JP2009095755A - Reaction treatment apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction treatment apparatus capable of carrying out temperature control at high precision, with respect to a reaction treatment apparatus provided with a plurality of reaction regions. <P>SOLUTION: The reaction treatment apparatus (1) includes a plurality of reaction regions (A) and heating parts installed for the individual reaction regions, and the heating parts respectively have a heat source, a heat interference detection part for detecting the heat interference among the reaction regions (A) and a heating control part for controlling the heating quantity of the heat source based on the temperature information detected by the heat interference detection part. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a reaction processing apparatus. More specifically, the present invention relates to a technique capable of performing reaction control with high accuracy in a reaction processing apparatus including a plurality of reaction regions.

  2. Description of the Related Art Reaction processing apparatuses that are provided with a plurality of reaction regions and can perform predetermined reactions in the respective reaction regions are used in various fields. An exhaustive analysis can be performed by using a reaction processing apparatus that can perform a predetermined reaction in a plurality of reaction regions. It can be used for a wide range of applications. It is expected to be applied to a wide range of fields including medical care, environmental science, and food analysis. As such a reaction processing apparatus, for example, it is considered to provide a plurality of reaction regions on the same substrate and perform various reactions using the substrate.

  On the other hand, in such a reaction processing apparatus, it is important that accurate temperature control can be performed in each reaction region. In this regard, Patent Document 1 and Patent Document 2 disclose techniques related to temperature control of a reaction processing apparatus. In addition, in order to heat a minute region, a semiconductor element is used.

JP2003-298068A. Japanese Patent Application Laid-Open No. 2004-025426.

  Accordingly, the main object of the present invention is to provide a reaction processing apparatus capable of performing temperature control with high accuracy in a reaction processing apparatus having a plurality of reaction regions.

First, the present invention includes a plurality of reaction regions and a heating unit provided for each reaction region, the heating unit including a heat source and a thermal interference detection unit that detects thermal interference between the reaction regions, There is provided a reaction processing apparatus including at least heating control means for controlling a heating amount of the heat source based on temperature information detected by the thermal interference detection unit. Thus, the thermal interference between reaction regions can be detected by providing the thermal interference detector between the reaction regions. Considering the influence of this thermal interference, more accurate temperature control becomes possible.
Next, the present invention provides a reaction processing apparatus in which the thermal interference detection unit detects thermal interference between the reaction regions using a temperature measuring element disposed between adjacent reaction regions. By disposing at least the temperature measuring element between the adjacent reaction regions as the thermal interference detection unit, it is possible to detect thermal interference with higher accuracy.
Subsequently, according to the present invention, the plurality of reaction regions and the heating units corresponding to the reaction regions are arranged in a matrix, and each heat source of the heating unit generates heat connected to a scanning line and a data line. The heating element is capable of generating heat by being sequentially scanned, and each temperature measuring element of the heating unit provides a reaction processing device disposed on the scanning direction side of the corresponding heating element. . Heat source control can be easily performed by arranging reaction regions and corresponding heating sections in a matrix and controlling each heat source using scanning lines and data lines. In addition, more accurate thermal interference can be detected by disposing the temperature measuring elements corresponding to the respective reaction regions on the heating direction side where they are heated. And this invention provides the reaction processing apparatus in which the said temperature sensing element is provided with the PIN diode at least.
Further, according to the present invention, the heating unit further includes a temperature detection unit that measures the temperature of the reaction region, and the heating control unit includes the temperature information detected by the thermal interference detection unit, and the temperature detection unit. Provided is a reaction processing apparatus that controls the amount of heating of the heat source based on detected temperature information. By providing not only the thermal interference detection unit but also a temperature detection unit for measuring the temperature of the reaction region, temperature control of each reaction region can be performed with higher accuracy.
Further, the present invention provides a reaction processing apparatus further comprising: an optical unit that irradiates light of a predetermined wavelength to the reaction region; and a light detection unit that can detect measurement target light generated by the light irradiation. provide. By providing such an optical system as a reaction processing apparatus, real-time analysis becomes possible. The present invention can be a gene amplification device in which a gene amplification reaction is performed in the reaction region.

  According to the present invention, highly accurate temperature control can be performed in a reaction processing apparatus having a plurality of reaction regions. As a result, the predetermined reaction performed in the reaction region can be controlled with high accuracy.

  Hereinafter, preferred embodiments of a reaction processing apparatus according to the present invention will be described with reference to the accompanying drawings. Each embodiment shown in the accompanying drawings shows an example of a typical embodiment according to the present invention, and the scope of the present invention is not interpreted narrowly. In the drawings used below, for the convenience of explanation, the configuration of the apparatus is shown in a simplified manner.

  FIG. 1 is a conceptual diagram of a first embodiment of a reaction processing apparatus according to the present invention, and FIG. 2 is a conceptual diagram for explaining a heating unit in the same embodiment. The size, apparatus structure, and the like of the reaction processing apparatus 1 can be appropriately selected depending on the application and the like, and can be designed or changed within the scope of the object of the present invention.

  The code | symbol 1 in FIG. 1 has shown the reaction processing apparatus which concerns on this invention. The reaction processing apparatus 1 includes a plurality of reaction regions A, each reaction region A having a heating unit, and the heating unit includes a heat source H and a temperature measuring body S used for a heat interference detection unit. I have.

  When the heat source H generates heat, the corresponding reaction region A can be heated. However, thermal interference occurs between the reaction regions A due to the heating at this time. In the reaction processing apparatus 1, this thermal interference can be detected by the temperature measuring body S of the thermal interference detection unit. Furthermore, the heating amount of the heat source H can be controlled based on the temperature information detected by the temperature measuring body S of the thermal interference detector by a heating control means (not shown).

  The structure and the like of the heating unit are not particularly limited, but preferably, the heat source H is formed by a thin film transistor (TFT) and is subjected to switching control. For example, the temperature control of each reaction region A can be performed individually using the switching function of the thin film transistor. In this temperature control, the voltage applied to the thin film transistor may be controlled to vary the current value between the source and the drain, or the current between the source and the drain may be controlled as a constant current power source. In particular, it is desirable to use thin film transistors as the heat source H and thin film transistors as switching elements, and to form these thin film transistors in the same circuit. Thereby, the heat source H and the switching control can be collectively controlled in the same circuit. A thin film transistor substrate (TFT substrate) is preferably used, which will be described later.

  Alternatively, the heat source H of the heating unit may be a heater that is formed of a heating resistor and that is switching-controlled by a thin film transistor. That is, the thin film transistor may be used only for switching. As the heating resistor, platinum, molybdenum, tantalum, tungsten, silicon carbide, molybdenum silicide, nickel-chromium alloy, iron-chromium-aluminum alloy, or the like can be used. In this case, the temperature can be controlled by controlling the value of the current flowing through the heating resistor. The type of thin film transistor used in the present invention is not particularly limited, and for example, a type such as polysilicon or α-silicon can be used as appropriate.

  The thermal interference detection unit only needs to be able to detect thermal interference that may occur between the reaction regions A, and the method and the like are not limited. For example, various sensors capable of detecting heat can be used. And like the reaction processing apparatus 1, the temperature measuring body S can be used as a thermal interference detection part, and this temperature measuring body S can be arrange | positioned between the reaction regions A adjacent. As the temperature measuring element S, for example, one having a certain correlation between temperature and electrical signal value (voltage, resistance, current, etc.) can be used. In this case, the temperature measuring body S can detect temperature information based on this value because the value of the detected electrical signal varies depending on the temperature. As the temperature measuring element S, for example, an element having a diode characteristic can be used, and a PIN diode is more preferably used.

  The heating control means only needs to be able to control the heating amount of each heat source H based on the temperature information detected by the thermal interference detection unit, and the configuration and method thereof are not limited. For example, although not shown, the detected electrical signal can be converted into a digital signal by an analog-digital converter (ADC) and taken into the CPU as digital data. Then, the CPU performs arithmetic processing to output a digital signal to the digital potentiometer as much as possible to obtain an optimum current value as a target temperature. In this way, it is possible to feed back to the heating control based on the temperature information detected by the thermal interference detector.

  In the present invention, it is desirable to further provide a temperature detector for detecting the temperature of the reaction region A. It is desirable to control the heating by the heating control means based on both the temperature information of the reaction region A detected by the temperature detection unit and the temperature information on the thermal interference obtained by the thermal interference detection unit. As a result, temperature control can be performed more accurately.

  Here, the configuration of each heating unit in each reaction region A will be described with reference to FIG. FIG. 2 further includes a temperature detection unit for detecting the temperature of the reaction region A. A temperature sensing element Hs is used as the temperature detection unit. Based on both the temperature information of the reaction region A detected by the temperature measuring body Hs and the temperature information on the thermal interference obtained by the temperature measuring body S of the thermal interference detecting unit, the heating control means controls the heating. be able to.

  In this case, the configurations of the heating element H and the temperature measuring element Hs are not limited, and a suitable configuration can be appropriately set in consideration of the purpose and the device structure. For example, a temperature measuring element may be provided adjacent to the heating element H (see symbols I and II in FIG. 2), or a temperature measuring element Hs may be provided in the central region of the heating element H (in FIG. 2). (See symbol III).

  FIG. 3 is a conceptual diagram of a second embodiment of the reaction processing apparatus according to the present invention. The code | symbol 2 of FIG. 3 has shown the reaction processing apparatus based on this invention. The reaction processing apparatus 2 has a structure in which a plurality of reaction regions A are arranged in a matrix, and corresponding heating units are arranged in a matrix. Hereinafter, the difference from the first embodiment will be mainly described, and the description of the common parts will be omitted.

  The reaction processing apparatus 2 includes a plurality of scanning lines and data lines for each heating unit in each reaction region A, and a heater unit of the heating unit is disposed at each of these intersections. The heater units are also arranged in a matrix along the gate lines (X direction) and the data lines (Y direction). By setting it as such a structure, each heat source can be collectively controlled.

  The scanning drive circuit sequentially selects the scanning lines (that is, lowers the scanning line), and the data line driving circuit applies a signal current to each data line in synchronization therewith, so that each heater unit is scanned in units of rows (scanning). The heating amount information can be written by lines 1, 2,. After writing, if each scanning line is not selected (that is, high level), the drive current having the same current value as the signal current continues to flow through each unit. In this way, a current of each desired magnitude can be passed. In this way, each heating unit (the heater unit) can be individually controlled.

  In the present invention, the temperature measuring element S is further used as a thermal interference detection unit, which is disposed on the heating direction side (see the arrow in FIG. 3) of the corresponding heat source of each heating unit. It is said. In the case of FIG. 3, the temperature information detected by the temperature measuring body S on the right side of the reaction region A can be reflected in the heating control of the reaction region A located on the right side of the temperature measuring body S.

  Temperature measuring bodies S are arranged as temperature sensing devices between the heater units, and the temperature measuring bodies S can detect the degree of influence of thermal interference. And it is desirable for the temperature measuring element S to be arranged on the scanning direction side of the corresponding heating element. Since heating control of each heating element is sequentially performed along the scanning direction, the heating is sequentially performed along this direction (see the arrow in FIG. 3).

  By arranging in a matrix like this, the heating operation of each heating unit can be stopped in units of scanning lines, and as a result, the temperature can be easily and quickly lowered. This is particularly remarkable when the reaction region A is a micro space. Further, since the heat generation time can be controlled, minute heat generation control can be easily performed.

  In addition to these, in the present invention, since the above-described thermal interference detection is performed using the temperature measuring element S or the like, the temperature detection can be performed with high accuracy, and the writing amount of the heating amount can be corrected. Furthermore, in addition to the thermal interference detection, the temperature of the reaction region A can be detected using the temperature measuring element Hs or the like, so that temperature control can be performed with higher accuracy.

  And about the reaction area | region A, the heating element H as said heat source and the temperature measuring body Hs used as said temperature detection part can be integrated in the same circuit shape. Furthermore, the temperature measuring body S can also be incorporated on the same circuit. As a result, these can be collectively controlled on the same circuit, which can contribute to the miniaturization of the device configuration.

  For example, by using at least a PIN diode as the temperature sensing element S and also using a PIN diode as the temperature sensing element Hs, scan sensing is performed in the column direction, and correction is performed for lin at the time of the next current writing. It is possible to perform heating control (current control) while controlling the degree of influence of thermal interference in real time.

  According to the present invention, even when the reaction areas A are arranged in a matrix, it is not necessary to lengthen the installation interval between the reaction areas A in order to eliminate the influence of thermal interference. As a result, it is possible to save more space as a device, and as a result, it can be used as a reaction processing apparatus that can perform comprehensive analysis more efficiently.

  FIG. 4 is a schematic side view of a third embodiment of the reaction processing apparatus according to the present invention. The code | symbol 3 in FIG. 4 has shown the reaction processing apparatus which concerns on this invention. The size and layer structure of the reaction processing apparatus 3 can also be appropriately selected according to the purpose, and the configuration of the reaction processing apparatus 3 can also be designed or changed within the scope of the object of the present invention. The reaction processing apparatus 3 is further characterized by having an optical measurement system.

  A configuration of the reaction processing apparatus 3 will be described. The reaction processing apparatus 3 includes a reaction substrate 31, a heating unit 32 that heats the reaction substrate 31, a Peltier element 33, a light source 34, and a guide that guides the light L 1 and L 2 emitted from the light source 34 to the reaction substrate 31. An optical plate 35, a light detection unit 36 that detects the measurement target light L3, a filter film 37 that transmits only light of a specific wavelength, and a measurement substrate 38 are provided. The heating unit 32 includes at least a thermal interference detection unit 321.

  For the optical measurement system that can be performed by the reaction processing apparatus according to the present invention, various spectroscopic measurements can be adopted, and the measurement target light is not limited. For example, a suitable spectroscopic measurement such as fluorescence measurement, scattered light measurement, IR measurement, UV measurement, etc. can be adopted as appropriate. For example, if fluorescence measurement is performed in the reaction processing apparatus 3, the sample is labeled with a fluorescent substance, excitation light having a specific wavelength is irradiated from the light source 34, and the fluorescence emitted by this is detected by the light detection unit 36. it can.

  The reaction substrate 31 includes a plurality of reaction regions A1, and a predetermined reaction can be performed in the reaction region A1. In the present invention, the shape or the like of the reaction region A1 is not particularly limited, and can be suitably set to a suitable shape and capacity.

  The volume of the reaction region A1 is not particularly limited, but is preferably a micro space, and is preferably 1 μL or less. By using such a micro space, the amount of the reaction solution required for the reaction region A1 is small, so that temperature control and the like can be performed with high accuracy and the reaction time can be shortened.

  For example, when the reaction area A1 is 300 μm × 300 μm × 300 μm (reaction area capacity: 27 nL), even if about 40,000 reaction areas A1 are installed on the reaction substrate 31, a device having an area of about 6 cm square may be used. . Since the device can be miniaturized in this way, a number of reaction regions A1 equal to the number of human genes can be arranged on the reaction substrate 31 in a matrix, making exhaustive analysis easier and easier. (See, for example, FIGS. 1 and 2).

  The material or the like of the reaction substrate 31 is not particularly limited as long as it is a material that can transmit the light L1, and can be appropriately selected in consideration of the measurement purpose, processability, and the like. For example, a transparent resin such as polymethyl methacrylate (PMMA), glass, or the like can be used. If fluorescence detection is to be performed, a low fluorescent light emitting plastic or the like can be used as the material of the reaction substrate 31.

  The reaction substrate 31 may be detachable from the heating unit 32, the light detection unit 36, the filter film 37, and the like. Or although not shown in figure, the reaction substrate 31 and the heating part 32 are made into integral structure, and this may be made removable.

  The heating unit 32 heats each reaction region A1 in the reaction substrate 31. Thereby, the temperature control of the reaction zone A1 is performed individually. Furthermore, since the thermal interference detection unit 321 is also provided, the heating control of the heating unit 32 can be performed with high accuracy. By controlling the heating temperature and the heating time for each reaction region A1 by the heating unit 32, the amplification reaction and the like in the reaction region A1 can be controlled with higher accuracy. As the configuration of the heating 32 (and the thermal interference detection unit 321 and the like), the configuration described above can be adopted as appropriate.

  In the present invention, it is desirable to provide the Peltier element 33 in order to control the temperature of the reaction region A1. The temperature of the entire reaction region A1 can be controlled by the Peltier element, and the temperature of each reaction region A1 can be controlled more precisely in each heating unit 32. Thereby, it is possible to accurately control minute temperature variations and deviations in the reaction regions A1. Thus, by providing the Peltier element 33, temperature control can be performed easily. As a result, highly accurate temperature control can be performed.

  For example, some conventional reaction processing apparatuses such as a PCR apparatus perform temperature control using a thermal cycler or the like, and a gradient mechanism may be added to this. However, there is a problem that the temperature control of the sample in each reaction region cannot be performed individually and with high accuracy. On the other hand, in the reaction processing apparatus according to the present invention, such a problem can be solved by providing each of the reaction regions A1 with the heating unit 32 and further including the thermal interference detection unit 321.

  As an example, when the PCR reaction is performed by the reaction processing apparatus 3, it is necessary to control the temperature according to the steps of “thermal denaturation, annealing (primer hybridization), extension reaction” as a PCR cycle. For example, the temperature in the reaction region A1 can be maintained in advance at the lowest temperature of the PCR cycle (for example, 55 ° C.). The temperature cycle of the entire reaction region A1 can be controlled by the Peltier element 33, and the temperature variation and deviation of each reaction region A1 can be individually corrected and controlled by each heating unit 32. Thus, more accurate temperature control can be performed by performing temperature control of the whole reaction region A1 and temperature control of each reaction region A1.

  In the present invention, the light source 34 and the light guide plate 35 for introducing the light L1 and L2 into each reaction region A1 are used as optical means capable of irradiating the light L1 having a specific wavelength to all of the plurality of reaction regions A1. be able to.

  The light source 34 is not particularly limited as long as it emits light of a specific wavelength, and it is preferable to use a white or monochromatic LED or LD. Thereby, the light which does not contain an unnecessary ultraviolet-ray and infrared rays can be obtained easily. And the installation location of the light source 34 and the number of light sources are not particularly limited. That is, although not shown, a plurality of light sources 34 may be provided so as to correspond to each reaction region A1, and light L1 and L2 may be directly irradiated toward each reaction region A1 corresponding to each light source 34. In this case, for example, since each reaction area A1 can be directly irradiated with the light source 34, it is possible to uniformly irradiate all the reaction areas A with light L1 and L2 by taking a larger amount of light and individually controlling the amount of light. it can.

  The light guide plate 35 is for guiding the light L <b> 1 emitted from the light source 34 to each reaction region A <b> 1 in the reaction substrate 31. Light L <b> 1 emitted from the light source 34 is introduced into the spacer 351 inside the light guide plate 35. A reflective film 352 is provided on the light guide plate 35, and the light L2 can be introduced into the reaction substrate 31 by using, for example, a dichroic mirror.

  Further, in the present invention, it is desirable to provide a filter film 353 that transmits only the light beams L1 and L2 at the bottom of the light guide plate 35. Thereby, the light L2 can be efficiently extracted from the light emitted from the light source 34 and guided to the reaction region A1. As the filter film 353, for example, a polarizing filter can be used.

  The light detection unit 36 detects the measurement target light L3 emitted in response to the lights L1 and 2 irradiated to the reaction region A1. For example, in the case of performing fluorescence measurement, fluorescence emitted by excitation is detected and measured. In the present invention, the configuration of the light detection unit 36 is not limited, and for example, a photodiode can be used.

  The light detection unit 36 is formed on the same substrate as the heating unit 32, but may be formed on a different substrate, or the heating unit 32 and the light detection unit 36 may be stacked.

  In the present invention, it is desirable to provide a filter film 37 that transmits only the wavelength light of the measurement target light L3 between each reaction region A1 and each corresponding light detection unit 36. By providing a filter film 37 that transmits only light of a predetermined wavelength between the reaction region A1 and the light detection unit 36, the detected measurement target light L3 can be efficiently extracted, so that a more accurate analysis can be performed. Can be done. As the filter film 37, for example, a polarizing filter or the like can be used.

  Thus, since the reaction processing apparatus 3 according to the present invention has the light detection unit 36, real-time analysis is possible. And the light detection part 36 is individually provided corresponding to each reaction area | region A1, and it becomes possible for each reaction area | region A1 to detect individually and with high precision.

  The heating unit 32 and the light detection unit 36 can be provided on the measurement substrate 38. In the present invention, the position where the measurement substrate 38 is provided is not limited to the position below the heating unit 32 or the light detection unit 36, and can be appropriately provided at a suitable location. The material of the measurement substrate 38 used in the present invention is not particularly limited, and for example, a glass substrate or various resin substrates can be used.

  Furthermore, in the reaction processing apparatus according to the present invention, it is desirable to use a TFT substrate having optical transparency. For example, when sensing a predetermined reaction (for example, a biological reaction) by photodetection, in order to construct a photodetection unit in the vicinity of the reaction region, it is more desirable to configure a TFT substrate having optical transparency. Factors that block the target light (for example, fluorescence detection) can be suppressed. And since the restriction | limiting with respect to the structure layout of a photon detection part is relieve | moderated, it can contribute greatly also to compactization as an apparatus device. Furthermore, when a circuit configuration for executing thermal control on a TFT substrate is used, for example, by forming a current mirror circuit, a current copy circuit, or the like, variation in characteristics (of each heat source) can be suppressed and high accuracy can be achieved. The advantage that the current limitation for supplying the calorific value can be realized can also be realized.

  Usually, when light to be measured such as fluorescence is detected on the upper and lower portions of a thin film transistor substrate (TFT substrate) as a heating means, a heater matrix is formed on a relatively large transparent insulating substrate having light transmittance. It can be considered to constitute This has the advantage of being excellent in terms of manufacturing cost and manufacturing process. However, the TFT has a problem that manufacturing variation and change with time are larger than those of a single crystal semiconductor element. However, since the reaction processing apparatus according to the present invention can perform temperature control in consideration of thermal interference and the like, such a problem can be solved.

  FIG. 5 is a conceptual view of a fourth embodiment of the reaction processing apparatus according to the present invention as viewed from the side. Hereinafter, differences from the above-described embodiments will be mainly described, and descriptions of common parts will be omitted.

  The reaction processing apparatus indicated by reference numeral 4 in FIG. 5 includes a reaction substrate 41, a heating unit 42 for heating the reaction substrate 41, a Peltier element 43, a light source 44, and light L1 and L2 on the reaction substrate 41. The light guide plate 45 to guide, the light detection part 46 which detects the measurement object light L3, and the measurement board | substrate 47 are provided. The heating unit 42 includes a thermal interference detection unit 421.

  This reaction processing apparatus 4 is common to the third embodiment and the like in that a heating unit 42 and a light detection unit 46 are provided for each reaction region A2. However, there is a difference in that the measurement target light L3 is detected by irradiating the light L2 from below the reaction substrate 41 and reflecting it within the reaction region A2. Here, the shape of the reaction region A2 has a curved surface portion for reflecting the measurement target light L3.

  In the reaction processing apparatus 4, the light L <b> 1 emitted from the light source 44 is irradiated to the reaction region A <b> 2 by the light guide plate 45. In the light guide plate 45, the light L 1 passes through the spacer 451, and the light L 2 is introduced into the reaction substrate 41 by the reflective film 452 and the filter film 453. And the measurement object light L3 is emitted by irradiating the sample etc. in the reaction liquid in reaction area | region A2 with this light L2. The measurement target light L3 is reflected by the wall surface in the reaction region A2, and is detected and measured by the light detection unit 46 provided below the reaction region A2.

  The reaction treatment apparatus according to the present invention described so far can be used in a wide range of applications because it can control the temperature with high accuracy in a plurality of reaction regions. For example, it performs various chemical reactions and biochemical reactions. It can be used as a possible reaction processing apparatus. Among these, in particular, it can be suitably used as a reaction processing apparatus for performing a gene amplification reaction.

  In order to carry out the gene amplification reaction, it is necessary to amplify the target gene. At this time, highly accurate temperature control is required. If the temperature control is insufficient, problems other than the target gene may be amplified or a uniform amplification rate may not be obtained. In contrast, according to the reaction processing apparatus of the present invention, high-precision temperature control can be performed, comprehensive analysis is possible, and it is possible to contribute to the reduction of the required sample volume. Is preferred.

  In the present invention, various methods can be performed as a gene amplification reaction, and in particular, it can be suitably used in the case of a PCR (polymerase chain reaction) method. The PCR method is a method for amplifying a small amount of DNA, and uses a heat-resistant DNA synthase to perform the reaction by utilizing the fact that the structure of DNA as a substrate is changed by heating. For example, (1) target DNA to be amplified, (2) at least two oligonucleotide primers that specifically bind to the target DNA, (3) buffer, (4) enzyme, (5) dATP, dCTP, dGTP , By using a deoxyribonucleotide triphosphate such as dTTP, etc., and repeating the cycle of “thermal denaturation → annealing (primer hybridization) → extension reaction” to amplify the target DNA to a desired amount, etc. it can.

  At this time, in order to control gene amplification with high accuracy, control of the reaction temperature is important. Of course, a so-called RT-PCR reaction (Reverse Transcriptase Polymerase Chain Reaction) can be performed in the same manner.

  In the PCR method, a temperature cycle of “95 ° C. (thermal denaturation) → 55 ° C. (primer hybridization) → 72 ° C. (DNA extension)” is performed, and for each cycle, the amount of cDNA in each reaction region is doubled. It will be amplified. And the temperature in each reaction region can be controlled to the optimal value of the designed primer reaction by the heating part 32 installed in each reaction region. In addition, since the primer hybridization time and the polymerase reaction time can be controlled, generation of unnecessary reaction byproducts can also be controlled. As a result, since the amplification rate of the gene (cDNA) in each reaction region can be made constant, a highly accurate PCR reaction can be performed.

  That is, according to the present invention, the difference in nucleic acid amplification rate in each reaction region can be corrected by individually controlling the reaction temperature in each reaction region. Thereby, the reaction efficiency of the nucleic acid amplification reaction in each reaction region can be made constant. As a result, even when a plurality of nucleic acid amplification reactions are performed in a plurality of reaction regions, the amount of nucleic acid amplification in each reaction region can be quantitatively known from one calibration curve. Therefore, the nucleic acid amplification apparatus according to the present invention can quantitatively analyze the expression level.

  Furthermore, even when the appropriate reaction temperature condition is unknown when performing a predetermined nucleic acid amplification reaction, the reaction efficiency of the nucleic acid amplification reaction is fed back by feeding back the appropriate reaction temperature from the initial result of the amplification reaction. Can be made constant. First, the correlation between the amount of amplification and the temperature at that time is detected within a fixed time from the start of the amplification reaction. Then, by feeding back the result to the temperature control mechanism of the heating unit, it is possible to determine an appropriate reaction temperature condition for performing the amplification reaction.

  In a general real-time PCR apparatus, a reaction time of 25 to 30 minutes is required to perform about 30 cycles consisting of “thermal denaturation → annealing → extension reaction”. At that time, the temperature is controlled at about 2 ° C./second. On the other hand, in the apparatus of the present invention, the temperature can be controlled at 20 ° C. or more / second, so the time can be shortened by about 40 seconds per cycle, and the reaction time of about 25 minutes or less in the entire 30 cycles. Can be achieved.

  In addition, according to the present invention, since a heating unit and a light detection unit can be installed for each reaction region (see FIGS. 4 and 5 etc.), the heating temperature and the heating time can be controlled independently for each reaction region. And can be detected individually. As a result, the expression level of genes and the like can be analyzed with high accuracy and in real time in a short time.

1 is a conceptual diagram of a first embodiment of a reaction processing apparatus according to the present invention. It is a conceptual diagram for demonstrating the structure of the heating part used in this invention. It is a conceptual diagram of 2nd Embodiment of the reaction processing apparatus which concerns on this invention. It is a schematic side view of 3rd Embodiment of the reaction processing apparatus which concerns on this invention. It is a schematic side view of 4th Embodiment of the reaction processing apparatus which concerns on this invention.

Explanation of symbols

1, 2, 3, 4 Reaction processing device 31, 41 Reaction substrate 32, 42 Heating unit 321, 421 Thermal interference detection unit 33, 43 Peltier element 34, 44 Light source 35, 45 Light guide plate 36, 46 Light detection unit 37 Filter film 38, 47 Measurement board H Heat source Hs Temperature sensor S Temperature sensor A Reaction region

Claims (7)

A plurality of reaction regions, and a heating unit provided for each reaction region,
The heating unit is
A heat source,
A thermal interference detector for detecting thermal interference between reaction regions;
Heating control means for controlling the heating amount of the heat source based on the temperature information detected by the thermal interference detector;
A reaction processing apparatus comprising at least   The reaction processing apparatus according to claim 1, wherein the thermal interference detection unit detects thermal interference between the reaction regions by a temperature measuring body arranged between adjacent reaction regions. The plurality of reaction regions and the heating parts corresponding to the reaction regions are arranged in a matrix, respectively.
Each heat source of the heating unit is a heating element connected to a scanning line and a data line, and the heating element can generate heat by sequentially scanning, and each temperature measuring body of the heating unit corresponds to The reaction processing apparatus according to claim 2, wherein each of the heating elements is arranged on a scanning direction side.   4. The reaction processing apparatus according to claim 3, wherein the temperature measuring body includes at least a PIN diode.   The heating unit further includes a temperature detection unit that measures the temperature of the reaction region, and the heating control unit includes temperature information detected by the thermal interference detection unit, and temperature information detected by the temperature detection unit. The reaction processing apparatus according to claim 1, wherein the heating amount of the heat source is controlled based on the above. Optical means for irradiating the reaction region with light of a predetermined wavelength;
A light detection unit capable of detecting the measurement target light generated by the light irradiation;
The reaction processing apparatus according to claim 1, further comprising:   The reaction processing apparatus according to claim 6, wherein the reaction processing apparatus is a gene amplification apparatus that performs a gene amplification reaction in the reaction region.

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