling hole (32), wherein a colloidal gold solution layer, a 2019-nCoV antibody layer and a sealing layer are coated in the sampling hole (32).
3. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: the detection signal input module is a square wave pulse input module and/or an alternating current input module.
4. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 3, wherein: the alternating current input module applies variable frequency alternating current between a working electrode and a counter electrode of the electrochemical immunosensor test paper; the square wave pulse input module applies a square wave pulse signal between a working electrode and a counter electrode of the electrochemical immunosensor test paper.
5. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 4, wherein: the size of the variable frequency alternating current is-0.005V, and the amplitude is 0.005V; the square wave pulse signal is initially-0.4V, the voltage increment is 0.004V, the pulse amplitude is 0.025V, and the frequency is 15 Hz.
6. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: the detection apparatus may further comprise a detector for detecting the presence of the analyte,
the temperature input module detects the current ambient temperature and outputs the current ambient temperature into a feedback signal;
and the data processing module compares the feedback signal obtained by the detection signal input module and the feedback signal obtained by the temperature input module with a standard curve of a database for calculation to obtain a detection result.
7. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: the electrode layer further comprises an actuation electrode (22); the electrochemical immunosensor test paper is provided with an identification unit which is a resistance element (24) arranged between a working electrode (23) and a starting electrode (22); or the starting electrode (22) and the working electrode (23) are connected, or the starting electrode (22) and the counter electrode (21) are connected in different connection modes.
8. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: also comprises a reagent cup (4) for containing the detection solution, which comprises a reagent cup main body (42) and a reagent cup sealing film (41) packaged on the reagent cup main body (42).
9. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 1, wherein: the kit also comprises a reagent tube (5) for containing the detection solution, and the kit comprises a tube cap (51), a tube body (52), a sealing film (53) arranged on the top opening of the tube body (52), and a tube cap sealing element (512).
10. The system for electrochemical immunodetection of novel coronaviruses 2019-nCoV according to claim 9, wherein: an L-shaped groove (521) is formed in the outer wall of the pipe body (52), a protruding block (511) capable of being clamped with the L-shaped groove (521) is formed in the inner surface of the pipe cover (51), and the pipe cover sealing piece (512) is arranged on the upper portion of the protruding block (511).
Priority Applications (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114441612A (en) * | 2021-12-30 | 2022-05-06 | 广州市赛特检测有限公司 | Electrochemical biosensor, detection method and application |
| CN114441615A (en) * | 2021-12-30 | 2022-05-06 | 广州市赛特检测有限公司 | Modification method of electrical impedance biosensor electrode for detecting new coronavirus |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114441612A (en) * | 2021-12-30 | 2022-05-06 | 广州市赛特检测有限公司 | Electrochemical biosensor, detection method and application |
| CN114441615A (en) * | 2021-12-30 | 2022-05-06 | 广州市赛特检测有限公司 | Modification method of electrical impedance biosensor electrode for detecting new coronavirus |
| CN114441615B (en) * | 2021-12-30 | 2022-12-02 | 广州市赛特检测有限公司 | Modification method of electrical impedance biosensor electrode for detecting new coronavirus |
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