CN104502427A - Microbiological detection device and microbiological detection method - Google Patents
Microbiological detection device and microbiological detection method Download PDFInfo
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- CN104502427A CN104502427A CN201510005659.0A CN201510005659A CN104502427A CN 104502427 A CN104502427 A CN 104502427A CN 201510005659 A CN201510005659 A CN 201510005659A CN 104502427 A CN104502427 A CN 104502427A
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Abstract
The invention relates to the field of microculture and detection, and provides a microbiological detection device and a microbiological detection method, aiming at solving the problems of time consuming, high cost and poor biocompatibility and the like in the existing microbiological detection method. The microbiological detection device comprises a reaction chamber used for containing nutrient solution, a sensitive element and an organic thin-film field effect transistor, wherein the sensitive element is communicated with the reaction chamber, is contacted with the nutrient solution and is used for sensing pH value variation of the nutrient solution and producing an electric charge according to the pH value variation; and the organic thin-film field effect transistor is electrically communicated with the sensitive element and is used for determining whether microorganisms exist or not according to charge of the sensitive element, and kinds of the microorganisms are also determined. The organic thin-film field effect transistor is adopted as a detecting element, so that microorganism growth condition in the nutrient solution can be detected in real time according to variation of channel current of the transistor, a detection result can be rapidly obtained, and the microbiological detection method has the advantages that the cost is low and biocompatibility is good compared with the existing detection method.
Description
Technical field
The present invention relates to microorganism to cultivate and detection field, a kind of microbial detection device and method are specifically provided.
Background technology
Microorganism is extensively present in occurring in nature.Main detection method used in the prior art is microorganism cultivation.In incubation, sample is placed in medium component and the controlled environment of condition of culture, thus accelerate growth of microorganism.At present, market exists multiple-microorganism culture apparatus, these devices can detect the existence of microorganism in sample in 12-48 hour, and can get rid of any infectious microorganism of existence in 5 days.In addition, by carrying out second incubation and test to sample, these devices can again by the character of the time Identifying micro-organisms of 12-48 hour.But needing the application scenario obtaining testing result fast, the time span needed for above-mentioned detection method is completely unacceptable.
International patent application WO2013096404 describes the method utilizing ISFET to carry out infectious microorganism detection.This detection method can shorten detection time to a certain extent, but also there are some problems, and such as cost is high, bio-compatibility is poor, sensitivity is low.
Therefore, need to develop a kind of microbial detection device and method, it can detect microorganism rapidly and accurately, and can avoid the problems referred to above of the prior art.
Summary of the invention
The present invention is intended to solve the above-mentioned problems in the prior art.Specifically, the present invention is intended to solve the problems such as existing microorganism detection method cannot detect in real time, detection time is long and cost is high, bio-compatibility is poor, sensitivity is low.
For this purpose, the invention provides a kind of microbial detection device.This microbial detection device comprises: reaction chamber, and it is for holding sample to be detected; Sensitive element, its be communicated with described reaction chamber and with described sample contact to be detected, for sensing the pH value change of described sample to be detected and producing electric charge according to the change of described pH value; And detector, whether itself and described sensitive element electric connection, exist microorganism and there is which kind of microorganism for determining according to the electric charge of described sensitive element, wherein said detector is organic film FET in described sample to be detected.
In the preferred implementation of mentioned microorganism pick-up unit, described organic film FET comprises substrate, the grid be arranged on substrate, be arranged on dielectric layer on grid and arrange source electrode on the dielectric layer, drain electrode and semiconductor layer, and described sensitive element adopts at least one in following setting: described sensitive element arrange over the substrate and with described grid electric connection or the part being arranged to described grid; Described sensitive element be arranged to described dielectric layer a part and with described grid, described source electrode, described drain electrode or described semiconductor layer electric connection; And described sensitive element and described source electrode, described drain electrode or described semiconductor layer electric connection or be arranged to the part of described source electrode, described drain electrode or described semiconductor layer.
In the preferred implementation of mentioned microorganism pick-up unit, described reaction chamber is arranged on the top of described sensitive element.
In the preferred implementation of mentioned microorganism pick-up unit, the volume size of described reaction chamber is arranged so that the state of single microbial in described reaction chamber changes the pH value change caused and also can be detected, and makes described reaction chamber can hold microorganism to prevent again larger mammalian cell from entering described reaction chamber.
In the preferred implementation of mentioned microorganism pick-up unit, the volume of described reaction chamber is received in the scope of liter to 1 at 1 microlitre.
In the preferred implementation of mentioned microorganism pick-up unit, described dielectric layer is also provided with controller, this controller is for controlling the parameter of described organic film FET to adapt to different detected objects and detection demand.
According to another aspect of the present invention, a kind of microorganism detection method is provided.This microorganism detection method comprises the following steps: to provide microbial detection device as above, and sample to be detected is put into described reaction chamber; And determine whether there is microorganism in described sample to be detected and there is which kind of microorganism according to the channel current change of described organic film FET.
In the preferred implementation of mentioned microorganism detection method, determine in described sample to be detected, whether to there is microorganism according to the change of the channel current of described organic film FET and the step that there is which kind of microorganism comprises further: the growth of microorganism in described sample to be detected causes the pH value of described sample to be detected to change; Described pH value change causes described sensitive element to produce electric charge; The electric charge of described sensitive element causes the induced charge on described grid, described source electrode, described drain electrode or described semiconductor layer to redistribute; And the induced charge redistribution on described grid, described source electrode, described drain electrode or described semiconductor layer causes the electromotive force between described source electrode and described drain electrode to change by the effect of electrode coupling electric capacity.
In the preferred implementation of mentioned microorganism detection method, determine in described sample to be detected, whether to there is microorganism according to the change of the channel current of described organic film FET and the step that there is which kind of microorganism also comprises: the potential change between described source electrode and described drain electrode causes the channel current of described organic film FET to change; And determine whether there is microorganism in described sample to be detected and there is which kind of microorganism according to the change curve of described channel current.
In the preferred implementation of mentioned microorganism detection method, described sample to be detected is microbial culture medium.
As can be seen from description above, the behavior that the present invention makes full use of organic film FET (organic thin-film transistor, OTFT) high electron mobility, highly sensitive advantage detects microorganism fast.Specifically, sample is placed in the condition being suitable for sample growth of microorganism, the index change (the pH value change caused by the change of such as gas concentration lwevel) of microorganism in sample can be detected in real time fast, thus determine whether there is microorganism in sample fast.In addition, can also according to this Plotting data microbial growth curve.The growth curve obtained by contrast and known standard microorganism growth curve, can also identify which kind of microorganism the microorganism in sample is.Moreover, also can monitor sample composition interpolation (as microbiotic) the change of the rate of growth that causes or metabolic activity in cells.
Those skilled in the art it is easily understood that, owing to adopting organic film FET as detector, pick-up unit of the present invention and method have multiple advantage: 1) can Real-Time Monitoring microorganism cultivation and determine whether microorganism exists and the kind of microorganism fast; 2) self have both signal to transform and enlarging function, be convenient to follow-up signal processing circuit integrated; 3) be easy to microminiaturized, may be used for preparing the chip that integrated level is high, measuring ability is complicated; 4) utilize the organic semiconductor of solubility, insulating layer material, by low temperature typography processing preparation, and the backing material selecting poor heat resistance, generalization can be conducive to, thus can significantly reduce costs and energy consumption, enhance productivity.
Accompanying drawing explanation
Fig. 1 shows the structural drawing according to microbial detection device of the present invention.
Fig. 2 shows the schematic diagram that multiple microbial detection device according to the present invention integrates.
Fig. 3 shows the process flow diagram according to microorganism detection method of the present invention.
Embodiment
First it should be noted that, describe technical scheme of the present invention below in conjunction with preferred implementation.But those skilled in the art are it is easily understood that technical scheme of the present invention is obviously not limited to the various details in described embodiment.Do not departing under principle of the present invention-namely uses OTFT to carry out the prerequisite of microorganism detection, can make various change to described embodiment, these do not need to pay any creative work, therefore will fall within protection scope of the present invention yet.
Specifically, as shown in Figure 1, microbial detection device 230 of the present invention comprises: reaction chamber 18, and it is for holding nutrient solution 10; Sensitive element 11, it is communicated with reaction chamber 18 and contacts with nutrient solution 10, for sensing the pH value change of nutrient solution 10 and producing electric charge according to described pH value change; And detector, whether itself and sensitive element 11 electric connection, exist microorganism for determining according to the electric charge of sensitive element 11 and there is which kind of microorganism in nutrient solution 10.According to technical scheme of the present invention, described detector is organic film FET-i.e. OTFT.
More specifically, described organic film FET comprises substrate 12, the grid 13 be arranged on substrate, is arranged on the dielectric layer 14 on grid 13 and is arranged on source electrode 17 on dielectric layer 14, drain electrode 16 and semiconductor layer 19.As shown in the figure, sensitive element 11 also arrange on the substrate 12 and with grid 13 electric connection.Although it should be pointed out that sensitive element 11 is positioned at the right side of grid 13 in FIG, this position relationship is only exemplary, and those skilled in the art can make adjustment to this according to embody rule, and this adjustment does not deviate from principle of the present invention.Similarly, although the sensitive element in Fig. 1 11 is shown as the bottom being positioned at reaction chamber 18, this position relationship is also exemplary, those skilled in the art can make adjustment to this according to embody rule, as long as sensitive element 11 can sense sample to be detected in reaction chamber 18-namely, the pH value of nutrient solution changes.Preferably, described reaction chamber 18 can be integrated into discrete component with sensitive element 11, and described sensitive element 11 can be arranged to the reaction chamber 18 of the part of described grid 13-namely and be set directly at the top of grid 13.Alternatively, described sensitive element 11 can be arranged to dielectric layer 14 a part (such as its right part) and with grid 13, source electrode 17, drain 16 or semiconductor layer 19 electric connection, or described sensitive element 11 can with source electrode 17, drain 16 or semiconductor layer 19 electric connection or be arranged to source electrode 17, a part for drain 16 or semiconductor layer 19.Generally speaking, as long as sensitive element 11 can sense the pH value change of the sample to be detected in reaction chamber 18 and can therefore make the channel current of described organic film FET change.Certainly, as shown in Figure 1, most preferred setting be sensitive element 11 be arranged to grid 13 a part or with grid 13 electric connection, thus changed the channel current of described organic film FET by grid 13.
In addition, the material of sensitive element 11 can be metal, also can be semiconductor, as silicon, silicon dioxide.When the pH value of the nutrient solution in reaction chamber 18 changes, the transfer of intramolecular charge can be caused, thus cause the electric charge on sensitive element 11 to redistribute, according to sensitive element 11 setting position, described electric charge is finally coupled in described transistor and makes channel current that corresponding change occur.Can determine whether there is microorganism in nutrient solution according to the change curve of channel current, and by the kind of microorganism can be determined compared with existing microbial growth curve.
Preferably, the volume size of described reaction chamber 18 is arranged so that the state of single microbial in described reaction chamber changes the pH value change caused and also can be detected by described organic film FET, and making described reaction chamber 18 can hold microorganism can prevent again larger mammalian cell from entering described reaction chamber 18, thus avoid obtaining the testing result that makes mistake.More preferably, the volume of described reaction chamber 18 can be received in the scope of liter to 1 at 1 microlitre.In addition, as shown in Figure 1, dielectric layer 14 is also provided with controller 15, this controller 15 is for controlling the parameter of described organic film FET to adapt to different detected objects and detection demand.Described controller 15 can be such as control gate, and its position can be selected arbitrarily as required, and need not be confined to shown in Fig. 1.
Next consult Fig. 2, the figure shows the schematic diagram that multiple microbial detection device according to the present invention integrates.As shown in Figure 2, multiple microbial detection device 230 according to the present invention has been integrated on same substrate, thus defines OTFT transistor array 220.
Consult Fig. 3 below, the figure shows the process flow diagram according to microorganism detection method 200 of the present invention.In step 100, first provide mentioned microorganism pick-up unit, this pick-up unit utilizes OTFT as detector.In a step 101, nutrient solution 10 is put into reaction chamber 18.In a step 102, the growth of microorganism in nutrient solution 10 causes the pH value of nutrient solution 10 to change.In step 103, described pH value change causes sensitive element 11 to produce electric charge.At step 104, the electric charge of sensitive element 11 causes the induced charge redistribution on grid 13, source electrode 17, drain electrode 16 or semiconductor layer 19.Next in step 105, the induced charge redistribution on grid 13, source electrode 17, drain electrode 16 or semiconductor layer 19 causes the electromotive force between source electrode 17 and drain electrode 16 to change by the effect of electrode coupling electric capacity.Afterwards in step 106, the potential change between source electrode 17 and drain electrode 16 causes the channel current of described organic film FET to change.In step 107, method of the present invention determines whether there is microorganism in nutrient solution and there is which kind of microorganism according to the change curve of described channel current finally.
Those skilled in the art, it is easily understood that the cross section of reaction chamber 18 and opening can be any shapes, include but not limited to rectangle, cube or cylindrical.The wall of reaction chamber 18 can be vertical, tilt or any other moulding.As mentioned above, one, two or more microorganisms can be held by the size optimizing reaction chamber.Also can increase the sensitivity of detection by the quantity controlling microorganism and shorten detection time.
From the angle of concrete reaction, be placed into nutrient solution 10 in reaction chamber 18, external environment can be utilized to control condition of culture, when to enter in reaction chamber when there being microorganism alive and grow, the equation of its aerobic respiration is C
6h
12o
6+ 6O
2=6H
2o+6CO
2+ ATP, carbon dioxide solubility generates carbonic acid in water, and the generation of carbonic acid causes the pH value of nutrient solution to change.The proton produced in metabolic process or in each fission process between different biosomes or CO
2amount and the fissional time be all different.Once sensitive element senses electric charge because of the change of nutrient solution pH value, the redistribution of induced charge will be produced on grid 13, by the effect of electrode coupling electric capacity, cause the electromotive force of grid 17,16 to change.If grid 17,16 produces potential change because detecting bioelectrical signals, so channel current just may produce the change of hundreds and thousands of times.That is, bio signal can amplify doubly a lot by electrical method through OTFT and be detected.OTFT OTFT is because carrier mobility is higher than existing silicon-based transistor doubly a lot, its sensitivity is corresponding raising also, and then can just carry out detection supervision in early days in cell division, and such as infectious microorganism etc. can be identified within the shorter time, for follow-up decision provides sufficient guarantee.
More specifically, compared with prior art, be can the incubation of Real-Time Monitoring microorganism for technique effect of the present invention.The vital movement process of microorganism can change the pH value of external environment automatically, and wherein PH changes has souring and becomes alkali two kinds of processes, often preponderates with souring in the cultivation of general microorganism.Therefore, along with incubation time extends, the pH value of nutrient culture media can decline gradually.Under existing cultivating system, cannot monitor the pH value of microbial cultivation process, one-shot measurement can only be carried out at set intervals, simultaneously because cultivating system is very large, therefore be difficult to just monitor in the cultivation firm incipient stage, (such as 4 hours) just can only detect after a period of time is carried out in cultivation later.Such problem is, is difficult to know that whether cultivation is in generation.If just finding to cultivate after the long period does not occur, then the loss of time can be caused.Utilize OTFT to carry out microorganism detection can effectively address this problem.First under miniflow system microorganism grow each time all comparatively large on the impact of the pH value of system, secondly the high sensitive of OTFT again can Sensitive Detection to the change of this pH value, thus the Real-Time Monitoring that just can realize microorganism cultivation in the initial period.
So far, shown by reference to the accompanying drawings preferred implementation describes technical scheme of the present invention, but those skilled in the art are it is easily understood that protection scope of the present invention is obviously not limited to these embodiments.Under the prerequisite not departing from principle of the present invention, those skilled in the art can make equivalent change or replacement to correlation technique feature, and these changes or the technical scheme after replacing it all will fall within protection scope of the present invention.
Claims (10)
1. a microbial detection device, is characterized in that comprising:
Reaction chamber, it is for holding sample to be detected;
Sensitive element, its be communicated with described reaction chamber and with described sample contact to be detected, for sensing the pH value change of described sample to be detected and producing electric charge according to the change of described pH value; And
Detector, whether itself and described sensitive element electric connection, exist microorganism for determining according to the electric charge of described sensitive element and there is which kind of microorganism in described sample to be detected,
Wherein said detector is organic film FET.
2. microbial detection device as claimed in claim 1, it is characterized in that, described organic film FET comprises substrate, the grid be arranged on substrate, be arranged on dielectric layer on grid and arrange source electrode on the dielectric layer, drain electrode and semiconductor layer, and
Described sensitive element adopts at least one in following setting:
Described sensitive element arrange over the substrate and with described grid electric connection or the part being arranged to described grid;
Described sensitive element be arranged to described dielectric layer a part and with described grid, described source electrode, described drain electrode or described semiconductor layer electric connection; And
Described sensitive element and described source electrode, described drain electrode or described semiconductor layer electric connection or be arranged to the part of described source electrode, described drain electrode or described semiconductor layer.
3. microbial detection device as claimed in claim 2, it is characterized in that, described reaction chamber is arranged on the top of described sensitive element.
4. microbial detection device as claimed any one in claims 1 to 3, it is characterized in that, the volume size of described reaction chamber is arranged so that the state of single microbial in described reaction chamber changes the pH value change caused and also can be detected, and makes described reaction chamber can hold microorganism to prevent again larger mammalian cell from entering described reaction chamber.
5. microbial detection device as claimed in claim 4, is characterized in that, the volume of described reaction chamber is received in the scope of liter to 1 at 1 microlitre.
6. microbial detection device as claimed any one in claims 1 to 3, it is characterized in that, described dielectric layer is also provided with controller, and this controller is for controlling the parameter of described organic film FET to adapt to different detected objects and detection demand.
7. a microorganism detection method, is characterized in that comprising the following steps:
Microbial detection device according to any one of claim 1 to 6 is provided,
Sample to be detected is put into described reaction chamber; And
Channel current change according to described organic film FET determines whether there is microorganism in described sample to be detected and there is which kind of microorganism.
8. microorganism detection method as claimed in claim 7, is characterized in that, determines whether to there is microorganism in described sample to be detected and the step that there is which kind of microorganism comprises further according to the channel current change of described organic film FET:
The growth of microorganism in described sample to be detected causes the pH value of described sample to be detected to change;
Described pH value change causes described sensitive element to produce electric charge;
The electric charge of described sensitive element causes the induced charge on described grid, described source electrode, described drain electrode or described semiconductor layer to redistribute; And
Induced charge redistribution on described grid, described source electrode, described drain electrode or described semiconductor layer causes the electromotive force between described source electrode and described drain electrode to change by the effect of electrode coupling electric capacity.
9. microorganism detection method as claimed in claim 8, is characterized in that, determines whether to there is microorganism in described sample to be detected and the step that there is which kind of microorganism also comprises according to the channel current change of described organic film FET:
Potential change between described source electrode and described drain electrode causes the channel current of described organic film FET to change; And
Determine whether there is microorganism in described sample to be detected and there is which kind of microorganism according to the change curve of described channel current.
10. the microbial detection device according to any one of claim 7 to 9, is characterized in that, described sample to be detected is microbial culture medium.
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Cited By (1)
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| CN107189937A (en) * | 2017-04-14 | 2017-09-22 | 漯河医学高等专科学校 | A kind of microbial detection device and microorganism detection method |
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Application publication date: 20150408 |