WO2019007062A1 - 流感病毒检测芯片和使用其检测流感病毒的方法 - Google Patents

流感病毒检测芯片和使用其检测流感病毒的方法 Download PDF

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WO2019007062A1
WO2019007062A1 PCT/CN2018/075642 CN2018075642W WO2019007062A1 WO 2019007062 A1 WO2019007062 A1 WO 2019007062A1 CN 2018075642 W CN2018075642 W CN 2018075642W WO 2019007062 A1 WO2019007062 A1 WO 2019007062A1
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pad
influenza virus
graphene oxide
oxide film
substrate
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PCT/CN2018/075642
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English (en)
French (fr)
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庞凤春
车春城
薛海林
邵喜斌
蔡佩芝
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京东方科技集团股份有限公司
北京京东方光电科技有限公司
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Priority to US16/086,322 priority Critical patent/US11585812B2/en
Priority to EP18766112.9A priority patent/EP3650861A4/en
Publication of WO2019007062A1 publication Critical patent/WO2019007062A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6432—Quenching
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/11—Orthomyxoviridae, e.g. influenza virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/26—Infectious diseases, e.g. generalised sepsis

Definitions

  • the present disclosure relates to the field of influenza virus detection, and more particularly to an influenza virus detection chip and a method of detecting influenza virus using an influenza virus detection chip.
  • Influenza is an acute respiratory infection caused by influenza virus, and it is also a disease that is highly contagious and spreads rapidly. The general autumn and winter season is its high incidence, and the complications and deaths caused by it are very serious.
  • the disease is caused by influenza virus and can be divided into three types: A, B, and C.
  • the type A virus often undergoes antigenic variation, is highly contagious, spreads rapidly, and is prone to widespread epidemics.
  • Influenza, infants, the elderly, and patients with cardiopulmonary diseases are prone to complications such as pneumonia.
  • influenza virus detection is mainly carried out by colloidal gold immunochromatography, ELISA, PCR and the like.
  • Colloidal gold immunochromatography requires a large amount of influenza virus to accumulate at the detection line until color development. This method has low detection sensitivity and cannot be detected when the content of influenza virus is low. Other methods are complicated in detection and longer in detection time. higher cost. Many researchers are currently working on new bio-influenza virus sensors to replace traditional influenza virus detection.
  • the present disclosure provides an influenza virus detection chip having high detection sensitivity and reduced detection complexity and a method for detecting influenza virus using an influenza virus detection chip.
  • influenza virus detecting chip comprising:
  • influenza virus detecting chip further includes a first substrate, and the graphene oxide film is bonded to the first substrate by a function of a functional group.
  • influenza virus detecting chip further includes a second substrate disposed opposite to the first substrate, the graphene oxide film, the bonding pad, the first electrode, and the second electrode are disposed at Between the first substrate and the second substrate.
  • influenza virus detection chip further includes a sample pad for absorbing the sample solution, wherein the sample pad, the bond pad, and the graphene oxide film are sequentially disposed in the first direction.
  • influenza virus detecting chip further includes an absorbent pad for absorbing excess sample solution, wherein the bonding pad, the graphene oxide film, and the absorbent pad are sequentially disposed in the first direction.
  • the first electrode and the second electrode are respectively adjacent to and in contact with both ends of the graphene oxide film in a second direction perpendicular to the first direction.
  • the first substrate and the second substrate comprise at least one of a glass substrate, a silicon substrate, or an organic polymer substrate.
  • one of the first substrate and the second substrate is transparent.
  • influenza virus detection chip further includes a sample pad for absorbing the sample solution and an absorption pad for absorbing the excess sample solution, wherein the sample pad, the bonding pad, the graphene oxide film, and the absorption pad are along The first direction is sequentially set.
  • the sample pad and the bond pad are each a separate or integral film layer made of polyester fibers.
  • the sample pad, bond pad, and absorbent pad have the same thickness.
  • the sample pad and the bonding pad are combined in at least one of the following ways: in a manner overlapping each other, in a manner overlapping each other, and in a manner overlapping each other.
  • a method of detecting influenza virus using the above influenza virus detecting chip comprising: sequentially flowing a sample solution through a bonding pad and a graphene oxide film; and irradiating the graphene oxide with a laser The film simultaneously detects the current across the graphene oxide film by using the first electrode and the second electrode; and determines that the influenza virus is contained in the sample solution when the current across the graphene oxide film changes.
  • influenza virus detection chip further includes a sample pad for absorbing the sample solution and an absorption pad for absorbing the excess sample solution, wherein the sample pad, the bonding pad, the graphene oxide film, and the absorption pad are along The first direction is sequentially set.
  • the sample pad and the bond pad are each a separate or integral film layer made of polyester fibers.
  • the sample pad and the bonding pad are combined in such a manner as to overlap each other
  • the bonding pad and the graphene oxide film are combined in such a manner as to overlap each other
  • the graphene oxide film and the absorption pad overlap each other. The way to combine.
  • FIG. 1 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure
  • FIG. 2 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure
  • FIG. 3 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure
  • FIG. 4 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure.
  • spatially relative terms such as “lower”, “above”, “upper”, “below”, etc., may be used to describe one element or feature and other elements or features as illustrated in the figures. Relationship. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation.
  • FIG. 1 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure.
  • an influenza virus detecting chip 100 includes an upper substrate 101, a lower substrate 102, and a layered layer between the upper substrate 101 and the lower substrate 102 in a first direction (ie, from left to right).
  • the bonding pad 103 and the graphene oxide film 104 are sequentially disposed.
  • both the upper substrate 101 and the lower substrate 102 may be a glass substrate, a silicon substrate, or an organic polymer substrate.
  • the upper substrate 101 and the lower substrate 102 may each be selected from a silicon substrate, a glass substrate, a PET substrate, a PMMA substrate, or the like.
  • the bonding pad 103 is a film layer made of, for example, polyester fiber, but in the present disclosure, the material for making the bonding pad 103 is not limited thereto.
  • the first electrode 105 and the second electrode 105' may be made of a general electrode material (for example, a metal, an alloy, a conductive metal oxide, a conductive metal nitride, or the like).
  • the first electrode 105 and the second electrode 105' may be made of a metal such as titanium, platinum, rhodium, gold, silver, molybdenum, aluminum, tungsten, copper, ruthenium, chromium, rhenium, or an alloy including the above, or conductive oxidation.
  • the substance is formed such as indium zinc oxide, aluminum zinc oxide, indium tin oxide, gallium zinc oxide or zinc tin oxide.
  • an electrode material may be deposited on the lower substrate 102, and then the first electrode 105 and the second electrode 105' may be formed by a photolithography process. Then, a graphene oxide solution may be dropped between the first electrode 105 and the second electrode 105' formed on the lower substrate 102 and dried to form a graphene oxide film 104 on the lower substrate 102.
  • the surface of the lower substrate 102 may be subjected to, for example, amino modification, so that the modified amino group on the surface of the lower substrate 102 can be on the surface of the graphene oxide film 104.
  • a group such as a carboxyl group or an epoxy group reacts, and therefore, the graphene oxide film 104 is bonded to the lower substrate 102 by the action of these functional groups.
  • the monoclonal antibody can also be bound to the graphene oxide film 104 by the action of a functional group.
  • the bonding pad 103 and the graphene oxide film 104 may be partially overlapped at positions in contact with each other, that is, the two may be combined in such a manner as to overlap each other, which may be advantageous for the sample solution. flow.
  • the upper substrate 101 and the lower substrate 102 may be bonded together by, for example, an adhesive, and the bonding pad 103 and the graphene oxide film 104 are sandwiched between the upper substrate 101 and the lower substrate 102.
  • the first electrode 105 and the second electrode 105' are respectively adjacent to and in contact with both ends of the graphene oxide film 104 in the second direction perpendicular to the first direction, and are disposed on the upper substrate 101 and Between the lower substrates 102.
  • the first electrode 105 and the second electrode 105' may be disposed in other manners as long as the current across the graphene oxide film 104 (or the resistance of the graphene oxide film 104 is detected) can be detected.
  • the sample solution to be detected for example, saliva or blood, etc.
  • the sample solution to be detected may be added to the influenza virus detection chip 100, so that the sample solution to be detected first flows to Combine the pad.
  • the sample solution to be detected contains an influenza virus (ie, an antigen)
  • the antigen specifically reacts with the fluorescently labeled monoclonal antibody in the binding pad, and then the sample solution continues to flow to the graphene oxide film 104 by capillary action. Specifically, it reacts specifically with the monoclonal antibody thereon to generate a double-antibody sandwich influenza virus complex.
  • one of the upper substrate 101 and the lower substrate 102 may be transparent, and of course, the upper substrate 101 and the lower substrate 102 may both be transparent.
  • the photon energy generated by the fluorescent label is absorbed by the graphene oxide, resulting in a decrease in the electrical resistance of the graphene oxide.
  • the electric resistance of the graphene oxide is decreased, the current change across the graphene oxide film 104 can be detected by using the first electrode 105 and the second electrode 105'. That is, when the current across the graphene oxide film 104 changes, it can be determined that the influenza virus is contained in the sample solution.
  • the fluorescently labeled monoclonal antibody and the monoclonal antibody on the graphene oxide film in the binding pad may be antibodies capable of specifically reacting to the same antigen.
  • the fluorescently labeled monoclonal antibody in the binding pad and the monoclonal antibody on the graphene oxide film may be the same antibody, for example, against the avian influenza virus NP protein, and the fluorescently labeled monoclonal antibody may be fluorescently labeled 1G11.
  • the monoclonal antibody on the graphene oxide film may also be 1G11, but no fluorescent labeling is performed, that is, the two antibodies are the same.
  • the present disclosure is not limited thereto, and the two antibodies may also be different antibodies capable of specifically reacting to the same antigen.
  • FIG. 2 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure.
  • the influenza virus detecting chip 200 further includes a sample pad 106 for absorbing a sample solution.
  • the sample pad 106 is disposed on the left side of the bonding pad 103 and is in contact with the bonding pad 103. That is, the sample pad 106, the bonding pad 103, and the graphene oxide film 104 are sequentially disposed in the first direction (ie, from left to right).
  • the sample pad 106 may also be made of polyester fiber, but is not limited thereto.
  • the sample pad 106 and the bonding pad 103 are made of the same material, they may be formed separately or integrally. That is, the sample pad 106 and the bonding pad 103 may be formed as separate or integrated film layers.
  • the fluorescently labeled monoclonal antibody can be bound to a region of the single film layer close to the graphene oxide film 104 without being bound away from the single film layer. On the area of the graphene oxide film 104.
  • a single membrane layer has two regions, one containing a fluorescently labeled monoclonal antibody and the other containing no fluorescently labeled monoclonal antibody.
  • the sample pad 106 and the bonding pad 103 may have the same thickness so as to be well sealed by the upper substrate 101 and the lower substrate 102.
  • the sample pad 106 and the bonding pad 103 when the sample pad 106 and the bonding pad 103 are two mutually independent components, the sample pad 106 and the bonding pad 103 may be combined in such a manner as to overlap each other to facilitate the sample solution to be detected. Smooth flow.
  • the bonding pad 103 and the graphene oxide film 104 may be combined in such a manner as to overlap each other.
  • the sample solution to be detected will first flow to the sample pad 106, and then flow to the bonding pad 103 by capillary action, and the subsequent history is the same as in the foregoing embodiment.
  • the process described in the above is similar and will not be described again.
  • FIG. 3 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure.
  • the influenza virus detecting chip 300 further includes an absorbent pad 107 for absorbing excess sample solution.
  • the absorption pad 107 is disposed on the right side of the graphene oxide film 104 and is in contact with the graphene oxide film 104. That is, the bonding pad 103, the graphene oxide film 104, and the absorption pad 107 are sequentially disposed in the first direction (ie, from left to right).
  • the absorbent pad 107 may be made of a composite fiber material, but is not limited thereto, and may be made of any material capable of absorbing a fluid.
  • the absorbent pad 107 and the bonding pad 103 may have the same thickness so as to be well sealed by the upper substrate 101 and the lower substrate 102.
  • the excess sample solution can be absorbed by the absorption pad 107.
  • the absorbent pad 107 and the graphene oxide film 104 may be combined in such a manner as to overlap each other to facilitate the smooth absorption of the excess sample solution by the absorbent pad 107.
  • FIG. 4 is a schematic exploded perspective view showing an influenza virus detecting chip according to an embodiment of the present disclosure.
  • the influenza virus detecting chip 400 further includes an absorbent pad 107 for absorbing excess sample solution.
  • the absorption pad 107 is disposed on the right side of the graphene oxide film 104 and is in contact with the graphene oxide film 104. That is, the sample pad 106, the bonding pad 103, the graphene oxide film 104, and the absorption pad 107 are sequentially disposed in the first direction (ie, from left to right).
  • the absorbent pad 107 can be formed and arranged in the same manner as described in the foregoing embodiments.
  • the sample pad 106, the bonding pad 103, and the absorption pad 107 have the same thickness so as to be well sealed by the upper substrate 101 and the lower substrate 102. Further, in order to facilitate the smooth flow of the sample solution, the sample pad 106 and the bonding pad 103 may be combined in such a manner as to overlap each other, and the bonding pad 103 and the graphene oxide film 104 may be combined in such a manner as to overlap each other, and / Or the graphene oxide film 104 and the absorbent pad 107 may be combined in such a manner as to overlap each other.
  • the sample solution to be detected will first flow to the sample pad 106, and then flow to the bonding pad 103 by capillary action, and the subsequent process is the same as in the foregoing embodiment.
  • the process described in the above is similar.
  • excess sample solution can be absorbed by the absorbent pad 107.
  • influenza virus detecting chip of the present disclosure and a method using the same for detecting influenza virus have high detection sensitivity, and detection is simple, quick, and convenient.
  • the presence or absence of the influenza virus can be detected by detecting the change of the current, the detection sensitivity is increased, and early detection and prevention can be performed to prevent the continuous infection of the influenza virus.

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Abstract

提供了一种流感病毒检测芯片和一种使用流感病毒检测芯片检测流感病毒的方法。所述流感病毒检测芯片包括：氧化石墨烯薄膜；结合垫，沿第一方向设置在所述氧化石墨烯薄膜的一侧；第一电极和第二电极，分别连接到所述氧化石墨烯膜的在与第一方向垂直的第二方向上的两端，其中，在所述结合垫中包含带有荧光标记的第一单克隆抗体，在氧化石墨烯膜上具有第二单克隆抗体，其中荧光标记具有C＝C-C＝C共轭双键。

Description

流感病毒检测芯片和使用其检测流感病毒的方法
相关申请的交叉引用
本申请要求于2017年07月05日递交的中国专利申请第201710541533.4号的优先权，在此全文引用上述中国专利申请公开的内容以作为本申请的一部分。
技术领域
本公开涉及流感病毒检测领域，且更具体地涉及一种流感病毒检测芯片和一种使用流感病毒检测芯片检测流感病毒的方法。
背景技术
流行性感冒是流感病毒引起的急性呼吸道感染，也是一种传染性强、传播速度快的疾病。一般秋冬季节是其高发期，所引起的并发症和死亡现象非常严重。该病是由流感病毒引起，可分为甲、乙、丙三型，甲型病毒经常发生抗原变异，传染性大，传播迅速，极易发生大范围流行。流感在婴幼儿、老年人和存在心肺基础疾病的患者容易并发肺炎等严重并发症而导致死亡。
目前，主要采用胶体金免疫层析法、ELISA、PCR等方法来进行流感病毒的检测。胶体金免疫层析法需要大量流感病毒累积到检测线处直到显色，这方法检测灵敏度低，当样品流感病毒的含量较低时无法进行检测；而其他方法检测过程复杂，检测时间较长，成本较高。目前许多研究人员正在努力研究新型生物流感病毒传感器，以便替代传统流感病毒检测。
应当注意，提供在上述背景部分中公开的信息仅用于更好地理解本公开的背景，并且因此可以包含未形成那些本领域技术人员已知的现有技术的信息。
发明内容
本公开提供了一种具有高检测灵敏度且检测复杂度减小的流感 病毒检测芯片和一种使用流感病毒检测芯片检测流感病毒的方法。
根据本公开的一方面，提供了一种流感病毒检测芯片，所述流感病毒检测芯片包括：
氧化石墨烯薄膜；
结合垫，沿第一方向设置在所述氧化石墨烯薄膜的一侧；
第一电极和第二电极，分别连接到所述氧化石墨烯膜的在与第一方向垂直的第二方向上的两端，
其中，在所述结合垫中包含带有荧光标记的第一单克隆抗体，在氧化石墨烯膜上具有第二单克隆抗体，其中荧光标记具有C＝C-C＝C共轭双键。
在一个实施例中，流感病毒检测芯片还包括第一基板，所述氧化石墨烯膜通过官能团的作用与所述第一基板结合。
在一个实施例中，流感病毒检测芯片还包括第二基板，与所述第一基板相对设置，所述氧化石墨烯薄膜、所述结合垫、所述第一电极和所述第二电极设置在所述第一基板与所述第二基板之间。
在一个实施例中，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫，其中，样品垫、结合垫和氧化石墨烯膜沿第一方向顺序地设置。
在一个实施例中，所述流感病毒检测芯片还包括用于吸收多余的样品溶液的吸收垫，其中，结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
在一个实施例中，第一电极和第二电极分别与氧化石墨烯膜的在与第一方向垂直的第二方向上的两端相邻并接触。
在一个实施例中，所述第一基板和所述第二基板包括玻璃基板、硅基板或有机聚合物基板中的至少一个。
在一个实施例中，所述第一基板和所述第二基板中的一个是透明的。
在一个实施例中，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫和用于吸收多余的样品溶液的吸收垫，其中，样品垫、结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
在一个实施例中，样品垫和结合垫均是由聚酯纤维制成的彼此独立的或一体化的膜层。
在一个实施例中，样品垫、结合垫和吸收垫具有相同的厚度。
在一个实施例中，样品垫和结合垫以以下结合方式中的至少一种：以彼此交叠的方式相结合、以彼此交叠的方式相结合，和以彼此交叠的方式相结合。
根据本公开的另一方面，提供了一种使用上述流感病毒检测芯片检测流感病毒的方法，所述方法包括：使样品溶液顺序地流过结合垫和氧化石墨烯膜；利用激光照射氧化石墨烯膜，同时利用第一电极和第二电极来检测检测石墨烯氧化物膜两端的电流；以及当石墨烯氧化物膜两端的电流发生变化时，确定在样品溶液中含有流感病毒。
在一个实施例中，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫和用于吸收多余的样品溶液的吸收垫，其中，样品垫、结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
在一个实施例中，样品垫和结合垫均是由聚酯纤维制成的彼此独立的或一体化的膜层。
在一个实施例中，样品垫和结合垫以彼此交叠的方式相结合，结合垫和氧化石墨烯膜以彼此交叠的方式相结合，和/或氧化石墨烯膜和吸收垫以彼此交叠的方式相结合。
应当理解，前面的一般描述和以下详细描述都仅是示例性和说明性的，而不是用于限制本公开。
本节提供本公开中描述的技术的各种实现或示例的概述，并不是所公开技术的全部范围或所有特征的全面公开。
附图说明
图1是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图；
图2是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图；
图3是示出根据本公开的一个实施例的流感病毒检测芯片的示 意性分解透视图；
图4是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图。
具体实施方式
将理解的是，当元件或层被称作在另一元件或层“上”或者“连接到”另一元件或层时，该元件或层可以直接在另一元件或层上、直接连接到或直接结合到另一元件或层，或者也可以存在中间元件或中间层。相反，当元件被称作“直接”在另一元件或层“上”或者“直接连接到”另一元件或层时，不存在中间元件或中间层。同样的标号始终指示同样的元件。如在这里使用的，术语“和/或”包括一个或多个相关所列的项目的任意组合和所有组合。
为了便于描述，在这里可使用空间相对术语，如“下”、“在…上方”、“上”、“在…下方”等来描述如图中所示的一个元件或特征与其它元件或特征的关系。将理解的是，空间相对术语意在包含除了在附图中描述的方位之外的装置在使用或操作中的不同方位。
如这里所使用的，除非上下文另外明确指出，否则单数形式的“一个(种)”和“所述(该)”也意图包括复数形式。还将理解的是，当在本说明书中使用术语“包含”和/或“包括”时，说明存在所述特征、整体、步骤、操作、元件和/或组件，但不排除存在或附加一个或多个其它特征、整体、步骤、操作、元件、组件和/或它们的组。
在下文中，将参照附图详细地解释本公开。
图1是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图。
参见图1，根据本实施例的流感病毒检测芯片100包括上基板101、下基板102以及在上基板101与下基板102之间同层地且沿第一方向(即，从左到右的方向)顺序地设置的结合垫103和氧化石墨烯膜104。此外，流感病毒检测芯片100还包括分别连接到氧化石墨烯膜104的在与第一方向垂直的第二方向上的两端的第一电极105 和第二电极105′，其中，在结合垫103中包含带有荧光标记的单克隆抗体，在氧化石墨烯膜104上具有单克隆抗体，其中荧光标记具有C＝C-C＝C共轭双键。
例如，上基板101和下基板102均可以为玻璃基板、硅基板或有机聚合物基板。具体而言，上基板101和下基板102均可以选自于硅基板、玻璃基板、PET基板、PMMA基板等。
结合垫103是由例如聚酯纤维制成的膜层，但是在本公开中，用于制作结合垫103的材料不限于此。
在结合垫103中包含带有荧光标记的单克隆抗体，其中，荧光标记具有C＝C-C＝C共轭双键。
第一电极105和第二电极105′可以由通用的电极材料(例如，金属、合金、导电金属氧化物、导电金属氮化物等)制成。例如，第一电极105和第二电极105′可以由金属诸如钛、铂、钌、金、银、钼、铝、钨、铜、钕、铬、钽、或包括上述金属的合金、或导电氧化物诸如氧化铟锌、氧化铝锌、氧化铟锡、氧化镓锌或氧化锌锡等来形成。
例如，可以在下基板102上沉积电极材料，然后利用光刻工艺来制作第一电极105和第二电极105′。然后，可以在形成在下基板102上的第一电极105和第二电极105′之间滴入氧化石墨烯溶液并实施干燥，从而在下基板102形成氧化石墨烯膜104。
在本实施例中，在下基板102上形成氧化石墨烯膜104之前，可以对下基板102的表面进行例如氨基修饰，从而在下基板102的表面上修饰的氨基能够与氧化石墨烯膜104的表面上的诸如羧基、环氧基之类的基团发生反应，因此，氧化石墨烯膜104通过这些官能团的作用结合到下基板102。
单克隆抗体也可以通过官能团的作用结合在氧化石墨烯膜104上。
此外，为了使样品溶液顺利流动，结合垫103和氧化石墨烯膜104可以在彼此接触的位置部分地叠置，即，二者可以以彼此交叠的方式相结合，这样可以有利于样品溶液的流动。
上基板101和下基板102可以通过例如粘结剂结合在一起，结合垫103和氧化石墨烯膜104被夹持在上基板101和下基板102之间。
在本实施例中，第一电极105和第二电极105′分别与氧化石墨烯膜104的在与第一方向垂直的第二方向上的两端相邻并接触，并设置在上基板101和下基板102之间。当然，第一电极105和第二电极105′可以以其他方式设置，只要能够检测氧化石墨烯膜104两端的电流(或检测氧化石墨烯膜104的电阻)即可。
当利用本实施例的流感病毒检测芯片100进行流感病毒检测时，可以将待检测的样品溶液(比如，唾液或血液等)加入到流感病毒检测芯片100中，使待检测的样品溶液首先流到结合垫处。当待检测的样品溶液含有流感病毒(即，抗原)时，该抗原与结合垫中的带有荧光标记的单克隆抗体发生特异性反应，然后样品溶液通过毛细作用继续流到氧化石墨烯膜104处，与其上的单克隆抗体发生特异性反应，从而生成双抗夹心流感病毒复合物。
所生成的双抗夹心流感病毒复合物固定在氧化石墨烯膜104的表面上，当利用激光照射到这些复合物时，由于流到氧化石墨烯膜104的荧光标记与氧化石墨烯膜104均具有C＝C-C＝C共轭双键，而且距离很近，从而导致C＝C-C＝C共轭双键发生堆叠，进而发生荧光能量共振转移，即荧光淬灭。为了便于激光照射到复合物，上基板101和下基板102中的一个可以是透明的，当然，上基板101和下基板102也可以均是透明的。
当发生荧光能量共振转移时，荧光标记产生的光子能量被氧化石墨烯吸收，从而导致氧化石墨烯的电阻减小。当氧化石墨烯的电阻减小时，通过利用第一电极105和第二电极105′能够检测到氧化石墨烯膜104两端的电流变化。也就是，当石墨烯氧化物膜104两端的电流发生变化时，可以确定在样品溶液中含有流感病毒。
在本公开的一个实施例中，结合垫中的带有荧光标记的单克隆抗体和氧化石墨烯膜上的单克隆抗体可以是对同一抗原能够发生特异性反应的抗体。结合垫中的带有荧光标记的单克隆抗体和氧化石墨烯膜上的单克隆抗体可以是同种抗体，例如针对禽流感病毒NP蛋白， 带荧光标记的单克隆抗体可以是经过荧光标记的1G11，氧化石墨烯薄膜上的单克隆抗体也可以是1G11，但没有进行荧光标记，即两种抗体相同。然而本公开不限于此，两种抗体也可以是能够对同一抗原发生特异性反应的不同抗体。
图2是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图。
在本实施例中，除了包括前述实施例中的上述组件之外，流感病毒检测芯片200还包括一个用于吸收样品溶液的样品垫106。样品垫106设置在结合垫103的左侧并与结合垫103接触。也就是说，样品垫106、结合垫103和氧化石墨烯膜104沿第一方向(即，从左到右的方向)顺序地设置。
在本实施例中，样品垫106也可以由聚酯纤维制成，但也不限于此。当样品垫106和结合垫103由相同的材料制成时，它们可以分别独立地形成，或者一体地形成。即，样品垫106和结合垫103可以形成为彼此独立的或者一体化的膜层。
当样品垫106和结合垫103形成为单个膜层时，带有荧光标记的单克隆抗体可以结合在单个膜层的靠近石墨烯氧化物膜104的区域上，而未结合在单个膜层的远离石墨烯氧化物膜104的区域上。在此情况下，单个膜层具有两个区域，其中一个区域上含有带有荧光标记的单克隆抗体，而另一区域上不含带有荧光标记的单克隆抗体。
在本实施例中，样品垫106和结合垫103可以具有相同的厚度，以便于被上基板101和下基板102良好地密封。
此外，在本实施例中，当样品垫106和结合垫103是两个彼此独立的组件时，样品垫106和结合垫103可以以彼此交叠的方式相结合，以有利于待检测的样品溶液的顺利流动。当然，在本实施例中，结合垫103和氧化石墨烯膜104也可以以彼此交叠的方式相结合。
当利用本实施例的流感病毒检测芯片200进行流感病毒检测时，待检测的样品溶液将首先流到样品垫106处，然后通过毛细作用流到结合垫103处，后续的历程与在前述实施例中描述的历程类似，因此不再赘述。
图3是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图。
在本实施例中，除了包括前述实施例中的上述组件之外，流感病毒检测芯片300还包括一个用于吸收多余的样品溶液的吸收垫107。吸收垫107设置在氧化石墨烯膜104的右侧并与氧化石墨烯膜104接触。也就是说，结合垫103、氧化石墨烯膜104和吸收垫107沿第一方向(即，从左到右的方向)顺序地设置。
在本实施例中，吸收垫107可以由复合纤维材料制成，但也不限于此，其可以由能够吸收流体的任何材质制成。
在本实施例中，吸收垫107和结合垫103可以具有相同的厚度，以便于被上基板101和下基板102良好地密封。
当利用本实施例的流感病毒检测芯片300进行流感病毒检测时，多余的样品溶液可以被吸收垫107吸收。此外，吸收垫107和氧化石墨烯膜104可以以彼此交叠的方式相结合，以有利于多余的样品溶液顺利地被吸收垫107吸收。
图4是示出根据本公开的一个实施例的流感病毒检测芯片的示意性分解透视图。
在本实施例中，除了包括前述实施例中的上述组件之外，流感病毒检测芯片400还包括一个用于吸收多余的样品溶液的吸收垫107。吸收垫107设置在氧化石墨烯膜104的右侧并与氧化石墨烯膜104接触。也就是说，样品垫106、结合垫103、氧化石墨烯膜104和吸收垫107沿第一方向(即，从左到右的方向)顺序地设置。
吸收垫107可以按照与在前述实施例中描述的方式相同的方式形成并设置。
在本实施例中，样品垫106、结合垫103和吸收垫107具有相同的厚度，以便于被上基板101和下基板102良好地密封。此外，为了有助于样品溶液的顺利流动，样品垫106和结合垫103可以以彼此交叠的方式相结合，结合垫103和氧化石墨烯膜104可以以彼此交叠的方式相结合，和/或氧化石墨烯膜104和吸收垫107可以以彼此交叠的方式相结合。
当利用本实施例的流感病毒检测芯片300进行流感病毒检测时，待检测的样品溶液将首先流到样品垫106处，然后通过毛细作用流到结合垫103处，后续的历程与在前述实施例中描述的历程类似。此外，多余的样品溶液可以被吸收垫107吸收。
本公开的流感病毒检测芯片和利用其来检测流感病毒的方法具有高检测灵敏度，并且检测简单、快捷和方便。此外，可以在样品中含有少量病毒时通过检测电流变化来检测流感病毒的存在与否，检测灵敏度增加，并可以进行早期检测预防，避免流感病毒的持续感染。
已经针对附图给出了对本公开的特定示例性实施例的前面的描述。这些示例性实施例并不意图是穷举性的或者将本公开局限于所公开的精确形式，并且明显的是，在以上教导的启示下，本领域普通技术人员能够做出许多修改和变化。因此，本公开的范围并不意图局限于前述的实施例，而是意图由权利要求和它们的等同物所限定。

Claims (16)

  1. 一种流感病毒检测芯片，所述流感病毒检测芯片包括：
    氧化石墨烯薄膜；
    结合垫，沿第一方向设置在所述氧化石墨烯薄膜的一侧；
    第一电极和第二电极，分别连接到所述氧化石墨烯膜的在与第一方向垂直的第二方向上的两端，
    其中，在所述结合垫中包含带有荧光标记的第一单克隆抗体，在氧化石墨烯膜上具有第二单克隆抗体，其中荧光标记具有C＝C-C＝C共轭双键。
  2. 根据权利要求1所述的流感病毒检测芯片，还包括第一基板，所述氧化石墨烯膜通过官能团的作用与所述第一基板结合。
  3. 根据权利要求2所述的流感病毒检测芯片，还包括第二基板，与所述第一基板相对设置，所述氧化石墨烯薄膜、所述结合垫、所述第一电极和所述第二电极设置在所述第一基板与所述第二基板之间。
  4. 根据权利要求1所述的流感病毒检测芯片，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫，其中，样品垫、结合垫和氧化石墨烯膜沿第一方向顺序地设置。
  5. 根据权利要求1所述的流感病毒检测芯片，所述流感病毒检测芯片还包括用于吸收多余的样品溶液的吸收垫，其中，结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
  6. 根据权利要求1所述的流感病毒检测芯片，其中，第一电极和第二电极分别与氧化石墨烯膜的在与第一方向垂直的第二方向上的两端相邻并接触。
  7. 根据权利要求3所述的流感病毒检测芯片，其中，所述第一基板和所述第二基板包括玻璃基板、硅基板或有机聚合物基板中的至少一个。
  8. 根据权利要求7所述的流感病毒检测芯片，其中，所述第一基板和所述第二基板中的一个是透明的。
  9. 根据权利要求1所述的流感病毒检测芯片，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫和用于吸收多余的样品溶 液的吸收垫，其中，样品垫、结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
  10. 根据权利要求9所述的流感病毒检测芯片，其中，样品垫和结合垫均是由聚酯纤维制成的彼此独立的或一体化的膜层。
  11. 根据权利要求9所述的流感病毒检测芯片，其中，样品垫、结合垫和吸收垫具有相同的厚度。
  12. 根据权利要求9所述的流感病毒检测芯片，其中，样品垫和结合垫以以下结合方式中的至少一种：以彼此交叠的方式相结合、以彼此交叠的方式相结合，和以彼此交叠的方式相结合。
  13. 一种使用根据权利要求1所述的流感病毒检测芯片检测流感病毒的方法，所述方法包括：
    使样品溶液顺序地流过结合垫和氧化石墨烯膜；
    利用激光照射氧化石墨烯膜，同时利用第一电极和第二电极来检测检测石墨烯氧化物膜两端的电流；以及
    当石墨烯氧化物膜两端的电流发生变化时，确定在样品溶液中含有流感病毒。
  14. 根据权利要求13所述的方法，其中，所述流感病毒检测芯片还包括用于吸收样品溶液的样品垫和用于吸收多余的样品溶液的吸收垫，其中，样品垫、结合垫、氧化石墨烯膜和吸收垫沿第一方向顺序地设置。
  15. 根据权利要求14所述的方法，其中，样品垫和结合垫均是由聚酯纤维制成的彼此独立的或一体化的膜层。
  16. 根据权利要求14所述的方法，其中，样品垫和结合垫以彼此交叠的方式相结合，结合垫和氧化石墨烯膜以彼此交叠的方式相结合，和/或氧化石墨烯膜和吸收垫以彼此交叠的方式相结合。
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