CN106770566B - The detection method of Salmonella sensor, preparation method and salmonella concentration - Google Patents
The detection method of Salmonella sensor, preparation method and salmonella concentration Download PDFInfo
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Abstract
The invention discloses the detection methods of Salmonella sensor, preparation method and salmonella concentration, the Salmonella sensor includes: electrolytic cell, electrolyte in electrolytic cell is set, the organic electrochemistry transistor in electrolytic cell, and the gate electrode being arranged in electrolytic cell are set；The organic electrochemistry transistor includes: substrate, and the source electrode and drain electrode of substrate, and the organic semiconductor thin-film layer coated in substrate connection source electrode and drain electrode is arranged in；Sensitive function layer of the photoelectric activity semiconductor material as sensor is modified on the gate electrode；It is modified on the gate electrode of photoelectric activity semiconductor material and is fixed with for the antibody in conjunction with salmonella.Salmonella sensor of the present invention has high sensitivity and extremely low detectable limit, and equipment is simple, is easily miniaturized.The present invention is based on the detection methods of the salmonella concentration of the Salmonella sensor, and quick, easy highly sensitive salmonella Concentration Testing may be implemented.
Description
Technical field
The present invention relates to biosensor technique field more particularly to a kind of Salmonella sensors, preparation method and sramana
The detection method of Salmonella concentration.
Background technique
Salmonella belongs to Gram-negative bacteria, is a kind of enteric bacilli pathogenic bacteria, is the important disease for causing food poisoning
Opportunistic pathogen, widely distributed, easily polluted source and various kinds of foods, especially birds, beasts and eggs meat products.In all kinds of bacteroidal foods
In poisoning, the case of salmonellal food poisoning ranks among the best throughout the year, and in recent years, China's salmonella infection event is not
Disconnected to increase, the food posioning of more than half is by salmonellal, it is seen that salmonella-polluted serious prestige
Human health has been coerced, economic development is constrained, therefore, has developed easy, accurate, quick, sensitive Salmeterol fluticasone propionate technology
It is of great significance for ensuring food safety with human health.
Currently, the method for detection salmonella mainly has traditional standard detection method, molecular biology method, immunology side
Method, electrical impedance method, salmonella color culture medium method etc., wherein traditional standard detection method, molecular biology method, immunology
Method is three kinds of methods being most widely used.Traditional standard detection method needs pre- increasing bacterium, selective enrichment and separation etc.
The cumbersome step of many complexity, overall process at least need 4 ~ 7 days.Obviously, traditional standard detection method has been unable to satisfy quick inspection
Survey the requirement of salmonella.Method based on molecular biology mainly have amplified fragment length polymorphism technology (AFLP),
Polymerase chain reaction technique (PCR) nucleic acid probe, biochip technology etc., molecular biology method detect quick and sensitivity
Height, but need expensive instrument and those skilled in the art, and the pre-treatment of sample is more complex, therefore be not suitable for scene on the spot
Detection and monitoring.Immunization method includes enzyme linked immunosorbent assay (ELISA)), dot enzyme-linked immuno absorption method (Dot-ELISA),
Immunomagnetic isolation technology, immunofluorescence analysis etc., immunological method simplicity, quick and high sensitivity, but detect low concentration
Easily occur when object false negative or false positive as a result, and its result interpret and need certain professional knowledge, be not easy to base
Detection.
Therefore, the existing technology needs to be improved and developed.
Summary of the invention
In view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of Salmonella sensors, preparation side
The detection method of method and salmonella concentration, so that it is low to solve existing Salmeterol fluticasone propionate sensitivity, equipment is expensive, sample
The cumbersome problem of product treatment process.
Technical scheme is as follows:
A kind of Salmonella sensor, for detecting the concentration of salmonella, the Salmonella sensor includes: electrolysis
The electrolyte in the electrolytic cell is arranged in pond, the organic electrochemistry transistor in the electrolytic cell is arranged in, and be arranged in institute
State the gate electrode in electrolytic cell；The organic electrochemistry transistor includes: substrate, be arranged in the substrate source electrode and
Drain electrode, and the organic semiconductor thin-film layer coated in substrate connection source electrode and drain electrode；It is modified on the gate electrode
There is sensitive function layer of the photoelectric activity semiconductor material as sensor；It is modified on the gate electrode of photoelectric activity semiconductor material
It is fixed with for the antibody in conjunction with salmonella.
The Salmonella sensor, wherein the photoelectric activity semiconductor material is organic semiconducting materials, inorganic
The combination of semiconductor material or both.
The Salmonella sensor, wherein the substrate is made of glass, polymer flexibility material or silicon wafer.
The Salmonella sensor, wherein the source electrode, drain electrode and gate electrode are by metal material, metal
Oxide semiconductor material, alloy material are constituted.
The Salmonella sensor, wherein the organic semiconductor thin-film layer is by poly- (3,4-rthylene dioxythiophene)-
Polystyrolsulfon acid, polypyrrole, polythiophene, polyaniline, polycarbazole or poly- (3,4- ethene dioxythiophene)-polystyrene sulphur
At least one of acid, polypyrrole, polythiophene, polyaniline, two or more copolymer of polycarbazole are constituted.
The Salmonella sensor, wherein the source electrode and drain electrode with a thickness of 50-500nm；It is described organic
Semiconductor film layer with a thickness of 10-300nm.
A kind of preparation method of the Salmonella sensor as described in any of the above item, comprising steps of
A, thoroughly cleaning substrate and drying, prepares source electrode and drain electrode, between source electrode and drain electrode on substrate
Organic semiconductor film layer is prepared, organic electrochemistry transistor is obtained；
B, thoroughly cleaning gate electrode and drying modifies photoelectric activity semiconductor material as the quick of sensor on gate electrode
Feel functional layer, the antibody being fixed in conjunction with salmonella on the gate electrode for being modified with photoelectric activity semiconductor material obtains
To being fixed with the gate electrode after the modification of antibody；
C, organic electrochemistry transistor and the gate electrode being fixed with after the modification of antibody are placed in the electricity equipped with electrolyte
The Salmonella sensor is made in Xie Chizhong.
The preparation method of the Salmonella sensor, wherein in the step A, the source electrode and drain electrode
It is to be prepared by one of vacuum thermal evaporation, magnetron sputtering or vapor deposition method；Prepare the side of organic semiconductor film layer
Method is spin coating or ink jet printing.
A kind of detection method of the salmonella concentration based on above-described Salmonella sensor, the salmonella
The detection method of concentration includes:
Under illumination condition, by the combination of antibody and salmonella on gate electrode, make the photoelectricity rheology on gate electrode
Change, cause the variation of the channel current of organic electrochemistry transistor, by the change for measuring organic electrochemistry transistor channel electric current
Change to realize the detection to various concentration salmonella.
The detection method of the salmonella concentration, wherein the detection method of the salmonella concentration further include:
Gold nano grain is modified on salmonella.
The utility model has the advantages that Salmonella sensor of the present invention has high sensitivity and extremely low detectable limit, and equipment
Simply, easily micromation.The present invention is based on the detection method of the salmonella concentration of the Salmonella sensor, may be implemented fast
Speed, easy highly sensitive salmonella Concentration Testing.
Detailed description of the invention
Fig. 1 is the overall structure diagram of Salmonella sensor of the present invention.
Fig. 2 is the structural schematic diagram of organic electrochemistry transistor of the present invention.
Fig. 3 is the I for the device that gate electrode is modified with CdS QDs under illumination " on/off "dsT curve.
Fig. 4 is that (salmonella concentration is 10 to salmonella8Cells/ml) the I that front and back is surveyed in conjunction with antibodydsT curve
(a is the I of the gate electrode of CdS QDs modificationdsT curve, b are I of the sessile antibody on the gate electrode that CdS QDs is modifiedds- T is bent
Line, c be salmonella in conjunction with antibody after IdsT curve).
Fig. 5 is the result that various concentration salmonella is tested using photoelectrochemical assay method.
Fig. 6 is the result that the Salmonella sensor based on steric effect tests various concentration salmonella.
Fig. 7 is the knot that various concentration salmonella is tested based on exciton-plasma effect Salmonella sensor
Fruit.
Specific embodiment
The present invention provides the detection method of a kind of Salmonella sensor, preparation method and salmonella concentration, to make this
The purpose of invention, technical solution and effect are clearer, define, and the present invention is described in more detail below.It should be appreciated that this
Locate described specific embodiment to be only used to explain the present invention, be not intended to limit the present invention.
The present invention provide it is a kind of for detecting the Salmonella sensor of the concentration of salmonella, as shown in Figure 1 and Figure 2, packet
Include: the electrolyte 2 in the electrolytic cell 1 is arranged in electrolytic cell 1, and the organic electrochemistry transistor in the electrolytic cell 1 is arranged in
9, and the gate electrode 3 being arranged in the electrolytic cell；The organic electrochemistry transistor 9 includes: substrate 5, is arranged in the lining
Source electrode 7 and drain electrode 8 on bottom 5, and it is thin coated in the organic semiconductor for connecting source electrode 7 and drain electrode 8 on substrate 5
Film layer 6；Sensitive function layer of the photoelectric activity semiconductor material 4 as sensor is modified on the gate electrode 3；It is modified with photoelectricity
It is fixed on the gate electrode 3 of active semiconductor material for the antibody in conjunction with salmonella.
Further, in the embodiment of the present invention, the photoelectric activity semiconductor material is organic semiconducting materials, inorganic half
The combination of conductor material or both；Such as CdS, TiO2。
Further, in the embodiment of the present invention, the substrate is by glass, polymer flexibility material (such as PET) or silicon
Piece is made.
Further, in the embodiment of the present invention, the source electrode, drain electrode and gate electrode are by metal material, metal oxygen
Compound semiconductor material, alloy material are constituted；Such as Au, Ag, Pt, Cu, ITO etc..
Further, in the embodiment of the present invention, the organic semiconductor thin-film layer is poly- by poly- (3,4-rthylene dioxythiophene)-
((PEDOT:PSS), polypyrrole, polythiophene, polyaniline, polycarbazole or poly- (3,4- ethene dioxythiophene)-gather styrene sulfonic acid
Styrene sulfonic acid, polypyrrole, polythiophene, polyaniline, polycarbazole at least one of two or more copolymer structure
At.
Further, in the embodiment of the present invention, the source electrode and drain electrode with a thickness of 50-500nm.
Further, in the embodiment of the present invention, the organic semiconductor thin-film layer with a thickness of 10-300nm.
Further, in the embodiment of the present invention, source electrode, drain electrode and gate electrode are placed in the electrolysis equipped with electrolyte
It is detected in pond, the electrolyte is used to be used as electron donor.
Organic electrochemistry transistor (OECT) of the present invention is wherein important one in Organic Thin Film Transistors (OTFT)
Class, with it is at low cost, be easy that preparation, operating voltage low (< 1V), bio-compatibility are good, easy be miniaturized, can be made into flexible device
Many advantages, such as.Since OECT has the function of sensing and signal amplification simultaneously, have very in biomolecule detection
High sensitivity and low detectable limit.At the same time, OECT can be prepared into small size device, be conducive to the micromation of sensor
And integration.
The present invention by optical electro-chemistry (PEC) biosensor technique and organic electrochemistry transistor (OECT) perfect combination, and
Applied to the detection of salmonella, a kind of Salmonella sensor is developed, there is easy, accurate, highly sensitive, easy micromation etc.
Excellent properties, required equipment is simple, has a wide range of application.
The embodiment of the invention also provides a kind of preparation methods of Salmonella sensor as described above, including step
It is rapid:
S100, thoroughly cleaning substrate and drying, prepare source electrode and drain electrode on substrate, source electrode and drain electrode it
Between prepare organic semiconductor film layer, obtain organic electrochemistry transistor；
S200, thoroughly cleaning gate electrode and drying modify photoelectric activity semiconductor material as sensor on gate electrode
Sensitive function layer, be fixed on the gate electrode for being modified with photoelectric activity semiconductor material anti-in conjunction with salmonella
Body obtains being fixed with the gate electrode after the modification of antibody；
S300, organic electrochemistry transistor and the gate electrode being fixed with after the modification of antibody are placed in equipped with electrolyte
In electrolytic cell, the Salmonella sensor is made.
Preferably, in the step S100, the source electrode and drain electrode be by vacuum thermal evaporation, magnetron sputtering or
One of vapor deposition method preparation.
Preferably, in the step S100, the method for preparing organic semiconductor film layer is spin coating or ink jet printing；Annealing
Temperature is 100-250 DEG C, and annealing atmosphere is nitrogen, time 20-60min.
Salmonella sensor provided in an embodiment of the present invention, by organic electrochemistry transistor (OECT) and optical electro-chemistry
(PEC) analysis method combines, and the gate electrode of OECT is modified with light electroactive material, and antibody is fixed on gate electrode,
Under the action of illumination, the antibody on salmonella and gate electrode occurs to will lead to photoelectric current hair on gate electrode after specific binding
It is raw to change, the variation of associated electrical parameter in organic electrochemistry transistor (channel current, transfer curve etc.) is further caused, is led to
Crossing the variation of measurement organic electrochemistry transistor associated electrical parameter (channel current, transfer curve etc.), to can reach detection different dense
Spend the purpose of salmonella.
Since organic electrochemistry transistor has both the effect of sensing and signal amplification, so Salmonella sensor of the present invention
Sensor has high sensitivity and extremely low detectable limit, and equipment is simple, is easily miniaturized, is able to solve existing Salmonella
The disadvantages of bacterial examination survey technology medium sensitivity is low, and equipment is expensive, and sample handling processes are cumbersome.
The embodiment of the invention also provides a kind of salmonella concentration based on above-described Salmonella sensor
The detection method of detection method, the salmonella concentration includes:
Under illumination condition, by the combination of antibody and salmonella on gate electrode, make the photoelectricity rheology on gate electrode
Change, cause the variation of the channel current of organic electrochemistry transistor, by the change for measuring organic electrochemistry transistor channel electric current
Change to realize the detection to various concentration salmonella.
When it is implemented, with the gate electrode of light electroactive material modification OECT, and antibody is fixed on gate electrode, in light
According under the action of, pass through the I of measurement deviceds- T(channel current-time) curve reacts the variation of photoelectric current on gate electrode, by
In being connected with antibody on gate electrode, when antibody is specifically bound with salmonella, the steric effect of salmonella can make
The decline of gate electrode photoelectric current is obtained, and various concentration salmonella causes the drop-out value of photoelectric current different, it accordingly can be to difference
The salmonella of concentration is detected.
Preferably, the detection method of the salmonella concentration further include: (fixation) gold nano is modified on salmonella
Particle.
It is elaborated below with specific embodiment to the present invention:
Salmonella sensor of the embodiment 1 based on steric effect
Principle: gate electrode selects the ITO electrode for being assembled with cadmiumsulfide quantum dot (CdS QDs) to work as light under illumination condition
Energy when being greater than energy needed for electron transition in CdS, the electrons of valence band transit to conduction band in CdS, form electron-hole
It is right.When in the electron injection electrode in conduction band, solution electron donor provide electronics to the hole in valence band, will form photoelectric current,
The generation of the electric current can reduce electrolyte/gate electrode interface current potential, to increase the effective grid electricity being applied on OECT device
Pressure.The channel current of OECT is as shown in following equation:
Wherein q represents electron charge, and μ represents hole mobility,The initial hole represented in organic semiconductor layer is close
Degree, W and L respectively represent the width and length of device channel, and t represents the thickness of organic semiconductor film, CiRepresent OECT device
Effective gate capacitance, VPPinch-off voltage is represented,Effective gate voltage is represented,Represent offset voltage, offset voltage and grid-
The voltage drop of electrolyte, the two interfaces of electrolyte-channel has relationship.
Due to the channel current I of organic electrochemistry transistordsBy gate voltage VGRegulation, can be seen by above equation
Out, the channel current I when effective gate voltage increasesdsIt can reduce.The step size that illumination " on/off " (off-on) is formed in Fig. 3
The size for generating photoelectric current on gate electrode is reflected indirectly, and the signal is exaggerated, therefore is generated when on gate electrode
Photoelectric current size change when, IdsIllumination " on/off " is formed by step size and can also change correspondingly in T curve, in gate electrode
Upper connection antibody, when antibody is specifically bound with salmonella, since the steric effect of salmonella can make grid electric
The decline of aurora electric current, and various concentration salmonella causes the drop-out value of photoelectric current different, it accordingly can be to various concentration
Salmonella is detected.
Fig. 4 is that concentration is 108The electrical signal variation diagram of cells/ml salmonella front and back in conjunction with antibody.Fig. 5 is to adopt
With traditional photoelectrochemical assay method test various concentration salmonella as a result, detectable limit is 103cells/ml.Fig. 6
It is the optical electro-chemistry sensor test various concentration salmonella based on organic electrochemistry transistor as a result, detectable limit is
102cells/ml.It can be seen that the sensitivity of the New Sensing Technology is higher than traditional optical electro-chemistry sensing technology.
The preparation process of Salmonella sensor based on steric effect
1. making source electrode, drain electrode and the organic semiconductor thin-film layer of organic electrochemistry transistor (OECT): will clean
Good glass is adjacent on the mask plate of design good pattern, by hot evaporation deposit metal electrodes, deposit respectively 10nm Cr and
The Au of 100nm is to obtain Au/Cr/ glass electrode, and one layer of spin coating mixed with poly- (the 3,4- of dimethyl sulfoxide (DMSO) on this electrode
Ethene dioxythiophene)-polystyrolsulfon acid (PEDOT:PSS) is clean by the place erasing for not needing covering PEDOT:PSS film；
In 180 DEG C of annealing 1h of nitrogen atmosphere, make PEDOT:PSS film is stronger to be attached to electrode surface and finally obtained OECT device
Part.
2. TGA(thioacetic acid) synthesis of the CdS QDs of modification: 50 mL, 0.01 M is added in three-necked flask
CdCl2Solution, stirring, is passed through nitrogen, and 250 μ L TGA are added after being warming up to 40 DEG C, reacts 30 min；During this period, using 1 M
NaOH solution adjust mixed liquor pH to 11；Then, 5.0mL 0.1M Na is added2S solution, lower 110 °C of nitrogen atmosphere add
Heat, flow back 4h, and after water (volume ratio 1:1) dilution, it is stand-by to be stored in 4 DEG C of refrigerators.
3. the preparation of the gate electrode of CdS QDs modification: it is (poly- that the ITO electrode after clean dry successively being immersed 2% PDDA
Polymer electrolyte, 0.5 M NaCl solution prepare) and CdS QDs solution in each 10 min, impregnated washed with water every time, the mistake
Journey is repeated 3 times, and obtains required multilayer film modified electrode, under light illumination, measures IdsT curve.
4. antibody is in the fixation of the CdS QDs surface gate electrode modified: passing through the NH on antibody2On group and CdS QDs
COOH group between coupling reaction carry out；The electrode that CdS QDs is modified immerses 20mg/ml EDC(1- (3- dimethylamino
Propyl) -3- ethyl-carbodiimide hydrochloride) and 10mg/ml NHS(N- HOSu NHS) solution in 1h, then use water
It is careful to rinse, by 25 μ L antibody (2mg/ml) drop after electrode surface and 4 DEG C of incubated overnights, carefully rushed using 10 mM PBS
It washes, to remove loose antibody.Then, using 1 mM MEA(ethanol amine) in 4 DEG C enclosed-electrode 2 hours, then with 10
MM PBS(phosphate buffer) it is careful rinse after, under light illumination, measure IdsT curve.
5. the combination of salmonella and antibody: being fixed with salmonella solution of the gate electrode in 1ml various concentration of antibody
In (preparation of 10mM PBS solution) impregnate 1h at room temperature so that it sufficiently combines, then carefully rinsed, removed not with 10mM PBS
In conjunction with salmonella, under light illumination, measure IdsT curve.
In this example, CdS QDs modification, connection antibody and the I with the gate electrode after salmonella combinationds- T is bent
Line measurement, V in 0.1M ascorbic acid (AA) solution (preparation of 0.1M PBS solution)G=0V, VDS=0.1V, excitation wavelength are
420nm。
Embodiment 2 is based on exciton-plasma effect salmonella immunosensor
Principle: it on the basis of steric effect, devises based on cadmiumsulfide quantum dot (CdS QDs) and gold nano grain
Exciton-plasma effect system between (Au NPs) further promotes the sensitivity of sensor, due to CdS QDs's
The ultra-violet absorption spectrum of fluorescence spectrum and Au NPs are overlapped, and the fluorescence of CdS QDs can excite Au NPs to occur under illumination condition
Surface plasma body resonant vibration, the interaction between them can change the exciton state in CdS QDs, cause the reduction of photoelectric current.
Therefore, Au NPs is modified on salmonella, and antibody is connected on gate electrode, can be excited after specifically binding sharp
Son-plasma effect leads to the reduction of photoelectric current, in IdsThe reduction that illumination " on/off " step is shown as on T curve, according to
The effect difference that various concentration salmonella causes photoelectric current to quench achievees the effect that detection various concentration salmonella, and Fig. 7 is
Based on exciton-plasma effect Salmonella sensor test various concentration salmonella as a result, detectable limit is
10cells/ml, it can be seen that, the sensitivity of the New Sensing Technology is higher than the Salmonella sensor based on steric effect.
Preparation process based on exciton-plasma effect Salmonella sensor
1. making source electrode, drain electrode and the organic semiconductor thin-film layer of organic electrochemistry transistor (OECT): will clean
Good glass is adjacent on the mask plate of design good pattern, by hot evaporation deposit metal electrodes, deposit respectively 10nm Cr and
The Au of 100nm is to obtain Au/Cr/ glass electrode, and one layer of spin coating mixed with poly- (the 3,4- of dimethyl sulfoxide (DMSO) on this electrode
Ethene dioxythiophene)-polystyrolsulfon acid (PEDOT:PSS) is clean by the place erasing for not needing covering PEDOT:PSS film；
In 180 DEG C of annealing 30min of nitrogen atmosphere, make PEDOT:PSS film is stronger to be attached to electrode surface and finally obtain
OECT device.
2. TGA(thioacetic acid) synthesis of the CdS QDs of modification: 50 mL, 0.01 M is added in three-necked flask
CdCl2Solution, stirring, is passed through nitrogen, and 250 μ L TGA are added after being warming up to 40 DEG C, reacts 30 min.During this period, using 1 M
NaOH solution adjust mixed liquor pH to 11.Then, 5.0mL 0.1M Na is added2S solution, the lower 110 °C of heating of nitrogen atmosphere, is returned
4h is flowed, after water (volume ratio 1:1) dilution, it is stand-by to be stored in 4 DEG C of refrigerators.
3. the synthesis of Au NPs: Au NPs passes through common NaBH4Restore HAuCl4Method carry out；0.6mL 0.1M
The NaBH that ice water is prepared4It is added to the 20mL 2.5 × 10 kept stirring4 M HAuCl4In solution；Solution rapidly goes to Chinese red
The formation of Au NPs is represented, 10 min are stirred in solution continuation in ice-water bath, then stir 3h under normal temperature conditions, herein mistake
Cheng Zhong, solution colour can gradually become claret；After stirring, it is stand-by that gold size solution is stored in 4 DEG C of refrigerators.
4. the preparation of the gate electrode of CdS QDs modification: it is (poly- that the ITO electrode after clean dry successively being immersed 2% PDDA
Polymer electrolyte, 0.5 M NaCl solution prepare) and CdS QDs solution in each 10 min, impregnated washed with water every time, the mistake
Journey is repeated 3 times, and obtains required multilayer film modified electrode, and CdS QDs drying measures I after stablizing under light illuminationdsT curve.
5. antibody is in the fixation of the CdS QDs surface gate electrode modified: passing through the NH on antibody2On group and CdS QDs
COOH group between coupling reaction carry out；The electrode that CdS QDs is modified immerses 20mg/ml EDC and 10mg/ml NHS
Solution in 1h, then carefully rinsed with water, 25 μ L antibody (2mg/ml) drop in electrode surface and after 4 DEG C of incubated overnights, made
It is carefully rinsed with 10 mM PBS, to remove loose antibody.Then, using 1 mM MEA(ethanol amine) it is closed in 4 DEG C
Electrode 2 hours, then with 10 mM PBS(phosphate buffers) it is careful rinse after, under light illumination, measure IdsT curve.
6. Au NPs(gold nano grain) to the label of salmonella: by 109Synthesized by cells salmonella and 2ml
Au NPs is directly mixed, and is centrifuged after concussion overnight, and bacterium is collected, and is added after 1ml 10mM PBS is uniformly mixed and is continued to be centrifuged
It collects, repeats the process 2 times, play cleaning action, remove unbonded Au NPs, finally repaired with 10mM PBS dilution Au NPs
The salmonella of decorations is to be measured at a series of concentration.
7. the combination of salmonella and antibody: the gate electrode for being fixed with antibody is molten in the salmonella of various concentration in 1ml
(preparation of 10mM PBS solution) impregnates one hour at room temperature so that it is sufficiently combined in liquid, is then carefully rinsed with 10mM PBS,
Unbonded salmonella is removed, under light illumination, measures IdsT curve.
In this example, CdS QDs modification, connection antibody and the I with the gate electrode after salmonella combinationdsT curve
Measurement, V in 0.1M ascorbic acid (AA) solution (preparation of 0.1M PBS solution)G=0V, VDS=0.1V, excitation wavelength 420nm.
Salmonella sensor provided by the present invention is a kind of novel biosensor technique, is caused by salmonella
The variation of photoelectric current on gate electrode further causes OECT associated electrical parameter (interfacial potential, effective gate voltage, channel current
Deng) variation, realize the detection to various concentration salmonella eventually by the variation of measurement OECT channel current.The present invention
The lowest detection limit using the Salmonella sensor of steric effect is 102Cells/ml utilizes exciton-plasma effect
Salmonella sensor the lowest detection limit up to 10cells/ml, fast and convenient highly sensitive salmonella may be implemented
It is actually detected.
The present invention combines optical electro-chemistry biosensor technique with organic electrochemistry transistor, develops a kind of novel
Salmonella sensing technology can be to current signal faint on gate electrode since OECT has both the effect of sensing and signal amplification
Variation amplifies, therefore the sensor has high sensitivity, and inventive sensor structure is simple, device size is small, institute
There is component to can be integrated on a small substrate, easy of integrationization, micromation, array are suitble to large-scale production；The sensing
Device operating voltage is low (< 1V), and organic semiconductor thin-film layer and all optional biology of the semiconductor material being assembled on gate electrode are simultaneous
The good material of capacitive provides good stability for sensor；And equipment price needed for detecting is low, easy to operate, detects people
Without having too many professional knowledge, which solves sensitive in current certain Salmeterol fluticasone propionate technologies member to a certain extent
Spend low, equipment is expensive, and sample handling processes are cumbersome, and testing staff's professional knowledge requires the disadvantages of high.
Optical electro-chemistry biosensor technique based on organic electrochemistry transistor is applied to salmonella for the first time by the present invention
Detection in, and for the first time design be based on exciton-plasma effect Salmonella sensor, in addition to salmonella, should
Sensing technology also has very big application prospect in the detection and monitoring of other pathogenic bacterias.
It is further to note that the organic semiconductor thin-film layer in organic electrochemistry transistor of the present invention is also changed to it
His inorganic semiconductor thin-film material such as graphene.The present invention is the modification photoelectric activity material on OECT gate electrode, is drawn under illumination
Electrolyte/gate electrode phase boundary potential, which changes, reaches biomolecule detection purpose, and having in organic electrochemistry transistor
Photoelectric activity material is modified on machine semiconductor film layer, and electrolyte/channel interface potential change is caused also can equally to reach under illumination
To sensing detection purpose.
In conclusion Salmonella sensor of the present invention has high sensitivity and extremely low detectable limit, and equipment
Simply, easily micromation.The present invention is based on the detection method of the salmonella concentration of the Salmonella sensor, may be implemented fast
Speed, easy highly sensitive salmonella Concentration Testing.
It should be understood that the application of the present invention is not limited to the above for those of ordinary skills can
With improvement or transformation based on the above description, all these modifications and variations all should belong to the guarantor of appended claims of the present invention
Protect range.
Claims (10)
1. a kind of Salmonella sensor, for detecting the concentration of salmonella, which is characterized in that the Salmonella sensor
Include: electrolytic cell, the electrolyte in the electrolytic cell be set, the organic electrochemistry transistor in the electrolytic cell is set,
And the gate electrode in the electrolytic cell is set；
The organic electrochemistry transistor includes: substrate, and the source electrode and drain electrode of the substrate is arranged in, and is coated in
The organic semiconductor thin-film layer of substrate connection source electrode and drain electrode；
Sensitive function layer of the photoelectric activity semiconductor material as sensor is modified on the gate electrode；
It is modified on the gate electrode of photoelectric activity semiconductor material and is fixed with for the antibody in conjunction with salmonella；
The testing principle of the Salmonella sensor is to be based on: anti-on salmonella and gate electrode under the action of illumination
It will lead to photoelectric current on gate electrode after body generation specific binding to change, further cause in organic electrochemistry transistor
The variation of channel current, the variation by measuring organic electrochemistry transistor channel electric current reach detection various concentration salmonella
Purpose；
The Salmonella sensor combines organic electrochemistry transistor and photoelectrochemical assay method, and illumination is effect
In on the gate electrode.
2. Salmonella sensor according to claim 1, which is characterized in that the photoelectric activity semiconductor material is to have
The combination of machine semiconductor material, inorganic semiconductor material or both.
3. Salmonella sensor according to claim 1, which is characterized in that the substrate is by glass, polymer flexible
Property material or silicon wafer are made.
4. Salmonella sensor according to claim 1, which is characterized in that the source electrode, drain electrode and gate electrode
It is to be made of metal material, metal oxide semiconductor material, alloy material.
5. Salmonella sensor according to claim 1, which is characterized in that the organic semiconductor thin-film layer is by gathering
(3,4- ethene dioxythiophene)-polystyrolsulfon acid, polypyrrole, polythiophene, polyaniline, polycarbazole or poly- (3,4- ethylene two
Oxygen thiophene)-polystyrolsulfon acid, polypyrrole, polythiophene, polyaniline, polycarbazole two or more copolymer in
At least one is constituted.
6. Salmonella sensor according to claim 1, which is characterized in that the source electrode and drain electrode with a thickness of
50-500nm；The organic semiconductor thin-film layer with a thickness of 10-300nm.
7. a kind of preparation method of Salmonella sensor as claimed in any one of claims 1 to 6, which is characterized in that including step
It is rapid:
A, thoroughly cleaning substrate and drying, prepares source electrode and drain electrode on substrate, prepares between source electrode and drain electrode
Organic semiconductor thin-film layer obtains organic electrochemistry transistor；
B, sensitive function of the photoelectric activity semiconductor material as sensor is modified in thoroughly cleaning gate electrode and drying on gate electrode
Ergosphere, the antibody being fixed in conjunction with salmonella on the gate electrode for being modified with photoelectric activity semiconductor material, consolidate
Surely the gate electrode after having the modification of antibody；
C, organic electrochemistry transistor and the gate electrode being fixed with after the modification of antibody are placed in the electrolytic cell equipped with electrolyte
In, the Salmonella sensor is made.
8. the preparation method of Salmonella sensor according to claim 7, which is characterized in that described in the step A
Source electrode and drain electrode be by one of vacuum thermal evaporation, magnetron sputtering or vapor deposition method prepare；It prepares organic
The method of semiconductor film layer is spin coating or ink jet printing.
9. a kind of detection method of the salmonella concentration based on Salmonella sensor described in claim 1, feature exist
In the detection method of the salmonella concentration includes:
Under illumination condition, by the combination of antibody and salmonella on gate electrode, makes the photocurrent variations on gate electrode, draw
The variation for playing the channel current of organic electrochemistry transistor, by measuring the variation of organic electrochemistry transistor channel electric current come real
Now to the detection of various concentration salmonella.
10. the detection method of salmonella concentration according to claim 9, which is characterized in that the salmonella concentration
Detection method further include: gold nano grain is modified on salmonella.
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| PCT/CN2017/110984 WO2018095246A1 (en) | 2016-11-25 | 2017-11-15 | Salmonella sensor, preparation method, and salmonella concentration detection method |
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| CN106770566B (en) * | 2016-11-25 | 2019-04-16 | 深圳大学 | The detection method of Salmonella sensor, preparation method and salmonella concentration |
| CN107525833B (en) * | 2017-07-10 | 2020-07-24 | 深圳大学 | Amyloid β -protein sensor based on porous gate electrode and preparation method thereof |
| CN108205008B (en) * | 2017-12-28 | 2020-04-21 | 深圳大学 | Toxin sensor based on organic photoelectric chemical transistor and preparation method thereof |
| CN110398528B (en) * | 2019-06-10 | 2022-04-12 | 深圳大学 | Liquid gate type IGZO thin film transistor-based anti-tuberculosis drug screening device and method |
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| CN112903770B (en) * | 2019-12-04 | 2022-05-17 | 中国石油化工股份有限公司 | Flexible sensor for measuring bacterial quantity of sulfide producing bacteria and method for measuring SRB bacterial quantity in sewage by indirect method |
| CN111257560A (en) * | 2020-04-06 | 2020-06-09 | 东莞正大康地饲料有限公司 | Method for rapidly detecting salmonella in fish meal |
| CN111551717B (en) * | 2020-04-10 | 2023-04-07 | 深圳大学 | Gastrin-releasing peptide precursor sensor based on organic photoelectrochemical transistor and preparation method and application thereof |
| CN111999365A (en) * | 2020-08-27 | 2020-11-27 | 合肥海关技术中心 | Method for detecting salmonella in food based on DNA electrochemical sensor |
| CN112816639B (en) * | 2021-01-06 | 2023-04-11 | 江苏大学 | Construction method of photoelectrochemical aptamer sensor for sensitive detection of enrofloxacin |
| CN113036042B (en) * | 2021-03-05 | 2023-05-26 | 天津大学 | Photoelectric biochemical sensor based on organic thin film field effect transistor, and preparation method and application thereof |
| CN114113271B (en) * | 2021-10-20 | 2023-12-15 | 杭州富鼎检测有限公司 | Preparation method of photoelectrochemical organic transistor sensor for detecting pesticide malathion |
| CN114384134B (en) * | 2021-12-09 | 2023-08-22 | 江苏大学 | Construction method and application of organic photoelectrochemical transistor sensor based on zinc oxide nano rod array |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735734A (en) * | 2011-03-29 | 2012-10-17 | 香港理工大学 | Non-invasive glucose sensor |
| CN103558270A (en) * | 2013-10-18 | 2014-02-05 | 海南大学 | Marine microorganism adhesion sensor based on organic electrochemical transistor and preparation method of sensor |
| CN105190310A (en) * | 2013-03-11 | 2015-12-23 | 托莱多大学 | A biosensor device to target analytes in situ, in vivo, and/or in real time, and methods of making and using the same |
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| US7575939B2 (en) * | 2000-10-30 | 2009-08-18 | Sru Biosystems, Inc. | Optical detection of label-free biomolecular interactions using microreplicated plastic sensor elements |
| WO2010042196A1 (en) * | 2008-10-08 | 2010-04-15 | Massachusetts Institute Of Technology | Catalytic materials, photoanodes, and photoelectrochemical cells for water electrolysis and other electrochemical techniques |
| CN106018514B (en) * | 2016-07-05 | 2019-04-05 | 济南大学 | A kind of preparation method of the optical electro-chemistry diethylstilbestrol sensor based on Copper-cladding Aluminum Bar nanometer photoelectronic material |
| CN106770587B (en) * | 2016-11-25 | 2019-03-26 | 深圳大学 | A kind of Photoelectrochemistrbiosensor biosensor and preparation method thereof |
| CN106770566B (en) * | 2016-11-25 | 2019-04-16 | 深圳大学 | The detection method of Salmonella sensor, preparation method and salmonella concentration |
-
2016
- 2016-11-25 CN CN201611054808.3A patent/CN106770566B/en active Active
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735734A (en) * | 2011-03-29 | 2012-10-17 | 香港理工大学 | Non-invasive glucose sensor |
| CN105190310A (en) * | 2013-03-11 | 2015-12-23 | 托莱多大学 | A biosensor device to target analytes in situ, in vivo, and/or in real time, and methods of making and using the same |
| CN103558270A (en) * | 2013-10-18 | 2014-02-05 | 海南大学 | Marine microorganism adhesion sensor based on organic electrochemical transistor and preparation method of sensor |
Non-Patent Citations (1)
| Title |
|---|
| 基于有机电化学晶体管的癌胚抗原、鸟嘌呤及酚类传感研究;王静静;《湖南师范大学硕士学位论文》;20160415;正文第5-7，10-11页 |
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