CN203083965U - Mycobacterium tuberculosis DNA (deoxyribonucleic acid) electrochemical sensing and detecting device - Google Patents
Mycobacterium tuberculosis DNA (deoxyribonucleic acid) electrochemical sensing and detecting device Download PDFInfo
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- CN203083965U CN203083965U CN 201320070368 CN201320070368U CN203083965U CN 203083965 U CN203083965 U CN 203083965U CN 201320070368 CN201320070368 CN 201320070368 CN 201320070368 U CN201320070368 U CN 201320070368U CN 203083965 U CN203083965 U CN 203083965U
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- tubercle bacillus
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Abstract
The utility model discloses a mycobacterium tuberculosis DNA (deoxyribonucleic acid) electrochemical sensing and detecting device. The electrochemical sensing and detecting device comprises a mycobacterium tuberculosis DNA electrochemical sensor, an electrochemical workstation and a computer. The electrochemical sensing and detecting device is characterized in that the mycobacterium tuberculosis DNA electrochemical sensor is connected with the electrochemical workstation through electrode wires, and the electrochemical workstation is connected with the computer through a data wire. According to the mycobacterium tuberculosis DNA electrochemical sensing and detecting device, the electrochemical DNA biologic sensing and detection of the mycobacterium tuberculosis are realized, so that the detection and identifying speeds of the mycobacterium tuberculosis are improved. By utilizing the device, the isolated culture is avoided, the time consumption is greatly reduced, the detection repeatability of the sensor can be improved by utilizing a protein control and assembling interface, the detection specificity can be improved by designing complementary sequences of specific fragments of the mycobacterium tuberculosis into capturing probes, and the detection sensitivity can be multiplied by designing repeated sequences in gene sequences of the mycobacterium tuberculosis into signal probes; and the sensor is convenient to carry, can be applied to point-of-care test and is capable of realizing economic, fast, sensitive and accurate point-of-care test.
Description
Technical field
The utility model relates to
GalvanochemistrySensor and medical test technical field, particularly
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit
Background technology
Much's bacillus (
M. tuberculosis), the abbreviation tubercle bacillus (
Tubercle bacilli).As far back as 1882, German bacteriologist Robert Koch proved that Much's bacillus is a pathogen lungy.This bacterium can be invaded each histoorgan of whole body, sees at most with pulmonary infection.After the eighties in 20th century, because acquired immune deficiency syndrome (AIDS) and the appearance of Much's bacillus antibody-resistant bacterium, the factors such as application, drug abuse, poverty and movement of population of immunodepressant, epidemic situation lungy worsens suddenly in the global range.Along with the chemical sproof enhancing of tulase, the incidence of disease lungy improves day by day, and being widely current of other diseases such as the acquired immune deficiency syndrome (AIDS) of tuberculosis merging especially in recent years makes quick diagnosis lungy and treatment in time seem particularly important.Traditional tuberculosis detection method is mainly bacterial cultivation.Along with progressively carrying out of genome project, increasing genome sequence is measured, and a large amount of gene order information is that various biological analysiss and detection are laid a good foundation.Utilize the method for molecular Biological Detection tubercle bacillus to mainly contain round pcr and biochip technology at present, yet in the clinical practice sample detection, there are problems such as poor specificity, easy pollution respectively in round pcr and genetic chip, and detect the cost height, therefore press for and make up new quick, responsive, low-cost, easy-operating detection technique.
At present, the DNA electrochemical sensor that utilizes galvanochemistry to combine with enzyme technology to make up is paid close attention to widely, and the related gene that is applied to diseases such as tumour detects research.Its principle of protein control assembled interface makes up protein control assembled interface for adopting in gold electrode surfaces, make the BSA molecular layer of electrode surface assembling one deck sphere, between these spherical molecules, expose the carat (measure of the purity of gold) point that is not occupied, then capture probe DNA is fixed in an orderly manner the carat (measure of the purity of gold) point of exposure; Under proper condition, capture probe DNA and signal probe DNA, DNA to be detected form " sandwich " structure by hybridizing fully, at signal probe DNA end mark biotin, modify Avidin on the enzyme, the interaction by biotin and Avidin is attached on " sandwich " structure enzyme; Place substrate solution that redox reaction takes place sensor, under set potential, measure substrate for enzymatic activity generation redox reaction and produce current signal.If the sequence that detects can not hybridize with capture probe DNA or signal probe DNA, then can't form " sandwich " structure, promptly do not have enzyme to be attached on the sensor, so oxidation that can't catalysis TMB, the current signal of generation is very weak.Therefore can realize specific recognition and detection by signal intensity stool and urine to complementary series and base mismatch sequence.This Novel DNA electrochemica biological sensor is not only quick, sensitive, accurate, cost is low, also has functions such as molecular recognition, separation and purification gene, thereby in fields such as molecular biology and laboratory medicines great application prospect is arranged.
(district repeat, DR), the repetitive sequence in this zone has high conservative one section DR district in the gene order of tubercle bacillus.
Summary of the invention
It is a kind of that the purpose of this utility model is to provide
The mycobacterium tuberculosis dna electrochemical sensingPick-up unit.Can pass through
The mycobacterium tuberculosis dna electrochemical sensingThe purpose that the design realization of device detects tubercle bacillus nucleic acid in the clinical samples by electrochemical method, and pass through
The mycobacterium tuberculosis dna electrochemical sensingBeing connected of device, electrochemical workstation and computing machine, given full play to characteristics such as it is quick, sensitive, accurate, low cost, it is right to realize
Tubercle bacillusThe DNA electrochemical sensing detect.
The purpose of this utility model is achieved in that described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, comprising mycobacterium tuberculosis dna electrochemical sensor, electrochemical workstation and a computing machine, its design feature is that the mycobacterium tuberculosis dna electrochemical sensor is connected with electrochemical workstation by electrode wires, electrochemical workstation connects computing machine by data line.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics comprise working electrode, contrast electrode, auxiliary electrode, temperature heater, isothermal reaction pond for the mycobacterium tuberculosis dna electrochemical sensor; The isothermal reaction pond is positioned on the temperature heater, and working electrode, contrast electrode and auxiliary electrode all are positioned in the reactant liquor in the isothermal reaction pond.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are that working electrode adopts gold electrode, are assembled with bovine serum albumin(BSA) on gold electrode surfaces, are being assembled with specific tubercle bacillus capture probe DNA on the gold electrode surfaces, between bovine serum albumin(BSA).
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are for being attached to mycobacterium tuberculosis dna to be detected on the specific tubercle bacillus capture probe DNA after by specific tubercle bacillus capture probe DNA and mycobacterium tuberculosis dna hybridization reaction to be detected on the gold electrode surfaces.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are for being attached to many times tubercle bacillus signal probe DNA on the mycobacterium tuberculosis dna to be detected after by mycobacterium tuberculosis dna to be detected and tubercle bacillus signal probe DNA hybridization reaction on the gold electrode surfaces.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are for behind the Avidin generation chemical reaction of being modified on institute's modified biotin and the horseradish peroxidase on by tubercle bacillus signal probe DNA on the gold electrode surfaces being attached to horseradish peroxidase on many times the tubercle bacillus signal probe DNA.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are that mycobacterium tuberculosis dna sequence to be detected is the repetitive sequence genetic fragment with high conservative.
Described
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, its characteristics are that tubercle bacillus signal probe DNA is the complementary series of the repetitive sequence genetic fragment in the mycobacterium tuberculosis dna sequence to be detected.
Advantage of the present utility model: the detection method that contrasts conventional tubercle bacillus, this method need not separation and Culture, consuming time significantly reducing (total about 2.5 hours consuming time), adopted the tubercle bacillus specific fragment to be designed to capture probe, the complementary series of repetitive sequence is designed to signal probe in tubercle bacillus self target sequence, this method for designing can make repeatability, specificity and the sensitivity of detection promote greatly, and this sensor is easy to carry, can be used for bedside fast, sensitive, accurately detect.
Description of drawings
Fig. 1 is a structural representation of the present utility model.
Fig. 2 is among Fig. 1
The mycobacterium tuberculosis dna electrochemical sensingThe structure for amplifying synoptic diagram of the working electrode of device.
Fig. 3 is Fig. 2's
The mycobacterium tuberculosis dna electrochemical sensingDevice detects
Tubercle bacillusThe structure for amplifying synoptic diagram of DNA.
Among the figure: 1 is the mycobacterium tuberculosis dna electrochemical sensor; 2 is electrochemical workstation; 3 is computing machine; 4 is data line; 5 is electrode wires; 6 is working electrode; 7 is contrast electrode; 8 is auxiliary electrode; 9 is that temperature heater, 10 is the isothermal reaction pond; 11 is bovine serum albumin(BSA); 12 is specific tubercle bacillus capture probe DNA; 13 is mycobacterium tuberculosis dna to be detected; 14 is tubercle bacillus signal probe DNA; 15 is horseradish peroxidase.
Embodiment
Below in conjunction with drawings and Examples the utility model is elaborated:
As Fig. 1, Fig. 2 and shown in Figure 3, described a kind of tubercle bacillus electrochemical DNA bio-sensing pick-up unit, comprise a mycobacterium tuberculosis dna electrochemical sensor 1, electrochemical workstation 2 and computing machine 3, its design feature is that mycobacterium tuberculosis dna electrochemical sensor 1 is connected with electrochemical workstation 2 by electrode wires 5, and electrochemical workstation 2 connects computing machine 3 by data line 4.Mycobacterium tuberculosis dna electrochemical sensor 1 comprises that 6 are working electrode; 7 is contrast electrode; 8 is auxiliary electrode; 9 is that temperature heater, 10 is the isothermal reaction pond.Isothermal reaction pond 10 is positioned on the temperature heater 9, and working electrode 6, contrast electrode 7, auxiliary electrode 8 are positioned in the reactant liquor in the isothermal reaction pond 10.Electrochemical workstation 2 is gathered on the mycobacterium tuberculosis dna electrochemical sensors 1 generation electric signal by working electrode 6, contrast electrode 7, auxiliary electrode 8 through electrode wires 5, and is transferred to computing machine 3 and carries out data analysis and handle.
Of the present utility model being produced as follows: comprise following 3 steps:
(1) at first make up working electrode 6 in the mycobacterium tuberculosis dna electrochemical sensor 1, the pre-service of earlier gold electrode being polished as working electrode, is immersed gold electrode temperature control reaction in the bovine serum albumin 11 then, takes out with the distilled water flushing, and nitrogen dries up.Drip in gold electrode surfaces and to be coated with specific tubercle bacillus capture probe DNA 12, take out with the distilled water flushing, nitrogen dries up.Gold electrode is immersed again and contain temperature control reaction among mycobacterium tuberculosis dna 13 to be detected and the tubercle bacillus signal probe DNA 14, take out with the distilled water flushing, nitrogen dries up.Drip in gold electrode surfaces and to be coated with horseradish peroxidase 15, with the distilled water flushing, to be measured.
(2) build mycobacterium tuberculosis dna electrochemical sensor 1, isothermal reaction pond 10 is positioned on the temperature heater 9, fixing working electrode 6, contrast electrode 7, auxiliary electrode 8 on the electrode suppor, regulate the support height, working electrode 6, contrast electrode 7, auxiliary electrode 8 are positioned in the reactant liquor in the isothermal reaction pond 10.
(3) building of electrochemical signals detection platform, mycobacterium tuberculosis dna electrochemical sensor 1 is connected with electrochemical workstation 2 by electrode wires 5, and electrochemical workstation 2 connects computing machine 3 by data line 4.
Experimentation:
(1) earlier tubercle bacillus electrochemical DNA bio-sensing pick-up unit of the present utility model is assembled opening power, computing machine 3 and electrochemical workstation 2 by above-mentioned steps.
(2) gold electrode is polished pre-service, 25 ℃ of temperature controls of BSA solution of putting into 0.1 % then react 15 min, and with the distilled water flushing, nitrogen dries up; Drip in gold electrode surfaces and to be coated with 25 ℃ of the specific tubercle bacillus capture probe of 4 μ l DNA and to place 60 min, with the distilled water flushing, nitrogen dries up; Then gold electrode is put into the mixed solution that contains mycobacterium tuberculosis dna to be detected and tubercle bacillus signal probe DNA and reacted 60 min in 44 ℃ of temperature controls, with the distilled water flushing, nitrogen dries up; Again gold electrode is put into 25 ℃ of temperature controls of 0.5 U/ml horseradish peroxidase solution and reacted 15 min, wash with distilled water; Gold electrode put into contain 0.05% tween-20 solution stirring to clean 5 min to be measured.
(3) will be immersed in through the working electrode 6 of finishing and contain H
2O
2Tmb substrate solution in, measure current value after the hybridization with electrochemical workstation 2, assay method is electric current-time curve method, condition determination is: initial potential is 0 v, sampling interval is 0.1s, experimental period is 120s.By the big or small discriminatory analysis of current value, after hybridization, detect current value and show then that greater than the current value of not hybridizing strand containing tubercle bacillus in the sample belongs to.
Claims (8)
1. one kind
Mycobacterium tuberculosis dna electrochemical sensing pick-up unitComprise a mycobacterium tuberculosis dna electrochemical sensor (1), electrochemical workstation (2) and computing machine (3), it is characterized in that: mycobacterium tuberculosis dna electrochemical sensor (1) is connected with electrochemical workstation (2) by electrode wires (5), and electrochemical workstation (2) connects computing machine by data line (4).
2. according to claim 1
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that: mycobacterium tuberculosis dna electrochemical sensor (1) comprises working electrode (6), contrast electrode (7), auxiliary electrode (8), temperature heater, isothermal reaction pond (10); Isothermal reaction pond (10) is positioned on the temperature heater (9), and working electrode (6), contrast electrode and auxiliary electrode (8) all are positioned in the interior reactant liquor in isothermal reaction pond (10).
3. according to claim 2
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that: working electrode adopts gold electrode, is assembled with bovine serum albumin(BSA) (11) on gold electrode surfaces, on the gold electrode surfaces, be positioned at be assembled with specific tubercle bacillus capture probe DNA(12 between the bovine serum albumin(BSA) (11)).
4. according to claim 3
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that on the gold electrode surfaces by behind specific tubercle bacillus capture probe DNA (12) and mycobacterium tuberculosis dna to be detected (13) hybridization reaction mycobacterium tuberculosis dna to be detected (13) being attached to specific tubercle bacillus capture probe DNA(12) on.
5. according to claim 4
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that on the gold electrode surfaces by mycobacterium tuberculosis dna to be detected (13) and tubercle bacillus signal probe DNA(14) behind the hybridization reaction with many times tubercle bacillus signal probe DNA(14) be attached on the mycobacterium tuberculosis dna to be detected (13).
6. according to claim 5
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that on the gold electrode surfaces by tubercle bacillus signal probe DNA(14) on horseradish peroxidase (15) is attached to behind the Avidin generation chemical reaction modified on institute's modified biotin and the horseradish peroxidase (15) on many times the tubercle bacillus signal probe DNA.
7. according to claim 5
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that mycobacterium tuberculosis dna to be detected (13) sequence is the repetitive sequence genetic fragment with high conservative.
8. according to claim 5
A kind of mycobacterium tuberculosis dna electrochemical sensing pick-up unit, it is characterized in that tubercle bacillus signal probe DNA(14) and be the complementary series of the repetitive sequence genetic fragment in mycobacterium tuberculosis dna to be detected (13) sequence.
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| CN 201320070368 CN203083965U (en) | 2013-02-06 | 2013-02-06 | Mycobacterium tuberculosis DNA (deoxyribonucleic acid) electrochemical sensing and detecting device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104483362A (en) * | 2014-12-30 | 2015-04-01 | 中国人民解放军第三军医大学第二附属医院 | Capture probe and signal probe modified electrode as well as preparation method and application thereof |
| CN104698043A (en) * | 2013-12-09 | 2015-06-10 | 重庆医科大学 | An electrochemical biosensor for detecting DNA of escherichia coli and a preparing method thereof |
| CN109722378A (en) * | 2018-12-20 | 2019-05-07 | 安徽中青检验检测有限公司 | A kind of food bacterial content detection device |
-
2013
- 2013-02-06 CN CN 201320070368 patent/CN203083965U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698043A (en) * | 2013-12-09 | 2015-06-10 | 重庆医科大学 | An electrochemical biosensor for detecting DNA of escherichia coli and a preparing method thereof |
| CN104483362A (en) * | 2014-12-30 | 2015-04-01 | 中国人民解放军第三军医大学第二附属医院 | Capture probe and signal probe modified electrode as well as preparation method and application thereof |
| CN109722378A (en) * | 2018-12-20 | 2019-05-07 | 安徽中青检验检测有限公司 | A kind of food bacterial content detection device |
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