CN202870030U - Legionella bacteria electrochemistry DNA (Deoxyribose Nucleic Acid) biological sensing and detecting device - Google Patents
Legionella bacteria electrochemistry DNA (Deoxyribose Nucleic Acid) biological sensing and detecting device Download PDFInfo
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- CN202870030U CN202870030U CN 201220536434 CN201220536434U CN202870030U CN 202870030 U CN202870030 U CN 202870030U CN 201220536434 CN201220536434 CN 201220536434 CN 201220536434 U CN201220536434 U CN 201220536434U CN 202870030 U CN202870030 U CN 202870030U
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Abstract
The utility model discloses a legionella bacteria electrochemistry DNA (Deoxyribose Nucleic Acid) biological sensing and detecting device. A DNA electrochemistry biosensing technique is applied to detection of the legionella bacteria; the purpose of detecting legionella bacteria nucleic acid in a sample through an electrochemistry method is achieved through the design of a legionella bacteria electrochemistry DNA sensor; and electrochemistry DNA biological sensing and detecting of the legionella bacteria are achieved through connection among the legionella bacteria electrochemistry DNA sensor, an electrochemistry workstation and a computer. The characteristics of rapidity, sensitivity, accuracy, low price and the like are fully played, so that the emergency response speed for monitoring and detecting the legionella bacteria is enhanced. The isolated culture is not needed through the utilization of the device, so that the time consumption is greatly reduced; through the utilization of the legionella bacteria specificity fragment as the probe, the specificity and sensitivity of the detection are greatly improved; and the sensor is convenient to carry, can be used for the field detection in a plurality of environments, is especially suitable for field qualitative detection and carries out rapid, sensitive and accurate detection in the field.
Description
Technical field
The utility model relates to
GalvanochemistrySensor and preventive medicine monitoring technical field, particularly
A kind of Legionella electrochemical DNA bio-sensing pick-up unit
Background technology
Once broke out the febris acuta breathing problem among the members of U.S. Philadelphia legion in 1976, after deliberation, found that a kind of malignant bacteria was also with it called after legionella pneumophilia.Have now found that to surpass 30 kinds of Legionnella, wherein at least 19 kinds is the cause of disease of people's parapneumonia.The most commonly encountered diseases substance is legionella pneumophilia (accounts for case 85% ~ 90%), and this bacterium is present in the water and soil, often sucks and causes that pneumonia type and non-pneumonia type infect through water system, solution cavity and atomizing.Also have obvious multiple organ injury except respiratory symptom, fatal rate is higher.Also can also can merge other infected by microbes up to 23% according to the nearest microbial hospital infection of investigation statistics legion, form refractory pneumonia, fatal rate is up to 45%.So the position of Legionella in opportunistic infect or inside-hospital infection paid attention to by people day by day.The immunoassay technology that detects at present Legionella has immunochromatographic method, enzyme immunoassay, radioimmunology, cytometry and Immunosensors Technology And.In addition, also has the check of fluorescein molecular hybridization in situ.But the not easily separated cultivation of Legionella, kind is various again, there is no at present simple and easy, the express laboratory method of inspection, so study the matter of utmost importance that new quick, special, responsive Legionella detection technique is current Legionella inspection technology research.And the DNA electrochemica biological sensor is a class Novel Biosensor of international hot research in recent years, it is that single stranded DNA (ssDNA) is modified electrode surface, consist of the dna modification electrode, because the height sequence selective of the probe ssDNA on the electrode and the hybridization of the complementary strand in the solution is so that this ssDNA modified electrode has extremely strong molecule distinguishability.Under proper condition, ssDNA probe molecule and sequence DNA to be measured (target sequence) hybridization, form double-stranded DNA (dsDNA) at electrode surface, thereby the change of the electrode surface structure before and after causing hybridizing, difference before and after this hybridization can be measured by the response change of electrochemical measuring technique, thereby reaches the detection target sequence.This Novel DNA electrochemica biological sensor is not only quick, sensitive, accurate, cheap, also has the functions such as molecular recognition, separation and purification gene, thereby in fields such as molecular biology and bioengineering medical science huge application prospect is arranged.This research comes up the detection that the electrochemica biological sensor technology is applied to Legionella, thereby has opened up new approaches and the method for quick, special, responsive Legionella detection technique.
Summary of the invention
The purpose of this utility model is to provide a kind of Legionella electrochemical DNA bio-sensing pick-up unit.The purpose that can detect Legionella nucleic acid in the sample by electrochemical method by the design realization of Legionella electrochemical DNA biosensor, and be connected connection with computing machine by Legionella electrochemical DNA biosensor, electrochemical workstation, the electrochemical DNA bio-sensing of realizing Legionella detects, given full play to its characteristics such as quick, sensitive, accurate, cheap, thus the emergency reaction speed that improves the Legionella monitoring and detect.
The purpose of this utility model is achieved in that
Described a kind of Legionella electrochemical DNA bio-sensing pick-up unit, comprising Legionella electrochemical DNA biosensor, electrochemical workstation and a computing machine, its characteristics are that the Legionella electrochemical DNA biosensor is connected with electrochemical workstation by electrode wires, electrochemical workstation connects computing machine by data line.
Described
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, its characteristics are that the Legionella electrochemical DNA biosensor comprises contrast electrode, auxiliary electrode, working electrode, reaction tank, magnetic stir bar and with the magnetic stirring apparatus of temperature control; Reaction tank is positioned on the magnetic stirring apparatus with temperature control, and magnetic stir bar is positioned over the reaction tank bottom, and contrast electrode, auxiliary electrode and working electrode are positioned in the reactant liquor in the reaction tank.
Described
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, its characteristics are that working electrode adopts glass-carbon electrode, are provided with specific Legionella dna probe on the glass-carbon electrode surface.
Described
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, its characteristics on the glass-carbon electrode surface by working electrode surface-the COOH group is connected with specific Legionella dna probe.
Advantage of the present utility model: contrast the conventional method that Legionella detects in the water sample in the past, this method need not to separate consuming time the greatly reducing of cultivation, having adopted the Legionella specific fragment is that probe can make the specificity of detection and sensitivity greatly promote, and this sensor is easy to carry, can be used for the Site Detection under the multiple environment, be particularly useful for on-the-spot qualitative detection, accomplish on-the-spot quick, sensitive, accurately detection.
Description of drawings
Fig. 1 is structural representation of the present utility model.
Fig. 2 is the structure for amplifying synoptic diagram of the working electrode of the Legionella electrochemical DNA biosensor among Fig. 1.
Fig. 3 is the structure for amplifying synoptic diagram that the working electrode of the Legionella electrochemical DNA biosensor of Fig. 2 is combined with Legionella dna fragmentation to be checked.
Among the figure: 1 is the Legionella electrochemical DNA biosensor; 2 is electrochemical workstation; 3 is computing machine; 4 is contrast electrode; 5 is auxiliary electrode; 6 is working electrode; 7 is reaction tank; 8 is magnetic stir bar; 9 are the magnetic stirring apparatus with temperature control; 10 is electrode wires; 11 is data line; 12 be working electrode surface-the COOH group; 13 is specific Legionella dna probe; 14 is Legionella dna fragmentation to be checked.
Embodiment
Below in conjunction with drawings and Examples the utility model is elaborated:
Such as Fig. 1, Fig. 2 and shown in Figure 3, described a kind of Legionella electrochemical DNA bio-sensing pick-up unit, comprise a Legionella electrochemical DNA biosensor 1, electrochemical workstation 2 and computing machine 3, its design feature is that Legionella electrochemical DNA biosensor 1 is connected with electrochemical workstation 2 by electrode wires 10, and electrochemical workstation 2 connects computing machine 3 by data line 11.Legionella electrochemical DNA biosensor 1 comprises contrast electrode 4, auxiliary electrode 5, working electrode 6, reaction tank 7, magnetic stir bar 8 and with the magnetic stirring apparatus 9 of temperature control.Reaction tank 7 is positioned on the magnetic stirring apparatus 9 with temperature control, and magnetic stir bar 8 is positioned over reaction tank 7 bottoms, and contrast electrode 4, auxiliary electrode 5 and working electrode 6 are positioned in the reactant liquor in the reaction tank 7.Electrochemical workstation 2 gathers on the Legionella electrochemical DNA biosensors 1 generation electric signal by contrast electrode 4, auxiliary electrode 5 and working electrode 6 through electrode wires 10, and is transferred to computing machine 3 and carries out data analysis and process.
Of the present utility model being produced as follows: comprise following 4 steps: (1) at first makes up the working electrode 6 in the Legionella electrochemical DNA biosensor 1, first glass-carbon electrode is carried out pre-service, then distilled water flushing after soaking with the coupling agent [5 mmol/L EDC (hydrochloric acid 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide)-8 mmol/L NHS (maloyl imines)] that 100mL is housed, N
2After drying up, directly drip specific Legionella dna probe 13 at electrode surface, with surveying its resistance value with electrochemical workstation after the 0.2%SDS washing; (2) build Legionella electrochemical DNA biosensor 1, reaction tank 7 is positioned on the magnetic stirring apparatus 9 with temperature control, magnetic stir bar 8 is positioned over reaction tank 7 bottoms, contrast electrode 4, auxiliary electrode 5 and working electrode 6 on electrode suppor is fixing, regulate the support height, contrast electrode 4, auxiliary electrode 5 and working electrode 6 are positioned in the reactant liquor in the reaction tank 7.(3) building of electrochemical signals detection platform, Legionella electrochemical DNA biosensor 1 is connected with electrochemical workstation 2 by electrode wires 10, and electrochemical workstation 2 connects computing machine 3 by data line 11.
Experimentation: first Legionella electrochemical DNA bio-sensing pick-up unit of the present utility model is assembled opening power, electrochemical workstation 2 and computing machine 3 by above-mentioned steps; Sample to be detected is put into reaction tank 7, be immersed in the sample to be tested solution working electrode 6 of the specific Legionella dna probe 13 of finishing behind 35 ℃ of hybridization 40min and hybridization connects upper Legionella dna fragmentation 14 to be checked, with 0.2%SDS washing 3 minutes, with electrochemical workstation 2 at 0.001mol/L iron cyanogen electricity to (K
3Fe (CN)
6/ K
4Fe (CN)
6) measure the resistance value after the hybridization in the solution, condition determination is: method is chosen as AC impedence method, and initial potential is chosen as instant open circuit potential, and high frequency is selected 100000Hz, and low frequency is selected 1Hz, and amplitude is 0.005V; Or 2 * 10
-5Do differential pulse voltammetry (DPV) scanning in 0.02mol/L Tris-HCl solution behind the enrichment 5min in the mol/L methylene blue solution, condition determination is: method is chosen as differential pulse voltammetry, initial potential is 1V for-1V stops current potential, the current potential increment is 0.004V, amplitude is 0.05V, and pulse width is 0.2 second.By AC impedance spectrogram discriminatory analysis, when then showing greater than strand resistance value before hybridizing, resistance value double-stranded after the hybridization contains Legionnella in the sample; By differential pulse voltammetry spectrogram discriminatory analysis, when then showing less than the DPV peak value of strand before hybridizing, DPV peak value double-stranded after the hybridization contains Legionnella in the sample.
Claims (4)
1.
A kind of Legionella electrochemical DNA bio-sensing pick-up unitComprise a Legionella electrochemical DNA biosensor (1), electrochemical workstation (2) and computing machine (3), it is characterized in that: Legionella electrochemical DNA biosensor (1) is connected with electrochemical workstation (2) by electrode wires (10), and electrochemical workstation (2) connects computing machine by data line (11).
2. according to claim 1
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, it is characterized in that: Legionella electrochemical DNA biosensor (1) comprises contrast electrode (4), auxiliary electrode (5), working electrode (6), reaction tank, magnetic stir bar (8) and with the magnetic stirring apparatus (9) of temperature control; Reaction tank (7) is positioned on the magnetic stirring apparatus (9) with temperature control, and magnetic stir bar (8) is positioned over reaction tank (7) bottom, and contrast electrode (4), auxiliary electrode (5) and working electrode are positioned in the reactant liquor in the reaction tank (7).
3. according to claim 2
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, it is characterized in that: working electrode (6) adopts glass-carbon electrode, is provided with specific Legionella dna probe (13) on the glass-carbon electrode surface.
4. according to claim 3
A kind of Legionella electrochemical DNA bio-sensing pick-up unit, it is characterized in that on the glass-carbon electrode surface by working electrode surface-COOH group (12) is connected with specific Legionella dna probe (13).
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201220536434 CN202870030U (en) | 2012-10-18 | 2012-10-18 | Legionella bacteria electrochemistry DNA (Deoxyribose Nucleic Acid) biological sensing and detecting device |
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| CN 201220536434 CN202870030U (en) | 2012-10-18 | 2012-10-18 | Legionella bacteria electrochemistry DNA (Deoxyribose Nucleic Acid) biological sensing and detecting device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104237354A (en) * | 2014-10-20 | 2014-12-24 | 中国人民解放军第三军医大学第一附属医院 | NDM-1 DNA probe modified electrode as well as preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104237354A (en) * | 2014-10-20 | 2014-12-24 | 中国人民解放军第三军医大学第一附属医院 | NDM-1 DNA probe modified electrode as well as preparation method and application thereof |
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| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130410 Termination date: 20151018 |
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