CN109580506A - A kind of fungal detection system and two Methods for Fungi Detection based on digital microfluidic technology - Google Patents
A kind of fungal detection system and two Methods for Fungi Detection based on digital microfluidic technology Download PDFInfo
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Abstract
The present invention provides a kind of fungal detection system based on digital microfluidic technology, including digital microcurrent-controlled chip, integrated circuit, photometric detection module, temperature control raising module, the digital microcurrent-controlled chip is connect with the integrated circuit, photometric detection module, temperature control raising module respectively, the temperature control raising module is set to below the photometric detection module, and the integrated circuit is connect with the photometric detection module, temperature control raising module respectively；A kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology is also provided.The beneficial effects of the present invention are: digital microcurrent-controlled realize control more flexible to drop, quick, automation and integrated aspect are with the obvious advantage, and reagent and sample requirement amount are smaller, and system bulk is smaller, and pipetting can multichannel carry out simultaneously, it is more efficient；Sample area, drop formation channel electrode array and hatching detection zone electrode use rectangle tooth shaped structure, can increase the contact area of drop and adjacent electrode, the more conducively movement of drop, help to reduce driving voltage.
Description
Technical field
The invention belongs to digital microfluidic technical fields, examine more particularly, to a kind of fungi based on digital microfluidic technology
The method of examining system and fungal detection.
Background technique
At present because fungus-caused disease incidence rises rapidly, and the hair of quick, effective, convenient and fast fungal detection system
Exhibition is also concerned.ELISA reaches its maturity as a kind of important two Methods for Fungi Detection application, utilizes the special of antibody and antigen
Property combine, through incubation washing after, be added substrate solution generate chromogenic reaction, determine its extinction under specific wavelength illumination
Degree, may be implemented the detection of fungi.
Electrowetting on dielectric technology changes drop by voltage and realizes the manipulation to drop in the wettability of dielectric surface,
Have many advantages such as that driving method is simple, driving force is strong, high degree of automation.It is micro-fluidic that a kind of micro-nano fluid is used as to handle skill
Art is concerned in recent years.Its is at low cost, flux is high, analysis speed is fast, reagent consumes few advantage, for fungi Bacteria Detection
System reduces that cost, device miniaturization are integrated etc. to be of great significance.Digital microcurrent-controlled based on dielectric wetting is used as one
Kind emerging discretization microlayer model manipulation means realize that discrete droplets accurately control (including movement, conjunction based on microelectrode array
And the operation such as division), the manipulation to microlayer model is realized using dielectric wetting phenomena of the drop on hydrophobizing surface, it can be real
Now the controllable of big flux discretization drop is precisely controlled.This mode of operation has the characteristics that high concurrency and full-automation, energy
The real-time controllable reaction of multi-path is enough realized, especially suitable for high integration, high-performance, micro- biochemical analysis system complicated for operation.
Summary of the invention
The problem to be solved in the present invention is to provide a kind of achievable fungi Bacteria Detections to minimize, integrated based on number
The fungal detection system and two Methods for Fungi Detection of microflow control technique.
In order to solve the above technical problems, the technical solution adopted by the present invention is that: it is a kind of based on the true of digital microfluidic technology
Bacterial examination examining system, including digital microcurrent-controlled chip, integrated circuit, photometric detection module, temperature control raising module, the number miniflow
Control chip is connect with the integrated circuit, photometric detection module, temperature control raising module respectively, and the temperature control raising module is set to institute
It states below photometric detection module, the integrated circuit is connect with the photometric detection module, temperature control raising module respectively, the collection
Computer is separately connected at circuit and photometric detection module.
In technical solution, it is preferred that the digital microcurrent-controlled chip includes the bottom crown, top crown and centre being placed in parallel
Gap, drop moves in the gap；It upper substrate that the top crown is set gradually from top to bottom, upper conductive layer and upper dredges
Water layer, the bottom crown include the lower hydrophobic layer set gradually from top to bottom, dielectric layer, lower conductiving layer and lower substrate；Under described
Conductive layer includes sample area, drop formation channel electrode array, reagent liquid storage unit and hatching detection zone, the sample area, liquid
Drop generates channel electrode array, reagent liquid storage unit is the effigurate metal electrode array of tool, the sample area, reagent
Liquid storage unit and hatching detection zone are set to around the drop formation channel electrode array.
In technical solution, it is preferred that the photometric detection module includes the light emitting diode set gradually from top to bottom, filter
Mating plate, aperture, silicon photocell.
In technical solution, it is preferred that the hatching detection zone is set between the aperture and the silicon photocell.
In technical solution, it is preferred that the sample area quantity is 2-4, and the hatching detection zone includes 5-10 subregion.
In technical solution, it is preferred that the sample area, drop formation channel electrode array and the electrode portion for hatching detection zone
Dividing is rectangle tooth shaped electrode.
In technical solution, it is preferred that the reagent liquid storage unit include label after antibody district, washing lotion area, substrate solution area,
Terminate liquid zone, critical value reference substance area, negative controls area, positive reference substance area and waste.
A kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology, includes the following steps:
Step 1, will label after antibody, washing lotion, substrate solution, terminate liquid, critical value reference substance, negative controls, the positive
Reference substance is separately added into each corresponding area of reagent liquid storage unit, and various kinds is originally separately added into each sample area；
Step 2, the control circuit coordination electrode driving circuit by integrated circuit carry out the on-off of electrode in a certain order
Electric control is moved to hatching detection zone by drop formation channel electrode array respectively by antibody reagent with single drops
Each subregion controls electrode in each subregion and is stirred to antibody reagent, reagent is then moved to waste；
Step 3, the droplet manipulation method according to step 2 are separately added into critical value control into each area of hatching detection zone
Product, negative controls, positive reference substance and each sample, are stirred respectively；
Step 4, integrated circuit control temperature control raising module are kept for 36.5-37.5 DEG C, 85-95min are incubated for, then by reagent
It is moved to waste；
Washing lotion is moved to hatching each area of detection zone, after being stirred respectively by step 5, the droplet manipulation method according to step 2
Waste is moved to, is repeated 4-8 times；
Substrate solution is moved to hatching each area of detection zone, is stirred respectively by step 6, the droplet manipulation method according to step 2
It mixes；
Step 7, control temperature control raising module, are kept for 36.5-37.5 DEG C, are incubated for 15-25min, will hatch each area of detection zone
Interior reagent is moved to waste；
Terminate liquid is moved to hatching each area of detection zone, is stirred respectively by step 8, the droplet manipulation method according to step 2
It mixes；
Step 9, integrated circuit control photometric detection module, make lumination of light emitting diode, measure hatching each area's light of detection zone
Signal power reads OD value, calculates sample index.
In technical solution, it is preferred that be marked with fluorocarbon oil in the gap.
In technical solution, it is preferred that the top crown, hatching detection zone are transparent.
The advantages and positive effects of the present invention are:
(1) digital microcurrent-controlled to realize control more flexible to drop, quick, automation and integrated aspect are with the obvious advantage,
Reagent and sample requirement amount are smaller, and system bulk is smaller, and pipetting can multichannel carry out simultaneously, it is more efficient；
(2) sample area, drop formation channel electrode array and hatching detection zone electrode use rectangle tooth shaped structure, can be with
Increase the contact area of drop and adjacent electrode, the more conducively movement of drop helps to reduce driving voltage；
(3) hatching detection zone and electrode channel are integrated, so that drop is mobile, hatch, detects highly integrated, improves and integrate
Degree simplifies liquid relief testing process；
(4) control group is set, and testing result is more accurate；
(5) hatching detection zone is up and down transparent configuration, and drop thickness is fixed, and light position is fixed, and can directly be shown
Photometric measurement after color, it is more efficient, more acurrate.
Detailed description of the invention
Fig. 1 is the overall system view of fungal detection system of the one embodiment of the invention based on digital microfluidic technology.
Fig. 2 is the side structure schematic diagram of fungal detection chip of the one embodiment of the invention based on digital microfluidic technology.
Fig. 3 is subregion and planar structure schematic diagram of the one embodiment of the invention in the digital microcurrent-controlled chip of Fig. 1.
Fig. 4 is the schematic diagram that one embodiment of the invention realizes fungal detection on the digital microcurrent-controlled chip of Fig. 1.
In figure:
The digital microcurrent-controlled chip of 1-, 2- integrated circuit, 3- photometric detection module, 4- temperature control raising module
11- bottom crown, 12- top crown, the gap 13-
Substrate under hydrophobic layer, 112- lower conductiving layer, 113- dielectric layer, 114- under 111-
The upper substrate of 121-, the upper conductive layer of 122-, the upper hydrophobic layer of 123-
The sample area 1121-, 1122- drop formation channel electrode array, 1123- reagent liquid storage unit, 1124- hatching detection
Area
31- light emitting diode, 32- optical filter, 33- aperture, 34- silicon photocell
Antibody district after C- label, D- washing lotion area, E- substrate solution area, F- terminate liquid zone, G- critical value reference substance area, H- yin
Property reference substance area, I- positive reference substance area, K- waste
Specific embodiment
Embodiment 1
With reference to the accompanying drawing by taking the Aspergillus galactomannan antigen for detection (ELISA method) on digital microcurrent-controlled chip 1 as an example,
Further careful description is carried out to the present invention.
As shown in Figure 1, a kind of fungal detection system based on digital microfluidic technology, including digital microcurrent-controlled chip 1, collection
At circuit 2, photometric detection module 3, temperature control raising module 4, digital microcurrent-controlled chip 1 respectively with integrated circuit 2, photometric detection mould
Block 3, temperature control raising module 4 connect, and temperature control raising module 4 is set to the lower section of photometric detection module 3, integrated circuit 2 respectively with luminosity
Detection module 3, temperature control raising module 4 connect, and integrated circuit 2 and photometric detection module 3 are separately connected computer, photometric detection mould
Block 3 includes light emitting diode 31, optical filter 32, aperture 33, the silicon photocell 34 set gradually from top to bottom.
As in Figure 2-4, digital microcurrent-controlled chip 1 is bipolar plate structure, including bottom crown 11, the top crown being placed in parallel
12 move in gap 13 with intermediate gap 13, drop；Upper substrate 121 that top crown 12 is set gradually from top to bottom, on lead
Electric layer 122 and upper hydrophobic layer 123, bottom crown 11 include set gradually from top to bottom lower hydrophobic layer 111, dielectric layer 113, under lead
Electric layer 112 and lower substrate 114,113 uniform ground of dielectric layer are covered on 112 upper surface of lower conductiving layer, upper substrate 121, lower base
Material is glass, and upper hydrophobic layer 123, lower hydrophobic layer 111 use plasma hydrophobic treatment, reduces other materials hydrophobic coating to electricity
The influence of characteristic is learned, the problem of hydrophobic layer easily punctures is improved.
Lower conductiving layer 112 includes 2-4 sample area 1121, drop formation channel electrode array 1122, reagent liquid storage unit
1123 are set to drop life with hatching detection zone 1124, sample area 1121, reagent liquid storage unit 1123 and hatching detection zone 1124
Around channel electrode array 1122, sample area 1121, drop formation channel electrode array 1122, reagent liquid storage unit 1123 are equal
To have effigurate metal or conductive glass electrode array, the electrode material of hatching detection zone 1124 is electro-conductive glass
(ITO), sample area 1121, drop formation channel electrode array 1122 and hatching 1124 electrode shape of detection zone are rectangle prong
Shape, rectangle tooth shaped electrode can increase the contact area of drop and adjacent electrode, peomote the movement of drop, reduce and drive
Dynamic voltage.
Reagent liquid storage unit 1123 includes antibody district, washing lotion area D, substrate solution area E, termination liquid zone F, critical value after label
Reference substance area G, negative controls area H, positive reference substance area I and waste K, convenient for storing, taking out each reagent.
Top crown 12, bottom crown 11 and luminosity hatching detection zone 3 are both designed as transparent configuration, and hatching detection zone designs up and down
For transparent configuration, drop thickness is fixed, and light position is fixed, and the photometric measurement after can directly being developed the color is more efficient, more quasi-
Really.
Hatch detection zone 1124 to be set between aperture 33 and silicon photocell 34, including 5 or more subregions, in preparation process
The coating process that fungal antibody corresponds to coating antigen is completed in hatching detection zone 1124.
A kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology, includes the following steps:
Step 1, will label after antibody, washing lotion, substrate solution, terminate liquid, critical value reference substance, negative controls, the positive
Reference substance is separately added into each corresponding area of reagent liquid storage unit 1123, various kinds is originally separately added into each sample area 1121；
Step 2, the control circuit coordination electrode driving circuit by integrated circuit 2 carry out the logical of electrode in a certain order
Power-off control, respectively by antibody reagent with single drops, is moved to hatching by drop formation channel electrode array 1122
Each subregion of detection zone 1124 controls electrode in each subregion and is stirred to antibody reagent, reagent is then moved to waste；
Step 3, the droplet manipulation method according to step 2 are separately added into critical value into each area of hatching detection zone 1124
Reference substance, negative controls, positive reference substance and each sample, are stirred respectively；
Step 4, integrated circuit 2 control temperature control raising module 4 and are kept for 36.5 DEG C, are incubated for 85min, are then moved to reagent
Waste；
Washing lotion is moved to the hatching each area of detection zone 1124, is stirred respectively by step 5, the droplet manipulation method according to step 2
It mixes and moves back to waste, be repeated 6 times；
Substrate solution is moved to hatching each area of detection zone, is stirred respectively by step 6, the droplet manipulation method according to step 2
It mixes；
Step 7, control temperature control raising module 4, are kept for 37.5 DEG C, are incubated for 20min, will be tried in the hatching each area of detection zone 1124
Agent is moved to waste；
Terminate liquid is moved to hatching each area of detection zone, is stirred respectively by step 8, the droplet manipulation method according to step 2
It mixes；
Step 9, integrated circuit 2 control photometric detection module 3, and light emitting diode 31 is made to shine, and it is each to measure hatching detection zone
Area's optical signal power reads OD value, calculates sample index；
Data processing:
The mean value of critical value control value: the OD value of 2 critical value reference substances is added, divided by 2；
Negative control index: by the OD value of negative control divided by the OD value mean value of critical value reference substance；
Positive control index: by the OD value of positive control divided by the OD value mean value of critical value reference substance；
Sample index: by the OD value of sample divided by the OD value mean value of critical value reference substance.
Quality control and result interpretation:
The OD value > 0.2 of critical value reference substance indicates that critical value control is effective；
Negative control index < 0.4 indicates that negative control is effective；
Positive control index > 1.5 indicates that positive control is effective；
Sample index >=0.5 is the positive, and sample index < 0.5 is feminine gender.
By the control circuit coordination electrode driving circuit of integrated circuit 2, the on-off for carrying out electrode in a certain order is automatically controlled
System generates drop from liquid storage electrode unit, then carries out antibody combination-sample-adding product-incubation-washing-incubation-plus substrate-incubation-
Termination-illumination photometric detection, when carrying out photometric detection, light emitting diode 31 issues optical signal, aperture 33 and lower section hatching
Detection zone 1124 and silicon photocell 34 are aligned, and optical signal is irradiated by optical filter 32 and aperture 33 with specific position, specific wavelength
Onto sample, light passing is irradiated on silicon photocell 34, is obtained optical power (luminosity) value, is transferred on computer.
Fluorocarbon oil is added in gap 13 in the volatilization of drop and surface adhesion in order to prevent, can promote the movement of drop.
Stirring device gymnastics is used as in region, is merged after repeating 20-30 drop movement, drop separation.
Fungal detection system whole process is in closing light protected environment.
Digital microcurrent-controlled manufacture craft of the invention is as follows:
The production of bottom crown 11:
(1) any insulation transparent material of 11 choice of the substrates of bottom crown, such as glass pass through the works such as photoetching, sputtering in substrate
Skill forms the metal electrode layer with certain figure, wherein hatching detection zone electrode material chooses the high material of light transmittance, such as ITO
Deng being formed by the chemical vapor deposition of plasma enhancing；
(2) dielectric layer 113 is with high dielectric constant, the insulation material that thickness is uniform, upper surface is smooth, breakdown characteristics are strong
Material, such as PDMS, PMMA, Paralyene-C, Si3N4 material, are formed, 2-5 μm of thickness by way of spin coating or deposition；
(3) hydrophobic layer is formed after being annealed by spin coating Teflon, can also by 113 upper surface of dielectric layer is carried out etc. from
Sub- hydrophobic treatment or silanizing solution processing are formed, and 2 μm of thickness or less；
(4) hatching detection zone is soaked in coating solution after completing, antibody is carried out and corresponds at the coating of coating antigen
Reason.
The production of top crown 12:
(1) any insulation transparent material of 12 choice of the substrates of top crown, such as glass；
(2) conductive layer chooses the high material of light transmittance, such as ITO, is formed by deposition；
(3) 12 hydrophobic layer of top crown is consistent with 11 hydrophobic layer of bottom crown.
It is 100 μm of gap 13 (optional 50-500 μm of spacing) between upper bottom crown 11, drop passes through corresponding bottom crown 11
Higher level's Board position aperture above liquid storage electrode unit is instilled from outside, and fluorocarbon oil oil sealing is added in gap 13, and it is dirty to play isolation
The effect for contaminating, preventing drop volatilization, promoting drop mobile.
Embodiment 2
A kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology, includes the following steps:
Step 1, will label after antibody, washing lotion, substrate solution, terminate liquid, critical value reference substance, negative controls, the positive
Reference substance is separately added into each corresponding area of reagent liquid storage unit 1123, various kinds is originally separately added into each sample area 1121；
Step 2, the control circuit coordination electrode driving circuit by integrated circuit 2 carry out the logical of electrode in a certain order
Power-off control, respectively by antibody reagent with single drops, is moved to hatching by drop formation channel electrode array 1122
Each subregion of detection zone 1124 controls electrode in each subregion and is stirred to antibody reagent, reagent is then moved to waste；
Step 3, the droplet manipulation method according to step 2 are separately added into critical value into each area of hatching detection zone 1124
Reference substance, negative controls, positive reference substance and each sample, are stirred respectively；
Step 4, integrated circuit 2 control temperature control raising module 4 and are kept for 37 DEG C, are incubated for 95min, are then moved to reagent useless
Liquid zone；
Washing lotion is moved to the hatching each area of detection zone 1124, is stirred respectively by step 5, the droplet manipulation method according to step 2
It mixes and moves back to waste, be repeated 4 times；
Substrate solution is moved to hatching each area of detection zone, is stirred respectively by step 6, the droplet manipulation method according to step 2
It mixes；
Step 7, control temperature control raising module 4, are kept for 36.5 DEG C, are incubated for 25min, will be tried in the hatching each area of detection zone 1124
Agent is moved to waste；
Terminate liquid is moved to hatching each area of detection zone, is stirred respectively by step 8, the droplet manipulation method according to step 2
It mixes；
Step 9, integrated circuit 2 control photometric detection module 3, and light emitting diode 31 is made to shine, and it is each to measure hatching detection zone
Area's optical signal power reads OD value, calculates sample index.
Data processing:
The mean value of critical value control value: the OD value of 2 critical value reference substances is added, divided by 2；
Negative control index: by the OD value of negative control divided by the OD value mean value of critical value reference substance；
Positive control index: by the OD value of positive control divided by the OD value mean value of critical value reference substance；
Sample index: by the OD value of sample divided by the OD value mean value of critical value reference substance.
Quality control and result interpretation:
The OD value > 0.2 of critical value reference substance indicates that critical value control is effective；
Negative control index < 0.4 indicates that negative control is effective；
Positive control index > 1.5 indicates that positive control is effective；
Sample index >=0.5 is the positive, and sample index < 0.5 is feminine gender.
Embodiment 3
A kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology, includes the following steps:
Step 1, will label after antibody, washing lotion, substrate solution, terminate liquid, critical value reference substance, negative controls, the positive
Reference substance is separately added into each corresponding area of reagent liquid storage unit 1123, various kinds is originally separately added into each sample area 1121；
Step 2, the control circuit coordination electrode driving circuit by integrated circuit 2 carry out the logical of electrode in a certain order
Power-off control, respectively by antibody reagent with single drops, is moved to hatching by drop formation channel electrode array 1122
Each subregion of detection zone 1124 controls electrode in each subregion and is stirred to antibody reagent, reagent is then moved to waste；
Step 3, the droplet manipulation method according to step 2 are separately added into critical value into each area of hatching detection zone 1124
Reference substance, negative controls, positive reference substance and each sample, are stirred respectively；
Step 4, integrated circuit 2 control temperature control raising module 4 and are kept for 37.5 DEG C, are incubated for 90min, are then moved to reagent
Waste；
Washing lotion is moved to the hatching each area of detection zone 1124, is stirred respectively by step 5, the droplet manipulation method according to step 2
It mixes and moves back to waste, be repeated 8 times；
Substrate solution is moved to hatching each area of detection zone, is stirred respectively by step 6, the droplet manipulation method according to step 2
It mixes；
Step 7, control temperature control raising module 4, are kept for 37 DEG C, are incubated for 15min, by reagent in the hatching each area of detection zone 1124
It is moved to waste；
Terminate liquid is moved to hatching each area of detection zone, is stirred respectively by step 8, the droplet manipulation method according to step 2
It mixes；
Step 9, integrated circuit 2 control photometric detection module 3, and light emitting diode 31 is made to shine, and it is each to measure hatching detection zone
Area's optical signal power reads OD value, calculates sample index.
Data processing:
The mean value of critical value control value: the OD value of 2 critical value reference substances is added, divided by 2；
Negative control index: by the OD value of negative control divided by the OD value mean value of critical value reference substance；
Positive control index: by the OD value of positive control divided by the OD value mean value of critical value reference substance；
Sample index: by the OD value of sample divided by the OD value mean value of critical value reference substance.
Quality control and result interpretation:
The OD value > 0.2 of critical value reference substance indicates that critical value control is effective；
Negative control index < 0.4 indicates that negative control is effective；
Positive control index > 1.5 indicates that positive control is effective；
Sample index >=0.5 is the positive, and sample index < 0.5 is feminine gender.
Several embodiments of the invention are described in detail above, but the content is only preferable implementation of the invention
Example, should not be considered as limiting the scope of the invention.It is all according to all the changes and improvements made by the present patent application range
Deng should still be within the scope of the patent of the present invention.
Claims (10)
1. a kind of fungal detection system based on digital microfluidic technology, it is characterised in that: including digital microcurrent-controlled chip, integrate
Circuit, photometric detection module, temperature control raising module, the digital microcurrent-controlled chip respectively with the integrated circuit, photometric detection
Module, the connection of temperature control raising module, the temperature control raising module are set to below the photometric detection module, the integrated circuit point
It is not connect with the photometric detection module, temperature control raising module, the integrated circuit and photometric detection module are separately connected calculating
Machine.
2. the fungal detection system according to claim 1 based on digital microfluidic technology, it is characterised in that: the number
Micro-fluidic chip includes that the bottom crown being placed in parallel, top crown and the gap of centre, drop move in the gap；On described
Upper substrate, upper conductive layer and the upper hydrophobic layer that pole plate is set gradually from top to bottom, the bottom crown include successively setting from top to bottom
Lower hydrophobic layer, dielectric layer, lower conductiving layer and the lower substrate set；The lower conductiving layer includes sample area, drop formation channel electrode
Array, reagent liquid storage unit and hatching detection zone, the sample area, drop formation channel electrode array, reagent liquid storage unit are equal
To have effigurate metal electrode array, the sample area, reagent liquid storage unit and hatching detection zone are set to the liquid
Drop generates around channel electrode array.
3. the fungal detection system according to claim 2 based on digital microfluidic technology, it is characterised in that: the luminosity
Detection module includes the light emitting diode set gradually from top to bottom, optical filter, aperture, silicon photocell.
4. the fungal detection system according to claim 3 based on digital microfluidic technology, it is characterised in that: the hatching
Detection zone is set between the aperture and the silicon photocell.
5. the fungal detection system according to claim 2 based on digital microfluidic technology, it is characterised in that: the sample
Area's quantity is 2-4, and the hatching detection zone includes 5-10 subregion.
6. the fungal detection system according to claim 2 based on digital microfluidic technology, it is characterised in that: the sample
The electrode section in area, drop formation channel electrode array and hatching detection zone is rectangle tooth shaped electrode.
7. the fungal detection system according to claim 2 based on digital microfluidic technology, it is characterised in that: the reagent
Liquid storage unit includes antibody district, washing lotion area, substrate solution area, termination liquid zone, critical value reference substance area, negative controls after label
Area, positive reference substance area and waste.
8. a kind of two Methods for Fungi Detection using the fungal detection system based on digital microfluidic technology, which is characterized in that including
Following steps:
Step 1, will label after antibody, washing lotion, substrate solution, terminate liquid, critical value reference substance, negative controls, positive control
Product are separately added into each corresponding area of reagent liquid storage unit, and various kinds is originally separately added into each sample area；
Step 2, the control circuit coordination electrode driving circuit by integrated circuit, the on-off for carrying out electrode in a certain order are automatically controlled
System is moved to each point of detection zone of hatching by drop formation channel electrode array respectively by antibody reagent with single drops
Area controls electrode in each subregion and is stirred to antibody reagent, reagent is then moved to waste；
Step 3, the droplet manipulation method according to step 2 are separately added into critical value reference substance, yin into each area of hatching detection zone
Property reference substance, positive reference substance and each sample, are stirred respectively；
Step 4, integrated circuit control temperature control raising module are kept for 36.5-37.5 DEG C, are incubated for 85-95min, then that reagent is mobile
To waste；
Step 5, the droplet manipulation method according to step 2, by washing lotion move to hatching each area of detection zone, be stirred respectively move back to
Waste repeats 4-8 times；
Substrate solution is moved to the hatching each area of detection zone 1124, is stirred respectively by step 6, the droplet manipulation method according to step 2
It mixes；
Step 7, control temperature control raising module, are kept for 36.5-37.5 DEG C, are incubated for 15-25min, will hatch and try in each area of detection zone
Agent is moved to waste；
Terminate liquid is moved to the hatching each area of detection zone 1124, is stirred respectively by step 8, the droplet manipulation method according to step 2
It mixes；
Step 9, integrated circuit control photometric detection module, make lumination of light emitting diode, measure the hatching each area's light of detection zone 1124
Signal power reads OD value, calculates sample index.
9. the two Methods for Fungi Detection of the fungal detection system according to claim 8 based on digital microfluidic technology, special
Sign is: being marked with fluorocarbon oil in the gap.
10. two Methods for Fungi Detection according to claim 8, it is characterised in that: the top crown, hatching detection zone are
It is bright.
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