Biochemical sensor array, method of manufacture, test strip and test kit

Abstract

The present disclosure relates to a biochemical sensor array, a preparation method, a detection test paper and a detection kit. The preparation method comprises: providing a substrate; forming a flexible layer on a second surface in a microelectronic printing manner, the flexible layer covering at least a through hole; forming a conductive layer on a first surface in a microelectronic printing manner, the conductive layer comprising a plurality of discrete conductive blocks, the conductive blocks covering a group of through holes, a first point and a second point; and forming a sensitive layer on a side of the flexible layer away from the substrate, the sensitive layer comprising a plurality of discrete film blocks, the film blocks at least partially overlapping the through holes. Thus, the flexible layer and the conductive layer are prepared in a microelectronic printing manner, the film thickness of the flexible layer and the conductive layer is accurately controlled, and the uniformity and repeatability of the biosensor are improved. The biochemical sensor array prepared comprises a plurality of biochemical sensors, and batch detection can be performed, thereby significantly improving the detection efficiency compared with a single biochemical sensor.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to the technical field of biosensor, and particularly relates to a biochemical sensing array, a preparation method, a detection test paper and a detection kit. BACKGROUND

[0002] The surface stress biosensor is a kind of biosensor, which can be applied to the fields of clinical medical diagnosis, biochemical detection, new drug discovery, etc. The detection principle of the surface stress biosensor is that the qualitative analysis of the to-be-detected biological molecules can be realized by modifying the sensor with a specific probe; after the to-be-detected biological molecules are combined with the corresponding probe, the sensitive part of the biosensor will produce a slight deformation, and the stress generated by the combination of the to-be-detected biological molecules with the probe of different concentrations is different, so that the quantitative analysis of the concentration of the to-be-detected biological molecules can be realized by detecting the electrical signals of different deformations.

[0003] In the related art, each film layer of the sensor is usually prepared by a spin coating method, which is difficult to accurately control the thickness of the film layer, resulting in poor uniformity and repeatability of the prepared biosensor. SUMMARY

[0004] In order to solve the above technical problems, the present disclosure provides a biochemical sensing array, a preparation method, a detection test paper and a detection kit.

[0005] In a first aspect, the present disclosure provides a preparation method of a biochemical sensing array, comprising:

[0006] providing a substrate; the substrate comprises a first surface and a second surface arranged oppositely, the first surface is provided with a plurality of through holes arranged in an array, a first trace and a plurality of second traces; the second trace corresponds to the through hole one by one, the periphery of the through hole is provided with a first point and a second point, the first trace is electrically connected with all the first points, and the second trace is electrically connected with the second point one by one;

[0007] forming a flexible layer on the second surface in a microelectronic printing manner; the flexible layer covers at least the through hole;

[0008] forming a conductive layer on the first surface in a microelectronic printing manner; the conductive layer comprises a plurality of discrete conductive blocks, and the conductive blocks cover a group of through holes, the first points and the second points;

[0009] forming a sensitive layer on the side of the flexible layer away from the substrate; the sensitive layer comprises a plurality of discrete film blocks, and the film blocks at least partially overlap with the through holes.

[0010] In a second aspect, the present disclosure further provides a biochemical sensing array, which is formed based on any one of the above preparation methods of the biochemical sensing array.

[0011] In a third aspect, the present disclosure further provides a detection test paper, comprising any one of the biochemical sensor arrays.

[0012] In a fourth aspect, the present disclosure further provides a detection test kit, comprising the detection test paper.

[0013] The technical solutions provided by the present disclosure have the following advantages compared with the prior art:

[0014] The biochemical sensor array, the preparation method, the detection test paper and the detection test kit provided by the present disclosure, the preparation method comprises: providing a substrate; the substrate comprises a first surface and a second surface arranged oppositely, the first surface is provided with a plurality of through holes arranged in an array, a first wire and a plurality of second wires; the second wire corresponds to the through hole one by one, the periphery of the through hole is provided with a first point and a second point, the first wire is electrically connected with all the first points, and the second wire is electrically connected with the second point one by one; a flexible layer is formed on the second surface in a microelectronic printing manner; the flexible layer covers at least the through hole; a conductive layer is formed on the first surface in a microelectronic printing manner; the conductive layer comprises a plurality of discrete conductive blocks, and the conductive block covers a group of through holes, first points and second points; a sensitive layer is formed on the side of the flexible layer away from the substrate; the sensitive layer comprises a plurality of discrete film blocks, and the film block at least partially overlaps with the through hole. Thus, the flexible layer and the conductive layer are prepared in a microelectronic printing manner, the film thickness of the flexible layer and the conductive layer is accurately controlled, which is beneficial to improving the uniformity and repeatability of the biochemical sensor; the prepared biochemical sensor array comprises a plurality of biochemical sensors, batch detection can be carried out, and compared with a single biochemical sensor, the detection efficiency is significantly improved. BRIEF DESCRIPTION OF DRAWINGS

[0015] The accompanying drawings, which are incorporated into and form part of the specification, illustrate embodiments consistent with the present disclosure and, together with the specification, serve to explain the principles of the present disclosure.

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present disclosure or the prior art, the accompanying drawings needed to be used in the embodiments or prior art description will be briefly introduced. Obviously, for those skilled in the art, other drawings can also be obtained from these drawings without creative labor.

[0017] Figure 1 A flowchart of a preparation method of a biochemical sensor array provided by the present disclosure is shown in the figure.

[0018] Figures 2-6 The structure diagram corresponding to each step in the preparation method of the biochemical sensor array provided by the present disclosure is shown in the figure.

[0019] Figure 7 The structure diagram of a substrate provided by the present disclosure is shown in the figure.

[0020] Figure 8 A detection result schematic diagram of the biochemical sensing array provided by the embodiment of the present disclosure. DETAILED DESCRIPTION

[0021] In order to enable a more clear understanding of the above-mentioned purposes, features and advantages of the present disclosure, the schemes of the present disclosure will be further described below. It should be noted that the embodiments of the present disclosure and the features in the embodiments can be combined with each other without conflict.

[0022] In the following description, a large number of specific details are set forth in order to facilitate a thorough understanding of the present disclosure, but the present disclosure can also be implemented in other manners different from those described herein; obviously, the embodiments described in the specification are only a part of the embodiments of the present disclosure, and not all the embodiments.

[0023] Figure 1 A flowchart of a preparation method of a biochemical sensing array provided by the embodiment of the present disclosure. Referring to Figure 1 , the preparation method of the biochemical sensing array comprises the following steps:

[0024] S110, providing a substrate.

[0025] In combination with Figures 2-3 , the substrate 10 comprises a first surface S1 and a second surface S2 arranged oppositely, the first surface S1 is provided with a plurality of through holes 11 arranged in an array, a first trace 12 and a plurality of second traces 13; the second trace 13 corresponds to the through hole 11 one by one, the periphery of the through hole 11 is provided with a first point 14 and a second point 15, the first trace 12 is electrically connected with all the first points 14, and the second trace 13 is electrically connected with the second points 15 one by one.

[0026] Exemplarily, the first surface S1 is the front surface of the substrate 10, and the second surface S2 is the back surface of the substrate 10.

[0027] Exemplarily, as Figure 2 shown, the first surface S1 of the substrate 10 is provided with 25 through holes 11, 1 first trace 12 and 25 second traces 13; the periphery of each through hole 11 is provided with 1 first point 14 and 1 second point 15, the first trace 12 is electrically connected with all the first points 14, and the second trace 13 is electrically connected with the second points 15 one by one.

[0028] In some embodiments, as Figure 2 shown, the first surface S1 of the substrate 10 is further provided with a signal interface 16, the signal interface 16 comprises a first signal interface 161 and a second signal interface 162 arranged in sequence along a first direction X, the first signal interface 161 is electrically connected with the first trace 12, and the second signal interface 162 is electrically connected with the second trace 13 one by one.

[0029] The substrate 10 comprises a flexible substrate, such as a flexible printed circuit (FPC).

[0030] S120, forming a flexible layer on the second surface in a micro-electronic printing manner.

[0031] In combination Figure 4 , the flexible layer 20 covers at least the through hole 11.

[0032] Exemplarily, the flexible layer 20 covers only the through hole 11.

[0033] Exemplarily, as shown in Figure 4 , the flexible layer 20 covers the through hole and the surface of the substrate 10 (i.e. part of the second surface S2) around the through hole 11.

[0034] Exemplarily, the flexible layer 20 covers the through hole 11 and the entire second surface S2 of the substrate 10.

[0035] In this step, the second surface S2 of the substrate 10 is a printing surface, the substrate 10 is placed at a predetermined position, and the printing position of the flexible layer 20 is set based on a printing coordinate system, which includes a printing start position and a printing end position.

[0036] The material of the flexible layer 20 is not limited in the embodiments of the present application, and all flexible materials known to those skilled in the art can be used, such as polydimethylsiloxane (PDMS), polyimide, polyethylene, ethylene-vinyl acetate copolymer, polyvinyl alcohol, polyethylene terephthalate and polymethyl methacrylate.

[0037] S130, forming a conductive layer on the first surface in a micro-electronic printing manner.

[0038] In combination Figure 5 , the conductive layer 30 comprises a plurality of discrete conductive blocks 31, and each conductive block 31 covers a group of through holes 11, first point positions 14 and second point positions 15. Figure 5 Exemplarily, as shown in , 25 through holes 11 are arranged on the substrate 10, one through hole 11 and the first point position 14 and the second point position 15 arranged around it form a group, and there are 25 groups in total, so the conductive layer 30 comprises 25 discrete conductive blocks 31, and each conductive block 31 covers a group of through holes 11, first point positions 14 and second point positions 15.

[0039] In this step, the first surface S1 of the substrate 10 is a printing surface, the substrate 10 is placed at a predetermined position, and the printing position of the conductive block 31 is set based on a printing coordinate system, so that the upper surface of each group of through holes 11, first point 14 and second point 15 covers the conductive block 31, and the conductive block 31 formed in contact with the first point 14 and the second point 15 forms an electrical connection, thereby turning on the first trace 12 and the second trace 13 of the group of sensors.

[0040] The preparation material of the conductive layer 30 is not limited in the embodiments of the present disclosure, including but not limited to silver nanowire, carbon nanotube, conductive silver paste, graphene and two-dimensional inorganic compound MXene.

[0041] S140, forming a sensitive layer on the side of the flexible layer away from the substrate.

[0042] In combination Figure 6 The sensitive layer 40 includes a plurality of discrete film blocks 41, and the film blocks 41 at least partially overlap the through holes 11. For example, as shown in Figure 6 The substrate 10 is provided with 25 through holes 11, and the sensitive layer 40 includes 25 discrete film blocks 41, and the film blocks 41 overlap the through holes 11.

[0043] In this step, the mask plate is overlapped with the second surface S2 of the substrate 10, and the mask plate is also provided with arrayed through holes, and the through holes of the mask plate at least partially overlap the through holes 11 of the substrate 10; then the sensitive layer 40 is formed on the side of the mask plate away from the substrate 10, and after the mask plate is removed, the discrete film blocks 41 are formed on the side of the flexible layer 20 away from the through holes 11. In some embodiments, the substrate 10 in the S110 step is used as a mask plate, and the substrate 10 is covered on another substrate 10 with a flexible layer 20, and the through holes 11 of the two substrates 10 overlap.

[0044] In the embodiments, the microelectronic printing has the advantage that the printing parameters are adjustable, the film layer thickness can be accurately controlled, the film layer can be standardized prepared, and the biochemical sensor can be standardized prepared. The obtained biochemical sensor array belongs to a surface stress biochemical sensor, which includes a plurality of arrayed biochemical sensors, each through hole corresponds to a biochemical sensor, and the biochemical sensors have good uniformity and repeatability, can be used for batch detection, and significantly improves the detection accuracy and detection efficiency.

[0045] The flexible layer 20 and the conductive layer 30 (or the conductive block 31) are printed by using a microelectronic printer, and the control accuracy of the film layer thickness can reach nanometer level.

[0046] In some embodiments, the steps of S130 and S140 can also be exchanged, that is, the sensitive layer 40 is prepared first, and then the conductive layer 30 is prepared.

[0047] It should be noted that,Figures 2-6 It is exemplarily illustrated that the biochemical sensor array includes 25 biochemical sensors and is arranged in a 5x5 array manner, and the first point 14 and the second point 15 are both square in shape, but this does not constitute a limitation to the preparation method of the biochemical sensor array provided in the embodiments of the present application. In other embodiments, a smaller or larger number of biochemical sensors can be arranged on the biochemical sensor array, for example, 32 or 20, and the first point 14 and the second point 15 can be arranged in other shapes, for example, rectangular, circular or triangular, which are not limited herein.

[0048] The preparation method of the biochemical sensor array provided in the embodiments of the present application includes: providing a substrate 10; the substrate 10 includes oppositely arranged first and second surfaces S1 and S2, and the first surface S1 is provided with a plurality of array-arranged through holes 11, a first trace 12 and a plurality of second traces 13; the second traces 13 correspond one-to-one to the through holes 11, the periphery of the through hole 11 is provided with a first point 14 and a second point 15, the first trace 12 is electrically connected to all the first points 14, and the second traces 13 are electrically connected to the second points 15 one-to-one; a flexible layer 20 is formed on the second surface in a microelectronic printing manner; the flexible layer 20 covers at least the through holes 11; a conductive layer 30 is formed on the first surface in a microelectronic printing manner; the conductive layer 30 includes a plurality of discrete conductive blocks 31, and the conductive blocks 31 cover a group of through holes 11, first points 14 and second points 15; a sensitive layer 40 is formed on the side of the flexible layer 20 away from the substrate 10; the sensitive layer 40 includes a plurality of discrete film blocks 41, and the film blocks 41 at least partially overlap the through holes 11. Thus, the flexible layer 20 and the conductive layer 30 are prepared in a microelectronic printing manner, the film thickness of the flexible layer 20 and the conductive layer 30 is precisely controlled, which is beneficial to improving the uniformity and repeatability of the biochemical sensor; the prepared biochemical sensor array includes a plurality of biochemical sensors, which can be used for batch detection, and compared with a single biochemical sensor, the detection efficiency is significantly improved.

[0049] In some embodiments, the flexible layer 20 includes a polydimethylsiloxane film layer; the step of "forming the flexible layer on the second surface in a microelectronic printing manner" includes the following steps:

[0050] The prepolymer and the curing agent are uniformly mixed at a mass ratio M:1 to obtain a mixed glue, and the mixed glue is vacuumized to remove bubbles; wherein 8≤M≤10;

[0051] The polydimethylsiloxane film layer is printed in a dispensing manner using the mixed glue as a raw material.

[0052] In this embodiment, before printing the polydimethylsiloxane film layer, the mixed glue needs to be prepared; the prepared mixed glue is poured into the raw material box of the printer, the second surface S2 of the substrate 10 is placed on the object table of the printing device, and the first printing parameter is set; after printing, when the polydimethylsiloxane film layer is solidified, the substrate 10 covered by the polydimethylsiloxane film layer is obtained.

[0053] The first printing parameter at least includes the first printing speed and the first printing air pressure.

[0054] In some embodiments, "printing the polydimethylsiloxane film layer by using the dispensing method" includes the following steps:

[0055] Printing the polydimethylsiloxane film layer by using the dispensing method based on the first printing speed and the first printing air pressure.

[0056] The first printing speed is 5-15 mm / s, and the first printing air pressure is 30-90 KPa.

[0057] The thickness of the printed polydimethylsiloxane film layer is negatively correlated with the first printing speed, that is, the faster the first printing speed, the smaller the thickness of the polydimethylsiloxane film layer, and the slower the first printing speed, the greater the thickness of the polydimethylsiloxane film layer. The thickness of the printed polydimethylsiloxane film layer is positively correlated with the first printing air pressure, that is, the greater the first printing air pressure, the greater the thickness of the polydimethylsiloxane film layer, and the smaller the first printing air pressure, the smaller the thickness of the polydimethylsiloxane film layer.

[0058] In this embodiment, the first printing speed is set to 5-15 mm / s, and the first printing air pressure is set to 30-90 KPa, so that the thickness of the polydimethylsiloxane film layer is controlled within an appropriate range, and the sensitivity and impact resistance are considered. If the first printing speed is greater than 15 mm / s and / or the first printing air pressure is less than 30 KPa, the thickness of the polydimethylsiloxane film layer is too thin, and the polydimethylsiloxane film layer is easy to break and damage. If the first printing speed is less than 5 mm / s and / or the first printing air pressure is greater than 90 KPa, the thickness of the polydimethylsiloxane film layer is too thick, and when the biological molecule to be detected is detected, the deformation amount of the sensor is reduced, and the sensitivity of the sensor is reduced.

[0059] For example, the first printing speed is set to 6 mm / s, or the first printing speed is set to 8 mm / s, or the first printing speed is set to 12 mm / s.

[0060] For example, the first printing air pressure is set to 90 KPa, or the first printing air pressure is set to 80 KPa.

[0061] In some embodiments, after the step of "printing the dimethicone film layer by dispensing", the preparation method further comprises the following steps:

[0062] heating the substrate at the first heating temperature until the dimethicone film layer solidifies.

[0063] In this embodiment, the just-printed dimethicone film layer still has certain viscosity, and the heating accelerates the solidification of the dimethicone film layer, thereby shortening the preparation time and avoiding damage to the dimethicone film layer in subsequent steps.

[0064] In this embodiment, the first heating temperature is set to 60-80℃, so as to shorten the solidification time of the dimethicone film layer as much as possible without damaging the structure of the dimethicone film layer. If the first heating temperature is too low (less than 60℃), the solidification speed of the dimethicone film layer is slow, and the waiting time is long. If the first heating temperature is too high (greater than 80℃), it exceeds the melting point of the un-solidified dimethicone, causing the dimethicone film layer to bubble and damage the structure of the film layer.

[0065] For example, the first heating temperature is 70℃, or the first heating temperature is 75℃.

[0066] It should be noted that all devices or equipment with heating function known to those skilled in the art can be used, such as a heating table or a drying box, which are not limited herein. There is a certain difference between the set temperature and the actual temperature of the heating device, and the actual temperature will fluctuate within the preset range of the set temperature. For example, the set temperature is 80℃, and the actual temperature is 80±5℃. For example, the set temperature is 60℃, and the actual temperature is 60±3℃.

[0067] For example, the first heating temperature is one of 65℃, 70℃ and 75℃.

[0068] In some embodiments, the conductive layer 30 comprises a silver paste layer; the step of "forming the conductive layer on the first surface by microelectronic printing" comprises the following steps:

[0069] The conductive silver paste is used as raw material, and the conductive blocks are printed by dispensing.

[0070] In this embodiment, the conductive silver paste is poured into the raw material box of the printer, the first surface S1 of the substrate 10 is placed on the object table of the printing device, and the second printing parameters are set. After printing, the silver paste layer is solidified to obtain a substrate 10 covered by a plurality of silver paste conductive blocks. The silver paste conductive blocks prepared in this embodiment have good conductivity, and each silver paste conductive block is in contact with and electrically connected to the same group of one point 14 and the second point, thereby connecting the first trace 12 and the second trace 13 of the sensor in the same group.

[0071] The second printing parameter comprises at least a second printing speed and a second printing air pressure.

[0072] In some embodiments, the printing the conductive block by the dispensing method comprises the following steps:

[0073] The conductive block is printed by the dispensing method based on the second printing speed and the second printing air pressure.

[0074] The second printing speed is 4-15 mm / s, and the second printing air pressure is 70-100 KPa.

[0075] The thickness of the printed conductive block 31 is negatively correlated with the second printing speed. The faster the second printing speed, the smaller the thickness of the conductive block 31. The slower the first printing speed, the greater the thickness of the conductive block 31. The thickness of the printed conductive block 31 is positively correlated with the second printing air pressure. The greater the second printing air pressure, the greater the thickness of the conductive block 31. The smaller the second printing air pressure, the smaller the thickness of the conductive block 31.

[0076] In this embodiment, the second printing speed is set to 4-15 mm / s, and the second printing air pressure is set to 70-100 KPa, so as to control the thickness of the conductive block within an appropriate range, taking into account the reliability and cost. If the first printing speed is greater than 15 mm / s and / or the first printing air pressure is less than 70 KPa, the thickness of the conductive block 31 is too thin, which leads to the first trace 12 and the second trace 13 not being conductive, affecting the reliability of the sensor. If the first printing speed is less than 4 mm / s and / or the first printing air pressure is greater than 100 KPa, the thickness of the conductive block 31 is too thick, increasing the preparation cost.

[0077] For example, the second printing speed is set to 4 mm / s, or the second printing speed is set to 5 mm / s, or the first printing speed is set to 10 mm / s.

[0078] For example, the second printing air pressure is set to 80 KPa, or the second printing air pressure is set to 100 KPa.

[0079] In some embodiments, after the printing of the conductive block by the dispensing method, the preparation method further comprises the following steps:

[0080] The substrate is heated at a second heating temperature until the conductive block solidifies.

[0081] The second heating temperature is 80-120°C, and the heating duration is 30-50 min.

[0082] In this embodiment, the freshly printed silver paste layer still has a certain fluidity, and heating is used to accelerate the solidification, thereby shortening the preparation time and avoiding the influence of subsequent operations on the uniformity of the film thickness and the breakage of the conductive block 31 structure.

[0083] The second heating duration is related to the thickness of the silver paste layer and the second heating temperature. When the thickness of the silver paste layer is fixed, the higher the second heating temperature, the shorter the second heating duration required for the silver paste layer to solidify; the lower the second heating temperature, the longer the second heating duration required for the silver paste layer to solidify. The second heating temperature in the embodiment is set to 80-120°C, which not only accelerates the solidification of the silver paste layer, but also does not affect the already solidified polydimethylsiloxane film layer.

[0084] For example, the second heating temperature is 80°C, or the second heating temperature is 100°C, or the second heating temperature is 120°C.

[0085] For example, the second heating duration is 30 min, or the second heating duration is 40 min, or the second heating duration is 50 min.

[0086] It should be noted that all devices or equipment with heating function known to those skilled in the art can be used, such as a heating table or a drying box, which is not limited herein. There is a certain difference between the set temperature and the actual temperature of the heating device, and the actual temperature will fluctuate within the preset range of the set temperature. For example, the set temperature is 100°C, and the actual temperature is 100±5°C. For example, the set temperature is 90°C, and the actual temperature is 90±5°C.

[0087] In some embodiments, the sensitive layer 40 includes a nano-gold layer; and the step of “forming a sensitive layer on the side of the flexible layer away from the substrate” includes the following steps:

[0088] The thin film block is formed on the side of the flexible layer away from the substrate by magnetron sputtering.

[0089] In the embodiment, the nano-gold layer is prepared by magnetron sputtering, which can accurately control the thickness and uniformity of the nano-gold layer. The prepared nano-gold layer is a thin film with high purity, density and uniformity, and has strong bonding force with the flexible layer 20 and high mechanical strength. In addition, the nano-gold layer has the advantages of high sputtering rate, simple operation, low operation temperature, low energy consumption, environmental protection, etc.

[0090] The magnetron sputtering conditions at least include a first sputtering duration.

[0091] The nano-gold layer is made of high-purity gold, and the particle size of a single particle reaches the nanometer level. Nano-gold has good biocompatibility and can combine with various biological molecules without affecting biological activity. The nano-gold layer prepared in the embodiment includes a plurality of discrete thin film blocks 41, and the thin film blocks 41 at least partially overlap the through holes 11.

[0092] In some embodiments, the step of “forming a thin film block on the side of the flexible layer away from the substrate by magnetron sputtering” includes the following steps:

[0093] sputtering a thin film block on the side of the flexible layer away from the substrate based on a first sputtering duration;

[0094] The first sputtering duration is 1-2 min.

[0095] In this embodiment, the thickness of the nano-gold layer is controlled by adjusting the first sputtering duration. The longer the first sputtering duration, the greater the thickness of the nano-gold layer. The shorter the first sputtering duration, the smaller the thickness of the nano-gold layer. Controlling the first sputtering duration in the range of 1-2 min ensures the binding performance of the nano-gold layer and the antibody, and prevents the cost from increasing due to the over-thickness of the film.

[0096] Exemplarily, the preparation method of the biochemical sensor array comprises the following steps:

[0097] (1) Configure a mixed glue of polydimethylsiloxane. Mix A glue (prepolymer) and B glue (curing agent) at a mass ratio of 10:1 and stir uniformly, and then vacuum to remove bubbles.

[0098] (2) Print a polydimethylsiloxane film layer. After pouring the prepared mixed glue into the raw material box of the micro-electronic printer, place the back of the flexible substrate on the stage, set the printing speed to 8 mm / s and the air pressure to 90 KPa; after printing, heat the flexible substrate at 80°C, and after the polydimethylsiloxane film layer solidifies, the flexible substrate with the back covered by the polydimethylsiloxane film layer is obtained.

[0099] (3) Print a silver paste layer. After pouring the conductive silver paste into the raw material box of the micro-electronic printer, place the front of the flexible substrate on the stage of the printer, set the printing speed to 5 mm / s and the air pressure to 80 KPa; after printing, heat the flexible substrate at 120°C for 30 min, and the flexible substrate with the front covered by the conductive layer is obtained.

[0100] (4) Form a nano-gold layer. Use a magnetron sputtering instrument to sputter a nano-gold layer on the surface of the polydimethylsiloxane film to obtain a nano-gold-polydimethylsiloxane (AuNPs-PDMS) composite film.

[0101] Exemplarily, the preparation method of the biochemical sensor array comprises the following steps:

[0102] (1) Configure a mixed glue of polydimethylsiloxane. Mix A glue (prepolymer) and B glue (curing agent) at a mass ratio of 10:1 and stir uniformly, and then vacuum to remove bubbles.

[0103] (2) printing the polydimethylsiloxane film layer. After the prepared mixture is poured into the raw material box of the micro-electronic printer, the back of the flexible substrate is placed on the stage, the printing speed is set to 6 mm / s, and the air pressure is set to 80 KPa; after printing is completed, the flexible substrate is heated at 80°C, and after the polydimethylsiloxane film layer is solidified, the flexible substrate with the back covered by the polydimethylsiloxane film layer is obtained.

[0104] (3) printing the silver paste layer. After the conductive silver paste is poured into the raw material box of the micro-electronic printer, the front of the flexible substrate is placed on the stage of the printer, the printing speed is set to 4 mm / s, and the air pressure is set to 100 KPa; after printing is completed, the flexible substrate is heated at 120°C for 30 min, and the flexible substrate with the front covered by the conductive layer is obtained.

[0105] (4) forming the nano-gold layer. The nano-gold layer is sputtered on the surface of the polydimethylsiloxane film by using a magnetron sputtering instrument, and a nano-gold-polydimethylsiloxane (AuNPs-PDMS) composite film is obtained.

[0106] In some embodiments, the preparation method further comprises the following steps:

[0107] antibodies are modified on the side of the film block away from the flexible layer;

[0108] The antibodies are used for specific binding with the biological molecules to be detected.

[0109] In the embodiment, the antibody solution is dropped on each film block 41 of the sensitive layer 40, and a preset time period is maintained, so that the antibody molecules are modified on the film block 41. When the biochemical sensor array prepared by using the embodiment is used for detection, different concentrations of the biological molecule solution to be detected are dropped on different biochemical sensors, so that the biological molecules to be detected are specifically combined with the antibodies; the flexible layer and the film of the sensitive layer are deformed, so that the distance between the conductive silver paste in the approximately conductive block becomes larger, the conductive path is reduced, and the resistance of the sensor is increased. The change value of the resistance can be detected by using a biological detector, and the detection of the biological molecules to be detected is realized.

[0110] For example, the sensitive layer 40 is a nano-gold layer, the antibody solution is dropped on each film block of the nano-gold layer, and a preset time period is maintained, so that the antibody molecules are modified on the nano-gold layer; then the biochemical sensor array is washed with a buffer solution to inhibit non-specific adsorption.

[0111] The preparation method of the biochemical sensor array provided in the embodiment of the application uses green and environmentally friendly raw materials, does not involve toxic materials such as fluorescent reagents, does not cause environmental pollution, and meets the green development concept of China. At the same time, the ecological benefits that may be generated are quantitatively analyzed and scientifically evaluated, which shows that the ecological benefits have a great promoting effect on economic sustainable development and full use of resources.

[0112] On the basis of the above-mentioned embodiments, the biochemical sensor array is formed based on any one of the preparation methods of the biochemical sensor array, has corresponding beneficial effects, and the same parts can be understood with reference to the above, and the following will not be described in detail.

[0113] In some embodiments, as shown in the biochemical sensor array includes a substrate 10, a flexible layer 20, a conductive layer 30 and a sensitive layer 40. Figures 2-6

[0114] The substrate 10 includes a first surface S1 and a second surface S2 arranged opposite to each other, the first surface S1 is provided with a plurality of through holes 11 arranged in an array, a first trace 12 and a plurality of second traces 13; the second trace 13 corresponds to the through hole 11 one by one, the circumference of the through hole 11 is provided with a first point 14 and a second point 15, the first trace 12 is electrically connected with all the first points 14, and the second trace 13 is electrically connected with the second point 15 one by one.

[0115] The flexible layer 20 is located on the second surface S2 of the substrate 10 and covers at least the through hole 11.

[0116] The sensitive layer is located on the side of the flexible layer 20 away from the substrate 10, and the sensitive layer 40 includes a plurality of discrete film blocks 41, and the film block 41 at least partially overlaps the through hole 11.

[0117] The conductive layer 30 is located on the first surface S1 of the substrate 10, and the conductive layer 30 includes a plurality of discrete conductive blocks 31, and the conductive block 31 covers a group of through holes 11, first points 14 and second points 15, and the conductive block 31 is in contact with the first point 14 and the second point 15 to form an electrical connection, thereby conducting the first trace 11 and the second trace 13 of the group of sensors.

[0118] The number of biochemical sensors included in the biochemical sensor array is equal to the number of through holes 11. For example, as shown in any one of the figures, the number of through holes 11 is 25, and the biochemical sensor array includes 25 biochemical sensors. Figures 2-6

[0119] It should be noted that, Figures 2-6 For example, it is exemplarily explained that the biochemical sensor array includes 25 biochemical sensors and is arranged in a 5x5 array manner, and the shapes of the first point 14 and the second point 15 are both squares, but this does not constitute a limitation on the biochemical sensor array provided by the embodiments of the present application. In other embodiments, a smaller or larger number of biochemical sensors can be arranged on the biochemical sensor array, for example, 32 or 20, and the first point 14 and the second point 15 can be arranged in other shapes, for example, rectangular, circular or triangular, which are not limited herein.

[0120] ​​In some embodiments, such as Figure 2 As shown, the first surface S1 of the substrate 10 is also provided with a signal interface 16. The signal interface 16 includes a first signal interface 161 and a second signal interface 162 arranged in sequence. The first signal interface 161 is electrically connected to the first trace 12, and the second signal interface 162 is electrically connected to the second trace 13 in a one-to-one correspondence.

[0121] The biochemical sensor array includes multiple signal access channels. Each signal access channel includes a first signal interface 161, a second signal interface 162, and a biochemical sensor connected between the first signal interface 161 and the second signal interface 162.

[0122] When measuring the biochemical sensor array using a biodetector, the signal interface 16 of the biochemical sensor array is inserted into the slot of the biodetector, so that the first signal interface 161 and the second signal interface 162 are electrically connected to the detection circuit inside the biodetector. The first signal interface 161 serves as a common signal interface, and the second signal interface 162 serves as a detection signal interface, detecting the signal of each signal access channel. For example, the signal of the signal access channel includes the resistance change value.

[0123] For example, such as Figure 2 As shown, 25 through holes 11 are provided on the substrate 10. The through holes 11 are arranged in a 5×5 array, corresponding to 25 sets of first points and second points, 1 first trace 12 and 1 first signal interface 161, 25 second traces 13 and 25 second signal interfaces 162. One end of the first trace 12 is electrically connected to the first signal interface 161, and the other end is electrically connected to all the first points 14. One end of the second trace 13 is electrically connected to the second signal interface 162 one by one, and the other end is electrically connected to the second point 15 one by one.

[0124] In some embodiments, the diameter of the through hole is 2.0 to 5.0 mm; the distance between the centers of adjacent through holes is 3.0 to 15 mm.

[0125] In some embodiments, the distance between the edge of the first point and the edge of the corresponding through hole is 0.3 to 1.0 mm; the distance between the edge of the second point and the edge of the corresponding through hole is 0.3 to 1.0 mm.

[0126] In some embodiments, the line width of the first trace is 0.2 to 0.5 mm; the line width of the second trace is 0.2 to 0.5 mm.

[0127] In some embodiments, the spacing between the first trace and the second trace, and between the second traces, is 0.2 to 0.5 mm.

[0128] In some embodiments, the width of the first signal interface is 0.3-0.7 mm, and the width of the second signal interface is 0.3-0.7 mm.

[0129] Exemplarily, as shown in Figure 7 , along the first direction X, the length L1 of the substrate 10 is 70 mm, the length L3 of the signal interface is 28.5 mm, the distance L5 between the centers of adjacent through holes 11 is 13.4 mm, the distance L6 between the edge of the first point 14 and the edge of the corresponding through hole 11 is 0.5 mm, and the distance L7 between the edge of the second point 15 and the edge of the corresponding through hole 11 is 0.5 mm; along the second direction Y, the width L2 of the array distribution area of the substrate 10 is 42 mm, and the width L4 of the tail area is 10 mm; the distance L5 between the centers of adjacent through holes 11 is 5 mm, and the line spacing L8 between the first trace 12 and the second trace 13 and between the second traces 13 is 0.5 mm; the diameter Φ of the through hole 11 is 3 mm; the line width of the first trace and the line width of the second trace are both 0.3 mm; along the first direction X, the width of the first signal interface 161 is 0.5 mm, and the width of the second signal interface 162 is 0.5 mm.

[0130] Exemplarily, the biochemical sensor array provided by the present disclosure is used to detect Escherichia coli O157:H7, and the specific detection process is as follows:

[0131] (1) Escherichia coli O157:H7 is diluted to 50, 100, 150, 200, and 250 CFU / mL, respectively.

[0132] (2) The above Escherichia coli O157:H7 with different concentrations is added dropwise on the biochemical sensor, and reacted for 30 min, so that the Escherichia coli O157:H7 specifically binds to the antibody.

[0133] (3) The resistance value is measured by using a portable in-vitro biological detector. In combination with Figure 5 , the biochemical sensor array has 26 signal interfaces, the first one on the left is the first signal interface (common signal interface), and the remaining 25 are the second signal interfaces (measurement signal interfaces); each second signal interface and the first signal interface are impedance test zones, and if the solution impedance of the test zone changes, the resistance measured by the two signal interfaces will change.

[0134] (4) The detection result is shown in Figure 8 , the resistance change value has a linear relationship with the concentration of Escherichia coli O157:H7, and the resistance change value increases with the increase of the concentration of Escherichia coli O157:H7, and the variance R 2 of the fitting formula is 0.98, indicating that the biochemical sensor array has good uniformity and repeatability.

[0135] On the basis of the above-mentioned embodiments, the embodiments of the present disclosure further provide a detection test paper, which comprises any one of the biochemical sensing arrays described above, and has corresponding beneficial effects. For the sake of avoiding repeated description, details are not described herein.

[0136] On the basis of the above-mentioned embodiments, the embodiments of the present disclosure further provide a detection test kit, which comprises the detection test paper described above, and has corresponding beneficial effects. For the sake of avoiding repeated description, details are not described herein.

[0137] In other embodiments, the detection test kit further comprises all constituent structures known to those skilled in the art, such as a shell, a detection reagent, a dropper, a sealed bag, and the like, which are not limited herein.

[0138] It should be noted that, in this document, relational terms such as "first" and "second", and the like, are used solely to distinguish one entity or action from another entity or action, without necessarily requiring or implying any actual such relationship or order between or among the entities or actions. Moreover, the terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without more limitations, an element preceded by "comprises... " does not, without more limitations, foreclose the existence of additional identical elements in the process, method, article, or apparatus that comprises the recited element.

[0139] The above description is merely one specific implementation of the present disclosure, which enables those skilled in the art to understand or implement the present disclosure. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present disclosure. Therefore, the present disclosure will not be limited to these embodiments described herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method of preparing a biochemical sensor array, characterized by, The application relates to a substrate and a preparation method thereof. The substrate comprises a first surface and a second surface arranged oppositely, the first surface is provided with a plurality of through holes arranged in an array, a first trace and a plurality of second traces, the second traces correspond to the through holes one by one, the through holes are provided with a first point and a second point, the first trace is electrically connected with all the first points, and the second traces are electrically connected with the second points one by one. A flexible layer is formed on the second surface in a microelectronic printing mode; the flexible layer covers at least the through holes. A conductive layer is formed on the first surface in a microelectronic printing mode; the conductive layer comprises a plurality of discrete conductive blocks, the conductive blocks cover a group of the through holes, the first points and the second points. A sensitive layer is formed on the side of the flexible layer away from the substrate; the sensitive layer comprises a plurality of discrete film blocks, and the film blocks at least partially overlap the through holes. The flexible layer comprises a polydimethylsiloxane film layer; the flexible layer is formed on the second surface in a microelectronic printing mode, which comprises the following steps:

2. The production method according to claim 1, characterized by, The prepolymer and the curing agent are uniformly mixed at a mass ratio M:1 to obtain mixed glue, and the mixed glue is vacuumized to remove bubbles; wherein 8<=M<=10. The polydimethylsiloxane film layer is printed in a dispensing mode by taking the mixed glue as raw material. The polydimethylsiloxane film layer is printed in a dispensing mode, which comprises the following steps:

3. The preparation method according to claim 2, characterized in that, The polydimethylsiloxane film layer is printed in a dispensing mode based on a first printing speed and a first printing air pressure; wherein the first printing speed is 5-15 mm / s, and the first printing air pressure is 30-90 KPa. After the polydimethylsiloxane film layer is printed in a dispensing mode, the preparation method further comprises the following steps:

4. The production method according to claim 2 or 3, characterized by, The substrate is heated at a first heating temperature until the polydimethylsiloxane film layer is solidified; wherein the first heating temperature is 60-80 DEG C. The conductive layer comprises a silver paste layer; the conductive layer is formed on the first surface in a microelectronic printing mode, which comprises the following steps:

5. The preparation method according to claim 1, characterized in that, The conductive blocks are printed in a dispensing mode by taking conductive silver paste as raw material. The conductive blocks are printed in a dispensing mode, which comprises the following steps:

6. The preparation method according to claim 5, characterized in that, The conductive blocks are printed in a dispensing mode based on a second printing speed and a second printing air pressure; wherein the second printing speed is 4-15 mm / s, and the second printing air pressure is 70-100 KPa. After the conductive blocks are printed in a dispensing mode, the preparation method further comprises the following steps:

7. The production method according to claim 5 or 6, characterized by, The substrate is heated at a second heating temperature until the conductive blocks are solidified; wherein the second heating temperature is 80-120 DEG C, and the heating duration is 30-50 min. The sensitive layer comprises a nano-gold layer; the sensitive layer is formed on the side of the flexible layer away from the substrate, which comprises the following steps:

8. The method of claim 1, wherein, The film blocks are formed on the side of the flexible layer away from the substrate in a magnetron sputtering mode. The film blocks are formed on the side of the flexible layer away from the substrate in a magnetron sputtering mode, which comprises the following steps:

9. The production method according to claim 8, characterized by, The film blocks are sputtered on the side of the flexible layer away from the substrate based on a first sputtering duration; wherein the first sputtering duration is 1-2 min. The application further relates to a substrate and a preparation method thereof.

10. The production method according to claim 8 or 9, characterized by, ​ The antibody is modified on the side of the thin film block away from the flexible layer; The antibody is used for specific binding with a biomolecule to be detected.

11. A biochemical sensor array, characterized in that, The biochemical sensing array is formed based on the preparation method of any one of claims 1 to 10.

12. The biochemical sensor array of claim 11, wherein, The first surface of the substrate is further provided with a signal interface, the signal interface comprising a first signal interface and a second signal interface arranged in sequence, the first signal interface being electrically connected with the first trace, and the second signal interface being electrically connected with the second trace one by one.

13. The biochemical sensing array according to claim 12, wherein, The diameter of the through hole is 2.0-5.0 mm; The distance between the centers of adjacent through holes is 3.0-15 mm; The distance between the edge of the first point and the edge of the corresponding through hole is 0.3-1.0 mm; The distance between the edge of the second point and the edge of the corresponding through hole is 0.3-1.0 mm; The line width of the first trace is 0.2-0.5 mm; The line width of the second trace is 0.2-0.5 mm; The line spacing between the first trace and the second trace and between the second traces is 0.2-0.5 mm; The width of the first signal interface is 0.3-0.7 mm; The width of the second signal interface is 0.3-0.7 mm.

14. A test strip, characterized in that The biochemical sensing array according to any one of claims 11 to 13. The biochemical sensing array according to any one of claims 11 to 13.

15. A test kit comprising, The test paper according to claim 14. The test paper according to claim 14.

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