CN111057642A - Temperature calibration device for PCR instrument - Google Patents
Temperature calibration device for PCR instrument Download PDFInfo
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- CN111057642A CN111057642A CN201911272909.1A CN201911272909A CN111057642A CN 111057642 A CN111057642 A CN 111057642A CN 201911272909 A CN201911272909 A CN 201911272909A CN 111057642 A CN111057642 A CN 111057642A
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- temperature
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- pcr instrument
- temperature sensor
- calibration device
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- 238000001514 detection method Methods 0.000 claims abstract description 31
- 238000009529 body temperature measurement Methods 0.000 claims abstract description 19
- 230000005540 biological transmission Effects 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 5
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 3
- 229910052802 copper Inorganic materials 0.000 claims description 3
- 239000010949 copper Substances 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 1
- 239000000741 silica gel Substances 0.000 claims 1
- 229910002027 silica gel Inorganic materials 0.000 claims 1
- 238000005259 measurement Methods 0.000 abstract description 3
- 238000003752 polymerase chain reaction Methods 0.000 description 43
- 238000001816 cooling Methods 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 230000003044 adaptive effect Effects 0.000 description 3
- 229920001296 polysiloxane Polymers 0.000 description 3
- 238000007859 qualitative PCR Methods 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical group [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229910052744 lithium Inorganic materials 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- -1 polyoxymethylene Polymers 0.000 description 2
- 229920006324 polyoxymethylene Polymers 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 238000002598 diffusion tensor imaging Methods 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229920003229 poly(methyl methacrylate) Polymers 0.000 description 1
- 239000004926 polymethyl methacrylate Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000005382 thermal cycling Methods 0.000 description 1
- 238000011522 transarterial infusion chemotherapy Methods 0.000 description 1
- 238000003466 welding Methods 0.000 description 1
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01K—MEASURING TEMPERATURE; MEASURING QUANTITY OF HEAT; THERMALLY-SENSITIVE ELEMENTS NOT OTHERWISE PROVIDED FOR
- G01K13/00—Thermometers specially adapted for specific purposes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01K—MEASURING TEMPERATURE; MEASURING QUANTITY OF HEAT; THERMALLY-SENSITIVE ELEMENTS NOT OTHERWISE PROVIDED FOR
- G01K7/00—Measuring temperature based on the use of electric or magnetic elements directly sensitive to heat ; Power supply therefor, e.g. using thermoelectric elements
- G01K7/16—Measuring temperature based on the use of electric or magnetic elements directly sensitive to heat ; Power supply therefor, e.g. using thermoelectric elements using resistive elements
- G01K7/18—Measuring temperature based on the use of electric or magnetic elements directly sensitive to heat ; Power supply therefor, e.g. using thermoelectric elements using resistive elements the element being a linear resistance, e.g. platinum resistance thermometer
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
Abstract
The invention discloses a temperature calibration device of a PCR instrument, which comprises a control host and a temperature detection assembly, wherein the control host is electrically connected with the temperature detection assembly, the temperature detection assembly comprises an upper cover and a lower cover, the upper cover is fixedly connected with the lower cover, a plurality of temperature measurement units are arranged on the lower cover, each temperature measurement unit comprises a temperature sensor, a temperature sensor connecting piece and a heat conducting part, the temperature sensor is positioned in a cavity of the heat conducting part, the heat conducting part is detachably connected with the temperature sensor connecting piece, the temperature sensor connecting piece is used for connecting the temperature sensor with the lower cover, and an opening is formed in the bottom of the heat conducting part. The invention has the advantages of convenient use, accurate measurement and wide application range of the PCR instrument.
Description
Technical Field
The invention relates to a temperature calibration device of a PCR instrument, belonging to the field of quality detection of medical equipment.
Background
The polymerase chain reaction analyzer is an instrument for simulating the copying process of DNA or RNA, specifically amplifying known sequences in the presence of template, primer, polymerase and the like, and detecting and analyzing the known sequences based on the PCR (polymerase chain reaction) technical principle.
According to the calibration standards of JJF 1527-2015 polymerase chain reaction analyzer: the polymerase chain reaction analyzer comprises a qualitative PCR analyzer and a quantitative PCR analyzer. The qualitative PCR instrument is generally composed of a sample carrier, a thermal cycling component, a control component, a power supply component and the like. The quantitative PCR instrument mainly comprises a sample carrier, a thermal cycle component, a control component, an optical detection component, a power supply component, a computer, application software and the like. Whether the qualitative PCR instrument or the quantitative PCR instrument is adopted, temperature indication error, temperature uniformity, average heating rate and average cooling rate are all parameters which need to be detected, so that the temperature detection is an important quality parameter for measuring and detecting the PCR instrument.
At present, PCR instruments have more market forms, the main difference is a sample carrier, and according to the sample carrier, 48-hole PCR instruments and 96-hole PCR instruments are applied more frequently.
The temperature calibration and detection device for the PCR instrument can meet the requirements of temperature measurement and detection of the 48-hole PCR instrument and the 96-hole PCR instrument, and meets the calibration standard JJF 1527-2015 polymerase chain reaction analyzer by using a control host, an interface converter and an automatic adaptive mode of two optional temperature detection modules.
Disclosure of Invention
The invention provides a temperature calibration device of a PCR instrument, which is convenient to use, accurate in measurement and wide in applicable range of the PCR instrument.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: the utility model provides a PCR appearance temperature calibrating device, includes main control system and temperature detect subassembly, main control system and temperature detect subassembly electrical connection, the temperature detect subassembly includes upper cover and lower cover, upper cover and lower cover fixed connection, set up a plurality of temperature measurement unit on the lower cover, temperature measurement unit includes temperature sensor, temperature sensor connecting piece and heat-conducting part, temperature sensor is located heat-conducting part's cavity, heat-conducting part can dismantle with the temperature sensor connecting piece and be connected, the temperature sensor connecting piece is used for being connected of temperature sensor and lower cover, the heat-conducting part lower part is provided with the trompil.
Preferably, the lower cover is a circuit board, and a lower cover flat socket is further arranged on the lower cover.
Preferably, an intermediate frame is further arranged between the upper cover and the lower cover, a plurality of temperature measurement unit mounting holes and a second flat socket hole are formed in the intermediate frame, the temperature measurement unit mounting holes correspond to the temperature sensor connecting pieces in position and are adaptive to the temperature sensor connecting pieces in size, and the second flat socket hole is adaptive to the lower cover flat socket in size.
Preferably, the control host is electrically connected with the temperature detection assembly through an interface converter, and the interface converter comprises a converter flat socket, a converter male head, an internal circuit and a flat cable; the internal circuit is used for converting signals of the flat socket to the male connector; the flat cable is used for connecting the lower cover flat socket and the converter flat socket.
Preferably, the flat cable is provided with a silicone shell.
Preferably, the interface converter is a 68-core connector, and the lower cover flat socket and the converter flat socket are 68-core flat sockets.
Preferably, the control host is provided with a connector female head and comprises a data processing unit, a power supply unit, a data transmission unit and a microprocessor, wherein the data processing unit, the power supply unit, the data transmission unit and the microprocessor are electrically connected.
Preferably, the data transmission unit is used for wireless data transmission; the data processing unit is used for acquiring temperature data measured by the temperature detection assembly, amplifying the temperature data and transmitting the temperature data to the microprocessor; and after the microprocessor calculates and processes the received data, the data after calculation and processing is uploaded through the data transmission unit.
Preferably, the temperature measuring unit is arranged on the lower cover according to the calibration specification of the PCR instrument.
Preferably, the heat conducting component is a conical hollow copper body component, and is connected with the temperature sensor connecting piece through threads.
The interface converter is adopted to detachably connect the control host and the temperature detection assembly, and the connecting wire is easy to replace, so that the temperature detection device is easy to maintain, update and operate. According to the invention, the temperature detection assembly is designed according to the detection standard of the PCR instrument to form a whole, when the PCR instrument to be detected needs to be replaced, the temperature detection assembly matched with the type of the PCR instrument is only required to be directly connected with the control host through the connecting wire, the operation is simple and convenient, and the detection efficiency is improved.
Drawings
FIG. 1 is a schematic structural diagram of a temperature calibration apparatus for a PCR instrument according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a temperature detecting assembly according to an embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a temperature measuring unit according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of the arrangement of a temperature measuring unit of a 96-well PCR instrument according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of the arrangement of a temperature measuring unit of a 48-well PCR instrument according to an embodiment of the present invention.
Wherein: 1-control host, 2-interface converter, 3-flat cable, 4-lower cover flat socket, 5-temperature detection component, 51-upper cover, 511-first flat socket hole, 52-lower cover, 53-middle frame, 531-temperature measurement unit mounting hole, 532-second flat socket hole, and 54-temperature measurement unit. 541-temperature sensor connection, 542-temperature sensor, 543-heat conducting part.
Detailed Description
For a better understanding of the nature of the invention, its description is further set forth below in connection with the specific embodiments and the drawings.
A temperature calibration device of a PCR instrument is suitable for temperature calibration of various types of PCR instruments, and the specific structure of the temperature calibration device is shown in figure 1, and the temperature calibration device comprises a control host 1, an interface converter 2 and a temperature detection assembly 5. The control host 1 is electrically connected with the temperature detection assembly 5 through the interface converter 2. The control host 1 is provided with a converter female head which is matched with a converter male head arranged on the interface converter 2.
As shown in fig. 2, the temperature detection assembly 5 includes an upper cover 51, a lower cover 52, and an intermediate frame 53. The intermediate frame 53 is located between the upper cover 51 and the lower cover 52. The upper cover 51, the lower cover 52 and the intermediate frame 53 are fixed by bonding with an insulating adhesive.
The lower cover 52 is preferably an ultra-thin circuit board of 120mm80mm 0.5mm (length/width/height). The lower cover 52 is provided with a lower-cover flat socket 4 and a plurality of temperature measuring units 54. The lower cover flat socket 4 is used for connection with the interface converter 2. The temperature measuring unit 54 is directly welded to the corresponding welding points of the lower cover 52, and is fixed and mounted through the middle frame 53 and the upper cover 51.
As shown in fig. 3, the temperature measuring unit 54 includes a temperature sensor connector 541, a temperature sensor 542, and a heat conductive member 543. The temperature sensor 542 is located in the cavity of the heat conducting component 543 and is fixed by the heat conducting silicone grease. The temperature sensor connector 541 is used for connection of the temperature sensor 542 to the lower cover 52. The temperature sensor 542 is preferably a PT100 temperature sensor. The heat conducting component 543 is provided with micro holes at its lower part, and the micro holes are used for conducting heat. The machined material of temperature sensor attachment 541 is preferably polyoxymethylene. The heat conducting member 543, which is preferably a conical hollow copper member, is detachably connected to the temperature sensor connection member 541 by a screw thread. The overall height of the temperature measuring unit 54 is 18.5 mm.
The middle frame 53 is provided with a plurality of temperature measuring unit mounting holes 531 and second flat socket holes 532. The temperature measuring unit mounting hole 531 is adapted to the size of the temperature sensor connector 541. The intermediate frame 53 is preferably a polyoxymethylene hard plate of 120mm80mm × 3mm (length/width/height). The temperature measuring unit mounting hole 531 is preferably a circular hole having a diameter of 11mm, and the center of the circular hole coincides with the center of the temperature measuring unit 54. The second flat socket hole 532 is sized to accommodate the lower cover flat socket 4. The second flat socket hole 532 is a square hole with a length/width of 40mm by 20mm, respectively.
The upper lid 51 is preferably a rigid sheet of polymethylmethacrylate with a length/width/height of 120mm80mm x 0.5mm, respectively. The upper cover 51 is provided with a first flat socket hole 511, and the first flat socket hole 511 is sized to fit the lower cover flat socket 4. The first flat socket holes 511 are square holes, and have a length/width of 40mm by 20mm, respectively. The lower cover flat socket 4 is placed in the first and second flat socket holes 511 and 532 with respect to the raised portion of the lower cover 52.
The arrangement of the temperature measuring cell 54 in the lower cover 52 is set according to the requirements of JJF 1527-2015 "calibration standards for polymerase chain reaction analyzers": the temperature detection assembly 5 is suitable for a 96-hole PCR instrument, and 15 temperature measurement units 54 are arranged; the temperature detection assembly 5 suitable for the 48-hole PCR instrument is provided with 8 temperature measurement units 54.
As shown in fig. 4, the spot layout suitable for the 96-well PCR instrument is 12 × 8. From top to bottom, from left to right, the intersections of the 1 st, 4 th and 8 th rows and the 1 st, 4 th, 7 th, 10 th and 12 th columns are selected in sequence, wherein the intersection of the 4 th row and the 4 th column is not taken, and the intersection of the 5 th row and the 4 th column is selected for substitution; the intersection of row 4 and column 10 is not taken and the intersection of row 5 and column 10 is selected instead. A total of 15 points were selected for the setting of the temperature measuring unit 54.
As shown in FIG. 5, the spotting mode suitable for the 48-well PCR instrument is a 6 × 8-spot array, and from top to bottom, from left to right, the 8-well temperature detection module sequentially selects the 1 st row, the 1 st, 5 th and 8 th columns of crossing points, the 3 rd row, the 4 th column, the 4 th row, the 6 th column, and the 6 th row, the 1 st, 4 th and 8 th columns of crossing points. A total of 8 points are selected for the setting of the temperature measuring unit 54.
The interface converter 2 comprises a converter flat socket, a converter male, an internal circuit and a flat cable 3. The internal circuitry is used to convert the signal from the flat jack to the converter male. The flat cable 3 is used to connect the lower cover flat socket 4 and the converter flat socket. The flat cable 3 is preferably a 68-core silicone sheath cable, which is resistant to high temperature and can be folded for use many times. The interface converter 2 is preferably a 68-core connector, and the lower cover flat socket 4 and the converter flat socket are preferably 68-core flat sockets.
The control host 1 is provided with a starting button, a working indicator light and a charging interface. The control host 1 includes a data processing unit, a power supply unit, a data transmission unit and a microprocessor. The data processing unit, the power supply unit, the data transmission unit and the microprocessor are electrically connected. The microprocessor control unit is preferably an STM32F103C8T6 processor and is used for controlling the whole system to work. The power supply unit is preferably a lithium battery charging circuit formed by a lithium battery charger TP4056 and a power supply conversion circuit formed by a power supply conversion chip SP 6203. The data transmission unit is preferably a wireless data transmission. The data processing unit is used for acquiring temperature data measured by the temperature detection component 5, amplifying the temperature data by an operational amplifier OPA333 and transmitting the amplified temperature data to the microprocessor; and after the microprocessor calculates and processes the received data, the data after calculation and processing is uploaded through the data transmission unit.
The method of use of the present invention is exemplified by a 96-well PCR apparatus. Before detection, the PCR instrument to be detected needs to be started and preheated for 30 minutes, a temperature detection assembly 5 containing 15 temperature measurement units is selected, and the control host 1, the interface converter 2 and the temperature detection assembly 5 are connected in sequence.
Setting a temperature control program according to the use instruction of the PCR instrument to be detected, wherein the specific program flow comprises the following steps: the initial temperature was 30 ℃ and held for 1 minute; heating to 95 ℃ and keeping for 3 minutes; cooling to 30 deg.C, and maintaining for 2 min; heating to 90 deg.C, and maintaining for 3 min; cooling to 50 deg.C, and maintaining for 3 min; heating to 70 ℃ and keeping for 3 minutes; cooling to 60 deg.C, and maintaining for 3 min; the temperature is reduced to 30 ℃ and kept for 1 minute.
And (3) directly inserting the temperature detection assembly 5 into a sample carrying platform of the PCR instrument to be detected, adjusting an upper cover of the PCR instrument to be detected, and tightly closing the upper cover. And when a starting button on the control host 1 is pressed, the work indicator lamp is always on. The 15 temperature sensors 542 collect temperature data and the control host uploads the collected temperature data until the whole temperature control program is finished. And correcting the PCR instrument to be detected according to the uploaded temperature data.
If need rectify 48 hole PCR appearance, then only need to change temperature-detecting element for containing 8 temperature measurement unit's temperature-detecting element to be connected can through flat cable and the flat socket of converter, the simple operation.
1. The temperature indicating value error is calculated according to the formula (1) and the formula (2):
in the formula: delta TdIndicating temperature error in a working area of the PCR instrument to be detected at DEG C; t issSetting the temperature value at DEG C in the working area of the PCR instrument to be detected;
is the average value of the measured values of the temperature measuring units, DEG C; t isiMeasured value of the ith temperature measuring unit in DEG C; and n is the number of temperature measurement units.
2. The temperature uniformity is calculated according to formula (3):
ΔTu＝Tmax-Tmin(3)
in the formula: delta TuTemperature uniformity, deg.C; t ismaxIs the maximum value of the measured value of the temperature measuring unit, DEG C; t isminIs the minimum value, deg.C, of the temperature measurement unit measurements.
3. Average rate of temperature rise
Collecting the constant temperature 10S when the temperature of the PCR instrument to be detected reaches the set temperature, taking a 50 ℃ temperature point, and recording the temperature as TA(ii) a Taking a temperature point of 90 ℃, and recording the temperature as TBPCR instrument from TATo TBIs recorded as t1The average rate of temperature rise is calculated according to equation (4):
in the formula: vUTThe average heating rate is DEG C/s; t is t1Is from TATo TBTime of (d), s.
4. The average cooling rate calibration result is calculated according to equation (5):
slave T of PCR instrumentBTo TAIs recorded as t2Calculating the average cooling rate according to the formula (5):
in the formula: vDTIs an averageCooling rate, DEG C/s; t is t2Is from TBTo TATime of (d), s.
It should be noted that while the invention has been described in terms of the above-mentioned embodiments, there are many other embodiments of the invention. It will be apparent to those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention, and it is intended that all such changes and modifications be covered by the appended claims and their equivalents.
Claims (10)
1. The utility model provides a PCR appearance temperature calibrating device which characterized in that: including main control system and temperature detect subassembly, main control system and temperature detect subassembly electrical connection, the temperature detect subassembly includes upper cover and lower cover, upper cover and lower cover fixed connection, set up a plurality of temperature measurement unit on the lower cover, the temperature measurement unit includes temperature sensor, temperature sensor connecting piece and heat-conducting part, temperature sensor is located heat-conducting part's cavity, heat-conducting part can dismantle with the temperature sensor connecting piece and be connected, the temperature sensor connecting piece is used for being connected of temperature sensor and lower cover, the heat-conducting part bottom is provided with the trompil.
2. The temperature calibration device for the PCR instrument according to claim 1, wherein: the lower cover is a circuit board, and a lower cover flat socket is further arranged on the lower cover.
3. The temperature calibration device for the PCR instrument according to claim 2, wherein: still be provided with the intermediate frame between upper cover and the lower cover, be provided with a plurality of temperature measurement unit mounting holes and the flat socket hole of second on the intermediate frame, the temperature measurement unit mounting hole corresponds and the size suits with the position of temperature sensor connecting piece, the flat socket hole size of second suits with the flat socket of lower cover.
4. The temperature calibration device for the PCR instrument according to claim 2, wherein: the control host is electrically connected with the temperature detection assembly through an interface converter, and the interface converter comprises a converter flat socket, a converter male head, an internal circuit and a flat cable; the internal circuit is used for converting signals of the flat socket to the male connector; the flat cable is used for connecting the lower cover flat socket and the converter flat socket.
5. The temperature calibration device for the PCR instrument according to claim 4, wherein: the flat cable is provided with a silica gel shell.
6. The temperature calibration device for the PCR instrument according to claim 4, wherein: the interface converter is a 68-core connector, and the lower cover flat socket and the converter flat socket are 68-core flat sockets.
7. The temperature calibration device for the PCR instrument according to claim 1, wherein: the control host is provided with a connector female head and comprises a data processing unit, a power supply unit, a data transmission unit and a microprocessor, wherein the data processing unit, the power supply unit, the data transmission unit and the microprocessor are electrically connected.
8. The temperature calibration device for the PCR instrument according to claim 7, wherein: the data transmission unit is used for wireless data transmission; the data processing unit is used for acquiring temperature data measured by the temperature detection assembly, amplifying the temperature data and transmitting the temperature data to the microprocessor; and after the microprocessor calculates and processes the received data, the data after calculation and processing is uploaded through the data transmission unit.
9. The temperature calibration device for the PCR instrument according to claim 1, wherein: the temperature measuring units are arranged on the lower cover according to the calibration specification of the PCR instrument.
10. The temperature calibration device for the PCR instrument according to claim 1, wherein: the heat conducting component is a conical hollow copper body component and is connected with the temperature sensor connecting piece through threads.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201911272909.1A CN111057642A (en) | 2019-12-12 | 2019-12-12 | Temperature calibration device for PCR instrument |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201911272909.1A CN111057642A (en) | 2019-12-12 | 2019-12-12 | Temperature calibration device for PCR instrument |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112945420A (en) * | 2021-02-02 | 2021-06-11 | 四川淯源仪器科技有限公司 | Temperature calibration system of wireless PCR instrument |
| CN115031871A (en) * | 2022-08-11 | 2022-09-09 | 北京林电伟业电子技术有限公司 | Multi-temperature-zone calibration method for constant-temperature rapid nucleic acid amplification detection analyzer |
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Application publication date: 20200424 |