Portable hydrogel resonance protein sensing detection device capable of being detected on site after being manufactured

Abstract

The utility model discloses an on-site preparation and on-site detection type hydrogel resonant protein sensing portable detection device, which comprises a step-by-step reaction module, a control module and a control module, the step-by-step reaction module comprises a first base, a cover plate and a plurality of flow channel units, and the flow channel units are separated into a plurality of grids for accommodating different reagents through separation units; different reagents are mixed for step-by-step reaction when different separation units are rotated; the hydrogel incubation module comprises an incubation reaction tank and a plurality of mold frames, the mold frames are arranged in the incubation reaction tank and are in one-to-one correspondence with the flow channel units, two split resonance rings with opposite opening directions are attached to the mold frames, and the mold frames are provided with containing grooves for containing reagents; after the gradually reacted reagent reacts with the reagent in the accommodating groove, a sample to be detected is dropped; and the detection module is used for forming an induction field with the split resonant ring of the corresponding mold frame for detection. The device realizes rapid and quantitative detection of portable protein markers of a sample to be detected, and has the advantages of simplicity, convenience and rapidness in operation, high sensitivity and simultaneous reading of multi-target results.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the biological detection technical field, concretely relates to a kind of water gel resonance protein sensing portable detection device of present preparation present measurement type. BACKGROUND

[0002] Currently, researchers mainly use time-consuming and complex optical detection methods to detect protein markers (such as in environmental liquid samples, saliva samples, or blood samples). Recently, a concept of digital enzyme-linked immunosorbent assay has emerged for multiplex detection of protein markers with attomolar sensitivity. However, this technology either still relies on large laboratory instruments, is complex to operate and requires professional personnel, and is time-consuming, or the small-sized portable devices used usually can only achieve semi-quantitative detection, with low accuracy, lack of standardization, and limited scope of application.

[0003] For example, Chinese Patent No. CN220854894U proposes a PCT and CRP detection kit, which relies on observation of color change of the detection piece for detection of PCT and CRP, and cannot quantitatively detect the CRP content in the sample to be tested. Chinese Patent No. CN220525831U proposes a MxA / CRP detection reagent card and kit, which requires the use of a large laboratory instrument, immunological quantitative analysis instrument, for quantitative detection of the sample to be tested. Chinese Patent No. CN118777610A proposes a time-resolved fluorescent microsphere, its preparation method, a fluorescent immunochromatography reagent plate, and a kit, which requires the use of a blood cell analyzer for detection of the sample to be tested by dropping the sample to be tested on the detection card. The minimum detection for CRP is 0.5 mg / mL, and the minimum detection for SAA is 2.5 mg / mL, with poor detection performance. Chinese Patent No. CN118879941A proposes a respiratory syncytial virus detection kit, and Chinese Patent No. CN119101724A proposes a freeze-dried PCR reagent and kit for respiratory virus nucleic acid detection. The detection of respiratory syncytial virus (RSV) requires the use of a fluorescent PCR detector for detection, which takes a long time and can only obtain positive or negative results, and cannot quantitatively detect.

[0004] In addition, the biomarker concentration in the diluted sample is low, which challenges the practical limit of many electrochemical sensors. In recent years, more and more hydrogel sensors manufactured by different methods are used in different fields to detect trace target. These hydrogel-based sensors have higher sensitivity, shorter response time and other characteristics, and are very suitable for developing point-of-care (POC) devices in complex biological fluid samples. However, the reagent kit in the prior art is usually prepared in advance, and then assembled and connected with other elements for detection, which may be affected by many factors during use, such as the limited storage time of the liquid frame stability after the preparation of the hydrogel, and the detection activity of the sensitive element antigen antibody in it is affected with time. Therefore, it is necessary to develop a rapid, high-sensitivity, hydrogel-based portable sample detection scheme. Utility model content

[0005] The utility model discloses a kind of present preparation present measurement type hydrogel resonance protein sensing portable detection devices for the above problems, to realize the portable protein marker rapid quantitative detection of sample to be measured, with the advantages of simple detection operation, fast, high sensitivity, multi-target result simultaneously read out.

[0006] To achieve the above object, the technical scheme adopted by the utility model is as follows:

[0007] The utility model discloses a kind of present preparation present measurement type hydrogel resonance protein sensing portable detection devices, including step-by-step reaction module, hydrogel incubation module and detection module, wherein:

[0008] Step-by-step reaction module includes first base, cover plate and several flow channel units, and first base and cover plate form several cavities, and flow channel unit is built-in in cavity one by one, and flow channel unit includes several partition units rotatably connected with first base, corresponding cavity is separated into several cells for containing different reagents by each partition unit, and different reagents are mixed to carry out step-by-step reaction when different partition units are rotated;

[0009] Hydrogel incubation module includes incubation reaction pool and several mold racks, and mold rack is built-in in incubation reaction pool and corresponds to flow channel unit one by one, and the upper wall and the lower wall of mold rack are respectively attached to first split resonant ring and second split resonant ring with opening direction opposite, and mold rack also has containing slot for containing reagent, after reagent after step-by-step reaction enters mold rack and reacts with reagent in containing slot, and drop into the containing sample to be measured in containing slot, the sample to be measured contains protein marker;

[0010] Detection module is used to form induction field with the first split resonant ring and the second split resonant ring of corresponding mold rack and carry out detection.

[0011] Preferably, the upper wall of the mold rack is sealed with a tearable film, which is located above the first split resonant ring.

[0012] Preferably, the hydrogel incubation module further comprises a plurality of conversion units, which are arranged above the mold racks one by one and used for pouring the sample to be tested into the corresponding mold racks.

[0013] Preferably, the conversion unit comprises a splicing frame, a conversion support and a key, the splicing frame comprises a plurality of turnover blocks flexibly connected with different side walls of the mold racks, the tearable film is further connected with the turnover blocks and provided with a tearable line, the conversion support comprises a split positioning ring and a plurality of support rods, each support rod is crosswise arranged in the split positioning ring, one end of each support rod is arranged opposite to the key, and the other end of each support rod is connected with the turnover blocks one by one.

[0014] Preferably, the hydrogel incubation module further comprises an upper cover and a mounting seat arranged above the incubation reaction tank in sequence, the upper cover is provided with a first through hole and a plurality of second through holes, the mounting seat is provided with a sample adding port and a plurality of through grooves, the sample adding port corresponds to the first through hole, the through grooves correspond to the second through holes one by one, the second through holes correspond to the mold racks one by one, and the conversion units are arranged in the corresponding second through holes and the through grooves in sequence.

[0015] Preferably, the incubation reaction tank is provided with a plurality of first flow channel inlets corresponding to the flow channel units in communication, and the mold rack is provided with a second flow channel inlet corresponding to the first flow channel inlet in communication, and the reagent after the step-by-step reaction enters the mold rack through the first flow channel inlet and the second flow channel inlet in sequence.

[0016] Preferably, the step-by-step reaction module comprises two flow channel units arranged side by side, which are respectively referred to as a first flow channel unit and a second flow channel unit, the first base is provided with a flow channel separation column and two baffles side by side, the cover plate comprises two shell plates, the flow channel separation column is located between the two baffles for isolation, the two shell plates are arranged on the baffles one by one and attached to the two sides of the flow channel separation column to form two cavities, the flow channel unit further comprises a flow channel bottom plate and a partition plate, the flow channel bottom plate is attached to the inner bottom wall of the corresponding cavity, the partition plate is connected to one end of the flow channel bottom plate away from the hydrogel incubation module, and is vertically attached to the first base and the shell plate to realize sealing of the corresponding cells, and a plurality of positioning blocks are further arranged side by side on each baffle, and the separation unit is rotationally connected with the positioning blocks one by one.

[0017] Preferably, the four partition units are arranged side by side, and each of the four partition units comprises a lever, a rotating shaft and two elastic pads, the rotating shaft is rotatably connected to the first base, the lever is connected to the rotating shaft and is arranged obliquely through the first base, one end of the lever abuts against the inner wall of the first base, and the other end of the lever extends out of the first base, the two elastic pads are attached to the lever and the first base and are located at two acute angle regions formed by the lever and the first base respectively, and the four partition units divide the corresponding cavity into four cells for containing different reagents, i.e., a first cell is formed between the first partition unit and the first base, a second cell is formed between the first partition unit and the second partition unit, a third cell is formed between the second partition unit and the third partition unit, and a fourth cell is formed between the third partition unit and the fourth partition unit.

[0018] Preferably, the detection module comprises a second base, a sealing plate, at least one coil, a connecting wire, an SMA interface and an SMA female head, the sealing plate is located between the incubation reaction pool and the second base, the coil, the connecting wire and the SMA interface are arranged on the sealing plate, the coil is connected to the SMA interface through the connecting wire for inductive detection, and the SMA female head is electrically connected to the SMA interface and is located on the second base.

[0019] Preferably, the detection module is electrically connected to the network analyzer through a cable to realize reading out of detection data.

[0020] Compared with the prior art, the utility model has the beneficial effects that:

[0021] This application presents an integrated device capable of rapid on-site fabrication and immediate multi-channel rapid detection. It comprises a step-by-step reaction module, a hydrogel incubation module, and a detection module, achieving multi-functional miniaturized assembly and integration of sensitive elements (such as hydrogels), sample incubation, and result readout. The hydrogel incubation module can be converted via a pressing conversion unit. The effect of deformation generated during hydrogel coupling incubation within the mold holder on the crack resonant ring is detected. Specifically, the raw materials for the preparation of the core sensitive element, the multi-target immunohydrogel, are assembled in the step-by-step reaction module. After step-by-step mixing of raw materials, gelation and coupling with the crack resonant ring are achieved within the mold holder, resulting in a core detection element with high sensitivity to the detection target. The incubation cell can be converted by pressing the conversion unit mold holder. Untreated test samples are dripped into the incubation cell through the sample dispensing port, and after a period of reaction, a network analyzer can be used to connect to the detection module to read the test results. In summary, this application is highly operable and integrates flexible storage, preparation, and detection into one unit. By storing it in the form of raw materials and preparing it rapidly for use when needed, it can improve the long-term stability of the immune activity of immune elements such as protein biomarkers, with fewer interfering factors. The on-demand design maximizes the long-term stability of the product, with a long shelf life, optimal sensitivity and specificity, and the advantages of simplicity, accuracy, and speed. The experimental results have good repeatability and can be used for the quantitative detection of protein biomarkers, etc. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the structure of the portable detection device for on-the-spot fabrication and testing of hydrogel resonant proteins according to this utility model;

[0023] Figure 2 This is a schematic diagram of the internal structure of the portable detection device for on-the-spot fabrication and testing of hydrogel resonant proteins according to this utility model.

[0024] Figure 3 This utility model Figure 2 Exploded view;

[0025] Figure 4 This is an exploded view of the hydrogel incubation module and detection module of this utility model;

[0026] Figure 5 This is a schematic diagram of the internal structure of the hydrogel incubation module of this utility model;

[0027] Figure 6 This utility model Figure 5 Exploded view;

[0028] Figure 7 This utility model Figure 5 A schematic diagram of the transition states;

[0029] Figure 8The utility model discloses a present preparation present measurement type hydrogel resonance protein sensing portable detection device's readout principle schematic diagram;

[0030] Figure 9 The utility model discloses a present preparation present measurement type hydrogel resonance protein sensing portable detection device's detection method flow chart;

[0031] Figure 10 The utility model discloses a hydrogel preparation characterization result chart, wherein, (a) is the scanning electron microscope image of gold nano particle distribution in hydrogel, (b) is the local enlarged view of (a), (c) is the energy dispersion X ray spectrum line sweep result chart of hydrogel preparation characterization result chart;

[0032] Figure 11 The utility model discloses the change diagram of the porous structure of hydrogel before and after adding the sample to be measured, wherein, (a) is the scanning electron microscope image of the porous structure of hydrogel before adding the sample to be measured, (b) is the scanning electron microscope image of the porous structure of hydrogel after adding the sample to be measured, the upper graph in (c) is the relationship diagram of the pore diameter and the number of hydrogel before adding the sample to be measured, and the lower graph in (c) is the relationship diagram of the pore diameter and the number of hydrogel after adding the sample to be measured.

[0033] Figure 12 The utility model discloses the response effect diagram of the first protein marker incubation different time;

[0034] Figure 13 The utility model discloses the standard curve diagram of the sample to be measured after the incubation of different first protein marker standard sample concentration for 5 minutes.

[0035] Figure 14 The utility model discloses the response effect diagram of the second protein marker incubation different time;

[0036] Figure 15 The utility model discloses the standard curve diagram of the sample to be measured after the incubation of different second protein marker standard sample concentration for 5 minutes.

[0037] Explanation of reference signs: 1, step-by-step reaction module; 2, hydrogel incubation module; 3, detection module; 11, first base; 12, first flow channel unit; 13, second flow channel unit; 14, cover plate; 111, flow channel separation column; 112, baffle; 113, positioning block; 121, flow channel bottom plate; 122, partition; 123, separation unit; 12a, first grid; 12b, second grid; 12c, third grid; 12d, fourth grid; 123a, lever; 123b, pivot; 123c, elastic pad; 21, incubation reaction pool; 22, mold rack; 23, conversion unit; 24, upper cover; 25, mounting seat; 26, button; 27, first split-port resonant ring; 28, second split-port resonant ring; 211, first flow channel inlet; 221, second flow channel inlet; 231, splicing rack; 232, easy-to-tear film; 233, conversion support; 233a, support rod; 233b, movable opening positioning ring; 251, sample addition port; 31, second base; 32, sealing plate; 33, coil; 34, connecting wire; 35, SMA interface; 36, SMA female head; 10, detection device; 20, network analyzer; 30, cable. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present application.

[0039] It should be noted that when a component is referred to as being "connected" with another component, it can be directly connected with the other component or there can be a component in the middle. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0040] Embodiment 1

[0041] As shown in Figures 1-8 A kind of hydrogel resonant protein sensing portable detection device of present preparation present measurement type, including step-by-step reaction module 1, hydrogel incubation module 2 and detection module 3, wherein:

[0042] The step-by-step reaction module 1 includes a first base 11, a cover plate 14, and several flow channel units. The first base 11 and the cover plate 14 form several cavities. The flow channel units are built into the cavities one by one. Each flow channel unit includes several partition units 123 that are rotatably connected to the first base 11. Each partition unit 123 divides the corresponding cavity into several grids for holding different reagents. When the different partition units 123 are rotated, the different reagents are mixed to carry out the step-by-step reaction.

[0043] The hydrogel incubation module 2 includes an incubation reaction pool 21 and several mold racks 22. The mold racks 22 are built into the incubation reaction pool 21 and correspond one-to-one with the flow channel units. The upper and lower walls of the mold racks 22 are respectively attached to the first slit resonant ring 27 and the second slit resonant ring 28 with opposite opening directions. The mold racks 22 also have a reagent receiving tank. After the reagents after the step-by-step reaction enter the mold racks 22 and react with the reagents in the receiving tank to form a hydrogel, the sample to be tested is dripped into the receiving tank. The sample to be tested contains protein markers.

[0044] The detection module 3 is used to form an induction field with the first crack resonant ring 27 and the second crack resonant ring 28 of the corresponding mold frame 22 for detection.

[0045] Among them, such as Figure 1 As shown, the hydrogel incubation module 2 is located directly above the detection module 3, and the number of flow channel units in the step-by-step reaction module 1 can be adjusted according to actual needs. The opening directions of the first slit resonant ring 27 and the second slit resonant ring 28 are opposite. The cover plate 14 can be an integral structure or a split structure, such as multiple flow channel units sharing one or each flow channel unit corresponding to one. The corresponding reagents can be pre-placed in each corresponding cavity during the production process. When the separator unit 123 rotates, the reagents will be mixed and enter the mold frame 22 in the hydrogel incubation module 2. After the reagent reaction is completed and gel is formed, it can be used to contact the sample to be tested to achieve on-site detection. The detection device 10 is the abbreviation of the on-the-spot hydrogel resonant protein sensing portable detection device of this application. The sample to be tested can be an environmental liquid sample, saliva sample, serum sample, etc. containing protein markers, such as containing inflammatory factors and viral antigens at least one of them.

[0046] The present application discloses a portable detection device for the present-preparation and present-measurement type hydrogel resonance protein sensor, which is an integrated device capable of rapid on-site preparation and immediate multiplex rapid detection, and comprises a step-by-step reaction module, a hydrogel incubation module and a detection module, so as to realize the multifunctional miniaturized assembly integration of sensitive element (such as hydrogel) preparation, sample addition and incubation, result reading and the like, wherein the hydrogel incubation module can be transformed by pressing a transformation unit, and the influence of the deformation of the hydrogel generated by the coupling incubation in the mold frame on the split resonant ring can be detected, that is, the raw materials for preparing the core sensitive element multi-target immune hydrogel are assembled in the step-by-step reaction module, the raw materials are mixed step by step, and then the hydrogel is formed in the mold frame and coupled with the split resonant ring, so that the core detection element with high sensitivity to the detection target is obtained. By pressing the transformation unit, the mold frame can be transformed into an incubation tank, and the untreated sample to be detected can be added from the sample addition port and reacted in the incubation tank for a period of time. Then, the network analyzer is connected to the detection module to realize the reading of the detection result. The device has the advantages of simple operation, high accuracy and rapidity, and can maximize the long-term stability of the product, has a long shelf life, high sensitivity and specificity, and is simple, accurate and rapid.

[0047] In an embodiment, the upper wall of the mold frame 22 is sealed with a tearable film 232, and the tearable film 232 is located above the first split resonant ring 27. The tearable film 232 can realize sealing protection and facilitate the reaction of the reagent after the step-by-step reaction with the reagent in the containing groove of the mold frame 22 to form the hydrogel.

[0048] In an embodiment, the hydrogel incubation module 2 further comprises a plurality of transformation units 23, which are arranged one by one above the mold frame 22 and are used to guide the sample to be detected into the corresponding mold frame 22. The transformation unit 23 facilitates the sample to be detected to flow into the corresponding mold frame 22, such as tearing the tearable film 232 to expose the immune response hydrogel formed in the mold frame 22.

[0049] In an embodiment, the transformation unit 23 comprises a splicing frame 231, a conversion support 233 and a button 26, the splicing frame 231 comprises a plurality of flip blocks flexibly connected with different side walls of the mold frame 22, the tearable film 232 is further connected with the flip blocks and provided with a tearable line, the conversion support 233 comprises a split positioning ring 233b and a plurality of support rods 233a, each support rod 233a is crosswise arranged in the split positioning ring 233b, and one end of each support rod 233a is arranged opposite to the button 26, and the other end of each support rod 233a is connected with the flip blocks one by one.

[0050] The three supporting rods 233a are connected with the turnover blocks on the mold frame 22 in a one-to-one correspondence, and the intersection of the three supporting rods 233a is flexibly positioned by the split positioning ring 233b. The upper end of the supporting rod 233a is movably connected with the button 26, and the button 26 is located above the supporting rod 233a. After the button 26 is pressed, the intersection angle of the three supporting rods 233a increases, the split positioning ring 233b at the intersection moves upward, and the lower end of the supporting rod 233a pushes the turnover blocks outward, thereby completing the demolding of the sandwiched resonant sensor and exposing the sandwiched resonant sensor in the incubation reaction tank 21. The sandwiched hydrogel resonant sensor is composed of the first split resonant ring 27, the second split resonant ring 28, and the immune response hydrogel coupled in the middle of the mold frame 22. The rapid demolding and exposure can be realized by the conversion unit 23, so as to facilitate the full reaction of the sample to be measured.

[0051] It is easy to understand that the upper end of the supporting rod 233a is movably connected with the button 26, which can be that the button 26 is not connected with the upper end of the three supporting rods 233a or is slidably connected with the upper end of the three supporting rods 233a. When the button 26 is pressed, the three supporting rods 233a diverge (the included angle increases), and the lower end of the three supporting rods 233a diverges and the distance between the lower ends increases, thereby pushing the turnover blocks on the mold frame 22 and tearing the easy-to-tear film 232. Alternatively, the button 26 located above the supporting rod 233a can be connected with other devices, such as the mounting seat 25, to realize the pressing function.

[0052] In an embodiment, the hydrogel incubation module 2 further comprises an upper cover 24 and a mounting seat 25 which are sequentially stacked above the incubation reaction tank 21. The upper cover 24 is provided with a first through hole and a plurality of second through holes. The mounting seat 25 is provided with a sample adding port 251 and a plurality of through grooves. The sample adding port 251 corresponds to the first through hole, the through grooves correspond to the second through holes in a one-to-one correspondence, and the second through holes correspond to the mold frame 22 in a one-to-one correspondence. The conversion unit 23 is sequentially arranged in the corresponding second through holes and the through grooves.

[0053] The number of sample adding ports 251 can be adjusted according to actual needs, and each sample adding port 251 can correspond to a flow channel unit, or a plurality of flow channel units can share one sample adding port 251. The upper cover 24 and the mounting seat 25 are arranged to avoid accidental touching of the conversion unit 23.

[0054] In an embodiment, the incubation reaction tank 21 is provided with a plurality of first flow channel inlets 211 which communicate with the flow channel units in a one-to-one correspondence. The mold frame 22 is provided with a second flow channel inlet 221 which communicates with the corresponding first flow channel inlet 211. The reagents after the step-by-step reaction enter the mold frame 22 through the first flow channel inlet 211 and the second flow channel inlet 221 in sequence. The step-by-step reaction module 1 is located on the side of the first flow channel inlet 211 of the hydrogel incubation module 2.

[0055] In an embodiment, the step-by-step reaction module 1 comprises two flow channel units arranged side by side, respectively denoted as a first flow channel unit 12 and a second flow channel unit 13, the first base 11 is provided with a flow channel partition column 111 and two baffles 112 arranged side by side, the cover plate 14 comprises two shell plates, the flow channel partition column 111 is located between the two baffles 112 for isolation, the two shell plates are respectively arranged on the baffles 112 and attached to the two sides of the flow channel partition column 111 to form two cavities, the flow channel unit further comprises a flow channel bottom plate 121 and a partition plate 122, the flow channel bottom plate 121 is arranged on the inner bottom wall of the corresponding cavity, the partition plate 122 is connected to one end of the flow channel bottom plate 121 away from the hydrogel incubation module 2, and is vertically attached to the first base 11 and the shell plate to realize sealing of the corresponding cells, a plurality of positioning blocks 113 are arranged side by side on each baffle 112, and the partition unit 123 is rotationally connected with the positioning blocks 113 one by one.

[0056] The partition plate 122 is used to ensure the one-way flow of the reagent liquid in the first flow channel unit 12 and the second flow channel unit 13, and the reagent liquid does not leak out from the partition plate 122. The flow channel bottom plate 121 is used to increase the height of the bottom of the first flow channel unit 12 and the second flow channel unit 13, to form better close contact with the partition unit 123, to ensure the relative sealing of each cell, and to prevent the reagent from mixing when the partition unit 123 is not rotated. At the same time, the surface of the flow channel bottom plate 121 has hydrophobic properties, so that the reagent does not remain in the flow channel and completely enters the next chamber (such as a cell or a mold rack containing groove). The partition unit 123 is rotationally connected with the positioning blocks 113 on the baffle 112 one by one, and the partition units 123 on each flow channel unit are arranged side by side. When the step-by-step reaction module 1 comprises two flow channel units arranged side by side, the partition units 123 of the two flow channel units can be arranged in an eight-character shape, and are arranged at an acute angle with the flow channel partition column 111 to facilitate the flow of the reagent.

[0057] In an embodiment, the four partition units 123 are arranged side by side, including a lever 123a, a rotating shaft 123b and two elastic pads 123c. The rotating shaft 123b is rotatably connected to the first base 11. The lever 123a is connected to the rotating shaft 123b and is arranged obliquely through the first base 11. One end of the lever 123a abuts against the inner wall of the first base 11, and the other end extends out of the first base 11. The two elastic pads 123c are attached to the lever 123a and the first base 11, and are respectively located in the two acute angle regions formed by the lever 123a and the first base 11. The four partition units 123 divide the corresponding cavities into four cells for containing different reagents, i.e., the first cell 12a is formed between the first partition unit 123 and the first base 11 away from the hydrogel incubation module 2, the second cell 12b is formed between the first partition unit 123 and the second partition unit 123 away from the hydrogel incubation module 2, the third cell 12c is formed between the second partition unit 123 and the third partition unit 123 away from the hydrogel incubation module 2, and the fourth cell 12d is formed between the third partition unit 123 and the fourth partition unit 123 away from the hydrogel incubation module 2.

[0058] In an embodiment, the four partition units 123 are arranged side by side, including a lever 123a, a rotating shaft 123b and two elastic pads 123c. The rotating shaft 123b is rotatably connected to the first base 11. The lever 123a is connected to the rotating shaft 123b and is arranged obliquely through the first base 11. One end of the lever 123a abuts against the inner wall of the first base 11, and the other end extends out of the first base 11. The two elastic pads 123c are attached to the lever 123a and the first base 11, and are respectively located in the two acute angle regions formed by the lever 123a and the first base 11. The four partition units 123 divide the corresponding cavities into four cells for containing different reagents, i.e., the first cell 12a is formed between the first partition unit 123 and the first base 11 away from the hydrogel incubation module 2, the second cell 12b is formed between the first partition unit 123 and the second partition unit 123 away from the hydrogel incubation module 2, the third cell 12c is formed between the second partition unit 123 and the third partition unit 123 away from the hydrogel incubation module 2, and the fourth cell 12d is formed between the third partition unit 123 and the fourth partition unit 123 away from the hydrogel incubation module 2.

[0059] In an embodiment, the detection module 3 includes a second base 31, a cover plate 32, at least one coil 33, a connecting line 34, an SMA interface 35 and an SMA female head 36. The cover plate 32 is located between the incubation reaction pool 21 and the second base 31. The coil 33, the connecting line 34 and the SMA interface 35 are arranged on the cover plate 32. The coil 33 is connected to the SMA interface 35 through the connecting line 34 for inductive detection of the mold rack 22. The SMA female head 36 is electrically connected to the SMA interface 35 and is located on the second base 31.

[0060] In an embodiment, the detection module 3 is further electrically connected to the network analyzer 20 through the cable 30 for reading out the detection data. The network analyzer 20 is electrically connected to the SMA female head 36 of the detection module 3 through the cable 30 for displaying the detection data, which is helpful for intuitive observation of the detection results and improvement of work efficiency.

[0061] Working Principle: The portable hydrogel resonant protein sensor detection device for on-demand testing contains several sandwich-shaped resonant sensors. Each sandwich-shaped resonant sensor consists of a first slit resonant ring 27, a second slit resonant ring 28, and an immunoresponsive hydrogel coupled between the mold frame 22. When in contact with free antigens in the sample, according to a competitive immunization strategy, the free protein marker antigens in the sample cause the hydrogel to expand and deform mechanically, thereby increasing the distance between the two slit resonant rings. The expansion of the hydrogel directly affects the capacitance of the resonant sensor, and thus its resonant frequency. At the output end, a network analyzer connected to the coil 33 of the detection module 3 enables wireless signal reading of the detection data and can transmit the detection data to a mobile terminal (such as a tablet, computer, etc.).

[0062] The following will elaborate on this point, such as Figures 9-15 The diagram illustrates the detection method using a portable, on-the-spot hydrogel resonant protein sensor in this embodiment, comprising the following steps:

[0063] S1. Following the order closest to the hydrogel incubation module 2, sequentially load the first reagent, second reagent, third reagent, and fourth reagent into the corresponding flow channel unit's grid, where:

[0064] The first reagent was ultrapure water and acrylamide;

[0065] The second reagent consisted of a vinylized product obtained by mixing 1 μg of protein marker with 0.5 μL of N-succinimide acrylate and then purifying it by water bath and ultrafiltration, as well as 0.5 μL of N,N'-methylenebisacrylamide and 0.75 μL of N,N,N',N'-tetramethylethylenediamine.

[0066] The third reagent is 0.2 μL-0.25 μL of ammonium persulfate;

[0067] The fourth reagent is a solution obtained by adding 16 ng of protein marker antibody to 1 mL of gold nanoparticles and incubating in the dark on a shaker.

[0068] Alternatively, the first reagent may be ultrapure water, acrylamide, 0.5 μL N,N'-methylenebisacrylamide and 0.75 μL N,N,N',N'-tetramethylethylenediamine;

[0069] The second reagent was a vinylized product obtained by mixing 1 μg of protein marker with 0.5 μL of N-succinimide acrylate and then purifying it by water bath and ultrafiltration.

[0070] The third reagent is 0.2 μL-0.25 μL of ammonium persulfate;

[0071] The fourth reagent is 25 μL of a solution obtained by adding 16 ng of a protein marker antibody to 1 mL of gold nanoparticles and incubating on a shaker in the dark.

[0072] Specifically, the split-ring resonator is also pre-processed before assembly, and the pre-processing includes sequentially performing ultrapure water cleaning, nitrogen blowing and oxygen plasma surface treatment. The oxygen plasma surface treatment time is 3 min-6 min. The ultrapure water is 31 μL, and the acrylamide is 7.5 mg-9 mg; the concentration of N,N'-methylene bisacrylamide is 20 mg / mL, and the concentration of N,N,N',N'-tetramethyl ethylenediamine is 0.6 mol / L-0.8 mol / L; the temperature of the shaker incubation in the dark is 4°C, and the time is 8 hours-14 hours; the concentration of ammonium persulfate is 2 mol / L. The temperature of the water bath is 35°C-37°C, and the time is 50 minutes-1 hour; the ultrafiltration purification is 14000 g centrifugation, and the time is 25 minutes-40 minutes at 2°C-6°C. Among them, RPM (revolutions per minute) and RCF (relative centrifugal force or g force) are usually used to describe the speed of centrifugation, that is, RCF refers to the acceleration applied to the sample to be tested, and 14000 g indicates that the centrifugal force is 14000 times the acceleration of gravity on earth.

[0073] S2, 1 μL-1.25 μL of ammonium persulfate is loaded into the mold rack 22.

[0074] S3, the on-site preparation and on-site measurement type hydrogel resonant protein sensing portable detection device is vertically placed, the first reagent and the second reagent are mixed and shaken to form a first solution, then the first solution and the third reagent are mixed and shaken to form a second solution after standing for a first predetermined time, and finally the remaining separation units 123 are actuated to mix the second solution and the fourth reagent into the mold rack 22 and vertically stand for a second predetermined time to obtain the reagent after the step-by-step reaction, that is, the hydrogel, and complete the coupling of the hydrogel and the split-ring resonator of the corresponding mold rack 22. The first predetermined time is preferably 20 seconds, and the second predetermined time is preferably 2 minutes-4 minutes.

[0075] S4, the on-site preparation and on-site measurement type hydrogel resonant protein sensing portable detection device is horizontally placed, and the switching unit 23 corresponding to the flow channel unit is pressed to tear the easy-to-tear film 232.

[0076] S5, the initial detection result f0 is read out by using the detection module 3.

[0077] S6, the sample to be tested is dropped into the hydrogel, and after standing for a third predetermined time, the secondary detection result f1 is read out by using the detection module 3. The third predetermined time is preferably 5 minutes.

[0078] S7, according to the initial detection result f0 and the secondary detection result f1 and combining the corresponding linear equation, the content detection result of the protein marker in the sample to be detected is calculated, that is, (f1-f0) / f0 is taken as the normalized resonance frequency offset of the corresponding linear equation to obtain.

[0079] The present application also provides a present-preparation present-detection type hydrogel resonance protein sensing portable detection device. The device comprises two flow channel units arranged side by side, one of which is used for detecting the content of a first protein marker, and the other is used for detecting a second protein marker. The two flow channel units are symmetrically arranged, and each flow channel unit comprises four lattices arranged side by side. The first protein marker is an inflammatory factor, and the second protein marker is a virus. The specific steps are as follows:

[0080] I. First protein marker content detection:

[0081] 1. After the split gap resonator is cleaned with ultrapure water and dried with nitrogen, the surface is treated with oxygen plasma for 3 minutes to enhance hydrophilicity. Then, the split gap resonator is assembled on the upper wall and lower wall of the mold frame, and the split gaps of the two split gap resonators are in opposite directions. The size of the split gap resonator can be selected according to actual needs. In this embodiment, the size of the split gap resonator is 10 mm x 10 mm.

[0082] 2. 31 μL of ultrapure water and 8.5 mg of acrylamide are assembled in the first lattice 12a of the first flow channel unit 12.

[0083] 3. 1 μg of the first protein marker is mixed with 0.5 μL of N-succinimidyl acrylate and placed in a 36°C water bath for one hour. The vinylated first protein marker is obtained by ultrafiltration purification and assembled in the second lattice 12b of the first flow channel unit 12.

[0084] 4. 0.5 μL of 20 mg / mL N,N'-methylene bisacrylamide and 0.75 μL of 0.8 mol / L N,N,N',N'-tetramethyl ethylenediamine are assembled in the second lattice 12b of the first flow channel unit 12.

[0085] 5. 0.25 μL of 2 mol / L ammonium persulfate is assembled in the third lattice 12c of the first flow channel unit 12.

[0086] 6. 16 ng of the first protein marker antibody is added to 1 mL of gold nanoparticles and incubated at 4°C on a shaking table in the dark for 12 hours. 25 μL of the incubated solution is assembled in the fourth lattice 12d of the first flow channel unit 12.

[0087] 7. 1 μL of 2 mol / L ammonium persulfate is assembled in the mold frame 22.

[0088] 8. Start using, the detection device is placed vertically, dial the first partition unit 123, mix the reagent in the first grid 12a and the second grid 12b, shake gently for 5 seconds, dial the second partition unit 123, mix the mixed solution in the second grid 12b and the reagent in the third grid 12c, stand still for 20 seconds after shaking gently for 5 seconds, dial the third and fourth partition units 123, the mixed solution in the third grid 12c and the reagent in the fourth grid 12d flow into the mold frame 22, and then stand vertically for 3 minutes, thus completing the preparation of the core response element first protein marker detection hydrogel and its coupling with the split resonant ring.

[0089] 9. Place the detection device horizontally, press the button 26 of the conversion unit 23 of the first flow channel unit 12, the conversion support 233 is linked to push the splicing frame 231 around, and the easy-to-tear film 232 is separated and separated, and the core response element (i.e. hydrogel) is demolded.

[0090] 10. The initial detection result f0 is read out by using the network analyzer connected with the detection module 3.

[0091] 11. The sample to be measured is added from the sample adding port 251, and the secondary detection result f1 is read out by using the network analyzer after standing for 5 minutes.

[0092] 12. The first protein marker content detection result in the sample to be measured is calculated according to the linear equation y=0.48188lg(x)+3.62886, wherein x is the concentration of the first protein marker in the sample to be measured, unit: pg / mL, and y is the normalized resonance frequency offset, which is the ratio of the difference between the initial detection result and the secondary detection result to the initial detection result, that is, (f1-f0) / f0.

[0093] The experimental results, as shown in Figures 10 to 13 , show the preparation and characterization of the core sensitive element hydrogel. Among them, Figure 10 show the distribution of gold nanoparticles in the hydrogel. The existence of Au, N and S elements in the hydrogel verifies the successful polymerization of gold nanoparticles and antibodies on the structure of the hydrogel, and this result verifies the feasibility of the utility model. 11 clearly shows the change of the porous structure of the hydrogel before and after adding the sample to be measured. The pore size increases from 62.55nm to 162.73nm after adding the sample to be measured, which verifies the mechanical deformation of the hydrogel after the protein in the sample to be measured is combined, the expansion increases the distance between the two split resonant rings of the up-down coupling, and then causes the change of the resonant frequency of the result reading, which verifies the mechanism of reading the frequency wireless signal from the detection target combined signal. Figure 12The response effect of the first protein marker incubated for different time (5 minutes, 10 minutes, 15 minutes) is shown, wherein the standard sample selects the first protein marker (100 pg / mL CRP) at a concentration of 100 pg / mL, and the blank control indicates that the control variable has no corresponding standard sample and other consistent detection solution. The results show that the detection device can detect the quantitative results of the first protein marker in 5 minutes at the fastest, indicating that it has great detection speed advantage for portable application. Figure 13 The standard curve of the sample to be tested after incubation for 5 minutes under different concentrations of the first protein marker standard sample is shown. R 2 represents the sum of squares of the difference between the sample points and the mean line, blank represents the response result in the blank saliva control group without standard sample, and the detection limit (LOD) is calculated according to the formula 3σ / S, wherein σ represents the sensor noise, and S represents the sensitivity. The results show that a very wide linear detection range (10 -2 pg / mL to 10 5 pg / mL) can be obtained in 5 minutes of detection, and a very low detection lower limit (24 fg / mL) is obtained, indicating that it has excellent detection high sensitivity advantage for portable application. The normalized resonance frequency shift is Figures 12 to 15 the normalized frequency shift in the formula.

[0094] II. Second protein marker content detection:

[0095] 1. After the ultra-pure water of each split resonator ring is cleaned and dried by nitrogen blowing, the surface is treated by oxygen plasma for 3 minutes to enhance the hydrophilicity, and then the split resonator ring is assembled on the upper wall and lower wall of the mold frame, and the split openings of the two split resonator rings are opposite. The size of the split resonator ring can be selected according to actual needs, such as the size of the split resonator ring in this embodiment, which is 10 mm x 10 mm in length x width.

[0096] 2. 31 μL of ultra-pure water and 8.5 mg of acrylamide are assembled in the first grid 12a of the second flow channel unit 13.

[0097] 3. 1 μg of the second protein marker antigen is mixed with 0.5 μL of N-succinimidyl acrylate and placed in a 36°C water bath for one hour, and the vinylated second protein marker antigen is obtained by ultrafiltration purification and assembled in the second grid 12b of the second flow channel unit 13.

[0098] 4. 0.5 μL of 20 mg / mL N,N'-methylene bisacrylamide and 0.75 μL of 0.8 mol / L N,N,N',N'-tetramethyl ethylenediamine are assembled in the second grid 12b of the second flow channel unit 13.

[0099] 5. 0.25 μL of 2 mol / L ammonium persulfate is assembled in the third grid 12c of the second flow channel unit 13.

[0100] 6. Add 16 ng of the second protein marker antibody into 1 mL of gold nanoparticles, and incubate at 4°C for 12 hours in the dark on a shaker. Take 25 μL and assemble in the fourth lattice 12d of the second flow channel unit 13.

[0101] 7. Assemble 1 μL of 2 mol / L ammonium persulfate in the mold rack 22.

[0102] 8. When in use, place the detection device vertically, and press the first separation unit 123 to mix the reagents in the first lattice 12a and the second lattice 12b. Gently shake for 5 seconds, and then press the second separation unit 123 to mix the mixed solution in the second lattice 12b with the reagents in the third lattice 12c. Gently shake for 5 seconds, and then stand still for 20 seconds. Press the third and fourth separation units 123 to make the mixed solution in the third lattice 12c flow into the mold rack 22, and then stand still vertically for 3 minutes. Thus, the preparation of the core responsive element second protein marker detection hydrogel and the coupling with the split ring resonator are completed.

[0103] 9. Place the detection device horizontally, press the button 26 of the conversion unit 23 of the second flow channel unit 13, and the conversion support 233 will push the splicing rack 231 and the easy-to-tear film 232 around to fall down and separate, respectively. Thus, the core responsive element (i.e., the hydrogel) is demolded.

[0104] 10. Read the initial detection result f0 by using the network analyzer connected with the detection module 3.

[0105] 11. Add more than 600 μL of the sample to be tested from the sample adding port 251, and stand still for 5 minutes. Then, read the second detection result f1 by using the network analyzer.

[0106] 12. Calculate the second protein marker content detection result of the sample to be tested according to the linear equation y = 0.44938lg(x) + 1.70794, wherein x is the concentration of RSV (the second protein marker) in the sample to be tested, in pg / mL, and y is the normalized resonance frequency shift, which is the ratio of the difference between the initial detection result and the second detection result and the initial detection result, i.e., (f1-f0) / f0.

[0107] The experimental results are shown in Figure 14 , Figure 15 . Figure 14The second protein marker incubated for different time (5 minutes, 10 minutes, 15 minutes) shows the response effect, and the standard sample selects 100 pg / mL concentration of the second protein marker (100 pg / mL RSV). The blank control indicates that the control variable has no corresponding standard sample and the other detection solutions are consistent. The results show that the detection device can detect the quantitative results of the second protein marker in 5 minutes at the fastest speed, indicating that it has great detection speed advantage for portable application. Figure 15 The standard curve of the second protein marker standard sample concentration under different sample incubation times is shown in the figure R 2 The difference between the sample point and the mean line is represented by the square sum of the blank, and the response result in the blank saliva control group without standard sample. The detection limit (LOD) is calculated according to the formula 3σ / S, where σ represents the sensor noise and S represents the sensitivity. The results show that a very wide linear detection range (10 -2 pg / mL to 10 5 pg / mL) can be obtained within 5 minutes of detection, and a very low detection lower limit (31 fg / mL) is obtained, indicating that it has excellent detection sensitivity advantage for portable application.

[0108] In addition, for the first reagent, ultrapure water, acrylamide, 0.5 μL N,N'-methylene bisacrylamide and 0.75 μL N,N,N',N'-tetramethyl ethylenediamine; the second reagent is 1 μg protein marker mixed with 0.5 μL N-succinimidyl acrylate, and then subjected to water bath and ultrafiltration purification to obtain the vinylated reactant; the third reagent is 0.2 μL-0.25 μL ammonium persulfate; the fourth reagent is 25 μL of the solution obtained by adding 16 ng of protein marker antibody to 1 mL of gold nanoparticles and incubating on a shaking bed in the dark. The specific detection method is the same as above, and the difference is only that the first and second lattices 12a and 12b are exchanged during pre-configuration, but it does not affect the specific detection steps and detection results, which will not be described here.

[0109] It is easy to understand that according to the structure of the present preparation and present measurement type hydrogel resonance protein sensing portable detection device (referred to as detection device), the type of protein marker of the detected sample can be adjusted adaptively, such as each flow channel unit for measuring different types of protein markers or different types of protein markers for separate detection or mixed detection. And the above parameter values can be adjusted arbitrarily within the specific limited range, which can be determined according to actual needs.

[0110] The in-situ preparation and in-situ measurement type hydrogel resonance protein sensing portable detection device has high sensitivity, high specificity, few interference factors, and the in-situ preparation and in-situ use design can maximize the long-term stability of the product, long shelf life, optimal sensitivity and specificity, and has the advantages of simplicity, precision and speed. The experiment is fast and accurate, the experimental results are reproducible, and can be used for quantitative detection of protein markers. It can be used to detect whether it contains protein markers and the content of protein markers, such as blood samples, saliva samples, protein-containing environmental liquid samples (river water, etc.), food (drinking water) detection, etc. It can also be combined with other instruments in the prior art to further evaluate.

[0111] The technical features of the above-described embodiments can be combined in any manner. To make the description concise, all possible combinations of the technical features in the above-described embodiments are not described, but as long as the combinations of the technical features do not contradict, they should be considered within the scope of the present disclosure.

[0112] The above-described embodiments only express the more specific and detailed embodiments described in the present application, but should not be construed as limiting the scope of the application. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of the present application should be subject to the appended claims.

Claims

1. A kind of present preparation present measurement type hydrogel resonance protein sensing portable detection device, it is characterized by: The present application discloses a portable detection device for instant preparation and detection of hydrogel resonance protein sensor, which comprises a step-by-step reaction module (1), a hydrogel incubation module (2) and a detection module (3). The step-by-step reaction module (1) comprises a first base (11), a cover plate (14) and a plurality of flow channel units, the first base (11) and the cover plate (14) form a plurality of cavities, and the flow channel units are correspondingly arranged in the cavities, and the flow channel units comprise a plurality of separation units (123) rotatably connected with the first base (11), each separation unit (123) separates the corresponding cavity into a plurality of cells for accommodating different reagents, and different reagents are mixed for step-by-step reaction when different separation units (123) are rotated. The hydrogel incubation module (2) comprises an incubation reaction pool (21) and a plurality of mold racks (22), the mold racks (22) are arranged in the incubation reaction pool (21) and correspond to the flow channel units, the upper wall and the lower wall of the mold rack (22) are respectively attached with a first split resonant ring (27) and a second split resonant ring (28) with opposite opening directions, and the mold rack (22) further has a receiving groove for accommodating reagents, after the reagents after the step-by-step reaction enter the mold rack (22) and react with the reagents in the receiving groove to form hydrogel, the sample to be detected is dropped into the receiving groove, and the sample to be detected contains protein markers. The detection module (3) is used for forming an induction field with the first split resonant ring (27) and the second split resonant ring (28) corresponding to the mold rack (22) to detect.

2. The in-situ hydrogel resonant protein sensor portable detection device of claim 1, wherein: The upper wall of the mold rack (22) is sealed with a tearable film (232), and the tearable film (232) is located above the first split resonant ring (27).

3. The in-situ hydrogel resonant protein sensor device according to claim 2, wherein the hydrogel is a poly (N-isopropylacrylamide) (PNIPAm) hydrogel. The hydrogel incubation module (2) further comprises a plurality of conversion units (23), the conversion units (23) are arranged above the mold racks (22) one by one, and are used for guiding the sample to be detected into the corresponding mold racks (22).

4. The in-situ hydrogel resonant protein sensor device according to claim 3, wherein the hydrogel is a poly (N-isopropylacrylamide) (PNIPAm) hydrogel. The conversion unit (23) comprises a splicing rack (231), a conversion support (233) and a key (26), the splicing rack (231) comprises a plurality of flip blocks flexibly connected with different side walls of the mold rack (22), the tearable film (232) is further connected with the flip blocks and provided with a tearable line, the conversion support (233) comprises a movable mouth positioning ring (233b) and a plurality of support rods (233a), each support rod (233a) is crossly arranged in the movable mouth positioning ring (233b), one end of each support rod (233a) is arranged opposite to the key (26), and the other end is connected with the flip blocks one by one.

5. The in-situ hydrogel resonant protein sensor device according to claim 3, wherein the hydrogel is a poly (N-isopropylacrylamide) (PNIPAm) hydrogel. The hydrogel incubation module (2) further comprises an upper cover (24) and a mounting base (25) arranged in sequence above the incubation reaction tank (21), the upper cover (24) is provided with a first through hole and a plurality of second through holes, the mounting base (25) is provided with a sample adding port (251) and a plurality of through grooves, the sample adding port (251) corresponds to the first through hole, the through grooves correspond to the second through holes one by one, and the second through holes correspond to the mold racks (22) one by one, and the conversion unit (23) is arranged in the corresponding second through hole and through groove in sequence.

6. The in-situ hydrogel resonant protein sensor device of claim 1, wherein: The incubation reaction tank (21) is provided with a plurality of first flow channel inlets (211) corresponding to the flow channel unit in communication, and the mold rack (22) is provided with a second flow channel inlet (221) corresponding to the first flow channel inlet (211) in communication, and the reagent after the step-by-step reaction enters the mold rack (22) through the first flow channel inlet (211) and the second flow channel inlet (221) in sequence.

7. The in-situ hydrogel resonant protein sensor (HRPS) portable detection device of claim 1, wherein: The step-by-step reaction module (1) comprises two flow channel units arranged side by side, which are respectively referred to as first flow channel unit (12) and second flow channel unit (13), the first base (11) is provided with flow channel separation column (111) and two baffles (112) side by side, the cover plate (14) comprises two shell plates, the flow channel separation column (111) is located between the two baffles (112) for isolation, two shell plates are respectively attached to both sides of the flow channel separation column (111) to form two cavities, the flow channel unit further comprises flow channel bottom plate (121) and partition plate (122), the flow channel bottom plate (121) is attached to the inner bottom wall of the corresponding cavity, the partition plate (122) is connected to one end of the flow channel bottom plate (121) away from the hydrogel incubation module (2), and is vertically attached to the first base (11) and the shell plate to realize the sealing of the corresponding grid, a plurality of positioning blocks (113) are further arranged side by side on each baffle (112), and the separation unit (123) is rotatably connected with the positioning blocks (113) one by one.

8. The in-situ hydrogel resonant protein sensor device of claim 1, wherein: The four separation units (123) are arranged side by side and include a lever (123a), a rotating shaft (123b) and two elastic pads (123c). The rotating shaft (123b) is rotatably connected to the first base (11). The lever (123a) is connected to the rotating shaft (123b) and is arranged obliquely through the first base (11). One end of the lever (123a) abuts against the inner wall of the first base (11), and the other end extends out of the first base (11). The two elastic pads (123c) are attached to the lever (123a) and the first base (11) and are located at two acute angle regions formed by the lever (123a) and the first base (11), respectively. The four separation units (123) divide the corresponding cavities into four grids for accommodating different reagents, i.e., a first grid (12a) is formed between the first separation unit (123) away from the hydrogel incubation module (2) and the first base (11), a second grid (12b) is formed between the first separation unit (123) away from the hydrogel incubation module (2) and the second separation unit (123), a third grid (12c) is formed between the second separation unit (123) away from the hydrogel incubation module (2) and the third separation unit (123), and a fourth grid (12d) is formed between the third separation unit (123) away from the hydrogel incubation module (2) and the fourth separation unit (123).

9. The in-situ hydrogel resonant protein sensor (HRPS) portable detection device of claim 1, wherein: The detection module (3) includes a second base (31), a cover plate (32), at least one coil (33), a connecting line (34), an SMA interface (35) and an SMA female head (36). The cover plate (32) is located between the incubation reaction tank (21) and the second base (31). The coil (33), the connecting line (34) and the SMA interface (35) are arranged on the cover plate (32). The coil (33) is connected to the mold rack (22) one by one for inductive detection and is connected to the SMA interface (35) through the connecting line (34). The SMA female head (36) is electrically connected to the SMA interface (35) and is located on the second base (31).

10. The in-situ hydrogel resonant protein sensor device according to any one of claims 1 to 9, wherein: The detection module (3) is also electrically connected to the network analyzer (20) through a cable (30) to realize reading out of detection data.

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