JP2014193134A - Nucleic acid analyzer, and temperature control method of the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a technique capable of realizing uniform and high-speed PCR in a plurality of reaction tanks arranged in a narrow region.SOLUTION: A nucleic acid analyzer has: a plurality of heating and cooling elements; a plurality of temperature measuring elements making a pair with each of the plurality of heating and cooling elements; and a control section which controls output amounts of the plurality of heating and cooling elements. The control section executes: processing of determining one of the plurality of heating and cooling elements as a reference element; processing of using temperature values detected by the temperature measuring elements, which make a pair with each of the plurality of heating and cooling elements, as an input, and controlling output amounts of the plurality of heating and cooling elements corresponding to a difference from a target temperature value; and processing of adjusting output amounts of one or the plurality of heating and cooling elements other than the reference element corresponding to temporal change amounts of a temperature value detected by the temperature measuring element making a pair with the reference element.

Description

  The present invention relates to a technique for controlling the temperature of a nucleic acid analyzer.

  For example, Japanese Unexamined Patent Publication No. 2010-502228 (Patent Document 1) is available as a document related to the background art. In the summary part of this publication, “The present invention relates to a device for performing a chemical or biological reaction in a reaction vessel receiving element for receiving a microtiter plate with several reaction vessels, The reaction vessel receiving element has several recesses arranged in a regular pattern for receiving each reaction vessel, a heating device for heating the reaction vessel receiving element, and cooling for cooling the reaction vessel The invention is characterized by the fact that the reactor receiving element is divided into several segments, each segment being thermally decoupled from each other, each segment being independent of the others A heating device may be assigned that may be actuated using the segmentation of the reactor receiving element to allow the zones to be set and held at different temperatures. Since the reaction vessel receiving element is suitable for receiving a standard microtiter plate, the device according to the present invention are described as well be. "Which is integrated into an existing process sequence.

  In addition, another document related to the background art is, for example, Japanese Patent Application Laid-Open No. 2007-280142 (Patent Document 2). The summary part of this publication states that “[Problem] Highly accurate gradient temperature control is possible even when the number of control points increases. [Solution] Temperature control blocks 24H, 24L-1, etc. for performing gradient temperature control. 24L-2 is hierarchized, and the average temperature GPV1 from the input temperature mode conversion block of the lower temperature control blocks 24L-1 and 24L-2 is used as the input temperature of the input temperature mode conversion block 20H of the upper temperature control block 24H. On the other hand, the control output from the pre-compensation block 23H of the upper temperature control block 24H is set as the target temperature of the target temperature mode conversion blocks 21L-1, 21L-2 of the lower temperature control blocks 24L-1, 24L-2, As a result, instead of individually controlling the gradient temperature for each of the lower temperature control blocks 24L-1 and 24L-2, both temperature control blocks 2 L-1,24L-2 so that a whole as gradient temperature control. "Is described as.

Special table 2010-502228 JP 2007-280142 A

  Patent Document 1 describes an apparatus specialized for heating and cooling processing in a temperature control apparatus for processing biological or chemical samples, for example, a method for controlling temperature in an insulated space. Has been. However, in the technique described in Patent Document 1, there is a problem that the temperature uniformity of the heating / cooling body cannot be sufficiently secured in an environment where the surroundings of the heating / cooling body cannot be sufficiently insulated. Further, the technique described in Patent Document 1 requires a heating element for controlling the heat radiation amount of the heating / cooling body, and there is a problem that it is not suitable for downsizing of the apparatus. In particular, the technique described in Patent Literature 1 may not be able to cope with temperature non-uniformity caused by a temperature change of a controlled object at high speed or individual differences of heating / cooling elements.

  Patent Document 2 discloses a technique aimed at eliminating a deviation between a plurality of heat sources. However, the technique described in Patent Document 2 is difficult to apply to “temperature control of PCR (Polymerase chain reaction)”. This is because the size of a sample that is a control target in PCR is smaller than the control target assumed in Patent Document 2, and thus the temperature of the control target easily changes. That is, in temperature control of PCR, temperature control with higher accuracy is required as compared with the technique described in Patent Document 2. In PCR, it is necessary to periodically change the temperature. Moreover, it is necessary to prevent the PCR time from becoming unexpected due to an unintended chemical reaction near the reaction temperature. For this reason, PCR requires control capable of temperature transition in a short time.

  In Patent Document 2, an average value of a plurality of devices is used as a representative temperature. For this reason, when the output amount of a heating / cooling element is close to the maximum output (100%) and the temperature of the heating / cooling element is less than the average value, the heating / cooling element is required after correction. Output amount may exceed the maximum output (100%), there is a problem that the correction does not work. Furthermore, in Patent Document 2, when a plurality of heat sources are arranged in a narrow region, there is a possibility that the influence between the heat sources becomes more obvious.

  Therefore, the present invention provides a technique capable of realizing uniform and high-speed PCR in a plurality of reaction vessels arranged in a narrow region.

  In order to solve the above problems, for example, the configuration described in the claims is adopted. The present application includes a plurality of means for solving the above-described problems. To give an example of the above, “a plurality of heating / cooling elements, a plurality of temperature measuring elements paired with each of the plurality of heating / cooling elements, A control unit that controls output amounts of the plurality of heating / cooling elements, wherein the control unit determines one of the plurality of heating / cooling elements as a reference element, and each of the plurality of heating / cooling elements; A process for controlling the output amount of the plurality of heating and cooling elements according to a difference from a target temperature value using a temperature value detected by a pair of temperature measuring elements as an input, and a temperature measuring element paired with the reference element A nucleic acid analyzer that executes a process of adjusting the output amount of one or a plurality of heating / cooling elements other than the reference element in accordance with the amount of change in temperature value to be detected.

  According to the present invention, uniform and high-speed PCR can be realized in a plurality of reaction vessels arranged in a narrow region. Problems, configurations, and effects other than those described above will be clarified by the following description of embodiments.

FIG. 1 is an example of a block diagram showing a configuration example of a system according to the present invention. FIG. 2 is a diagram showing an example of the structure of the temperature control unit according to the present invention. FIG. 3 is a cross-sectional view (A-A ′ cross-section) of FIG. 2 showing an example of the structure of the temperature control unit according to the present invention. FIG. 4 is a cross-sectional view (B-B ′ cross section) of FIG. 2 showing an example of the structure of the temperature control unit according to the present invention. FIG. 5 is an example of a flowchart for explaining preprocessing according to the present invention. FIG. 6 is an example of a flowchart for explaining the initial processing according to the present invention. FIG. 7 is an example of a flowchart showing an outline of the temperature control process according to the present invention. FIG. 8 is an example of a flowchart for explaining a correction value calculation process for a non-reference heating / cooling element. FIG. 9 is an example of a flowchart showing an outline of the temperature control process according to the present invention. FIG. 10 is an example of a flowchart showing an outline of the temperature control process according to the present invention.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. The embodiment of the present invention is not limited to the embodiments described later, and various modifications are possible within the scope of the technical idea.

[Example 1]
<System>
FIG. 1 is a block diagram illustrating a schematic configuration of the nucleic acid analysis system according to the present embodiment. Below, the case where temperature control of PCR is implement | achieved by the system shown in FIG. 1 is demonstrated.

  The nucleic acid analysis system shown in FIG. 1 includes a temperature measurement element 1, a measurement unit 2, an output control unit 3, a heating / cooling element 4, a heat radiating fan 5, and a control unit 6. The temperature measuring element 1 is used for measuring a sample or its surrounding temperature. The measuring unit 2 performs A / D (analog to digital) conversion on analog data of temperatures measured by the three temperature measuring elements 1 and outputs the data as digital data. The output control unit 3 performs D / A (digital to analog) conversion on the digital data supplied from the control unit 6 and outputs the digital data to the three heating / cooling elements 4 as analog data. The heating / cooling element 4 generates heat or absorbs heat according to an electric signal (analog data) given from the output control unit 3. The heat radiating fan 5 is used for efficiently radiating heat from the heat radiating portion. The control unit 6 executes arithmetic processing for calculating an output amount to be given to the output control unit 3 based on the information given from the measurement unit 2. The control unit 6 also performs output control of the heat dissipation fan 5. The control unit 6 includes a volatile memory and a nonvolatile memory inside, and executes writing and retrieving of information using them.

  In the case of the present embodiment, three temperature measuring elements 1 and three heating / cooling elements 4 are arranged, and one temperature measuring element 1 and one heating / cooling element 4 are treated as a pair. Corresponding to the pair, the measuring unit 2 holds information on three temperature measuring elements 1, and the output control unit 3 holds information on three heating / cooling elements 4.

  In the case of the present embodiment, the control unit 6 includes a sequence control unit 10, a variable setting unit 11, and an output amount calculation unit 12. The sequence control unit 10 holds, for example, information on the set temperature to be maintained in each processing stage (thermal denaturation, primer addition, extension) of PCR, and temperature maintenance time information for maintaining the set temperature for a certain period of time for promoting the reaction. And based on the input from the measurement part 2, it determines completion of each process step, and resets the preset temperature according to a step. Further, the sequence control unit 10 determines and controls whether or not the output of the heat dissipation fan 5 is necessary, for example, depending on the processing stage.

  The variable setting unit 11 sets parameters used when calculating the output amount to the output control unit 3. The parameters include PID calculation parameters, set temperatures, and the like.

  Based on the input from the measurement unit 2, the output amount calculation unit 12 periodically calculates the output amount to the output control unit 3 using the variables set by the variable setting unit 11. That is, the output amount calculation unit 12 can set independent outputs for each of the three heating / cooling elements 4 based on the inputs of the three temperature measuring elements 1.

<Configuration of temperature control unit>
In FIG. 2, the structure of the temperature control part used by a present Example is shown. The temperature control unit 100 is a member formed from a single material made of a metal having good thermal conductivity such as aluminum or silver. One surface of the temperature control unit 100 is provided with one or a plurality of dents used as a reaction tank 101 for performing PCR. In the case of the present embodiment, eight reaction vessels 101 are arranged for one temperature adjustment unit 100.

  FIG. 3 shows an A-A ′ cross section of the temperature control unit 100 shown in FIG. 2. On the upper surface side of the temperature control unit 100, a heat insulating unit 201 used as a lid of the reaction tank 101 is disposed. In the present embodiment, the heating / cooling element 4 is disposed on the lower surface side of the temperature control unit 100. On the lower surface side of the heating / cooling element 4, a heat dissipating part 202 that promotes heat dissipation of the heat accumulated in the heating / cooling element 4 is disposed. The heat radiating fan 5 is disposed on the lower surface of the heat radiating portion 202.

  As shown in FIG. 3, the temperature measuring element 1 is disposed immediately below the reaction vessel 101 (inside the temperature control unit 100). This is because, when the temperature measuring element 1 is disposed inside the reaction vessel 101, for example, if an unintended chemical reaction occurs between the sample and the temperature measuring element 1, it may affect PCR. For the temperature measuring element 1, for example, a resistance temperature detector is used. A resistance temperature detector is an element that utilizes the property that an electrical resistance value of a metal, a semiconductor, or the like varies with temperature. For the heating / cooling element 4, for example, a Peltier element is used. A Peltier element is a semiconductor element that utilizes the phenomenon (Peltier effect) in which heat is transferred from one metal to the other when a current is passed through a junction of two types of metal. Can be controlled. Moreover, the relationship between heating and cooling can be reversed by changing the direction of the current.

  FIG. 4 shows a B-B ′ cross section of the temperature control unit 100 shown in FIG. 2. As shown in FIG. 4, the reaction vessels 101 are arranged in a line on the upper surface of the temperature control unit 100. The three temperature measuring elements 1 are arranged in a line below the reaction vessel 101. Below the temperature measuring element 1, three heating / cooling elements 4 are arranged in a row so that one heating / cooling element 4 corresponds to each temperature measuring element 1.

  The structure of this example assumes that PCR is performed on a plurality of samples simultaneously in a narrow region. Eight reaction tanks 101 are arranged in a line because fine flow paths are provided on both sides of each reaction tank 101 (in the direction of AA ′ in FIG. 2), and eight reactions are performed through these flow paths. This is because the sample can flow into and out of the tank 101. When preparations such as mixing of reagents necessary for PCR are completed and a sample capable of performing PCR is obtained, the sample flows into the reaction tank 101 from the above-described fine flow path, and PCR is performed. When the PCR is completed, the sample flows out from the reaction vessel 101 to the region where the next step is performed through the fine flow path described above. By applying the above processing, it is possible to automate from reagent adjustment to PCR in the pretreatment process in the nucleic acid analyzer. As in this embodiment, by using a fine channel, the amount of reagent used can be minimized. As a result, the amount of reagent used for PCR can be reduced, and the cost required for operation of the nucleic acid analyzer can be reduced.

<Ingenuity to achieve uniform PCR>
Also in the case of the above structure, the output amount of the heating / cooling element 4 is determined by simply feeding back the temperature measured by the temperature measuring element 1 disposed closest to each of the three heating / cooling elements 4. Therefore, it is inevitable that a temperature difference occurs between the eight reaction vessels 101. If there is such a temperature difference, it is considered that there is a difference in PCR efficiency among the eight reaction vessels 101. In order to solve this problem, it is necessary to reduce the temperature difference between the heating and cooling elements 4. Therefore, in the present embodiment, the following preprocessing is executed.

<Pretreatment>
FIG. 5 is a flowchart showing an example of preprocessing for performing temperature control for reducing the temperature difference between the eight reaction vessels 101. This process is performed by the control unit 6 immediately after the activation of the nucleic acid analyzer and before the execution of the PCR temperature control.

  In this embodiment, one of the three heating / cooling elements 4 is used as a reference heating / cooling element. In FIG. 5, the reference heating / cooling element is referred to as a reference Peltier element.

  In the following description, the reference heating / cooling element is defined as an element measured by the temperature measuring element 1 among the three heating / cooling elements 4 and having the slowest temperature change rate. Factors that cause a slow temperature change rate include a plurality of factors such as individual differences in output performance of each heating / cooling element 4 and heat transfer conditions around the location.

  There are two specific methods for determining the reference heating / cooling element. One method is that when the information of the reference heating / cooling element acquired when the nucleic acid analyzer was used last time is stored in the nonvolatile memory of the control unit 6 in advance, the element performs the initial adjustment process. Necessary conditions ("Presence / absence of defective element system in previous use", "Elapsed period since last use", "Number of times PCR was executed continuously without executing initial adjustment process", "Used last time When it is confirmed that the number and position of the reaction tanks does not fall under the “user selection process” that is executed as necessary, the previous reference heating / cooling element is used as the next reference heating / cooling element. is there. Here, the element system means that the temperature measuring element 1 and the heating / cooling element 4 are a pair, and the defective element system means that either or both of the temperature measuring element 1 and the heating / cooling element 4 are used in the nucleic acid analyzer. It is defined as insufficient performance. The other method is a method of determining the reference heating / cooling element by executing the initial adjustment process. Details of the initial adjustment processing will be described later with reference to FIG. The controller 6 always executes one of these two methods to determine the reference heating / cooling element.

  The control unit 6 according to the present embodiment holds information used for determining the reference heating / cooling element in two types. One piece of information is information unique to the device configuration. This information includes, for example, the lower limit value of the heating / cooling power required for the heating / cooling element 4, the upper limit value of the number of times that the PCR has been executed continuously without executing the initial adjustment processing (cumulative measurement count), and the previous device There is an upper limit of the elapsed time since using. These pieces of information are used for determining whether or not the element system is defective or performing the initial adjustment process. Also, these pieces of information need to be set in advance before executing the initial adjustment process due to the nature thereof. The other information is information unique to each device, and is held on the non-volatile memory in the control unit 6 as “initial adjustment information”. This information includes, for example, the temperature change rate (temperature change rate table) in each temperature zone of each heating and cooling element 4, the date and time when the previous initial adjustment process was executed (the previous use date and time), and without performing the initial adjustment process. In order to keep the temperature of the temperature control unit 100 uniform, the number of times the PCR was performed (cumulative measurement count), the number and position of the reaction tanks used last time (previous use conditions), and the temperature of the temperature control unit 100 were always There are a coefficient (output value correction coefficient) used when correcting the force, and information on an element system determined to be defective in the previous initial adjustment process (performance defective element system information at the previous use).

  Information on the temperature change rate is created in the initial adjustment process, and is referred to in the control amount calculation process (FIG. 7) of the heating / cooling element described later. This information is read into the volatile memory in the control unit 6 immediately after the start of the apparatus and before the start of PCR execution, and is referred to at a constant cycle when calculating the output amount of the heating / cooling element 4 thereafter.

  Two types of information are held in the temperature change rate table. One information is temperature change rate information when the heating / cooling element 4 is operated alone, and the other is temperature change rate information when all the three heating / cooling elements 4 are operated simultaneously.

  Other information included in the initial adjustment information refers to each item as necessary when the control unit 6 determines the reference heating / cooling element. The contents executed in each step will be described below. In the following description, step is abbreviated as “S”.

・ S301
In 301, the control unit 6 confirms whether or not the initial adjustment information exists on the internal nonvolatile memory. The control unit 6 executes S302 when the initial adjustment information exists, and executes S309 when the initial adjustment information does not exist.

・ S302
In S302, the control unit 6 reads the initial adjustment information into the volatile memory of the control unit 6, and proceeds to S303.

・ S303
In S303, the control unit 6 extracts the “presence / absence of a defective device system at the previous use” in the initial adjustment information, and if there is no defective device system at the previous use, the control unit 6 proceeds to S304, If there is, the process proceeds to S309.

・ S304
In S304, the control unit 6 extracts the “previous use date and time” in the initial adjustment information, and compares the current date with the current date and time to calculate the elapsed time from the previous use. If the calculated elapsed time does not exceed the “specified elapsed time (elapsed period since last use)” in the initial adjustment information, the process proceeds to S305, and if the calculated elapsed time is equal to or greater than the “specified elapsed time”, the process proceeds to S309.
In S305, the control unit 6 extracts the “cumulative measurement count” in the initial adjustment information. If the cumulative measurement count is equal to or less than the specified number, the process proceeds to S306, and if it exceeds the specified number, the process proceeds to S309.

・ S306
In S306, the control unit 6 extracts the “previous use condition” in the initial adjustment information, and proceeds to S307 if the number and position of the reaction tanks used last time are the same as this time, and if the use conditions are different, the process proceeds to S309. Proceed to In this process, for example, when only a part of the eight reaction vessels 101 is used instead of all, the heating / cooling element 4 is selected to minimize the frequency of use of the heating / cooling element 4. Can do. Thereby, in addition to the effect of extending the life of the heating / cooling element 4, an effect of reducing excessive heat generation in the apparatus can be obtained.

・ S307
In S307, the control unit 6 inquires of the user whether or not the initial adjustment process needs to be performed. If the initial adjustment process is unnecessary, the process proceeds to S308, and if the initial adjustment process is necessary, the process proceeds to S309.

・ S308
In S308, since the control unit 6 does not execute the initial adjustment process, the “cumulative measurement count” in the initial adjustment information is incremented by 1, and the process proceeds to S310.

・ S309
In S309, the control unit 6 resets the “cumulative measurement count” in the initial adjustment information to 0, and proceeds to S311.

・ S310
In S310, the control unit 6 writes the current cumulative number of measurements in the initial adjustment information, and proceeds to S312.

・ S311
In S311, the control unit 6 performs initial adjustment processing, and proceeds to S312. Details of the initial adjustment processing are described in FIG.

・ S312
In S312, the control unit 6 selects one of the three heating / cooling elements 4 as the reference heating / cooling element 4 to be used during the current PCR.

  By executing the above-described processing, the heating / cooling element 4 having the slowest temperature change rate can be defined under the current environment. By adjusting the output amount of the other two heating / cooling elements 4 while monitoring the behavior of the reference heating / cooling element, the temperature difference between the three heating / cooling elements 4 can be reduced. PCR homogenization in the tank 101 can be achieved.

  FIG. 6 shows a specific example of the initial adjustment process S311 executed during the process of determining the reference heating / cooling element shown in FIG. This process is also executed by the control unit 6.

・ S401
In S401, the control unit 6 selects one element from the heating / cooling element 4, executes heating and cooling at the maximum output once, measures and records the temperature change rate within each temperature range. . This process is sequentially executed by the three heating / cooling elements 4, and the process proceeds to S402. The measured temperature change rate is stored in the initial adjustment information as a temperature change rate table. In this embodiment, the temperature change rate table (single) is defined as a table recording the temperature change rate of the heating / cooling element 4 within a certain temperature range.

・ S402
In S402, the control unit 6 refers to the result of the temperature change rate table created in S401 and confirms the presence or absence of a defective element system. The control unit 6 proceeds to S403 when determining that there is a defective element system “absent”, and proceeds to S404 when determining that there is a defective element system “present”. For example, in all three heating / cooling elements 4, the control unit 6 has a temperature change rate equal to or higher than the “minimum value of temperature change rate” held in the nonvolatile memory in the control unit 6 in any temperature range. Advances to S403, and if any one of the heating and cooling elements 4 is less than the “minimum value of temperature change rate”, the process advances to S404.

・ S403
In S403, the control unit 6 performs control for heating and cooling all three heating / cooling elements 4 simultaneously at 50% of the maximum output once, and measures the temperature change rate within each temperature range. Then, add to the temperature change rate table and proceed to S405. The measured temperature change rate is stored in the initial adjustment information as a temperature change rate table.

・ S404
In S404, the control unit 6 determines that the element system whose temperature change rate is less than the “minimum temperature change rate value” in the initial adjustment information is defective, and recommends the user to replace the corresponding element system. Make a notification. Further, in order not to shift to the subsequent processing, the information about the corresponding element system is described as “performance defect element system information at the time of previous use” in the initial adjustment information, and then the process ends.

・ S405
In S405, the control unit 6 selects the heating / cooling element 4 having the slowest temperature change rate based on the temperature change rate table created in S401 and S403, and sets it as the reference heating / cooling element.

・ S406
In S406, the control unit 6 outputs the following items as initial adjustment information. The items listed are “Last Use Date / Time”, “Cumulative Measurement Count”, “Last Use Condition”, “Temperature Change Rate Table”, “Output Value Correction Coefficient”, “Performance Element Information for Previous Use”. .

  By this processing, it becomes possible to detect the heating / cooling element 4 having the slowest temperature change speed as a system characteristic under the current apparatus conditions and select it as the reference heating / cooling element.

  By combining the behavior of the other two heating / cooling elements 4 with the behavior of this reference heating / cooling element, all three heating / cooling elements 4 can be output without requiring output exceeding the hardware performance. It is possible to make the behavior of the heating / cooling element 4 uniform.

  At the same time, when performing PCR in the present apparatus, it becomes possible to detect the heating / cooling element 4 having insufficient performance before performing PCR. This contributes to avoiding unforeseen circumstances such as wasting samples due to PCR failure and performing PCR under conditions such as temperature and time that are different from expectations.

<Temperature control process>
FIG. 7 shows an example of a control process for aligning the behaviors of the three heating / cooling elements 4 using the reference heating / cooling elements determined in the process of FIG. This process is also executed by the control unit 6.

・ S501
In S501, the control unit 6 uses the three temperature measuring elements 1, monitors the temperatures of the three heating / cooling elements 4 paired with each, and proceeds to S502. Processing of temperature monitoring, calculation of the output amount of the heating / cooling element 4 and output to the heating / cooling element 4 (S501 to S513) is executed at regular time intervals.

・ S502
In S502, the control unit 6 calculates three output amounts I by the PID calculation from the three temperatures measured in S501 and the set temperatures set by the variable setting unit 11, and the process proceeds to S503. . The output amount I obtained here is a value that takes into account only the current temperature and the set temperature at each location of the temperature measuring element 1. However, if the output as it is is output to the heating / cooling element 4, it is considered that a temperature difference occurs in the reaction tank 101 due to a difference in heat transfer conditions around each heating / cooling element 4. Therefore, the correction process described below is required.

・ S503
In S503, the control unit 6 calculates a correction value for the non-reference heating / cooling element based on the temperature change rate table and the three output amounts I, and proceeds to S504. A specific example of the correction value calculation process of the non-reference heating / cooling element is shown in the correction value calculation process of the non-reference heating / cooling element to be described later with reference to FIG.

・ S504
In S504, the control unit 6 calculates the output amount of the non-reference heating and cooling element based on the output amount I of the non-reference heating and cooling element calculated in S502 and the correction value of the non-reference heating and cooling element calculated in S503. Proceed to S505.

・ S505
In S505, the control unit 6 proceeds to S512 if the output amounts of the two non-reference heating / cooling elements calculated in S504 are both the maximum output (100%) or less, and the maximum output (100% ), The process proceeds to S506. In this control method, the heating / cooling element 4 having the smallest temperature change rate is used as the reference heating / cooling element. However, when two heating / cooling elements 4 having very similar temperature change rates are used, the output amount of the non-reference heating / cooling element may be required to be greater than the maximum output (100%). In order to cope with this, an example of a correction method for the reference heating / cooling element will be described below.

・ S506
In S506, the control unit 6 selects an output amount having a larger value from the output amounts of the two non-reference heating / cooling elements, holds the information, and proceeds to S507.

・ S507
In S507, the control unit 6 sets the output amount of the non-reference heating / cooling element having the larger value among the output amounts of the two non-reference heating / cooling elements to the maximum output (100%), and proceeds to S508.

・ S508
In S508, the control unit 6 calculates a correction value for the output amount of the reference heating / cooling element in accordance with the heating / cooling element 4 in which the output amount is set to the maximum output (100%) in S507, and proceeds to S509.

・ S509
In S509, the control unit 6 calculates the output amount of the reference heating / cooling element from the output amount I of the reference heating / cooling element calculated in S502 and the correction value of the reference heating / cooling element calculated in S508, and proceeds to S510.

・ S510
In S510, the control unit 6 calculates a correction value for the output amount of the non-reference heating / cooling element whose output amount is undetermined, based on the output amount of the reference heating / cooling element calculated in S509 and the measured temperature of the reference heating / cooling element. , Go to S511.

・ S511
In S511, the control unit 6 calculates the output amount of the non-reference heating / cooling element from the correction value of the non-reference heating / cooling element whose output amount calculated in S510 is undetermined and the output amount calculated in S502, and proceeds to S512.

・ S512
In S512, the control unit 6 outputs the output amounts of the three heating / cooling elements 4 calculated so far to the heating / cooling element 4, and proceeds to S513.

・ S513
In S513, the control unit 6 proceeds to S514 if the PCR is completed, and repeats the process again from S501 if it is not completed yet.

・ S514
In S514, the control unit 6 outputs “the number of corrections of the output amount of the reference heating / cooling element” as the initial adjustment information.

  With this processing, the reference heating / cooling element having the slowest temperature change speed in terms of hardware performance exhibits the most efficient performance as a whole system, while the other two heating / cooling elements 4 adjust the output amount. Is possible. This makes it possible to reduce the temperature difference between the heating / cooling elements 4 and between the reaction vessels 101 while maintaining high speed, and realize temperature control suitable for PCR for all eight reaction vessels 101. be able to.

  FIG. 8 shows a specific processing example of S503 of the non-reference heating / cooling element shown in FIG. The control unit 6 also executes this process.

・ S601
In S601, the control unit 6 refers to the temperature change rate table based on the temperature measured by the temperature measuring element 1, and sets the temperature change rate of the heating / cooling element 4 when the output amount is 50% for all the heating / cooling elements 4. Read and proceed to S602.

・ S602
In S602, the control unit 6 calculates a correction value (correction value I) required to eliminate the current temperature difference between the reference heating / cooling element and the non-reference heating / cooling element, based on the temperature change rate read in S601. Proceed to S603.

・ S603
In S603, the control unit 6 calculates a predicted value of the temperature reached by the reference heating / cooling element during the next scan based on the temperature change rate of the reference heating / cooling element, and proceeds to S604.

・ S604
In S604, the control unit 6 arrives at the non-reference heating / cooling element at the next scan based on the output amount I of the non-reference heating / cooling element calculated in S502, the correction value I of the output amount calculated in S602 and the temperature change rate table. The predicted value of the temperature to be calculated is calculated, and the process proceeds to S605.

・ S605
In S605, the control unit 6 calculates the difference between the temperature predicted value at the next scan of the reference heating / cooling element calculated at S603 and the temperature predicted value at the next scan of the non-reference heating / cooling element calculated at S604. Based on the above, a correction value (correction value II) of the output amount required to eliminate the difference between the arrival temperatures of the non-reference heating / cooling element and the reference heating / cooling element at the next scan is calculated, and the process proceeds to S606.

・ S606
In S606, the control unit 6 uses the correction value I calculated in S602 and the correction value II calculated in S605 to correct the non-reference heating / cooling element correction values required to make the temperatures of the eight reaction vessels 101 uniform during the next scan. Is calculated.

  If correction of the reference heating / cooling element is required, as in S508 and S510, use the non-reference heating / cooling element (a) (see Fig. 7) with the output amount set to the maximum output (100%) in S507. As described above, the output amounts of the reference heating / cooling element and the non-reference heating / cooling element (b) (see FIG. 7) are corrected.

  With this process, the control unit 6 can calculate a correction value for the output amount of the heating / cooling element 4 that is necessary for the control to keep the temperatures of the eight reaction vessels 101 uniform at all times.

<Effect>
As described above, by applying the temperature control of the present embodiment, even in a structure in which a plurality of heat sources are arranged in a compact region, the temperature is the highest as a system in which the temperature measuring element 1 and the heating / cooling element 4 are combined. By using the heating / cooling element 4 with a slow change rate as a reference and adjusting the output amount of the other heating / cooling elements 4, it is possible to make the temperature change behavior of each heating / cooling element 4 and thus each reaction tank uniform. Thereby, temperature control suitable for PCR requiring high-speed and high-precision control can be executed.

  In this embodiment, the output amount of the heating / cooling element 4 at the time of creating the temperature change rate table in S403 is 50% of the maximum output, but is not limited to this, and the heating / cooling element is arbitrarily set as necessary. The same effect can be obtained even if the output amount of 4 is selected. Accordingly, when the correction value calculation processing for the non-reference heating / cooling element shown in FIG. 8 is performed, the temperature change rate read from the temperature change rate table is calculated as a response to the output amount when the temperature change rate table is created. .

[Example 2]
In the first embodiment, the method for uniformly controlling the temperature when the output of the heating / cooling element 4 is always almost the upper limit value (maximum output) of the heating / cooling element 4 has been described. A control method assuming the case will be described. For example, temperature transition from annealing treatment to extension treatment in PCR with three stages of temperature transition, temperature transition from initial enzyme treatment and initial denaturation treatment to thermal denaturation treatment at the beginning of PCR execution, and further from extension reaction treatment to final extension reaction Consider a case where the temperature change width is narrow, such as processing, and the output required for the heating and cooling element 4 has a sufficient margin with respect to the upper limit value of the element.

<Temperature control process>
When there is a sufficient margin in the output of each heating / cooling element 4, the heating / cooling element 4 having the fastest temperature change rate is used as the reference heating / cooling element, and the output amount of the other heating / cooling elements 4 is corrected, thereby achieving higher speed. PCR becomes possible.

  In particular, PCR is characterized by the control of periodically changing several temperature ranges. Therefore, when the temperature range to be changed is large and the output required for the heating / cooling element 4 is close to the upper limit value, as shown in the first embodiment, the heating / cooling element 4 having the slowest temperature change rate is replaced with another heating element. The behavior of the cooling element 4 is matched. On the other hand, when the temperature range to be changed is small and the output required for the heating / cooling element 4 has a margin, the behavior of the other heating / cooling elements 4 can be matched with the heating / cooling element 4 having the fastest temperature change rate. Uniform and high-speed PCR can be executed in all reaction vessels 101.

  FIG. 9 shows an example of temperature control processing for reducing the temperature difference between the eight reaction vessels 101 while switching the reference heating / cooling element. This process is executed by the control unit 6. Moreover, this process adds the process which switches a reference | standard heating / cooling element to the process of FIG. Therefore, in FIG. 9, the same reference numerals are given to the corresponding parts to FIG. In the following, only the additions to FIG. 7 will be described.

・ S701
S701 is executed between S502 and S503. In S701, the control unit 6 proceeds to S702 if the difference between the current target temperature and the previous target temperature is 15 ° C. or more, and proceeds to S703 if the difference is less than 15 ° C.

・ S702
In S702, the control unit 6 sets the heating / cooling element 4 having the slowest temperature change speed as a reference heating / cooling element based on the temperature change speed table, and proceeds to S503. Here, the process when the temperature change width is large and the output amount required for the heating / cooling element 4 is close to the upper limit value is shown. As in the first embodiment, the heating / cooling element 4 having the slowest temperature change rate is shown. It is set as a standard.

・ S703
In S703, the control unit 6 sets the heating / cooling element 4 having the fastest temperature change rate as the reference heating / cooling element based on the temperature change rate table, and proceeds to S503. This shows processing when the temperature change width is small and the output amount required for the heating / cooling element 4 still has a margin to the upper limit.

  The temperature threshold (15 ° C) for switching control in S701 is an example, and the temperature change range where the output amount does not reach the upper limit depending on the specifications of the heating and cooling elements to be used and the positional relationship when multiple elements are arranged Can be set arbitrarily. Moreover, it is good also as variable according to the period (generally 20-30 cycles) of PCR.

  In another example, as the condition for switching the reference heating / cooling element, the output amount of each heating / cooling element 4 by PID calculation can be used instead of the temperature change width.

  FIG. 10 shows a flowchart showing an example of temperature control processing for reducing the temperature difference between the eight reaction vessels 101 while switching the reference heating / cooling element. This process is executed by the control unit 6. Moreover, this process adds the process which switches a reference | standard heating / cooling element to the process of FIG. Therefore, in FIG. 10, the same reference numerals are given to the portions corresponding to FIG. 7. Only the additions to FIG. 7 will be described below.

・ S801
S801 is also executed between S502 and S503. In S801, the control unit 6 proceeds to S802 if one of the heating / cooling elements 4 has an output amount of 80% or more of the maximum output, and the output amount of all the heating / cooling elements 4 is 80. If less than%, proceed to S803.

・ S802
In S802, the control unit 6 sets the heating / cooling element 4 having the slowest temperature change rate as the reference heating / cooling element based on the temperature change rate table, and proceeds to S503. Here, a process is shown in which the difference between the current temperature measured by the temperature measuring element 1 and the target temperature is large, and the output amount required for the heating / cooling element 4 is close to the upper limit value. In addition, the heating / cooling element 4 having the slowest temperature change rate is set as a reference.

・ S803
In S803, the control unit 6 sets the heating / cooling element 4 having the fastest temperature change rate as the reference heating / cooling element based on the temperature change rate table, and proceeds to S503. This shows processing when the difference between the current temperature measured by the temperature measuring element 1 and the target temperature is small and the output amount required for the heating / cooling element 4 still has a margin to the upper limit value.

  Note that the output amount threshold value for control switching in S801 is merely an example, and the same effect can be obtained with any setting. Moreover, it is good also as variable according to the period of PCR.

<Effect>
As described above, by applying the temperature control of this embodiment, priority is given to high speed within the range of the hardware configuration such as the number and arrangement of the heating and cooling elements 4 and the performance of each heating and cooling element 4. The heating / cooling element 4 can be appropriately selected and changed as the reference heating / cooling element from the plurality of heating / cooling elements 4 as necessary.

  In addition, when the output amount required for each heating / cooling element 4 has a margin with respect to the upper limit value as the element, the other heating / cooling element outputs the output amount to the heating / cooling element having the fastest temperature change rate. Since it becomes possible to raise and match | combine, PCR can be performed at high speed, maintaining the uniformity of the temperature distribution of the reaction tank 101. FIG.

  Further, the reference heating / cooling element is not limited to the actually measured value of the temperature change of the heating / cooling element 4. For example, time series data of an arbitrary temperature change prepared in advance can be used as a temperature change of the reference heating / cooling element in a pseudo manner. However, in this case, since it is impossible to correct the reference heating / cooling element, the temperature change curve creation is restricted by the performance related to the upper limit of the output amount of the element.

<Other embodiments>
In addition, this invention is not limited to the Example mentioned above, Various modifications are included.

  For example, in the above-described embodiment, the number of the heating / cooling elements 4 and the temperature measuring elements 1 has been described as three. However, the number is not limited to these, and for example, the present invention can be similarly applied when more elements are used. Thus, the same effect can be obtained. 1 shows an example in which one measuring unit 2 is arranged and one output control unit 3 is arranged, but there may be a plurality of each. For example, the same number of measuring units 2 as the temperature measuring elements 1 are arranged, and A / D conversion of analog data obtained from each temperature measuring element 1 is performed by dividing each measuring unit 2 to perform temperature measurement as in the first embodiment. Is possible.

  In this embodiment, the reference heating / cooling element is used by switching between the slowest heating / cooling element 4 and the fastest heating / cooling element 4, but the method of selecting the reference heating / cooling element is not limited to this, It is possible to select arbitrarily as required.

  In the above-described embodiments, some of the embodiments have been described in detail in order to explain the present invention in an easy-to-understand manner, and it is not always necessary to provide all the configurations and processes described. Further, a part of a certain embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of a certain embodiment. Moreover, it is also possible to add, delete, or replace another configuration for a part of the configuration of each embodiment.

  Moreover, you may implement | achieve some or all of each structure, a function, a process part, a process means, etc. which were mentioned above as an integrated circuit or other hardware, for example. Each of the above-described configurations, functions, and the like may be realized by the processor interpreting and executing a program that realizes each function. That is, it may be realized as software. Information such as programs, tables, and files for realizing each function can be stored in a memory, a hard disk, a storage device such as an SSD (Solid State Drive), or a storage medium such as an IC card, an SD card, or a DVD.

  Control lines and information lines indicate what is considered necessary for the description, and do not represent all control lines and information lines necessary for the product. In practice, it can be considered that almost all components are connected to each other.

1: Temperature measurement element 2: Measurement unit 3: Output control unit 4: Heating / cooling element 5: Heat radiation fan 6: Control unit 10: Sequence control unit 11: Variable setting unit 12: Output amount calculation unit 100: Temperature adjustment unit 101: Reaction tank 201: heat insulation part 202: heat radiation part

Claims (14)

A plurality of heating and cooling elements;
A plurality of temperature measuring elements paired with each of the plurality of heating and cooling elements;
A control unit for controlling the output amount of the plurality of heating and cooling elements,
The control unit inputs a process for determining one of the plurality of heating / cooling elements as a reference element, and a temperature value detected by a temperature measuring element paired with each of the plurality of heating / cooling elements, as a target temperature value A process for controlling the output amounts of the plurality of heating and cooling elements according to the difference between them and one time other than the reference element according to the time variation of the temperature value detected by the temperature measuring element paired with the reference element Alternatively, a nucleic acid analyzer that performs a process of adjusting the output amount of a plurality of heating and cooling elements. The nucleic acid analyzer according to claim 1,
The controller is
The plurality of heating / cooling elements are simultaneously controlled with the same output to compare the temperature values detected by the plurality of temperature measuring elements paired with the heating / cooling elements, and the heating / cooling element exhibiting the lowest rate of temperature change is A nucleic acid analyzer characterized by being selected as a reference element. The nucleic acid analyzer according to claim 2,
The controller is
When the lowest temperature change rate falls below a certain value, the temperature control device is not selected as the reference element. The nucleic acid analyzer according to claim 1,
The controller is
The plurality of heating / cooling elements are simultaneously controlled with the same output to compare the temperature values detected by the plurality of temperature measuring elements each paired with the heating / cooling element, and the heating / cooling element exhibiting the highest temperature change rate is A nucleic acid analyzer characterized by being selected as a reference element. The nucleic acid analyzer according to claim 1,
The controller is
When the difference between the current target temperature and the previous target temperature is greater than a certain value, the plurality of heating and cooling elements are simultaneously controlled with the same output, and a plurality of temperature measurements that respectively pair with the heating and cooling elements. When comparing the temperature value detected by the element, select the heating and cooling element that showed the lowest rate of temperature change as the reference element,
When the difference between the current target temperature and the previous target temperature is not greater than the predetermined value, the plurality of heating and cooling elements are simultaneously controlled with the same output, and a plurality of pairs that respectively pair with the heating and cooling elements. A nucleic acid analyzer characterized by selecting, as the reference element, a heating / cooling element that exhibits the highest rate of temperature change when comparing temperature values detected by a temperature measuring element. The nucleic acid analyzer according to claim 1,
The controller is
When even one of the output amounts of the plurality of heating / cooling elements is larger than a certain value, the plurality of heating / cooling elements are simultaneously controlled with the same output, and a plurality of temperature measuring elements respectively paired with the heating / cooling elements are controlled. When comparing the detected temperature value, select the heating and cooling element showing the lowest rate of temperature change as the reference element,
When all of the output amounts of the plurality of heating / cooling elements are smaller than the certain value, the plurality of heating / cooling elements are simultaneously controlled with the same output, and the plurality of temperature measuring elements respectively paired with the heating / cooling elements are controlled. A nucleic acid analyzer characterized by selecting, as the reference element, a heating / cooling element that exhibits the highest temperature change rate when comparing detected temperature values. The nucleic acid analyzer according to claim 1,
The controller is
When the output amount of one or a plurality of heating / cooling elements other than the reference element is determined and the output possible amount is exceeded, the output amount of the reference element is adjusted. Controlling the temperature of the nucleic acid analyzer having a plurality of heating / cooling elements, a plurality of temperature measuring elements paired with each of the plurality of heating / cooling elements, and a control unit for controlling the output amount of the plurality of heating / cooling elements In the way to
A process in which the control unit determines one of the plurality of heating and cooling elements as a reference element;
The control unit controls an output amount of the plurality of heating / cooling elements according to a difference from a target temperature value, using a temperature value detected by a temperature measuring element paired with each of the plurality of heating / cooling elements as an input. Processing,
The control unit includes a process of adjusting the output amount of one or a plurality of heating / cooling elements other than the reference element according to a time change amount of a temperature value detected by a temperature measuring element paired with the reference element. A temperature control method for a nucleic acid analyzer. The temperature control method according to claim 8,
The controller is
The plurality of heating / cooling elements are simultaneously controlled with the same output to compare the temperature values detected by the plurality of temperature measuring elements paired with the heating / cooling elements, and the heating / cooling element exhibiting the lowest rate of temperature change is A temperature control method, wherein the temperature control method is selected as a reference element. The temperature control method according to claim 9, wherein
The controller is
When the lowest temperature change rate falls below a certain value, the temperature control method is not selected as the reference element. The temperature control method according to claim 8,
The controller is
The plurality of heating / cooling elements are simultaneously controlled with the same output to compare the temperature values detected by the plurality of temperature measuring elements each paired with the heating / cooling element, and the heating / cooling element exhibiting the highest temperature change rate is A temperature control method, wherein the temperature control method is selected as a reference element. The temperature control method according to claim 8,
The controller is
When the difference between the current target temperature and the previous target temperature is greater than a certain value, the plurality of heating and cooling elements are simultaneously controlled with the same output, and a plurality of temperature measurements that respectively pair with the heating and cooling elements. When comparing the temperature value detected by the element, select the heating and cooling element that showed the lowest rate of temperature change as the reference element,
When the difference between the current target temperature and the previous target temperature is not greater than the predetermined value, the plurality of heating and cooling elements are simultaneously controlled with the same output, and a plurality of pairs that respectively pair with the heating and cooling elements. A temperature control method comprising: selecting a heating / cooling element that exhibits the highest rate of temperature change when comparing temperature values detected by a temperature measuring element as the reference element. The temperature control method according to claim 8,
The controller is
When even one of the output amounts of the plurality of heating / cooling elements is larger than a certain value, the plurality of heating / cooling elements are simultaneously controlled with the same output, and a plurality of temperature measuring elements respectively paired with the heating / cooling elements are controlled. When comparing the detected temperature value, select the heating and cooling element showing the lowest rate of temperature change as the reference element,
When all of the output amounts of the plurality of heating / cooling elements are smaller than the certain value, the plurality of heating / cooling elements are simultaneously controlled with the same output, and the plurality of temperature measuring elements respectively paired with the heating / cooling elements are controlled. A temperature control method comprising: selecting a heating / cooling element that exhibits the highest rate of temperature change when comparing detected temperature values as the reference element. The temperature control method according to claim 8,
The controller is
When the output amount of one or a plurality of heating / cooling elements other than the reference element is determined, and the output possible amount is exceeded, the output amount of the reference element is adjusted.

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