CN102517399B - Heat transfer detection method based on DNA amplification - Google Patents
Heat transfer detection method based on DNA amplification Download PDFInfo
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- CN102517399B CN102517399B CN 201210006323 CN201210006323A CN102517399B CN 102517399 B CN102517399 B CN 102517399B CN 201210006323 CN201210006323 CN 201210006323 CN 201210006323 A CN201210006323 A CN 201210006323A CN 102517399 B CN102517399 B CN 102517399B
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- 238000001514 detection method Methods 0.000 title claims abstract description 74
- 230000004544 DNA amplification Effects 0.000 title abstract description 5
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 72
- 238000006243 chemical reaction Methods 0.000 claims abstract description 42
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 36
- 238000000034 method Methods 0.000 claims abstract description 32
- 230000008569 process Effects 0.000 claims abstract description 29
- 239000000835 fiber Substances 0.000 claims abstract description 16
- 239000000523 sample Substances 0.000 claims abstract description 14
- 238000010367 cloning Methods 0.000 claims description 23
- 238000012360 testing method Methods 0.000 claims description 10
- 238000013499 data model Methods 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 claims description 5
- 230000008859 change Effects 0.000 claims description 5
- 230000004044 response Effects 0.000 claims description 5
- 230000011514 reflex Effects 0.000 claims description 4
- 238000004088 simulation Methods 0.000 claims description 4
- 239000012295 chemical reaction liquid Substances 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 2
- 239000013307 optical fiber Substances 0.000 abstract description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating Or Analyzing Materials Using Thermal Means (AREA)
Abstract
The invention relates to a heat transfer detection method based on DNA amplification, which utilizes a fiber Bragg grating sensor, a conventional platinum resistance sensor and a process detection module to establish a detection system on a DNA amplification module, the detection system comprises a fiber Bragg grating detection subsystem and a platinum resistance sensor detection subsystem, the fiber bragg grating detection subsystem comprises a fiber bragg grating sensor and a signal demodulation system, the optical fiber Bragg grating sensor comprises a sensing probe, the signal demodulation system is connected with the sensing probe and inserts the sensing probe into the reaction hole, the platinum resistance sensor detection subsystem comprises a control unit and a platinum resistance sensor, the flow detection module is connected with the fiber Bragg grating detection subsystem and the platinum resistance sensor detection subsystem through an interface module.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of heat passage detection method based on DNA cloning.
Background technology
The step of archaeal dna polymerase being introduced PCR is: the DNA reaction solution is placed in the test tube, and the test tube that reaction solution is housed is placed in each hole of PCR module of reaction chamber, and the gene amplification process is finished automatically by sequence of control.Up to the present, Chinese scholars mainly utilizes automatic control technology that the temperature variation of PCR module is controlled on sequence of control, the immanent structure expression of gene that Stephanie J.Culler seminar has utilized the PCR device analysis, but most achievement in research concentrates on some index of improving round pcr and improving PCR equipment.E.T.Lagally adopts the method that improves temperature rate that the DNA cloning process is studied as far back as calendar year 2001, Grover J. and Juncosa R.D studied the elevation rate of conventional PCR equipment in 2008, and employing Digital Control modes such as T.M.H.Lee have improved the reaction control accuracy of PCR.In the clinical application of PCR, a ubiquitous problem is: the state that is arranged in the DNA reaction solution of each hole test tube of module changes with the difference of step of reaction, in this process, the chemical bond of DNA is fracture or generation constantly, the suction that is accompanied by, the heat release state is also different, how to analyze these change of state rules, to improving the DNA cloning quality and accurately controlling and have a direct impact to being placed in each hole of module the temperature variation of test-tube reaction liquid, the prior-art devices complicated operation, system's property safe and reliable to operation is low, prior art is undesirable to detecting effect, and being applied in has certain limitation in the actual engineering.
Summary of the invention
The present invention overcomes the deficiencies in the prior art, a kind of heat passage detection method based on DNA cloning has been proposed, described method is utilized fiber Bragg grating sensor and distribution character model thereof, utilize this characteristic that the data rule of DNA reaction solution is gathered and analyzed, the heat passage rule of analyzing DNA amplification.
Technical scheme of the present invention is: a kind of heat passage detection method based on DNA cloning, described detection method is utilized fiber Bragg grating sensor, conventional platinum sensor and flow process detection module are set up detection system on the DNA cloning module, described DNA cloning module is planeform, be positioned in the airtight reaction compartment, its top is provided with reacting hole, described detection system comprises Fiber Bragg Grating FBG detection subsystem and platinum sensor detection subsystem, described Fiber Bragg Grating FBG detection subsystem comprises fiber Bragg grating sensor and signal demodulating system, described fiber Bragg grating sensor comprises sensing probe, described signal demodulating system is connected with described sensing probe, described sensing probe is inserted in the described reacting hole, the platinum sensor detection subsystem comprises control unit and platinum sensor, described control unit is connected with platinum sensor, described platinum sensor closely is arranged on the bottom of DNA cloning module, described flow process detection module is connected with the platinum sensor detection subsystem with described Fiber Bragg Grating FBG detection subsystem by interface module, and test procedure is as follows:
1) starts the platinum sensor detection subsystem, utilize control unit to receive the temperature value that platinum sensor sends, be transformed into the temperature control data, pass to the flow process detection module through after the analog-to-digital conversion;
2) start the Fiber Bragg Grating FBG detection subsystem, the data that Bragg grating sensor is transmitted are transformed into the reaction chamber data through behind the signal demodulating system, pass to the flow process detection module;
3) the flow process detection module through after the normalized, is set up the one-to-one relationship data model to temperature control data and reaction chamber data;
4) according to the data model of having set up, the flow process detection module is determined DNA step of reaction and relevant border condition, COMPREHENSIVE CALCULATING, and be heat absorption or exothermic process and revise according to the data judging of Prague transmitter, the hot-fluid that calculating is captured in reaction solution in the fracture of chemical bond or the generative process changes, and the heat enthalpy value analysis temperature of conversion changes and heat flow density.
5) according to the 4th data construct DNA response data simulation model that goes on foot.
Described platinum sensor detection subsystem comprises three road detection signals, and every road comprises a platinum sensor.
Described DNA cloning module is provided with 96 reacting holes.
The present invention has following beneficial effect
1) the present invention adopts according to digital simulation model construction data control model, and the whole amplification procedure of analog D NA discloses the influence factor to dna sample, for the objective law that discloses the DNA internal-response provides foundation.
2) the present invention sets up the temperature variant distribution character of Fiber Bragg Grating FBG, discloses the suction law of heat release in DNA cloning stage, discloses the objective law of DNA internal-response, provides a whole set of Fiber Bragg Grating FBG temperature variant theory and technology basic data.
3) the present invention has set up the practicable data acquisition modes of DNA cloning process.
4) the present invention has disclosed the reaction rule of DNA, will help the exploitation of PCR equipment, and market outlook and social benefit are huge.
Embodiment
The present invention utilizes fiber Bragg grating sensor, conventional platinum sensor and flow process detection module to set up detection system on the DNA cloning module, described DNA cloning module is planeform, be positioned in the airtight reaction compartment, its top is provided with reacting hole, and common described DNA cloning module is provided with 96 reacting holes.Described detection system comprises Fiber Bragg Grating FBG detection subsystem and platinum sensor detection subsystem, described Fiber Bragg Grating FBG detection subsystem comprises fiber Bragg grating sensor and signal demodulating system, described fiber Bragg grating sensor comprises sensing probe, described signal demodulating system is connected with described sensing probe, several described sensing probes are inserted respectively in some hole of described 96 reacting holes, the sensor detecting subsystem comprises control unit and platinum sensor, described control unit is connected with platinum sensor, comprise three road detection signals, every road comprises a platinum sensor.Described platinum sensor closely is arranged on the bottom of DNA cloning module, and described flow process detection module is connected with the platinum sensor detection subsystem with described Fiber Bragg Grating FBG detection subsystem by interface module.
When utilizing described fiber Bragg grating sensor detected temperatures, according to coupled mode theory, the foveal reflex wavelength of Fiber Bragg Grating FBG can be expressed as
λ
B＝2n
effΛ (1)
N in the formula
EffBe the effective refractive index of guided mode, Λ is the cycle of grating.By (1) formula as can be seen, the foveal reflex wavelength is relevant with the grating cycle with effective refractive index;
When grating was subjected to variation of temperature and influences, its effective refractive index and grating cycle can change thereupon, thereby reflection wavelength also can change, and relational expression is:
Δλ
B＝2Δn
effΛ+2n
effΔΛ
Following formula substitution (1) is obtained
(2)
The grating cycle varying type and the thermo-optical coeffecient that are caused by thermal expansion effects cause that the effective refractive index varying type is respectively:
ΔΛ＝α·Λ·ΔT
Δn
eff＝ξ·n
eff·ΔT
Wherein α and ξ are respectively the thermal expansivity and the thermo-optical coeffecient of optical fiber.
Above-mentioned two formula substitution (2) Shi Kede:
Δλ
B＝k
T·λ
B·ΔT
Make α
T=k
Tλ
B
Then following formula can be written as:
Δλ
B＝α
T·ΔT (3)
The foveal reflex wavelength X of common optical fiber
B=1200nm, the temperature sensitivity coefficient k
T=7.5x10
-6/ C, therefore, α
TIt is the function of temperature.Formula (3) is a linear relationship to certain temperature, but when temperature variation was big, above-mentioned linear formula showed nonlinear characteristic.Be adapted at the bigger polymeric material of outer wrap temperature sensitivity coefficient, as, k
T=87x10
-6/ C, at this moment, α
T=9.88, that is, for the spatial resolution of every degree between 0-9.88.The present invention adopts the conventional modular unit in 96 holes, DCU data control unit divides three tunnel placement sensor groups, every road is provided with four module sensors, be used to test the control data collecting unit, can be under the situation of condition permission, in the reacting hole space on module top, arrange 12 reaction chamber inner module transmitters with the correspondence position of module sensors, be called for short the reaction chamber transmitter, the homogeneous and the harmony that are used for the acquisition module temperature, the data acquisition unit of setting up is used for experimental system is carried out data analysis, and as the auxiliary basic data of Bragg grating data collecting system.
Suppose that module sensors is A
Ij(i=1-3, j=1-4), the reaction chamber transmitter is W
Ij(i=1-3, j=1-4),
Wherein, A
Ij(i=1-3, j=1,2) and W
Ij(i=1-3 j=1-4) is analog quantity, and establishing the function that analog quantity transfers data volume (decimal system temperature value) to is G, can get by experiment with after the The Coupling: t at any one time, and the temperature data that its module sensors obtains is:
Wherein, i=1-3, what j=1-4, following formula represented is the actual temperature value of certain test point of automatic control system loading.
T at any one time, the λ of optical fiber
BValue is constantly to change, the data of the DNA reaction solution of the dna sample that foundation is measured, and foundation (3) Shi Kede:
Δλ
B(t)＝α
T·ΔT(t) (5)
From (5) formula as can be known, the temperature variation Δ T of the DNA reaction solution in (Δ t) can pass through to calculate Δ λ according to the demodulated equipment of Bragg grating between t moment partial zones
B(t) obtain, and temperature value can be by the three-way sensor combined calculation of module temperature constantly at arbitrary t.
So, t at any one time
0, the actual temperature data that its automatic control system loads can be calculated by (4) formula,
Wherein, i=1-3, j=1-4.
From the above analysis, to t
0In between the partial zones constantly, the variable quantity of the actual temperature of the corresponding test point in its reaction chamber can be calculated by (5) formula:
ΔT(t
0)|
ij＝Δλ
B(t
0)/α
T (α
T≠0)
Comparison through (4) formula and (5) formula can accurately calculate at arbitrary t
0Constantly, the precise temp changing value Δ η of DNA reaction chamber internal reaction liquid
Ij| t
0, that is,
Δη
ij|t
0＝|ΔT(t
0)|
ij-T
ij(t
0)| (6)
Can extrapolate the heat absorption of DNA reaction solution reaction process and the concrete rule of heat release from (6) formula, and can accurately simulate DNA biological respinse characteristic, and then set up the data model of DNA reaction, according to the data T that obtains according to above-mentioned formula
Ij(t), by determining the DNA step of reaction and inhale the heat release state with the contrast of Δ T.
Test procedure is as follows:
1) starts the platinum sensor detection subsystem, utilize control unit to receive the temperature value that platinum sensor sends, be transformed into the temperature control data, pass to the flow process detection module through after the analog-to-digital conversion;
2) start the Fiber Bragg Grating FBG detection subsystem, the data that Bragg grating sensor is transmitted are transformed into the reaction chamber data through behind the signal demodulating system, pass to the flow process detection module;
3) the flow process detection module through after the normalized, is set up the one-to-one relationship data model to temperature control data and reaction chamber data;
4) according to the data model of having set up, the flow process detection module is determined DNA step of reaction and relevant border condition, COMPREHENSIVE CALCULATING, and be heat absorption or exothermic process and revise according to the data judging of Prague transmitter, the hot-fluid that calculating is captured in reaction solution in the fracture of chemical bond or the generative process changes, and the heat enthalpy value analysis temperature of conversion changes and heat flow density.
5) according to the 4th data construct DNA response data simulation model that goes on foot.
Claims (3)
1. heat passage detection method based on DNA cloning, it is characterized in that: described detection method is utilized fiber Bragg grating sensor, conventional platinum sensor and flow process detection module are set up detection system on the DNA cloning module, described DNA cloning module is planeform, be positioned in the airtight reaction compartment, its top is provided with reacting hole, described detection system comprises Fiber Bragg Grating FBG detection subsystem and platinum sensor detection subsystem, described Fiber Bragg Grating FBG detection subsystem comprises fiber Bragg grating sensor and signal demodulating system, described fiber Bragg grating sensor comprises sensing probe, described signal demodulating system is connected with described sensing probe, described sensing probe is inserted in the described reacting hole, the platinum sensor detection subsystem comprises control unit and platinum sensor, described control unit is connected with platinum sensor, described platinum sensor closely is arranged on the bottom of DNA cloning module, described flow process detection module is connected with the platinum sensor detection subsystem with described Fiber Bragg Grating FBG detection subsystem by interface module, and it is as follows to detect step:
1) starts the platinum sensor detection subsystem, utilize control unit to receive the temperature value that platinum sensor sends, be transformed into the temperature control data, pass to the flow process detection module through after the analog-to-digital conversion;
2) start the Fiber Bragg Grating FBG detection subsystem, the data that Bragg grating sensor is transmitted are transformed into the reaction chamber data through behind the signal demodulating system, pass to the flow process detection module;
3) the flow process detection module temperature control data and reaction chamber data according to formula Δ λ
B=α
TAfter Δ T carries out normalized, set up the one-to-one relationship data model, wherein, Δ λ
BBe foveal reflex wavelength change amount, α
TBe the function of temperature, Δ T is the temperature variation of DNA reaction solution;
4) through after the processing of the 3rd step, determine DNA step of reaction and relevant border condition in conjunction with the flow process detection module, the arbitrary t in flow process detection module testing process
0Constantly, according to formula Δ η
Ij| t
0=| Δ T (t
0) |
Ij-T
Ij(t
0) | after carrying out COMPREHENSIVE CALCULATING and revising, and according to this variation of calculation result analysis temperature and heat flow density; Wherein, Δ η
Ij| t
0Expression t
0The precise temp changing value of DNA reaction chamber internal reaction liquid constantly, i=1-3, j=1-4, following formula represent is the actual temperature value by the test point that value determined of the value of i and j that automatic control system loads, Δ T (t
0) |
IjBe the t of DNA reaction solution by the test point that value determined of the value of i and j
0Moment temperature variation, T
Ij(t
0) be illustrated in t
0The time be engraved in the temperature data that the module sensors by the test point that value determined of the value of i and j obtains;
5) according to the 4th data construct DNA response data simulation model that goes on foot.
2. the heat passage detection method based on DNA cloning according to claim 1 is characterized in that: described platinum sensor detection subsystem comprises three road detection signals, and every road comprises a platinum sensor.
3. the heat passage detection method based on DNA cloning according to claim 1 and 2 is characterized in that: described DNA cloning module is provided with 96 reacting holes.
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| CN105718757B (en) * | 2016-01-25 | 2018-03-20 | 青岛理工大学 | Data modeling method for DNA amplification process |
| CN105740645B (en) * | 2016-01-29 | 2019-02-26 | 青岛理工大学 | Rapid Simulated Annealing Algorithm Based on Gene Regulation |
| CN105511269B (en) * | 2016-01-29 | 2018-08-17 | 青岛理工大学 | Gene regulation nonlinear power system control model |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101526405A (en) * | 2009-04-20 | 2009-09-09 | 青岛理工大学 | Calibration method for temperature test of PCR instrument |
| CN101539778A (en) * | 2009-04-30 | 2009-09-23 | 青岛理工大学 | General basic control platform of PCR appearance |
| CN102288147A (en) * | 2011-05-10 | 2011-12-21 | 大连理工大学 | Device for measuring scouring depth monitored based on active temperature control distributed temperature |
| CN102517205A (en) * | 2012-01-09 | 2012-06-27 | 青岛理工大学 | Heat transfer detection device based on DNA amplification |
| CN202390442U (en) * | 2012-01-09 | 2012-08-22 | 青岛理工大学 | Heat transfer detection device based on DNA amplification |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101526405A (en) * | 2009-04-20 | 2009-09-09 | 青岛理工大学 | Calibration method for temperature test of PCR instrument |
| CN101539778A (en) * | 2009-04-30 | 2009-09-23 | 青岛理工大学 | General basic control platform of PCR appearance |
| CN102288147A (en) * | 2011-05-10 | 2011-12-21 | 大连理工大学 | Device for measuring scouring depth monitored based on active temperature control distributed temperature |
| CN102517205A (en) * | 2012-01-09 | 2012-06-27 | 青岛理工大学 | Heat transfer detection device based on DNA amplification |
| CN202390442U (en) * | 2012-01-09 | 2012-08-22 | 青岛理工大学 | Heat transfer detection device based on DNA amplification |
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