CN107727276A - A kind of calibration method for gene-amplificative instrament temp measuring system - Google Patents
A kind of calibration method for gene-amplificative instrament temp measuring system Download PDFInfo
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- CN107727276A CN107727276A CN201710952294.1A CN201710952294A CN107727276A CN 107727276 A CN107727276 A CN 107727276A CN 201710952294 A CN201710952294 A CN 201710952294A CN 107727276 A CN107727276 A CN 107727276A
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- G01—MEASURING; TESTING
- G01K—MEASURING TEMPERATURE; MEASURING QUANTITY OF HEAT; THERMALLY-SENSITIVE ELEMENTS NOT OTHERWISE PROVIDED FOR
- G01K15/00—Testing or calibrating of thermometers
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Abstract
The present invention relates to the technical field of gene-amplificative instrament thermometric, disclose a kind of calibration method for gene-amplificative instrament temp measuring system, the porous isothermal block separated using the liquid inside the temperature measurement module and thermostat that make gene-amplificative instrament temp measuring system to be measured is calibrated to gene-amplificative instrament temp measuring system, and the step of determined according to the comparison of measurement data and normal data.The porous isothermal block includes the receiving storehouse of closing, a side for accommodating storehouse is provided with the first opening, and the edge of the first opening is connected with substrate, and porous block is provided with substrate, multiple instrument connections are evenly arranged with porous block, porous block is extended into by the first opening and accommodated in storehouse；The top surface being connected with the side for accommodating storehouse is provided with the second opening, and the edge of the second opening is connected with one end of long cylinder.The present invention isolates temperature measurement module with the liquid in thermostat by porous isothermal block, and the submergence of temperature measurement module and standard meter is reached alignment requirements, improves measurement accuracy.
Description
Technical field
The invention belongs to the technical field of gene-amplificative instrament thermometric, and in particular to one kind is used for gene-amplificative instrament temp measuring system
Calibration method.
Background technology
Gene-amplificative instrament is also referred to as Polymerized human serum albumin (hereinafter referred to as " PCR instrument "), is that a kind of outward appearance is new, structure is special
Different high-precision temperature meter tool, is mainly used as the standard of PCR instrument temperature performance calibration, the country there is no specifically for such
The calibration method of temp measuring system.With the continuous development of biotechnology, the calibration requirements of PCR instrument are also presented geometric progression and increased therewith
Long, the calibration requirements of PCR instrument temp measuring system are also predictable.
Because the length of the temperature sensor of PCR instrument temp measuring system only has 1.4cm or so, the degree of accuracy but requires to reach ±
0.10 DEG C, magnitude tracing is carried out with comparison method, higher level's measurement criteria is first-class RTD standard set-up.If only use liquid constant temperature
Groove provides temperature source, has following 3 major issues.
First, the thermometric standard as PCR instrument should be clean, pollution-free, and thermostat is contacted during calibration sensor
0~120 DEG C of liquid medium, will certainly cause the pollution of PCR instrument temp measuring system.
Second, the temperature sensor of the PCR instrument temp measuring system insertion depth deficiency in thermostat, under the liquid level of thermostat
Temperature homogeneity and fluctuation degree at 1.5cm all do not reach 0.03 DEG C, and such performance can not meet temperature correction demand.
3rd, during the comparison method amount of progress biography, the insertion depth of standard platinum resistance should with by school temp measuring system TEMP
Device is identical, the problem of equally also facing insertion depth deficiency.
At present, our countries do not advise also specifically for the calibrating standard of PCR instrument temp measuring system, company standard and industry
Model.With the enhancing that PCR instrument manufacturer and user realize to quality control, the calibration method of PCR instrument temp measuring system is filled up
Blank seems extremely urgent.
The content of the invention
The present invention provides a kind of calibration method for gene-amplificative instrament temp measuring system, and solving existing calibration method can be right
Gene-amplificative instrament temp measuring system pollutes, due to the temperature sensor and standard platinum resistance temperature of gene-amplificative instrament temp measuring system
The insertion depth deficiency of meter, it is impossible to the problem of meeting temperature correction requirement.
The present invention can be achieved through the following technical solutions：
A kind of calibration method for gene-amplificative instrament temp measuring system, using making gene-amplificative instrament thermometric system to be measured
The porous isothermal block that liquid inside the temperature measurement module and thermostat of system separates is carried out to described gene-amplificative instrament temp measuring system
The step of calibrating, and being determined according to the comparison of measurement data and normal data.
Further, the porous isothermal block includes substrate, sets porous block on the substrate, on the porous block
Even to be provided with multiple instrument connections, the substrate is connected with one end of long cylinder, and the porous block is enclosed in inside long cylinder, described
The side of substrate is provided with storage tube for placing standard platinum resistance thermometer in the one side of long cylinder.
Further, the porous isothermal block includes the receiving storehouse of closing, and a side for accommodating storehouse is provided with first
Opening, the edge of first opening are connected with substrate, porous block are provided with the substrate, is uniformly arranged on the porous block
There are multiple instrument connections, the porous block is extended into by the first opening and accommodated in storehouse；It is described accommodate storehouse with the side phase
A top surface even is provided with the second opening, and the edge of second opening is connected with one end of long cylinder.
Further, the side that the porous isothermal block using closing is calibrated to gene-amplificative instrament temp measuring system to be measured
Method includes：Step I, the temperature measurement module of gene-amplificative instrament temp measuring system to be calibrated is correspondingly placed into porous isothermal block, and done
It is good heat-insulated；Step II, porous isothermal block is put among thermostat, is submerged to required depth, and by standard platinum resistance temperature
Meter is submerged in the placement pipe of identical depth or the porous isothermal block of insertion, is stood a period of time, is reached required corrector strip
Part；Step III, the performance parameter to gene-amplificative instrament temp measuring system to be calibrated：The temperature error of indication, temperature measurement range and
Temperature resolution measures, and is determined according to the comparison of measurement data and normal data.
Further, the step I includes each temperature sensor of temperature measurement module directly is correspondingly placed into porous isothermal block
Each instrument connection in, according to the weight of temperature measurement module itself decide whether pressurize strip of paper used for sealing, cover adiabatic cotton；
Or each temperature sensor of temperature measurement module is correspondingly placed into each instrument connection of porous isothermal block first, root
Decide whether the strip of paper used for sealing that pressurizes according to the weight of temperature measurement module itself, adiabatic cotton is covered, then by gene-amplificative instrament thermometric to be calibrated
The data wire of system is extended into the long cylinder being connected with the second opening through the first opening, and porous block finally is fixed into first
The edge of opening.
Further, the standing of step II time more than half an hour, required depth are described not less than 180 millimeters
Calibration condition is less than 0.01 DEG C/30 minutes for the temperature fluctuation of standard platinum resistance thermometer measured value.
The present invention is beneficial to be had technical effect that：
Can be by the temperature measurement module and thermostat of gene-amplificative instrament temp measuring system to be calibrated by porous isothermal block
Liquid is isolated, and avoids and it is polluted, and the submergence of temperature measurement module and standard platinum resistance thermometer is reached school
Alignment request, the precision of measured value is improved, ensured the reliability of calibration process, be the value of existing gene-amplificative instrament thermometric system
Demarcation provides unified examination criteria, has ensured the reliability of its parameters value.
Brief description of the drawings
Fig. 1 is the overall construction drawing of the porous isothermal block of the horizontal plug-in type of the present invention, and grey lines are base to be calibrated
The temperature measurement module of gene-amplification instrument temp measuring system；
Fig. 2 is the overall construction drawing of the porous isothermal block of the vertical insertion type of the present invention, and grey lines are base to be calibrated
The temperature measurement module of gene-amplification instrument temp measuring system；
Fig. 3 is integrally formed the substrate of formula structure and the top schematic diagram of porous block for the present invention's；
Fig. 4 is integrally formed the substrate of formula structure and the schematic bottom view of porous block for the present invention's；
Wherein, 1- substrates, 2- porous blocks, the long cylinders of 3-, 4- storage tubes, 5- accommodate storehouse, 6- support handles, 7- projections.
Embodiment
Below in conjunction with the accompanying drawings and preferred embodiment describes the embodiment of the present invention in detail.
1, Fig. 2 and Fig. 3 referring to the drawings, according to the TEMP of the temperature measurement module of gene-amplificative instrament temp measuring system to be calibrated
The direction of insertion of device, porous isothermal block of the invention are divided into two types：Vertical insertion type and horizontal plug-in type, it is described vertical slotting
Entering the porous isothermal block of formula includes substrate 1, sets porous block 2 and long cylinder 3 on substrate 1, porous block 2 is enclosed in long cylinder 3
Inside, the storage tube for placing standard platinum resistance thermometer is provided with the one side of the side of substrate 1 or remote long cylinder 3
4, so, multiple temperature sensors of temperature measurement module, which can both correspond to, to be inserted perpendicularly into each instrument connection of porous block 2, its data wire
Directly interface corresponding to temp measuring system is extended to along the long inwall of cylinder 3；The porous isothermal block of the horizontal plug-in type includes closing
Storehouse 5 is accommodated, the first opening is provided with a side for accommodating storehouse 5, the edge of the first opening is connected with substrate 1, on substrate 1
Porous block 2 is provided with, porous block 2 is extended into by the first opening and accommodated inside storehouse 1；In receiving storehouse 1 and above-mentioned side phase
A top surface even is provided with the second opening, and the edge of the second opening is connected with one end of long cylinder 3, so, temperature measurement module it is more
Individual temperature sensor can both be corresponded to and was horizontally inserted in each instrument connection of porous block, and its data wire extends to survey along the long inwall of cylinder 3
Interface corresponding to warm system.
The porous block 2, using formula structure is integrally formed, is evenly arranged with substrate 1 in substrate 1 backwards to the one side of porous block 2
Multiple projections 6, the position of the projection 6 correspond with the test hole site of porous block 2 and set, and the instrument connection on porous block 2 can
With using blind hole structure.
Two opposite flanks of the long one end of the cylinder 3 away from porous block 2 respectively set the support hand that a position can be adjusted
Handle 7, the notch of the support handle 7 and thermostat coordinate.Long lumbar hole can be provided with along its length in the side of long cylinder 3,
One end of support handle 7 sets external thread rod, and the external thread rod coordinates through long lumbar hole and nut, it is possible to fixed support handle
In the position of long lumbar hole, the position adjustments of support handle 7 are realized, so as to adjust the survey of gene-amplificative instrament temp measuring system to be calibrated
Warm module is submerged into the depth of thermostat.
The substrate 1 is made with porous block 2 or substrate 1, porous block 2 with accommodating storehouse 5 of same material, such as copper or aluminium
Metal.Gene-amplificative instrament temp measuring system is calibrated using above-mentioned porous isothermal block, including：
First, calibration condition
1 environmental condition
1.1 environment temperature：(15~35) DEG C；Ambient humidity：≤ 85%RH.
The environmental condition of 1.2 Electrical Measuring Instrument equipment should meet its corresponding requirements.
During 1.3 calibration, the temp measuring system supply voltage of gene-amplificative instrament should be in the range of normal work, for thermostatic equipment
It should meet corresponding requirement with the Power Supplies Condition that electrical measurement is used together.
2 measurement standards and corollary equipment
Measurement see the table below with standard and corollary equipment.
The desk-top thermostat of 7321 deep-wells of FLUKE companies has can be selected in thermostat, and the thermostat is tied for rectangular opening
Structure, opening size are 120 millimeters * 172 millimeters, 457/482 millimeter of liquid level, 16 liters of volume.- 20 DEG C of temperature range covering~
150 DEG C, thermostatic medium from low temperature silicone oil in 0 DEG C~120 DEG C temperature-measuring ranges, the stability of the thermostat reach 0.005 DEG C/
10min, uniformity reach 0.005 DEG C.
2nd, the preparation before calibrating：
A) open electric logging equipment to be preheated, preheating time at least 20min or the corresponding requirements for meeting its operation instructions.
B) standard platinum resistance thermometer is put into the water triple point bottle frozen make in advance, measures its water triple point value.Will
The water triple point value newly measured inputs supporting electric logging equipment and used.
C normal operating conditions) is at by the requirement of thermostat and operation instructions, and ensures the liquid level of working region
In defined position.
D) gene-amplificative instrament temp measuring system main frame to be calibrated start preheating, into measuring state.
E the temperature measurement module of gene-amplificative instrament temp measuring system to be calibrated) is put into porous isothermal block, standard platinum resistance temperature
Meter is inserted in the storage tube of porous isothermal block, and the temperature sensing part and porous isothermal block bottom for ensureing standard platinum resistance thermometer are kept
Level, then be submerged into thermostat, submergence otherwise less than 180 millimeters.Thermostat is set in the temperature spot of required calibration,
By porous isothermal block together with thermostat after temperature stabilization half an hour, gene magnification that the standard platinum resistance thermometer of monitoring is shown
The temperature fluctuation of instrument temp measuring system calibrating installation is less than 0.01 DEG C/30 minutes, can be calibrated.
Wherein, can be directly corresponding by each temperature sensor of temperature measurement module for the porous isothermal block of vertical insertion type
It is put into each instrument connection of porous block, the strip of paper used for sealing that pressurizes is decided whether according to the weight of temperature measurement module itself, covers adiabatic cotton；
For the porous isothermal block of horizontal plug-in type, each temperature sensor of temperature measurement module is correspondingly placed into first porous
In each instrument connection of block, the strip of paper used for sealing that pressurizes is decided whether according to the weight of temperature measurement module itself, adiabatic cotton is covered, then will treat school
The data wire of accurate gene-amplificative instrament temp measuring system is extended into the long cylinder being connected with the second opening through the first opening, finally
Porous block is fixed to the edge of the first opening.
4th, calibration process：
1st, the temperature error of indication
The temperature spot of the 1.1 measurement temperature errors of indication is typically chosen 30 DEG C, 50 DEG C, 60 DEG C, 70 DEG C, 90 DEG C, 95 DEG C.
1.2 are set in thermostat on 30 DEG C of temperature spots, thermostat actual temperature maximum deviation be no more than ± 0.20 DEG C (with
Secondary standard platinum resistance thermometer indicating value is defined), stablize more than 30 minutes, read standard platinum resistance thermometer respectively and gene expands
Increase the temperature value of each passage of instrument temp measuring system, continuous 4 measurements, and record.
In measurement process, thermostat temperature change is no more than 0.01 DEG C.
1.3 measurement request according to above-mentioned 1.2, measure successively under 50 DEG C, 60 DEG C, 70 DEG C, 90 DEG C, 95 DEG C of states
The temperature value of standard platinum resistance thermometer and each passage of gene-amplificative instrament temp measuring system to be calibrated, and record.
Note：Temperature spot can separately select according to the requirement of client, measure according to the method described above.
2nd, temperature measurement range
Requirement in the 1 of calibration process as described above, gene-amplificative instrament temp measuring system is measured respectively and is surveyed in temperature
Measure the error of indication under 30 DEG C of conventional lower limit and conventional 95 DEG C of states of the upper limit.
Note：The temperature measurement range that the temperature survey upper limit and lowest temperature can also mark according to manufacturer, according to upper
The method of stating measures.
3rd, temperature resolving power
3.1) requirement in the 1 of calibration process as described above, the temperature measurement module of gene-amplificative instrament temp measuring system are put
Enter in porous isothermal block, then entirety is put into thermostat, submergence is not less than 180 millimeters, and thermostat temperature is set in into 90
DEG C, constant temperature more than 30 minutes, the measurement gene-amplificative instrament temp measuring system stable indicating value t1 under 90 DEG C of states of display, and record this
When standard platinum resistance thermometer indicating value T1℃。
3.2) the temperature resolving power D1 nominally shown according to the temp measuring system of gene-amplificative instrament to be calibrated, slowly adjustment is permanent
The design temperature of warm groove, until the display value stabilization of the temp measuring system of gene-amplificative instrament to be calibrated at (t1+D1) DEG C, records this
When standard platinum resistance thermometer indicating value T2℃。
3.3) 3.1 and 3.2 operation, continuous 4 measurements, and record data are repeated.
4th, sample frequency
Sample frequency is gene-amplificative instrament temp measuring system refreshing frequency per second, can be risen in thermostat temperature from 50 DEG C
During 95 DEG C, with the manual methods of range estimation or manual time-keeping, record what system refresh temperature data in the 1s times was shown
Number.If gene-amplificative instrament temp measuring system manufacturer sample frequency is not illustrated when, can be determined by customer demand
Whether the calibration result of sample frequency is provided.
Note：It can be measured according to the method described above according to specific requirement of the client to sample frequency.
3rd, the processing of calibration result
1st, the calculating of the temperature error of indication：
1.1 regulation according to JJG160, scale merit platinum resistance thermometer measure obtained temperature 4 times under 30 DEG C of states
Value, according to following equation, calculates its average value
Wherein, N represents pendulous frequency；
1.2 according to following equation, calculates the temp measuring system of gene-amplificative instrament to be calibrated each quilt under 30 DEG C of states
The average value of 4 measurements of measured value of school passage
Wherein, i represents to represent pendulous frequency by school passage, N i-th of the temp measuring system of gene-amplificative instrament to be calibrated；
The temp measuring system of 1.3 calculating gene-amplificative instrament to be calibrated is under 30 DEG C of states each by the measured value of school passage
The temperature error of indication；
It is accurate to 0.1 DEG C
1.4 computational methods described according to step 1.1-1.3, the temp measuring system of gene-amplificative instrament to be calibrated is calculated successively
The error of indication under 50 DEG C, 60 DEG C, 70 DEG C, 90 DEG C, 95 DEG C of states.
2nd, the calculating of temperature resolving power：
2.1 according to following equation, and calculating the stable indicating value under 90 DEG C of states of gene-amplificative instrament temp measuring system display is
During t1, the average value of 4 indicating values of standard platinum resistance thermometer；
2.2, according to following equation, calculate gene-amplificative instrament temp measuring system and show value stabilization in (t1+D1) DEG C when, standard platinum
The average value of 4 indicating values of resistance thermometer；
2.3 according to below equation, temperature resolving power is calculated；
Wherein, σ is expressed as temperature resolution, should be accurate to 0.1 DEG C.
The present invention can be by the temperature measurement module of gene-amplificative instrament temp measuring system to be calibrated by the porous isothermal block of closing
Isolate with the liquid in thermostat, avoid and it is polluted, and make the leaching of temperature measurement module and standard platinum resistance thermometer
Do not have depth to reach alignment requirements, improve the precision of measured value, ensured the reliability of calibration process, be existing gene-amplificative instrament
The value demarcation of thermometric system provides unified examination criteria, has ensured the reliability of its parameters value.
Although the foregoing describing the embodiment of the present invention, it will be appreciated by those of skill in the art that these
It is merely illustrative of, on the premise of without departing substantially from the principle and essence of the present invention, a variety of changes can be made to these embodiments
More or change, therefore, protection scope of the present invention is defined by the appended claims.
Claims (6)
- A kind of 1. calibration method for gene-amplificative instrament temp measuring system, it is characterised in that：Expand using gene to be measured is made Increase the porous isothermal block that the liquid inside the temperature measurement module and thermostat of instrument temp measuring system separates to survey described gene-amplificative instrament Warm system is calibrated, and the step of determined according to the comparison of measurement data and normal data.
- 2. the calibration method according to claim 1 for gene-amplificative instrament temp measuring system, it is characterised in that：It is described porous Isothermal block includes substrate, sets porous block on the substrate, and multiple instrument connections are evenly arranged with the porous block, described Substrate is connected with one end of long cylinder, and the porous block is enclosed in inside long cylinder, in the side of the substrate or backwards to long cylinder One side on be provided with storage tube for placing standard platinum resistance thermometer.
- 3. the calibration method according to claim 1 for gene-amplificative instrament temp measuring system, it is characterised in that：It is described porous Isothermal block includes the receiving storehouse of closing, and a side for accommodating storehouse is provided with the first opening, the edge of first opening It is connected with substrate, porous block is provided with the substrate, multiple instrument connections, the porous block is evenly arranged with the porous block Extended into and accommodated in storehouse by the first opening；The top surface being connected with the side for accommodating storehouse is provided with second and opened Mouthful, the edge of second opening is connected with one end of long cylinder.
- 4. the calibration method for gene-amplificative instrament temp measuring system according to Claims 2 or 3, it is characterised in that the profit The method calibrated with the porous isothermal block of closing to gene-amplificative instrament temp measuring system to be measured includes：Step I, will be to be calibrated The temperature measurement module of gene-amplificative instrament temp measuring system be correspondingly placed into porous isothermal block, and carry out heat-insulated；Step II, will be porous etc. Warm block is put among thermostat, is submerged to required depth, and by standard platinum resistance thermometer be submerged to identical depth or Insert in the placement pipe of porous isothermal block, stand a period of time, reach required calibration condition；Step III, to base to be calibrated The performance parameter of gene-amplification instrument temp measuring system：The temperature error of indication, temperature measurement range and temperature resolution measure, and root Determined according to the comparison of measurement data and normal data.
- 5. the calibration method according to claim 4 for gene-amplificative instrament temp measuring system, it is characterised in that：The step I includes directly being correspondingly placed into each temperature sensor of temperature measurement module in each instrument connection of porous isothermal block, according to thermometric The weight of module itself decides whether the strip of paper used for sealing that pressurizes, and covers adiabatic cotton；Or each temperature sensor of temperature measurement module is correspondingly placed into each instrument connection of porous isothermal block first, according to survey The warm module weight of itself decides whether the strip of paper used for sealing that pressurizes, and adiabatic cotton is covered, then by gene-amplificative instrament temp measuring system to be calibrated Data wire extended into through the first opening with the second long cylinder for being connected of opening, porous block finally being fixed into first and is open Edge.
- 6. the calibration method according to claim 4 for gene-amplificative instrament temp measuring system, it is characterised in that：The step II standing time more than half an hour, required depth are not less than 180 millimeters, and the calibration condition is standard platinum resistance temperature The temperature fluctuation for measuring value is less than 0.01 DEG C/30 minutes.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110487445A (en) * | 2019-03-22 | 2019-11-22 | 中国计量科学研究院 | A kind of means for correcting of PCR instrument temperature calibration instrument and bearing calibration |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080212643A1 (en) * | 2007-03-02 | 2008-09-04 | Mcgahhey D David | Temperature monitoring device |
| CN202471301U (en) * | 2012-01-10 | 2012-10-03 | 宁波工程学院 | Optical fiber heat transmission detecting device |
-
2017
- 2017-10-13 CN CN201710952294.1A patent/CN107727276A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080212643A1 (en) * | 2007-03-02 | 2008-09-04 | Mcgahhey D David | Temperature monitoring device |
| CN202471301U (en) * | 2012-01-10 | 2012-10-03 | 宁波工程学院 | Optical fiber heat transmission detecting device |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110487445A (en) * | 2019-03-22 | 2019-11-22 | 中国计量科学研究院 | A kind of means for correcting of PCR instrument temperature calibration instrument and bearing calibration |
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