CN117074661A - Biosensor, detection kit and detection device - Google Patents
Biosensor, detection kit and detection device Download PDFInfo
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- CN117074661A CN117074661A CN202310921361.9A CN202310921361A CN117074661A CN 117074661 A CN117074661 A CN 117074661A CN 202310921361 A CN202310921361 A CN 202310921361A CN 117074661 A CN117074661 A CN 117074661A
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
- G01N33/5438—Electrodes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
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- G01N2333/08—RNA viruses
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Abstract
The present application relates to a biosensor, a detection kit and a detection device. The biosensor includes: a substrate; a channel layer on a surface of the substrate; the source electrode and the drain electrode are alternately arranged on the surface of the channel layer, which is far away from the substrate, at intervals; the dielectric layer covers the surface of the channel layer, on which the source electrode and the drain electrode are not arranged, and the surface of the source electrode and the drain electrode, which is far away from the substrate; the gate electrodes are arranged on the surface of the dielectric layer far away from the channel layer; the gate electrodes, the source electrodes and the drain electrodes are all arranged at intervals and are positioned between the source electrodes and the drain electrodes; the gate electrode is provided with an extension end, and the extension end protrudes out of the source electrode, the drain electrode, the channel layer and the dielectric layer in the length direction of the gate electrode; and the biological probe is fixed on the surface of the gate electrode, which is far away from the dielectric layer. When the biosensor detects a target detection object, the detection speed is high and the operation is simple.
Description
Technical Field
The application relates to the technical field of sensors, in particular to a biosensor, a detection kit and detection equipment.
Background
Virus classification detection has been used in many fields such as biology and medicine. At present, virus classification detection mainly depends on a common polymerase chain reaction or a fluorescent quantitative polymerase chain reaction, and the detection methods need longer detection time and have higher requirements on instrument operation and laboratory environment. Therefore, there is a need for developing a method for detecting a target object which can be classified rapidly and is easy to operate.
Disclosure of Invention
Based on this, it is necessary to provide a biosensor, a detection kit and a detection device. When the biosensor detects a target detection object, the detection speed is high and the operation is simple.
In a first aspect, the present application provides a biosensor comprising:
a substrate;
a channel layer on a surface of the substrate;
the source electrode and the drain electrode are alternately arranged on the surface, far away from the substrate, of the channel layer at intervals;
a dielectric layer covering surfaces of the channel layer where the source electrode and the drain electrode are not provided, and surfaces of the source electrode and the drain electrode away from the substrate;
the number of the gate electrodes is a plurality of, and the gate electrodes are positioned on the surface of the dielectric layer far away from the channel layer; a plurality of gate electrodes are arranged at intervals with the source electrode and the drain electrode and are positioned between the source electrode and the drain electrode; the gate electrode is provided with an extension end, and the extension end protrudes out of the source electrode, the drain electrode, the channel layer and the dielectric layer in the length direction of the gate electrode;
and the biological probe is fixed on the surface of the extending end, which is far away from the medium layer.
In some embodiments, the gate electrode further has a connection portion detachably connected to the extension end, and the bio-probe is fixed on a surface of the connection portion.
In some embodiments, the biosensor further comprises a sample well for receiving a sample to be measured, the sample well being disposed on a surface of the dielectric layer, the connection being located within the sample well.
In some embodiments, each of the extension ends has immobilized thereon the biological probe capable of specifically binding to a different target detection object.
In some embodiments, the dielectric layer covers a surface of the substrate where the channel layer is not disposed.
In some embodiments, a surface of the gate electrode proximate to the substrate and a surface of the dielectric layer distal from the substrate are flush.
In some embodiments, the biological probe comprises at least one of an antibody, an antigen, an enzyme, a RNA, PNA, DNA single strand, a DNA double strand, a two-dimensional DNA nanostructure, and a three-dimensional DNA nanostructure.
In some embodiments, the material of the channel layer includes at least one of graphene, transition metal dichalcogenide, silicon, an oxide semiconductor, and a polymer semiconductor.
In some embodiments, the material of the dielectric layer includes at least one of zirconium oxide and hafnium oxide.
In some embodiments, the substrate comprises one of glass, silica, sapphire, and mica.
In some embodiments, the materials of the source electrode, the drain electrode, and the gate electrode are each independently selected from at least one of gold, silver, copper, titanium, chromium, a carbon material, and a conductive polymer material.
In some embodiments, the thicknesses of the source electrode, the drain electrode, and the gate electrode are each independently selected from 20nm to 500nm.
In a second aspect, the present application provides a test kit comprising: the biosensor of any of the above claims.
In a third aspect, the present application provides a detection apparatus comprising: the biosensor of any one of the above or the above detection kit.
The biosensor comprises a substrate, a channel layer, a source electrode, a drain electrode, a dielectric layer, gate electrodes and biological probes, wherein the number of the gate electrodes is multiple, the gate electrodes are provided with extension ends, and the biological probes are fixed on the surface of each extension end, which is far away from the dielectric layer. The biosensor is based on a field effect transistor and has multiple channels, and a bio-probe is fixed on the surface of the extension end. When the biosensor is used to detect a target detection object, a specific biological probe can be immobilized on the surface of the extension end. When a sample to be detected is dripped into a region where the biological probe is fixed on the surface of the gate electrode, if a target detection object which can be specifically identified by the biological probe exists in the sample, the biological probe can be specifically combined with the target detection object, an electric signal between the source electrode and the drain electrode is monitored, if the biological probe is specifically combined with the target detection object, the electric signal can be obviously changed immediately when the specific combination occurs, if no specific combination occurs, the electric signal cannot be obviously changed. When the biosensor is used for detecting a target detection object, the detection speed is high, and the operation is simple. Meanwhile, the multichannel transistor can be suitable for fixing a plurality of different probes on a plurality of gate electrodes and detecting different target detection objects. And the gate electrode is provided with an extension end, and when the sample is added, the sample to be measured is only required to be added to the area attached with the probe on the extension end, so that the risk of pollution to the source electrode, the drain electrode and the channel is reduced.
Drawings
FIG. 1 is a schematic view showing a partial structure of a biosensor according to an embodiment of the present application;
FIG. 2 is a schematic diagram of a biosensor according to an embodiment of the present application;
FIG. 3 is a schematic view of a biosensor according to another embodiment of the present application;
FIG. 4 is a graph showing the detection results of the novel coronavirus provided in example 1 of the present application;
FIG. 5 is a schematic diagram of the detection result of the A-stream virus provided in example 1 of the present application;
fig. 6 is a schematic diagram of the detection result of the b-stream virus provided in embodiment 1 of the present application.
Description of the reference numerals
100. A substrate; 200. a channel layer; 300. a drain electrode; 400. a source electrode; 500. a dielectric layer; 600. a gate electrode; 620. an extension end; 630. a connection part; 700. a biological probe.
Detailed Description
In order that the above objects, features and advantages of the application will be readily understood, a more particular description of the application will be rendered by reference to the appended drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. The present application may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the application, whereby the application is not limited to the specific embodiments disclosed below.
Unless defined otherwise, 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 herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Referring to fig. 1 to 2, the direction indicated by the arrow is the length direction of the gate electrode 600. It will be appreciated that fig. 2 is a top view of the biosensor, in which the channel layer 200 is blocked by the dielectric layer 500, since the dielectric layer 500 covers the surfaces of the channel layer 200 where the active electrode 400 and the drain electrode 300 are not disposed and the surfaces of the source electrode 400 and the drain electrode 300 away from the substrate 100. An embodiment of the present application provides a biosensor including: a substrate 100; a channel layer 200, the channel layer 200 being located on a surface of the substrate 100; the source electrode 400 and the drain electrode 300 are alternately arranged on the surface of the channel layer 200 away from the substrate 100 at intervals; a dielectric layer 500, the dielectric layer 500 covering surfaces of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed, and surfaces of the source electrode 400 and the drain electrode 300 away from the substrate 100; the number of gate electrodes 600 is plural, and the gate electrodes 600 are located on the surface of the dielectric layer 500 remote from the channel layer 200; the plurality of gate electrodes 600 are spaced apart from the source electrode 400 and the drain electrode 300 and are positioned between the source electrode 400 and the drain electrode 300, the gate electrode 600 has extension ends 620, and the extension ends 620 protrude from the source electrode 400, the drain electrode 300, the channel layer 200 and the dielectric layer 500 in the length direction of the gate electrode 600; biological probe 700 is immobilized on the surface of gate electrode 600 remote from dielectric layer 500.
The biosensor includes a substrate 100, a channel layer 200, a source electrode 400, a drain electrode 300, a dielectric layer 500, a gate electrode 600 and bio-probes 700, wherein the gate electrode 600 has a plurality of extended ends 620, and the bio-probes 700 are fixed on the surface of each extended end 620 away from the dielectric layer 500. The biosensor is based on a field effect transistor and has multiple channels, and a bio-probe 700 is fixed on the surface of the gate electrode 600. In detecting a target detection object using the biosensor, a specific bio-probe 700 may be fixed to the surface of the extension 620. When a sample to be detected is dripped into a region where the biological probe 700 is fixed on the surface of the gate electrode 600, if a target detection object which can be specifically identified by the biological probe 700 exists in the sample, the biological probe 700 can be specifically combined with the target detection object, an electric signal between the source electrode 400 and the drain electrode 300 is monitored, if the biological probe 700 is specifically combined with the target detection object, the electric signal can be obviously changed immediately when the specific combination occurs, and the detection speed is high; if no specific binding occurs, no significant change in the electrical signal occurs. When the biosensor is used for detecting a target detection object, the detection speed is high, the operation is simple, and the specificity is high. Meanwhile, the multi-channel transistor can be adapted to immobilize a plurality of different bio-probes 700 on a plurality of gate electrodes 600 while detecting different target detection objects. And the gate electrode 600 has an extension end 620, and during sample loading, only the sample to be measured needs to be added to the area attached with the probe on the extension end, so that the risk of pollution to the source electrode, the drain electrode and the channel is reduced, the utilization rate of the biosensor is improved, and the repeated use of the biosensor is realized.
It is understood that the biological probe 700 can specifically bind to a target detection object such as a nucleic acid biomarker, an antibody, an antigen, a metabolic small molecule or an ion of a virus, a bacterium, a fungus, a chlamydia.
It is understood that the biological probe 700 immobilized on the surface of the gate electrode 600 may be one or more, and the plurality of biological probes 700 may refer to biological probes 700 capable of specifically identifying different target detection objects, and when the biological probe 700 on the surface of the gate electrode 600 is one, identification and detection may be performed on a single target detection object. When the number of the biological probes 700 on the surface of the gate electrode 600 is plural, it is also possible to detect whether or not the sample to be detected contains several corresponding target detection objects at the same time.
For example, when the target detection object is a single strand of DNA, the biological probe may be a single strand of DNA complementary thereto. When the target analyte is a bacterium or virus, the biological probe may be an antibody to the bacterium or virus. When the target detection object is an antibody, the biological probe may be a secondary antibody capable of specifically recognizing the antibody.
Compared with the traditional detection platform, the biosensor of the application constructs a field effect transistor biosensor which can be used for detecting target detection objects and can detect various target detection objects at the same time. The detection of multiple target detection objects is realized by modifying and fixing multiple biological probes 700 on the gate electrode 600 and by the change of the conductivity of the device caused by the complementary hybridization of the nucleic acid molecular base sequences between the biological probes 700 and the corresponding target detection objects, and the method has the advantages of high stability, high integration level, simple operation, high sensitivity, short response time, high flux, low cost and the like. The application can solve the problems of long detection time and complex operation of the common polymerase chain reaction or the fluorescent quantitative polymerase chain reaction, and can overcome the defect that the biological probe 700 is directly fixed and modified on the surface of the semiconductor channel in the liquid gate field effect transistor.
In some embodiments, the gate electrode 600 further has a connection portion 630, the connection portion 630 is detachably connected to the extension 620, and the bio-probe 700 is fixed on a surface of the connection portion 630. By providing the detachable connection portion 630, it is possible to add only the sample to be measured to the connection portion 630 for detection when the biosensor is used for detection, reducing the contamination to other parts of the biosensor. Meanwhile, the connection part 630 is convenient to replace after detection is completed, and the repeated utilization of the biosensor can be realized. Further, it is also possible to realize detection of different target detection objects by replacing the connection part 630 to which different biological probes 700 are fixed.
In one embodiment, the connection portion 630 is a detection piece to which the bio-probe 700 is attached, and the detection piece is connected to the extension 620 through a wire point.
In some embodiments, the biosensor further includes a sample well for receiving a sample to be measured, the sample well being disposed on a surface of the medium layer 500, and the connection portion 630 being located in the sample well. The sample groove is arranged to enable the connecting portion 630 to be located in the sample groove, the sample to be detected can be only contacted with the connecting portion 630 and detected, pollution of the sample to be detected to other parts of the biosensor can be reduced, and the biosensor can be reused only by replacing the connecting portion 630.
In some embodiments, the sample cell is a liquid cell or microfluidic channel.
Referring to fig. 3, in some embodiments, a dielectric layer 500 covers a surface of the substrate 100 where the channel layer 200 is not disposed. It will be appreciated that fig. 3 is a top view of the biosensor, in which the lower layers are all blocked by the dielectric layer 500, since the dielectric layer 500 covers the surface of the cover substrate 100 where the channel layer 200 is not provided. The lower layers are all covered by the medium layer, so that the risk of pollution to each layer of the lower layers during sample loading is further reduced.
In some embodiments, the surface of gate electrode 600 proximate substrate 100 is flush with the surface of dielectric layer 500 distal from substrate 100.
In some embodiments, the biological probe 700 includes at least one of an antibody, an antigen, an enzyme, a RNA, PNA, DNA single strand, a DNA double strand, a two-dimensional DNA nanostructure, and a three-dimensional DNA nanostructure. It is understood that a DNA nanostructure is an artificial DNA structure that is designed artificially and produces useful nucleic acid structures.
In some embodiments, the biological probe 700 is immobilized on the surface of the gate electrode 600 by at least one of covalent bonding, adsorption, entrapment, and cross-linking.
The covalent bonding method is one of the methods for modifying the electrode surface, and the substrate electrode surface is first pretreated to introduce bonding groups, and a predetermined functional group is attached to the electrode surface by chemical bonding organic synthesis reaction of the electrode surface. The modified electrode has many excellent characteristics: such as molecular selective recognition function, selective response, high stability, etc. The chemical modifier with different expected functional groups can be covalently bonded to the pretreated surfaces of different electrodes by the modification method.
The adsorption method is a method of forming a film by fixing a modifier on the surface of a solid base electrode by using the non-covalent bond between the base electrode and the modifier.
The principle of entrapment is to entrap biomolecules in the network space of water insoluble gel polymer pores. The embedding method is simple to operate, has small influence on the activity of the biomolecules, and the prepared immobilized biomolecules have high strength.
The crosslinking method uses a bifunctional or multifunctional reagent to directly crosslink with reactive groups such as amino groups, carboxyl groups and the like on the surface of the biomolecules to form covalent bonds for fixing the biomolecules, and the binding force is the covalent bond. The biological molecule fixed by the method is tightly combined with the carrier, and common cross-linking agents are glutaraldehyde, toluene diisocyanate, bis-azo biphenyl and the like.
In some embodiments, the material of the channel layer 200 includes at least one of graphene, transition metal dichalcogenide, silicon, an oxide semiconductor, and a polymer semiconductor. The transition metal dichalcogenide means a compound containing a transition metal element and a sulfur element, and optionally, the transition metal dichalcogenide includes at least one of molybdenum disulfide and tungsten disulfide.
In some embodiments, the material of dielectric layer 500 includes at least one of zirconium oxide and hafnium oxide. Zirconium oxide and hafnium oxide are both dielectric materials with high dielectric coefficients.
In some embodiments, the substrate 100 comprises one of glass, silica, sapphire, and mica.
In some embodiments, the material of the source electrode 400 includes at least one of gold, silver, copper, titanium, chromium, carbon material, and conductive polymer material.
In some embodiments, the material of the drain electrode 300 includes at least one of gold, silver, copper, titanium, chromium, carbon material, and conductive polymer material.
In some embodiments, the material of the gate electrode 600 includes at least one of gold, silver, copper, titanium, chromium, carbon material, and conductive polymer material.
In some embodiments, the thickness of the source electrode 400 is 20nm to 500nm. Alternatively, the source electrode 400 has a thickness of 20nm, 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm, 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm, 200nm, 210nm, 220nm, 230nm, 240nm, 250nm, 260nm, 270nm, 280nm, 290nm, 300nm, 310nm, 320nm, 330nm, 340nm, 350nm, 360nm, 370nm, 380nm, 390nm, 400nm, 410nm, 420nm, 430nm, 440nm, 450nm, 460nm, 470nm, 480nm, 490nm or 500nm.
In some embodiments, the thickness of the drain electrode 300 is 20nm to 500nm. Alternatively, the drain electrode 300 has a thickness of 20nm, 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm, 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm, 200nm, 210nm, 220nm, 230nm, 240nm, 250nm, 260nm, 270nm, 280nm, 290nm, 300nm, 310nm, 320nm, 330nm, 340nm, 350nm, 360nm, 370nm, 380nm, 390nm, 400nm, 410nm, 420nm, 430nm, 440nm, 450nm, 460nm, 470nm, 480nm, 490nm or 500nm.
In some embodiments, the gate electrode 600 has a thickness of 20nm to 500nm. Alternatively, the gate electrode 600 has a thickness of 20nm, 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm, 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm, 200nm, 210nm, 220nm, 230nm, 240nm, 250nm, 260nm, 270nm, 280nm, 290nm, 300nm, 310nm, 320nm, 330nm, 340nm, 350nm, 360nm, 370nm, 380nm, 390nm, 400nm, 410nm, 420nm, 430nm, 440nm, 450nm, 460nm, 470nm, 480nm, 490nm or 500nm.
Still another embodiment of the present application provides a method for manufacturing a biosensor, including:
forming a channel layer 200 on the substrate 100;
preparing source electrodes 400 and drain electrodes 300 on the surface of the channel layer 200, the source electrodes 400 and the drain electrodes 300 being alternately arranged at intervals;
preparing a dielectric layer 500 on the surface of the channel layer 200, the dielectric layer 500 covering the surface of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed, and the surfaces of the source electrode 400 and the drain electrode 300;
preparing gate electrodes 600 on the surface of the dielectric layer 500, the number of the gate electrodes 600 being plural, the gate electrodes 600 being located on the surface of the dielectric layer 500; the plurality of gate electrodes 600 are spaced apart from the source electrode 400 and the drain electrode 300, and are positioned between the source electrode 400 and the drain electrode 300; the gate electrode 600 has an extension 620, and the extension 620 protrudes from the source electrode 400, the drain electrode 300, the channel layer 200, and the dielectric layer 500 in the length direction of the gate electrode 600;
the biological probe 700 is immobilized on the surface of the gate electrode 600 remote from the dielectric layer 500.
In some embodiments, a method of making a biosensor, comprising:
(1) A channel layer 200 is formed on the substrate 100.
(2) The position of the patterned source electrode 400 and the patterned drain electrode 300 are defined on the surface of the channel layer 200 by adopting a photolithography method, the source electrode 400 and the patterned drain electrode 300 are deposited, the source electrode 400 and the patterned drain electrode 300 are alternately arranged at intervals, the materials of the source electrode 400 and the patterned drain electrode 300 are respectively and independently selected from at least one of gold, silver, copper, titanium, chromium, carbon materials and conductive polymer materials, and the thicknesses of the source electrode and the patterned drain electrode are respectively and independently selected from 20 nm-500 nm.
(3) A dielectric layer 500 is deposited on the surface of the channel layer 200, the dielectric layer 500 covering the entire surface of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed and the surfaces of the source electrode 400 and the drain electrode 300, and the material of the dielectric layer 500 includes at least one of zirconium oxide and hafnium oxide.
(4) Defining the position of the patterned gate electrode 600 on the surface of the dielectric layer 500 by adopting a photolithography method, and depositing the gate electrodes 600 again, wherein the number of the gate electrodes 600 is a plurality of, and the gate electrodes 600 are positioned on the surface of the dielectric layer 500; the plurality of gate electrodes 600 are spaced apart from the source electrode 400 and the drain electrode 300, and are positioned between the source electrode 400 and the drain electrode 300. The material of the gate electrode 600 is at least one selected from gold, silver, copper, titanium, chromium, carbon materials and conductive polymer materials, and the thickness of the gate electrode 600 is 20 nm-500 nm; the gate electrode 600 has extension ends 620, the extension ends 620 protrude from the source electrode 400, the drain electrode 300, the channel layer 200 and the dielectric layer 500 in the length direction of the gate electrode 600, and the gate electrode 600 further has connection portions 630, the connection portions 630 being detachably connected to the extension ends 620.
(5) The bio-probe 700 is fixed on the surface of the connection portion 630 of the gate electrode 600, which is remote from the dielectric layer 500.
In one embodiment, the method of preparing the biosensor comprises the steps of:
(1) A channel layer 200 is formed on the substrate 100.
(2) The position of the patterned source electrode 400 and the patterned drain electrode 300 are defined on the surface of the channel layer 200 by adopting a photolithography method, the source electrode 400 and the patterned drain electrode 300 are deposited, the source electrode 400 and the patterned drain electrode 300 are alternately arranged at intervals, the materials of the source electrode 400 and the patterned drain electrode 300 are respectively and independently selected from at least one of gold, silver, copper, titanium, chromium, carbon materials and conductive polymer materials, and the thicknesses of the source electrode and the patterned drain electrode are respectively and independently selected from 20 nm-500 nm.
(3) A dielectric layer 500 is deposited on the surface of the channel layer 200, the dielectric layer 500 covering the entire surface of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed and the surfaces of the source electrode 400 and the drain electrode 300, and the material of the dielectric layer 500 includes at least one of zirconium oxide and hafnium oxide.
(4) Defining the position of the patterned gate electrode 600 on the surface of the dielectric layer 500 by adopting a photolithography method, and depositing the gate electrodes 600 again, wherein the number of the gate electrodes 600 is a plurality of, and the gate electrodes 600 are positioned on the surface of the dielectric layer 500; the plurality of gate electrodes 600 are spaced apart from the source electrode 400 and the drain electrode 300, and are positioned between the source electrode 400 and the drain electrode 300. The material of the gate electrode 600 is at least one selected from gold, silver, copper, titanium, chromium, carbon materials and conductive polymer materials, and the thickness of the gate electrode 600 is 20 nm-500 nm; the gate electrode 600 has extension ends 620, the extension ends 620 protrude from the source electrode 400, the drain electrode 300, the channel layer 200 and the dielectric layer 500 in the length direction of the gate electrode 600, and the gate electrode 600 further has connection portions 630, the connection portions 630 being detachably connected to the extension ends 620.
(5) The bio-probe 700 is fixed on the surface of the connection portion 630 of the gate electrode 600, which is remote from the dielectric layer 500.
Yet another embodiment of the present application provides a detection kit comprising: the biosensor of any of the above.
A further embodiment of the present application provides a detection apparatus including: the biosensor of any one of the above or the above detection kit.
The following are specific embodiments
Example 1
Biosensor
Referring again to fig. 1 to 3, the biosensor in the present embodiment has the following structure: a substrate 100; a channel layer 200, the channel layer 200 being located on a surface of the substrate 100; the source electrode 400 and the drain electrode 300 are alternately arranged on the surface of the channel layer 200 away from the substrate 100 at intervals; a dielectric layer 500, the dielectric layer 500 covering the surface of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed, and covering the surfaces of the source electrode 400 and the drain electrode 300, and covering the surface of the substrate 100 where the channel 200 is not disposed; the plurality of gate electrodes 600 are positioned on the surface of the dielectric layer 500 away from the channel layer 200 and alternately spaced apart from the source electrode 400 and the drain electrode 300, the gate electrodes 600 are positioned between the source electrode 400 and the drain electrode 300, and the surface of the gate electrode 600 close to the substrate 100 is flush with the surface of the dielectric layer 500 away from the substrate 100. The gate electrode 600 has extension ends 620, the extension ends 620 protrude from the source electrode 400, the drain electrode 300, the channel layer 200 and the dielectric layer 500 in the length direction of the gate electrode 600, the gate electrode 600 further has connection portions 630, the connection portions 630 are detachably connected to the extension ends 620, and three bio-probes 700 are fixed on the surfaces of the connection portions 630. Biological probe 700 is immobilized on the surface of gate electrode 600 remote from dielectric layer 500. In the application, the substrate 100 is a silicon substrate 100 with a silicon dioxide layer on the surface, the channel layer 200 is made of a single-layer graphene film, the medium layer 500 is made of zirconia, the source electrode 400 and the drain electrode 300 are both composite electrodes of 5nm chromium-40 nm gold, the gate electrode is a 40nm gold electrode, three biological probes 700 are used, three biological probes 700 with mercapto groups modified at the tail ends are used, and the three biological probes 700 can be respectively connected with nucleic acid molecules of three target detection objects including a novel coronavirus, an A-stream virus and an B-stream virus through a base complementation principle. And further includes a sample groove provided on the surface of the connection part 630.
Preparation of biosensors
(1) A single-layer graphene film is prepared on a copper foil by a chemical vapor deposition method, and the prepared graphene film is transferred to a clean silicon dioxide/silicon substrate 100 by an electrochemical stripping method.
(2) The patterned source electrode 400 and drain electrode 300 are defined by ultraviolet lithography and oxygen ion etching, and then a composite electrode of 5nm chromium-40 nm gold is prepared as the source electrode 400 and drain electrode 300 by thermal evaporation.
(3) Zirconium oxide is deposited on the surface of the graphene film as a dielectric layer 500, and the dielectric layer 500 covers the surface of the channel layer 200 where the source electrode 400 and the drain electrode 300 are not disposed and the surfaces of the source electrode 400 and the drain electrode 300.
(4) A patterned gate electrode 600 is deposited on the surface of the dielectric layer 500, the gate electrode 600 is a gold electrode with a thickness of 40nm, the gate electrode 600 has an extension end 620, the extension end 620 protrudes from the source electrode 400, the drain electrode 300, the channel layer 200 and the dielectric layer 500 in the length direction of the gate electrode 600, the gate electrode 600 further has a connection portion 630, and the connection portion 630 is detachably connected to the extension end 620.
(5) After the surfaces of the connection portions 630 of the three different gate electrodes 600 were respectively contacted with three solutions containing the bio-probes 700 capable of being specifically bound to the new coronavirus, the alphavirus, and the ethylvirus, respectively, for 10 minutes, and after each contact, the gate electrodes 600 were rinsed with ultra pure water, the three bio-probes 700 were fixedly connected to the surfaces of the connection portions 630 of the gate electrodes 600 through gold-thiol covalent bonds.
Detection of target detection object
And (3) treating the upper respiratory tract pharyngeal swab sample containing the new coronavirus and not containing the A-flow virus and the B-flow virus to extract virus nucleic acid. The treatment method comprises extracting 200 microliters of sample from the collecting tube of the upper respiratory tract pharyngeal swab sample, placing into a nucleic acid extraction kit, and extracting to obtain a solution containing the novel coronavirus nucleic acid molecules. The solution is dropped onto the surface of the connection part 630 in the sample cell, and the current between the source electrode 400 and the drain electrode 300 is detected.
Referring to fig. 4 to 6, when a sample to be tested containing a new coronavirus but not containing a new coronavirus and a new b-stream virus is detected using a biosensor to which three types of bio-probes 700 for detecting a new coronavirus, a new a-stream virus and a new b-stream virus are immobilized, it can be seen that the combination of the gate electrode 600 to which the new coronavirus detection probe is immobilized with the new coronavirus in the sample causes a significant change in the electrical signals between the source electrode 400 and the drain electrode 300, while the other two electrical signals are not significantly changed.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. The scope of the application is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted in accordance with the contents of the claims.
Claims (10)
1. A biosensor, comprising:
a substrate;
a channel layer on a surface of the substrate;
the source electrode and the drain electrode are alternately arranged on the surface, far away from the substrate, of the channel layer at intervals;
a dielectric layer covering surfaces of the channel layer where the source electrode and the drain electrode are not provided, and surfaces of the source electrode and the drain electrode away from the substrate;
the number of the gate electrodes is a plurality of, and the gate electrodes are positioned on the surface of the dielectric layer far away from the channel layer; a plurality of gate electrodes are arranged at intervals with the source electrode and the drain electrode and are positioned between the source electrode and the drain electrode; the gate electrode is provided with an extension end, and the extension end protrudes out of the source electrode, the drain electrode, the channel layer and the dielectric layer in the length direction of the gate electrode;
and the biological probe is fixed on the surface of the extending end, which is far away from the medium layer.
2. The biosensor of claim 1, wherein the gate electrode further has a connection portion detachably connected to the extension end, and the bio-probe is fixed to a surface of the connection portion.
3. The biosensor of claim 2, further comprising a sample well for receiving a sample to be measured, the sample well being disposed on a surface of the connection portion.
4. The biosensor of claim 1, wherein each of the extension ends has immobilized thereon the biological probe capable of specifically binding to a different target detection object.
5. The biosensor of claim 1, wherein the dielectric layer covers a surface of the substrate where no channel layer is provided.
6. The biosensor of claim 1, wherein a surface of the gate electrode proximate to the substrate and a surface of the dielectric layer distal from the substrate are flush.
7. The biosensor of any one of claims 1-6, wherein the material of the channel layer comprises at least one of graphene, transition metal dichalcogenide, silicon, oxide semiconductor, and polymer semiconductor;
and/or the material of the dielectric layer comprises at least one of zirconium oxide and hafnium oxide;
and/or the substrate comprises one of glass, silica, sapphire, and mica;
and/or the biological probe comprises at least one of an antibody, an antigen, an enzyme, a RNA, PNA, DNA single strand, a DNA double strand, a two-dimensional DNA nanostructure, and a three-dimensional DNA nanostructure.
8. The biosensor of any one of claims 1-6, wherein the source electrode, the drain electrode, and the gate electrode are each independently selected from at least one of gold, silver, copper, titanium, chromium, a carbon material, and a conductive polymer material;
and/or the thicknesses of the source electrode, the drain electrode and the gate electrode are respectively and independently selected from 20 nm-500 nm.
9. A test kit comprising: the biosensor of any one of claims 1 to 8.
10. A detection apparatus, characterized by comprising: the biosensor of any one of claims 1 to 8 or the detection kit of claim 9.
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