CN104165906B - A kind of Viral diagnosis instrument - Google Patents
A kind of Viral diagnosis instrument Download PDFInfo
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- CN104165906B CN104165906B CN201410334079.1A CN201410334079A CN104165906B CN 104165906 B CN104165906 B CN 104165906B CN 201410334079 A CN201410334079 A CN 201410334079A CN 104165906 B CN104165906 B CN 104165906B
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Abstract
The invention discloses a kind of Viral diagnosis instrument, the detector includes interdigitated array microelectrode (1), micro-fluidic detection pond (2) and impedance detection module (3)；When injecting micro-fluidic detection pond (2) containing virulent sample, the antibody or other bio-identification materials that embedded interdigitated array microelectrode (1) surface in micro-fluidic detection pond (2) is fixed will be with virus combinations, produce impedance variations, impedance measurement is carried out by impedance detection module (3), data processing is carried out using standard curve, you can Quantitative detection avian influenza virus content.A kind of Viral diagnosis instrument of the present invention has the advantages that fast, easy to operate detection speed, quantitative analysis and Site Detection.
Description
Technical field
The present invention relates to virus detection techniques field, a kind of Viral diagnosis instrument is more particularly to.
Background technology
In recent years, viral animal epidemic outburst is lasting occurs, such as H 5 N 1 avian influenza, H7N9.Counted according to WHO：2003
Since year, existing 62 national animals and 15 national people have been infected in H5N1 avian influenza virus, 649 people's cases of infection
385 death, the death rate nearly 60%.Highly pathogenic bird flu not only has resulted in huge economic losses, and seriously threatens society
Public health security.
The key of viral animal epidemic prevention and control is quick detection epidemic disease virus.It is existing by taking avian influenza virus as an example
There is avian flu virus detection method mainly to include isolation of virus, inverse transcription polymerase chain reaction method (RT-PCR) and collaurum
Immuno-chromatographic test paper strip.Isolation of virus is the goldstandard method of current diagnosis and identification, and sensitivity is high, but detection time is too
Length is, it is necessary to 7~10 days；RT-PCR was the detection method of world health organisation recommendations, it is necessary to 3~6 hours, and sensitivity is higher, but
Need the sample pre-treatments of complexity；The ELISA test strip time is short, and generally in 15 minutes, but sensitivity is relatively low, and false positive rate is inclined
It is high.Existing method for detecting virus existence time is long, complex operation or the low deficiency of accuracy, and this is limited to a certain extent
Basic animal control and prevention of disease institutions conduct animal epidemic rapid screening works.
The content of the invention
(1) technical problem to be solved
The technical problem to be solved in the present invention is how to improve the detection speed and accuracy rate of virus, while simplifying detection behaviour
Make.
(2) technical scheme
In order to solve the above-mentioned technical problem, the invention provides a kind of Viral diagnosis instrument, the detector includes interdigital battle array
Row microelectrode, micro-fluidic detection pond and impedance detection module；
The interdigitated array microelectrode surface is fixed with bio-identification material；
The micro-fluidic detection pond includes electrode fixture, micro-fluidic module and fastening pull-push piece；
Working chamber is set inside the electrode fixture, and the interdigitated array microelectrode is fixed in the working chamber, described
The upper surface being located at directly over the interdigitated array microelectrode of battery fixture sets a hole, and in micro- positioned at the interdigitated array
The position of the pad of electrode sets two wire guides；
The micro-fluidic module includes the detection cell being adapted to the hole, and the liquid flow being connected with the detection cell
Road；
The fastening pull-push piece fixes the electrode fixture, and drives the electrode fixture to move, and makes the electrode fixture
Hole be tightly connected and separate with the detection cell of the micro-fluidic module；
The impedance detection module includes impedance measuring circuit, and the impedance measuring circuit is produced by impedance measuring equipment to swash
Encourage signal and put on high-pass filter and be filtered, filtered pumping signal is entered by voltage follower to DC offset voltage
Row voltage-regulation is after-applied in the interdigitated array microelectrode, and the impedance measurement module is by controlling the range of range adjuster
Select to be acquired the response signal of the interdigitated array microelectrode.
Preferably, the interdigitated array microelectrode fixes bio-identification material using albumin A or covalent bonding, and uses ox blood
Albumin closes the residual binding sites of the interdigitated array microelectrode.
Preferably, the interdigitated array microelectrode include substrate, and in be etched with the substrate a pair of gold electrodes and
A pair of gold electrode pads；The gold electrode combines shape by the finger electrode of multiple identical sizes with identical spacing parallel connection
Into the pair of gold electrode is staggered, and is connected respectively with the gold electrode pad.
Preferably, the electrode fixture includes base and upper lid；The base, which is set, installs the interdigitated array microelectrode
Groove, groove, the pad that the upper lid is provided with for installing the interdigitated array microelectrode pad be connected with extraneous
Two wire guides and the hole that is connected with the detection cell；The base is tightly connected with upper lid.
Preferably, the base is additionally provided with four outer convex cylindricals for being used to lock the electrode fixture, and the upper lid is also
Four circular holes being adapted with the outer convex cylindrical are set.
Preferably, the rubber ring being adapted is installed in the hole.
Preferably, the flow channel for liquids includes two microchannels and two fluid connectors；In two microchannels
A microchannel be connected by a fluid connector with sample tube；Another microchannel is connected by another fluid connector and pump
Connect, extracted out liquid by the pump.
Preferably, the fastening pull-push piece, which includes one, is used to install the sliding block of the electrode fixture, one and the cunning
Groove that block matches, one be used for pusher, a rotary handle and a screw mandrel for the upward sliding block；It is described
Pusher is connected with the rotary handle by the screw mandrel；Rotate the rotary handle to move up, the pusher extruding
The sliding block, the interdigitated array microelectrode is combined closely with the detection cell；The rotary handle is rotated to move down, it is described
Pusher drives the sliding block slide downward, takes out the electrode fixture.
Preferably, the impedance detection module includes impedance measuring equipment, high-pass filter, voltage follower, range regulation
Device；The output end of the impedance measuring equipment connects the input of the voltage follower by high-pass filter, the voltage with
Connect the pad of the interdigitated array microelectrode respectively with the output end of device and the input of the impedance measuring equipment；The range
The input of adjuster is connected with the input of the impedance measuring equipment；The output end of the range adjuster is surveyed with the impedance
The feedback resistance end connection of measuring device, the impedance measurement module can control the range adjuster and carry out measurement range selection.
Preferably, the impedance detection module also includes microprocessor, touch display screen, voltage-stablizer；The voltage-stablizer is
The impedance measuring equipment is powered；The microprocessor carries out information exchange with the impedance measuring equipment, reads the impedance measurement
The impedance value of device simultaneously carries out man-machine interaction in the touch display screen.
(3) beneficial effect
The invention provides a kind of Viral diagnosis instrument, when injecting micro-fluidic detection pond containing virulent sample, electrode table
The antibody or other bio-identification materials that face is fixed are combined with virus, are produced impedance variations, are hindered by impedance detection module
Anti- measurement, carries out data processing, you can Quantitative detection avian influenza virus content using standard curve.A kind of virus of the invention
Detector has the advantages that fast detection speed, operation semi-automation, quantitative analysis and Site Detection.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the accompanying drawing used required in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, on the premise of not paying creative work, can be with
Other accompanying drawings are obtained according to these accompanying drawings.
Fig. 1 is a kind of structural representation of Viral diagnosis instrument of the preferred embodiment of the present invention；
Fig. 2 is a kind of Viral diagnosis instrument operation principle schematic diagram of the preferred embodiment of the present invention；
Fig. 3 is a kind of interdigitated array microelectrode structural representation of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；
Fig. 4 is a kind of structural representation in the micro-fluidic detection pond of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；
Fig. 5 a show for a kind of structure of the base of the electrode fixture of Viral diagnosis instrument of the preferred embodiment of the present invention
It is intended to；
Fig. 5 b show for a kind of structure of the upper lid of the electrode fixture of Viral diagnosis instrument of the preferred embodiment of the present invention
It is intended to；
Fig. 6 is a kind of structural representation of the impedance detection module of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；
Fig. 7 connects for a kind of circuit of the impedance detection module of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure.
Embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.Following examples are used to illustrate this hair
It is bright, but can not be used for limiting the scope of the present invention.
Fig. 1 is a kind of structural representation of Viral diagnosis instrument of the preferred embodiment of the present invention；The detector bag
Include interdigitated array microelectrode 1, micro-fluidic detection pond 2 and impedance detection module 3.The present embodiment is used for detecting avian influenza virus.
The interdigitated array microelectrode 1 is used for detecting that electrochemical impedance changes, and the micro-fluidic detection pond 2 is used for accurate control and flows through fork
Refer to the sample size and redox probe on the surface of Array microelectrode 1, the impedance detection module 3 is used for the resistance of accurate measuring electrode
Anti- amplitude and phase angle change are simultaneously converted into readable avian influenza virus concentration.
Fig. 3 is a kind of interdigitated array microelectrode structural representation of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；The interdigitated array microelectrode 1 include substrate 1.1, and in be etched with the substrate 1.1 a pair of gold electrodes 1.2,1.3 with
And a pair of gold electrode pads 1.4,1.5；The gold electrode 1.2,1.3 is by the finger electrode of multiple identical sizes with identical spacing
Parallel connection combines to be formed；The gold electrode 1.2,1.3 is staggered, and is connected respectively with the gold electrode pad 1.4,1.5.
Avian influenza virus antibody is modified with the gold electrode 1.2,1.3 surfaces, process is as shown in Figure 2：First pass through quiet
Electricity and hydrophobic effect adsorb albumin A on the surface of the gold electrode 1.2,1.3, recycle the immune ball of albumin A and antibody
The Fc sections of albumen occur specific binding and antibody are fixed on the gold electrode 1.2,1.3, are finally sealed using bovine serum albumin
The binding site for closing residual avoids nonspecific reaction.The interdigitated array microelectrode 1 is arranged on the micro-fluidic detection pond 2
In.During implementation, first the sample comprising avian influenza virus is injected in the micro-fluidic detection pond, avian influenza virus will be by the gold
Antibody on electrode 1.2,1.3 surfaces is captured, and reinjects redox probe (10mM [Fe (CN)6]3-/4-) the measurement gold
The impedance variations of electrode 1.2,1.3 carry out avian influenza virus content in judgement sample.
Fig. 4 is a kind of structural representation in the micro-fluidic detection pond of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；The micro-fluidic detection pond 2 includes electrode fixture 2.1, micro-fluidic module 2.2 and fastening pull-push piece 2.3.It is described micro-fluidic
The material of detection cell 2.2 is teflon or other biochemistry inert materials.
Fig. 5 a show for a kind of structure of the base of the electrode fixture of Viral diagnosis instrument of the preferred embodiment of the present invention
It is intended to；Fig. 5 b are a kind of structural representation of the upper lid of the electrode fixture of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；The electrode fixture 2.1 includes base 2.1.1 and upper lid 2.1.2；The base 2.1.1 is provided with the installation interdigital battle array
The groove 2.1.1.1 of row microelectrode 1, the upper lid 2.1.2 are provided with for installing the recessed of the pad of interdigitated array microelectrode 1
The hole that groove 2.1.2.3, the pad are connected with extraneous two wire guide 2.1.2.4 being connected and with the detection cell
2.1.2.1；The base 2.1.1 is tightly connected with upper lid 2.1.2, the hole 2.1.2.1 install the rubber ring that is adapted there is provided
Compensator or trimmer pressure.
The base 2.1.1 is additionally provided with the four outer convex cylindrical 2.1.1.2 and use for being used to lock the electrode fixture
Four and the outer convex cylindrical are also set up in the circular hole 2.1.1.3, the upper lid 2.1.2 for taking out the interdigitated array microelectrode 1
2.1.1.2 the circular hole 2.1.2.2 being adapted.
During implementation, first the interdigitated array microelectrode 1 is arranged in the groove 2.1.1.1, then by the upper lid
2.1.2 align and the base 2.1.1 and push, upper the lid 2.1.2 and the base 2.1.1 pass through the outer convex cylindrical
2.1.1.2 with the tight fit strong bonded of the circular hole 2.1.2.2, finally rubber ring is arranged in the circular hole 2.1.2.1.
The wire that the signal of the interdigitated array microelectrode 1 is welded in a gold medal thin slice by two one end passes through the wire guide 2.1.2.4
It is acquired.
The micro-fluidic module 2.2 includes the detection cell 2.2.1 that is adapted to the hole 2.1.2.1, and with the detection
The flow channel for liquids of pond 2.2.1 connections；The flow channel for liquids includes two microchannel 2.2.2 and two fluid connector 2.2.3；
A microchannel in two microchannel 2.2.2 is connected by a fluid connector 2.2.3 with sample tube；Another micro- logical
Road 2.2.2 is connected by another fluid connector 2.2.3 with pump, is extracted out liquid by the pump.
The fastening pull-push piece 2.3 fixes the electrode fixture 2.1, and drives the electrode fixture 2.1 to move, and makes described
The hole 2.1.2.1 of electrode fixture 2.1 is tightly connected and separated with the detection cell 2.2.1 of the micro-fluidic module 2.2.The fastening
Pull-push piece 2.3 include one be used for install the sliding block 2.3.1 of the electrode fixture 2.1, one match with the sliding block 2.3.1
Groove 2.3.2, one be used for the upward sliding block 2.3.1 pusher 2.3.3, one with pusher 2.3.3 match
The rotary handle 2.3.4 of a set and threaded screw mandrel 2.3.5；The sliding block 2.3.1 is provided with one and the electrode
The groove that fixture 2.1 is adapted, for installing the electrode fixture 2.1；The pusher 2.3.3 and the rotary handle
2.3.4 the screw mandrel 2.3.5 connections are passed through.Before detection, by rotating the rotary handle 2.3.4, the promotion is moved up
Device 2.3.3, extrudes the sliding block 2.3.1, the interdigitated array microelectrode 1 can be made to be combined closely with the detection cell 2.2.1, and two
Rubber ring is provided between person as pressure buffer；After detection, moved down by rotating the rotary handle 2.3.4, it is described
Pusher 2.3.3 drives the sliding block 2.3.1 slide downwards, takes out the electrode fixture 2.1.
The micro-fluidic module 2.2 and fastening pull-push piece 2.3 are locked by bolt 2.4 and nut 2.5.
Fig. 6 is a kind of structural representation of the impedance detection module of Viral diagnosis instrument of the preferred embodiment of the present invention
Figure；Fig. 7 is a kind of circuit connection diagram of the impedance detection module of Viral diagnosis instrument of the preferred embodiment of the present invention.
The impedance detection module 3 includes microprocessor, impedance measuring equipment, high-pass filter, voltage follower, range and adjusted
Save device；The output end of the impedance measuring equipment connects the input of the voltage follower, the voltage by high-pass filter
The input of the output end of follower and the impedance measuring equipment connects the pad of the interdigitated array microelectrode 1 respectively；It is described
The input of range adjuster is connected with the input of the impedance measuring equipment；The output end of the range adjuster and the resistance
The feedback resistance end connection of anti-measuring appliance；The range adjuster is connected with the microprocessor；The microprocessor with it is described
Impedance measuring equipment is connected.The impedance detection module 3 also includes display screen, voltage-stablizer and voltage-stabilized power supply circuit；The voltage stabilizing
Device is that the impedance measuring equipment is powered；The display screen is connected with the microprocessor, for man-machine interactive operation；The voltage stabilizing
Power circuit is that the microprocessor and display screen are powered.
The microprocessor uses S3C2440ARM9 processors, and the voltage-stabilized power supply circuit uses AMS1117-3.3 low pressure
Poor linear voltage stabilization chip uses touch display screen there is provided 3.3V voltages, the touch display screen.The impedance measuring equipment, voltage stabilizing
Device, high-pass filter, voltage follower and range adjuster composition impedance measuring circuit, for accurately measuring the interdigitated array
The impedance magnitude and phase angle of microelectrode 1.The impedance measuring equipment uses AD5933 or AD5934 impedance transducers, sharp for producing
Encourage voltage signal to put on the interdigitated array microelectrode 1, and gather the response signal of the interdigitated array microelectrode 1；Institute
The high-precision linear voltage regulator that voltage-stablizer uses ADP3303-3.3 is stated, for providing high-precision 3.3V power supplys to described
Impedance measuring equipment；The high-pass filter is RC high-pass filtering circuits, for filtering low noise signal；The voltage follower
Using 1/2AD8606 amplifiers, for by the response signal DC offset voltage of the interdigitated array microelectrode 1 adjust to
1.65V, eliminates bias voltage difference；The range adjuster uses ADG811 analog switches, for automatically selecting suitable mark
Directrix curve.During implementation, two gold-plated electrode clamps are connected by the impedance measuring circuit, the electrode clamp is clamped respectively
On two wires that the wire guide 2.1.2.4 of the electrode holder 1 comes out, the signal of the interdigitated array microelectrode 1 is obtained.
A kind of Viral diagnosis instrument that the present invention is provided, when injecting micro-fluidic detection pond containing virulent sample, electrode table
The bio-identification material that face is fixed is combined with virus, produces impedance variations, and impedance measurement is carried out by impedance detection module, is utilized
Standard curve carries out data processing, you can Quantitative detection avian influenza virus content.A kind of Viral diagnosis instrument of the present invention has
Detection speed is fast, operation semi-automation, the advantage of quantitative analysis and Site Detection.
Embodiment of above is merely to illustrate the present invention, rather than limitation of the present invention.Although with reference to embodiment to this hair
It is bright to be described in detail, it will be understood by those within the art that, to technical scheme carry out it is various combination,
Modification or equivalent substitution, without departure from the spirit and scope of technical solution of the present invention, the right that all should cover in the present invention is wanted
Ask among scope.
Claims (10)
1. a kind of Viral diagnosis instrument, it is characterised in that the detector includes interdigitated array microelectrode (1), micro-fluidic detection pond
And impedance detection module (3) (2)；
Bio-identification material is fixed with the surface of the interdigitated array microelectrode (1)；
The micro-fluidic detection pond (2) includes electrode fixture (2.1), micro-fluidic module (2.2) and fastening pull-push piece (2.3)；
Working chamber is set inside the electrode fixture (2.1), and the interdigitated array microelectrode (1) is fixed in the working chamber,
The electrode fixture (2.1) sets a hole (2.1.2.1) in the upper surface directly over the interdigitated array microelectrode (1),
And two wire guides (2.1.2.4) are set in the position of the pad positioned at the interdigitated array microelectrode (1)；
The micro-fluidic module (2.2) includes being adapted to the hole (2.1.2.1) that electrode fixture (2.1) upper surface is set
Detection cell (2.2.1), and the flow channel for liquids being connected with the detection cell (2.2.1)；
The fixed electrode fixture (2.1) of fastening pull-push piece (2.3), and drive the electrode fixture (2.1) to move, make institute
The hole (2.1.2.1) of electrode fixture (2.1) is stated to be tightly connected and divide with the detection cell (2.2.1) of the micro-fluidic module (2.2)
From；
The impedance detection module (3) includes impedance measuring circuit, and the impedance measuring circuit is produced by impedance measuring equipment to swash
Encourage signal and put on high-pass filter and be filtered, filtered pumping signal is entered by voltage follower to DC offset voltage
Row voltage-regulation is after-applied in the interdigitated array microelectrode (1), and the impedance measuring equipment is by controlling the amount of range adjuster
Journey selects to be acquired the response signal of the interdigitated array microelectrode (1).
2. a kind of detector according to claim 1, it is characterised in that the interdigitated array microelectrode (1) uses albumin A
Or covalent bonding fixes bio-identification material, and the residual for closing the interdigitated array microelectrode (1) with bovine serum albumin is combined
Site.
3. a kind of detector according to claim 1 or 2, it is characterised in that the interdigitated array microelectrode (1) includes base
Bottom (1.1), and in being etched with a pair of gold electrodes (1.2), (1.3) and a pair of gold electrode pads in the substrate (1.1)
(1.4)、(1.5)；The gold electrode (1.2), (1.3) are by the finger electrode of multiple identical sizes with identical spacing parallel connection
Combination is formed, and the gold electrode (1.2), (1.3) are staggered, and are connected respectively with the gold electrode pad (1.4), (1.5).
4. a kind of detector according to claim 1, it is characterised in that the electrode fixture (2.1) includes base
(2.1.1) and upper lid (2.1.2)；The base (2.1.1) sets the groove for installing the interdigitated array microelectrode (1), described
Groove, the pad that upper lid (2.1.2) is provided with for installing interdigitated array microelectrode (1) pad are connected with the external world
Two wire guides and the hole (2.1.2.1) being tightly connected with the detection cell；The base (2.1.1) and upper lid (2.1.2)
It is tightly connected.
5. a kind of detector according to claim 4, it is characterised in that the base (2.1.1) is additionally provided with four use
In the outer convex cylindrical (2.1.1.2) for locking the electrode fixture, the upper lid (2.1.2) also sets up four and the outer convex cylindrical
The circular hole (2.1.2.2) that (2.1.1.2) is adapted.
6. a kind of detector according to any one of claim 1,4,5, it is characterised in that the hole (2.1.2.1) is installed
The rubber ring being adapted.
7. a kind of detector according to claim 1, it is characterised in that the flow channel for liquids includes two microchannels
(2.2.2) and two fluid connectors (2.2.3)；A microchannel in two microchannels (2.2.2) passes through one stream
Body connector (2.2.3) is connected with sample tube；Another microchannel (2.2.2) passes through another fluid connector (2.2.3) and pump
Connection, is extracted out liquid by the pump.
8. a kind of detector according to claim 1, it is characterised in that the fastening pull-push piece (2.3) includes a use
In installing the sliding block (2.3.1) of the electrode fixture (2.1), the groove that matches with the sliding block (2.3.1)
(2.3.2), one be used for the pusher (2.3.3) of the upward sliding block (2.3.1), a rotary handle (2.3.4) with
And a screw mandrel (2.3.5)；The pusher (2.3.3) is connected with the rotary handle (2.3.4) by the screw mandrel (2.3.5)
Connect；Rotate the rotary handle (2.3.4) to move up, the pusher (2.3.3) extrudes the sliding block (2.3.1), described
Interdigitated array microelectrode (1) is combined closely with the detection cell (2.2.1)；The rotary handle (2.3.4) is rotated to move down,
The pusher (2.3.3) drives sliding block (2.3.1) slide downward, takes out the electrode fixture (2.1).
9. a kind of detector according to claim 1, it is characterised in that the impedance detection module (3) is surveyed including impedance
Measuring device, high-pass filter, voltage follower, range adjuster；The output end of the impedance measuring equipment is connected by high-pass filter
Connect the input of the voltage follower, the input of the output end of the voltage follower and the impedance measuring equipment connects respectively
Connect the pad of the interdigitated array microelectrode (1)；The input of the input of the range adjuster and the impedance measuring equipment
Connection；The output end of the range adjuster is connected with the feedback resistance end of the impedance measuring equipment, the impedance measurement module
(3) it can control the range adjuster and carry out measurement range selection.
10. a kind of detector according to claim 9, it is characterised in that the impedance detection module (3) also includes micro- place
Manage device, touch display screen, voltage-stablizer；The voltage-stablizer is that the impedance measuring equipment is powered；The microprocessor and the range
Adjuster, impedance measuring equipment connection, the microprocessor carry out information exchange with the impedance measuring equipment, read the resistance
The impedance value of anti-measuring appliance simultaneously carries out man-machine interaction in the touch display screen.
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| CN107389752A (en) * | 2017-07-31 | 2017-11-24 | 重庆微奥云生物技术有限公司 | A kind of bacterial antigens detecting system and detection method of quality control |
| CN109919922B (en) * | 2019-02-27 | 2023-02-03 | 重庆大学 | Quality detection method for silicon-silicon direct bonding based on combination of spatial domain and morphology |
| CN112547143B (en) * | 2019-09-26 | 2022-07-22 | 京东方科技集团股份有限公司 | Micro-fluidic chip and blood cell detection device |
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