PCR device

Abstract

A polymerase chain reaction (PCR) apparatus comprising: a transport module including a carrier; a track that is circular and enables the carrier to move along the track; at least one tube holder disposed on the carrier and removably including at least one reaction tube; and a temperature supply module including at least one temperature supply block for adjusting the temperature of the at least one reaction tube.

Description

Technical Field

[0001] The present disclosure relates to an experimental reaction apparatus and a reaction method, and particularly to a polymerase chain reaction (PCR) apparatus that reduces the waiting time for pre-reaction loading of a sample, makes the total heat supply time common, and makes the heat medium common.

Background Art

[0002] The polymerase chain reaction (PCR) technique involves repeatedly heating and cooling a sample solution containing nucleic acid, thereby exponentially increasing the nucleic acid having a specific base sequence region. Techniques for increasing nucleic acid are usually applied to life science, genetic engineering, and medicine for analysis and diagnosis.

[0003] To meet the needs of users, various PCR apparatuses are commercially available. In a basic laboratory, usually, the collected sample is added to a micro test tube, and then the micro test tube is placed in a PCR apparatus, and the sample undergoes a cycle of simultaneously and collectively increasing nucleic acid. Therefore, these types of batch reaction apparatuses are suitable for academic research that not only requires a large number of samples but also requires comparison and analysis of multiple samples.

[0004] However, medical tests performed for clinical treatment often face emergencies, requiring immediate test results to determine necessary treatment (STAT) actions. When samples are placed on a bench to initiate the reaction, and the reaction is performed in batches using a PCR instrument, the supplied STAT samples must wait their turn to be loaded for the reaction, resulting in a significant delay in obtaining STAT test results. Specifically, the total time required to obtain test results includes the time it takes for the STAT sample to initiate the reaction and the time it spends waiting its turn. In severe scenarios, the test results may not be received in time, delaying the treatment given to the patient owner of the sample. Furthermore, while STAT samples are often in urgent cases, they are rarely supplied simultaneously. Therefore, even though it involves reduced instrument efficiency and increased power consumption, only a small number of samples can be loaded to initiate the reaction performed in batches using a PCR instrument, from the perspective of obtaining test results as quickly as possible.

[0005] As mentioned above, obtaining test results for STAT samples is urgent, and since STAT samples arrive at various points in time, it is not possible to load all samples together once all sample collection is complete. To address these problems, conventional PCR devices are provided that have multiple sample chambers for holding and reacting samples separately. Specifically, it is possible not only to add samples to multiple sample chambers at different times, but also to set various heat supply temperatures, heat supply times, or the number of heat supply cycles depending on when the sample is added to the sample chamber and the type of reaction the sample undergoes. However, the large PCR devices described above in the disclosed prior art are equivalent to combining multiple small PCR devices. Since the multiple small PCR devices perform reactions separately, have different heat supply blocks, and have different heat supply modes, the multiple small PCR devices do not have a common heat supply module, resulting in the output of the large PCR device becoming too large. In addition, because the multiple small PCR devices perform reactions separately, quality control of them is difficult.

[0006] Therefore, it is essential to provide PCR equipment that allows for faster loading of reaction samples into the bench, reduces costs, and facilitates sample quality control. [Overview of the project]

[0007] In consideration of the aforementioned shortcomings of the prior art, the object of this disclosure is to provide a polymerase chain reaction (PCR) apparatus comprising at least one holder on which at least one reaction tube is separately and detachably arranged, thereby eliminating the need to arrange multiple reaction tubes on at least one holder for each batch. Furthermore, in order to enable the reaction tubes to share a heat source and thus reduce power consumption, at least one holder is arranged on a transport module for driving the reaction tubes on the holder to move synchronously, forming a cyclic device.

[0008] To achieve the above and other objectives, one embodiment of this disclosure is provided. A transport module, The bench and, A loop-shaped rail that allows the bench to move along the rail, A bench is placed on the bench and allows at least one reaction tube to be detachably positioned in at least one holder and A transport module including, A heat supply module including at least one heat supply block for regulating the temperature of at least one reaction tube We provide a PCR device equipped with the following features.

[0009] Preferably, the number of holders is 1 to 100.

[0010] Preferably, the heat supply module further comprises a heat transfer medium and an output unit.

[0011] Preferably, the output unit stores and / or outputs a heat transfer medium, the heat supply block regulates the temperature of the heat transfer medium, and the output unit is connected to the heat supply block by a pipeline to allow the flow of the heat transfer medium.

[0012] Preferably, the heat transfer medium is a gas or a liquid.

[0013] Preferably, some of the multiple heat supply blocks share the same output section.

[0014] Preferably, the heat supply module further comprises a valve for controlling contact between the heat transfer medium and at least one reaction tube.

[0015] Preferably, the PCR apparatus further comprises a control module for controlling the operating mode of the transport module and the heat supply mode of the heat supply module.

[0016] Preferably, the operating mode of the transport module includes the number of stations on which the first holder stops, and the stop time during which the first holder remains at each station before returning to its initial position at the end of one cycle of the transport module's operation.

[0017] Preferably, the number of stations at which at least one holder stops is between 0 and 100.

[0018] Preferably, the downtime during which at least one holder remains at each station is 1 to 300 seconds.

[0019] Preferably, at least one holder stops for the same number of stations for the same downtime.

[0020] Preferably, one PCR cycle is performed while at least one holder remains at each station.

[0021] Preferably, the heat supply mode includes a heat supply temperature and a heat supply time.

[0022] Preferably, the heat supply temperature is 92 - 96°C, 45 - 70°C, and 67 - 77°C, respectively, in the denaturation stage, annealing stage, and extension stage during one PCR cycle.

[0023] Preferably, the total heat supply time in the denaturation stage, annealing stage, and extension stage during one PCR cycle is 1 - 300 seconds.

[0024] Preferably, the heat supply module has at least one heat supply mode.

[0025] Preferably, the heat supply module adjusts the temperature of the first part of the holder at the first time point in the first heat supply mode, adjusts the temperature of the second part of the holder in the second heat supply mode, and at least one of the heat supply temperature and heat supply time is different between the first heat supply mode and the second heat supply mode.

[0026] Preferably, the PCR device further includes a drive module for supplying power required to drive the bench.

[0027] Preferably, the PCR device further includes an inspection module for inspecting the product content in at least one reaction tube.

[0028] The foregoing and other objects, features, and advantages of the present disclosure will become apparent by referring to the following detailed description, preferred embodiments, and the accompanying drawings.

Brief Description of the Drawings

[0029] To explain the present disclosure and enable those skilled in the art to fully understand the disclosed method for implementing the present disclosure, many specific details are provided in the following detailed description. However, clearly, one or more methods of implementing the present disclosure can be carried out without the aforementioned specific details. In other situations, well-known structures and process flows are schematically depicted in the accompanying drawings for the sake of brevity.

[0030] [Figure 1] This is a cross-sectional view of the PCR apparatus in this disclosure.

[0031] [Figure 2] This is a top view of the PCR apparatus disclosed herein.

[0032] [Figure 3] This is a schematic diagram of the heat supply module of the PCR apparatus of this disclosure. In the heat supply module shown in Figure 3, one holder is connected to one heat supply block. Figure 3 shows two heat supply blocks whose temperature changes to supply heat to two corresponding reaction tubes, respectively. Other heat supply blocks, holders, and reaction tubes are not shown in Figure 3.

[0033] [Figure 4] This is a schematic diagram of the heat supply module of the PCR apparatus of the present disclosure. In the heat supply module shown in Figure 4, one holder is connected to one heat supply block. Figure 4 shows that multiple heat supply blocks within each of the two heat supply block groups, although each having a different temperature, do not change in temperature and each supply heat at a first temperature (4°C) to the corresponding reaction tube. Other heat supply blocks, holders, and reaction tubes are not shown in Figure 4.

[0034] [Figure 5] This is a schematic diagram of the heat supply module of the PCR apparatus of this disclosure. In the heat supply module shown in Figure 5, multiple holders are connected to the same group of heat supply blocks. Figure 5 shows that although the multiple heat supply blocks within one group of heat supply blocks have different temperatures, their temperatures remain constant, supplying heat at a first temperature (4°C) to two reaction tubes. Other heat supply blocks, holders, and reaction tubes are not shown in Figure 5.

[0035] [Figure 6]This is a schematic diagram of the heat supply module of the PCR apparatus of this disclosure. In the heat supply module shown in Figure 6, multiple holders are connected to the same heat supply block group, and multiple output units are connected to different heat supply blocks. In Figure 6, one heat supply block group is the same as in Figure 5, but it is shown that it supplies heat at a second temperature (60°C) to two reaction tubes. Other heat supply blocks, holders, and reaction tubes are not shown in Figure 6.

[0036] [Figure 7] This is a schematic diagram of the heat supply module of the PCR apparatus of this disclosure. In the heat supply module shown in Figure 7, multiple holders are connected to the same heat supply block group. In Figure 7, one heat supply block group is the same as in Figure 5, but it supplies heat at a first temperature (4°C) to the first reaction tube (left side in the figure) and heat at a second temperature (60°C) to the second reaction tube (right side in the figure). Other heat supply blocks, holders, and reaction tubes are not shown in Figure 7. [Modes for carrying out the invention]

[0037] Embodiments of this disclosure are illustrated by the accompanying drawings and described in detail below. While embodiments can be implemented in any form, they are not the only forms in which specific embodiments of this disclosure can be implemented or applied. Therefore, the aforementioned forms relating to implementing specific embodiments of this disclosure should not be construed as limiting the aforementioned embodiments. Methods of implementing this disclosure encompass the features of a specific embodiment, the steps of a method for constructing and operating a specific embodiment, and the sequence of steps of such method. However, identical or equivalent functions and sequences of steps can also be achieved by any other specific embodiment. In contrast, the purpose of embodiments is to fully and completely disclose this disclosure and to adequately explain its essence to those skilled in the art. Similar reference numerals used in the accompanying drawings indicate similar components. In the following description, prior art functions or structures are not described in detail to avoid repetition of details unnecessary to the embodiments.

[0038] Unless otherwise defined, all technical terms or specialized terms used herein have the same meaning as those commonly understood by those skilled in the art. In the event of any conflict between the two, the definition contained in this specification shall prevail.

[0039] Unless otherwise inconsistent with the context, all singular nouns used herein encompass their plural forms, and all plural nouns used herein encompass their singular forms. The expressions “at least one” and “one or more” as used herein and in the appended claims are synonymous, meaning one, two, three or more.

[0040] To broaden the scope of the claimed disclosure, approximations are applied to all numerical ranges and parameters, although relevant numerical values ​​used in specific embodiments are expressed as accurately as possible herein. However, all numerical values ​​inherently and necessarily involve a standard deviation resulting from the relevant testing methods. As used herein, the term “around” typically refers to a particular numerical value or range, indicating that the actual numerical value or range is 10%, 5%, 1%, or 0.5% greater or less than the given numerical value or range. Alternatively, the term “around” suggests that the actual numerical value has an acceptable error of the mean, depending on the considerations taken into account by those skilled in the art. In addition to embodiments, or unless otherwise specified, all ranges, quantities, numerical values ​​and percentages used herein (e.g., descriptions of required material quantities, downtime, temperature, operating requirements, ratios, etc.) are modified by the term “around.” Thus, unless otherwise specified, all numerical values ​​and parameters disclosed herein and in the appended claims are approximate and may be modified as necessary. Numerical values ​​and parameters must be characterized by at least specific significant figures and interpreted as values ​​obtainable through a common carry system. In this regard, numerical ranges are expressed as starting from one endpoint and ending at another, or as being between two endpoints. Unless otherwise specified, all numerical ranges include the endpoints.

[0041] One embodiment of the present disclosure provides a polymerase chain reaction (PCR) apparatus comprising a transport module and a heat supply module. In one embodiment of the present disclosure, the transport module comprises a rail, a bench, and at least one holder.

[0042] In one embodiment, the rail is loop-shaped. A loop-shaped rail is a rail in which the start and end points of the rail are in the same position. In one embodiment, a bench moves along the rail. At least one holder is positioned on the bench and holds the reaction tube, and therefore the reaction tube moves along the rail together with the bench. The loop-shaped rail of the present disclosure allows the reaction tube to be positioned to correspond to different heat supply blocks as it moves along the rail, thereby achieving temperature control. In one embodiment, the rail is functionally divided into a reaction zone and a temporary storage zone. The reaction takes place in the reaction tube while it is passing through the reaction zone, but no reaction takes place while it is passing through the temporary storage zone. The temporary storage zone is further divided into a first temporary storage zone in which a sample can be placed to undergo a reaction, and a second temporary storage zone in which a sample that has finished reacting can be removed.

[0043] Conventional batch devices contain many samples to be reacted in batches in order to obtain results for many samples in a short period of time. However, new sample batches must wait their turn until the previous sample batch has completed several to tens of PCR cycles, and as a result, new sample batches cannot start reacting as soon as possible or immediately after the sample arrives. If the sample is a STAT sample, conventional batch devices require a lot of waiting time, delaying the acquisition of results. To address STAT samples, this disclosure provides holders, each containing a small number of samples. Thus, according to this disclosure, STAT samples can be loaded onto the bench and reacted as quickly as possible, provided that the number of samples in each batch is reduced, or even reduced to one.

[0044] In one embodiment, at least one holder is placed on the bench. In a preferred embodiment, the holders placed on the bench are spaced equally apart from each other. In one embodiment, the number of holders is 1 to 100. In one embodiment, the transport module has 60 holders, each holding one reaction tube, and therefore 60 reaction tubes are placed within the transport module. Thus, as many as 60 reaction tubes are undergoing PCR reactions in the PCR device simultaneously. Preferably, the number of holders is approximately 1, 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, or 100, depending on the number of samples undergoing PCR reactions in the PCR device.

[0045] In one embodiment, at least one reaction tube is detachably disposed in at least one holder. In a preferred embodiment, the holder has fixing elements that can fix the reaction tube in place and prevent the reaction tube from detaching from the holder or from vibrating within the holder. In one embodiment, the disclosure does not restrict the direction in which the reaction tube enters the transport module. In a preferred embodiment, for convenience, the reaction tube enters the transport module from the upper side of the transport module and is placed on or fixed to a holder on the transport module. In one embodiment, the reaction tube is inserted into the holder.

[0046] In one embodiment, the heat supply module is located on at least one side of the transport module, e.g., the top, bottom, left, and / or right side, but the disclosure is not limited thereto. In a preferred embodiment, for convenience, the heat supply module is located on the opposite side of the transport module and in a position corresponding to the reaction tube. In a specific embodiment, if the reaction tube enters the transport module from the top side of the transport module, the heat supply module is located on the bottom side of the transport module. In a feasible embodiment, the heat supply module is located in the center of the transport module and supplies heat to the holder completely and efficiently, thereby regulating the temperature of the reaction tube. In one embodiment of the disclosure, the heat supply block regulates the temperature of at least one reaction tube. For the heat supply module to supply heat to a reaction tube, it is necessary to supply heat at a temperature higher than the initial temperature of the reaction tube to raise its temperature, and to supply heat at a temperature lower than the initial temperature of the reaction tube to lower its temperature. Thus, the heat supply module supplying heat to a reaction tube is not limited to heating the reaction tube to raise its temperature, but also includes cooling the reaction tube to lower its temperature.

[0047] In one embodiment of this disclosure, the operating modes of the heat supply module include a heat supply block type, a liquid-cooled type, and an air-cooled type. The three aforementioned operating modes of the heat supply module are described below.

[0048] Heat supply block type

[0049] In one embodiment of the present disclosure, the heat supply block system refers to the heating technique used by the heating blocks of conventional PCR modules. In the heat supply block system, the heat supply block and the holder must be in direct contact in order to supply heat to the reaction tube. A heat supply module of the heat supply block system comprises at least one heat supply block. In one embodiment, one holder on the rail is positioned to correspond to one heat supply block, and the heat supply block changes temperature to cause the reactants in the reaction tube to undergo multiple steps. In another embodiment, one holder on the rail is positioned to correspond to a group of heat supply blocks, and the group of heat supply blocks comprises multiple heat supply blocks that have different but constant temperatures, allowing one heat supply block to cause the reactants in the reaction tube to undergo one single step.

[0050] Regarding a heat supply block type heat supply module, the PCR apparatus further comprises a vertical movement module for raising and lowering a transport module. When the vertical movement module raises the transport module, the holders positioned on the transport module move away from the heat supply block so that the heat supply block does not come into contact with the holders. In contrast, when the vertical movement module lowers the transport module, the holders positioned on the transport module move toward the heat supply block, causing the heat supply block to come into contact with the holders. Unlike the aforementioned PCR apparatus which comprises a vertical movement module for raising and lowering a transport module, another PCR apparatus in another embodiment comprises a vertical movement module for raising and lowering a heat supply module. These two PCR apparatuses operate on the same principle, and the contact between each holder and the corresponding heat supply block depends on whether the vertical movement module is repositioning the transport module or the heat supply module. In a specific embodiment, the vertical movement module is a robotic arm for gripping holders and bringing them into contact with or away from the heat supply block, and the robotic arm comprises a main arm and a sub-arm for fine-tuning its movement and position.

[0051] liquid cooled

[0052] A liquid-cooled heat supply module comprises a heat transfer medium and an output unit. The heat transfer medium is a liquid, such as water, oil, any thermally conductive liquid medium, or a combination thereof, adapted to contact a holder. In one embodiment, the output unit stores and / or outputs the heat transfer medium. In one embodiment, the output unit is connected to a heat supply block by a pipeline, and the heat supply block is connected to a holder by a pipeline.

[0053] In one embodiment, a liquid-cooled heat supply module comprises a single output unit connected by pipelines to heat supply blocks of different temperatures. For example, if the number of holders is 60, the heat transfer medium output from the output unit passes through a first pipeline, a second pipeline, a third pipeline, ... a 60th pipeline, and comes into contact with the first heat supply block, a second heat supply block, a third heat supply block, ... a 60th heat supply block, respectively.

[0054] In another embodiment, a liquid-cooled heat supply module comprises multiple output units, each connected by pipelines to a heat supply block at the same temperature. For example, if there are 60 holders and 4 heat supply blocks (groups of heat supply blocks) at different temperatures, the heat transfer medium output from the four output units and passing through the first, second, third, and fourth pipelines contact the first, second, third, and fourth heat supply blocks, respectively. In a specific embodiment, the output unit is a reservoir. Multiple output units in a liquid-cooled heat supply module have the advantage that the heat transfer medium within the same output unit reliably moves to a heat supply block at the same temperature, preventing convergence of heat transfer mediums from different output units, and making it easier to recycle and output the heat transfer medium without requiring significant temperature adjustment.

[0055] A liquid-cooled heat supply module comprises at least one heat supply block. In one embodiment, one holder is connected by a pipeline to one heat supply block, which changes the temperature of the reactants in a reaction tube to carry out multiple processes. In another embodiment, one holder is connected by a pipeline to a group of heat supply blocks, which include multiple heat supply blocks with different but constant temperatures, and one heat supply block can carry out one single process of the reactants in a reaction tube. In yet another embodiment, multiple holders are connected by a pipeline to the same heat supply block or group of heat supply blocks, and thus share a heat supply block or group of heat supply blocks.

[0056] The liquid-cooled heat supply module further comprises a valve. A heat transfer medium adapted to contact an object of a specific temperature contacts at least one reaction tube under the control of the valve. In one embodiment, the valve is shaped similarly to a notched knob on a pepper shaker and can be opened and closed in a manner similar to turning a knob. When the notched portion of the knob is in a position corresponding to the pipeline, the heat transfer medium contacts the holder. In contrast, when the non-notched portion of the knob is in a position corresponding to the pipeline, the heat transfer medium cannot contact the holder. In one embodiment, the valve is shaped similarly to a spring-loaded marble and is opened and closed by pressing the marble. When the holder is in a position corresponding to the marble, the holder presses the marble downwards, and the heat transfer medium contacts the holder. In contrast, when the holder is not in a position corresponding to the marble, the marble springs up, thereby blocking the pipeline and preventing the heat transfer medium from contacting the holder. Depending on its type, the valve is selectively adjustable to open and close under the control of a control module.

[0057] In one embodiment, the liquid-cooled heat supply module includes a heating unit. The heating unit is positioned between a valve and a holder and has a shape similar to a washbasin sink. Specifically, a discharge port is located at the bottom of the heating unit, and the heat transfer medium is discharged from the discharge port and accumulated in the sink, allowing the holder to be immersed in the accumulated heat transfer medium. In one embodiment, a pump is positioned at one end of the output section, apart from the other pipeline connection end of the output section, in order to output the heat transfer medium under pressurization. In another embodiment, a vacuum device is positioned at one end of the heating unit, apart from the other pipeline connection end of the heating unit, in order to output the heat transfer medium under negative pressure.

[0058] In the liquid-cooled embodiment, the heat transfer medium is output from the output unit under pressure applied by the pump, flows through a pipeline to a heat supply block, the temperature of the heat transfer medium is regulated by the heat supply block, flows through a pipeline to a valve, the valve is opened to allow the heat transfer medium to enter the heating unit and come into contact with the holder, and the heat transfer medium is recycled and returned to the output unit.

[0059] Air-cooled

[0060] An air-cooled heat supply module comprises a heat transfer medium and an output unit. The heat transfer medium is a gas, such as air, a noble gas, or a combination thereof, adapted to be in contact with a holder. In one embodiment, the output unit stores and / or outputs the heat transfer medium. In one embodiment, the output unit is connected to a heat supply block by a pipeline, and the heat supply block is connected to a holder by a pipeline.

[0061] In one embodiment, the air-cooled heat supply module has one output unit, which is connected by pipelines to heat supply blocks of different temperatures (with the same total amount of airflow per unit time). For example, if the number of holders is 60, the heat transfer medium output from the output unit passes through the first pipeline, the second pipeline, the third pipeline, ... the 60th pipeline, and contacts the first heat supply block, the second heat supply block, the third heat supply block, ... the 60th heat supply block, respectively. The air-cooled heat supply module with a single output unit ensures that all heat transfer medium moved through the pipelines is of equal quantity and quality, thus contributing to quality control of the PCR instrument; that only one temperature control valve is ON at the same time in a single station; that the total number of valve openings and closings is the same; and that the total amount of airflow consumed per unit time is constant.

[0062] In another embodiment, the liquid-cooled heat supply module comprises multiple output units, each connected by pipelines to heat supply blocks of the same temperature. For example, if there are 60 holders and 4 heat supply blocks (groups of heat supply blocks), and the heat supply blocks are at different temperatures, four output units are provided to output a heat transfer medium, allowing it to pass through the first, second, third, and fourth pipelines, respectively, so that it contacts the first, second, third, and fourth heat supply blocks. In a specific embodiment, the output unit is a fan, which rotates to output a gas that functions as a heat transfer medium. In one embodiment, the fan is located at one end of the heat supply block, as opposed to the other holder connection end of the heat supply block.

[0063] The pneumatic heat supply module comprises at least one heat supply block. In one embodiment, one holder is connected by pipeline to one heat supply block, which changes its temperature to allow the reactants in the reaction tube to carry out multiple steps. In another embodiment, one holder is connected by pipeline to a group of heat supply blocks, which include multiple heat supply blocks with different but constant temperatures, allowing one heat supply block to allow the reactants in the reaction tube to carry out one single step. In yet another embodiment, multiple holders are connected by pipeline to one heat supply block or group of heat supply blocks, sharing one heat supply block or group of heat supply blocks.

[0064] The air-cooled heat supply module further comprises a valve. A heat transfer medium adapted to contact an object of a specific temperature contacts at least one reaction tube under the control of the valve. In one embodiment, the valve is shaped similarly to a notched knob on a pepper shaker and can be opened and closed in a manner similar to turning a knob. When the notched portion of the knob is in a position corresponding to the pipeline, the heat transfer medium contacts the holder. In contrast, when the non-notched portion of the knob is in a position corresponding to the pipeline, the heat transfer medium cannot contact the holder. In one embodiment, the valve is shaped similarly to a spring-loaded marble and is opened and closed by pressing the marble. When the holder is in a position corresponding to the marble, the holder presses the marble downward, allowing the heat transfer medium to contact the holder. In contrast, when the holder is not in a position corresponding to the marble, the marble springs up, thereby blocking the pipeline and preventing the heat transfer medium from contacting the holder. Depending on its type, the valve is selectively adjustable to open and close under the control of a control module.

[0065] In one embodiment, a pump is located at one end of the output section, apart from the other pipeline connection end of the output section, to output the heat transfer medium under pressurized conditions. In another embodiment, a vacuum device is located at one end of the heating unit, apart from the other pipeline connection end of the heating unit, to output the heat transfer medium under negative pressure. In a specific embodiment, the fan of the output section may be replaced by an air extractor or vacuum device. The air extractor or vacuum device is connected to a holder to draw or extract air from the holder end, allowing air at the heat supply block end to pass through the heat supply block before moving toward the holder.

[0066] In the air-cooled embodiment, the heat transfer medium is output from the output unit, flows through a pipeline to the heat supply block, the temperature of the heat transfer medium is regulated by the heat supply block, flows through the pipeline to a valve, the valve is opened to allow the heat transfer medium to contact the holder, and the heat transfer medium is discharged or recycled. An air-cooled heat supply module with a single output unit ensures that all heat transfer medium moved through the pipeline is of equal quantity and quality, thus contributing to quality control of the PCR instrument, that only one temperature control valve is ON at a time in a single station, that the total number of valve openings and closings is the same, and that the total airflow consumption per unit time is constant.

[0067] In one embodiment of the present disclosure, the air-cooled heat supply module further comprises a control module for controlling the operating mode of the transport module. The operating mode of the transport module includes the number of stations (specifically, the total number of stations) at which the first holder stops, and the stop time at which the first holder remains at each station before returning to its initial position at the completion of one cycle of the transport module's operation. In one embodiment, the number of stations at which at least one holder stops is between 0 and 100. In one embodiment, the stop time at which at least one holder remains at each station is between 1 and 300 seconds.

[0068] The transport module operates in a continuous mode, provided that the number of stations on which at least one holder stops is zero. Thus, the transport module operates continuously so that no holder stops at any station. If the number of stations on which at least one holder stops is a positive integer greater than zero, the transport module operates in an intermittent mode. Thus, the transport module's operation is paused, and holders remain stopped at stations. This disclosure does not limit the feasible range of the number of stations on which at least one holder can stop before the same holder returns to its initial position at the completion of one cycle of the transport module's operation. The number of stations on which at least one holder can stop is 1, 5, 10, 15, 20, 30, 40, 50, 60, ... 100.

[0069] In one embodiment of the present disclosure, at least one holder stops for the same amount of time at each station, for the same duration at each station. The present disclosure employs a cyclic design to enable all holders positioned on a transport module to move with the transport module in operation. Thus, the holders spontaneously stop for the same amount of time at each station, for the same duration at each station.

[0070] Each PCR cycle includes a denaturation step, an annealing step, and an expansion step, in which the reaction takes place at different temperatures. In one embodiment, at least one holder remaining in a station undergoes one step of the PCR cycle and therefore maintains the same temperature while remaining in the station. For example, at least one holder remaining in a station undergoes pre-PCR. In another embodiment, at least one holder remaining in a station undergoes multiple steps of the PCR cycle and therefore changes its temperature to different temperatures while remaining in the station.

[0071] In yet another embodiment, at least one holder undergoes one PCR cycle in total while remaining at one station, and thus its temperature changes to the different temperatures of all stages of the PCR cycle while remaining at the station. The number of stations a holder stops at before one cycle of the transport module is completed and the holder returns to its initial position is equal to the number of PCR cycles that the reaction tubes placed on the holder undergo. Because all holders move synchronously along the rails, not only is the time a holder spends at each station the same, but the holders also undergo the reaction for the same stopping time at each station. Thus, the number of stations a holder stops at before one cycle of the transport module is completed and the holder returns to its initial position is greater than the number of PCR cycles that the reaction tubes placed on the holder undergo.

[0072] In one embodiment of a heat supply block, liquid-cooled, or air-cooled system, if one holder is positioned to correspond to one heat supply block, then performing one PCR cycle requires changing the temperature of the heat supply block to execute different stages. However, as disclosed in one embodiment of this disclosure, if one holder is positioned to correspond to one or more groups of heat supply blocks, the heat supply module comprises a first heat supply block, a second heat supply block, and a third heat supply block, respectively, to supply the heat required for the denaturation, annealing, and expansion stages of one PCR cycle, with differences between the first, second, and third temperatures. Thus, the holder can be quickly switched between different temperatures. Unlike the embodiment where only one heat supply block is positioned to correspond to the holder, this embodiment saves time that would otherwise be spent changing the temperature of a single heat supply block to execute different stages.

[0073] In one embodiment of the present disclosure, the PCR apparatus further comprises a control module for controlling the heat supply mode of the heat supply module, the heat supply mode including heat supply time and heat supply temperature. The heat supply mode is adjustable according to the characteristics of the sample and nucleic acid polymerase. In one embodiment, the heat supply temperatures required for the denaturation, annealing, and expansion phases in the PCR cycle are 92-96°C, 45-70°C, or 67-77°C, respectively. In one embodiment, the heat supply times for the denaturation, annealing, and expansion phases are 0.25-75 seconds, 0.25-150 seconds, and 0.5-75 seconds, respectively, and the total heat supply time for one PCR cycle is approximately 1-300 seconds.

[0074] In one embodiment, the heat supply module has at least one heat supply mode. The heat supply module adjusts the temperature of a first portion of the holder in a first heat supply mode at a first time point, and adjusts the temperature of a second portion of the holder in a second heat supply mode. The first and second heat supply modes differ from each other in at least one of the heat supply temperature and heat supply time. Specifically, at different stages of the PCR cycle, the first holder requires a heat supply temperature of 95°C, 60°C, or 72°C and a heat supply time of 15 seconds, 30 seconds, or 15 seconds, respectively, while the 60th holder requires the same heat supply temperature as the first holder, but the heat supply time is adjusted and changed to 10 seconds, 20 seconds, or 30 seconds, respectively. This is because the sample in the first reaction tube, placed on the first holder, stops at the first station to undergo the first PCR cycle reaction, then moves to the next station with the transport module in operation to undergo the PCR cycle, and continues until it stops at the 59th station to undergo the 59th PCR cycle. Therefore, the amount of nucleic acid in the nucleic acid sample in the first reaction tube that will immediately stop at the 60th station is a multiple of the amount of nucleic acid in the initial sample in the first reaction tube that remains at the first station, and the heat supply time for the denaturation stage can be further reduced. The heat supply mode of some holders of the heat supply module of the PCR apparatus of this disclosure can be adjusted as needed to provide a third heat supply mode that is different from the first and second heat supply modes, for example, in at least one of the heat supply temperature and heat supply time.

[0075] In one embodiment, the PCR apparatus further comprises a drive module for supplying the power necessary to drive the bench. In a feasible embodiment, the drive module comprises a motor and a power transmission component. The motor outputs power, which is then transmitted to the bench by a power transmission component, provided in the form of gears or belts, so that the bench can rotate.

[0076] In one embodiment, the PCR apparatus further comprises a testing module for testing the product content in at least one reaction tube. In a feasible embodiment, the testing module is attached to a transport module for testing the product content in at least one reaction tube. In a preferred embodiment, a self-contained testing unit is attached to each reaction tube to monitor the product content in real time.

[0077] Embodiment

[0078] Figure 1 is a cross-sectional view of a PCR apparatus 1 of the present disclosure. The PCR apparatus 1 comprises a transport module 11, a reaction tube 12, and a heat supply module 13. The transport module 11 comprises a rail 111, a bench 112, and 12 holders 113. The bench 112 receives power from a drive module 15 to operate and move along the rail 111. The holders 113 are positioned on the bench 112. The reaction tube 12 is detachably positioned on the holders 113. Below the transport module 11 is the heat supply module 13. The heat supply module 13 comprises 12 heat supply blocks 131, a heat transfer medium 132, an output unit 133, and a valve 134. The valve 134 controls the contact between the heat transfer medium 132 and the reaction tube 12. The PCR apparatus 1 further comprises a control module 14 for controlling the operating mode of the transport module 11 and the heat supply mode of the heat supply module 13.

[0079] Referring to Figure 2, a top view of the PCR apparatus of this disclosure is shown. The PCR apparatus comprises 12 holders 113 arranged on a bench 112.

[0080] Referring to Figures 3, 4, 5, 6, and 7, schematic diagrams of the heat supply modes in the heat supply module 13 of the PCR apparatus of this disclosure are shown. As shown in Figure 3, the heat supply module 13 comprises two heat supply blocks 131, two heat transfer media 132, and one output unit 133, each heat supply block 131 being able to change to a different temperature. For illustrative purposes, the drawings show two holders, each holding a reaction tube (not shown), hereafter referred to as the first holder on the left and the second holder on the right. The four different temperatures to which the heat supply block 131 changes are, respectively, 4°C (for use in preserving amplified nucleic acid products), 60°C (for use in primer annealing), 72°C (for use in DNA polymerization), and 94°C (for use in DNA denaturation). The heat transfer media 132 are output collectively from the output unit 133 and then contact the heat supply blocks 131 at different temperatures via pipelines. The heat transfer medium 132 is temperature-controlled by the heat supply block 131 and then flows through the pipeline toward the holders. Two valves 134, located at positions corresponding to the first and second holders, open to allow the heat transfer medium 132, temperature-controlled by the heat supply block 131, to pass through.

[0081] As shown in Figure 4, the heat supply module 13 comprises two heat supply block groups, each having four heat supply blocks 131 with different temperatures, four heat transfer media 132, and one output unit 133. For illustrative purposes, the drawing shows two holders, each holding a reaction tube (not shown), hereafter referred to as the first holder (left) and the second holder (right). The four different temperatures to which the heat supply block 131 changes are 4°C (for storage of amplified nucleic acid products), 60°C (for primer annealing), 72°C (for DNA polymerization), and 94°C (for DNA denaturation), respectively. The heat transfer media 132 are output collectively from the output unit 133 and then come into contact with the heat supply blocks 131 at different temperatures via pipelines. After the heat transfer media 132 are temperature-regulated by the heat supply blocks 131, they flow through pipelines towards the holders. Two valves 134 located in positions corresponding to the first and second holders open to allow heat transfer medium 132, which is temperature-controlled by the heat supply block 131 at a temperature of 4°C, to pass through, but close to block heat transfer medium 132, which is temperature-controlled by the heat supply block 131 at other temperatures.

[0082] As shown in Figure 5, the heat supply module 13 comprises four heat supply blocks 131 with different temperatures, four heat transfer media 132, and one output unit 133. For illustrative purposes, the drawing shows two holders, each holding a reaction tube (not shown), hereafter referred to as the first holder on the left and the second holder on the right. The four different temperatures to which the heat supply blocks 131 change are 4°C, 60°C, 72°C, and 94°C. The heat transfer media 132 are output collectively from the output unit 133 and then come into contact with the respective heat supply blocks 131 at different temperatures via pipelines. After the heat transfer media 132 are temperature-regulated by the heat supply blocks 131, they flow through pipelines towards the holders. Two valves 134 located in positions corresponding to the first and second holders open to allow heat transfer medium 132, which is temperature-controlled by the heat supply block 131 at a temperature of 4°C, to pass through, but close to block heat transfer medium 132, which is temperature-controlled by the heat supply block 131 at other temperatures.

[0083] In comparison with Figure 5, Figure 6 shows a notable technical feature in which the heat supply module comprises four output units 133 connected to heat supply blocks 131 at different temperatures, and two valves 134 located in positions corresponding to the first and second holders, which open to allow the passage of heat transfer medium 132 temperature-regulated by the heat supply block 131 at 60°C, but close to block the passage of heat transfer medium 132 temperature-regulated by the heat supply block 131 at other temperatures.

[0084] In comparison with Figure 5, Figure 7 shows a remarkable technical feature in which the valve 134 located in the position corresponding to the first holder opens to allow the heat transfer medium 132 temperature-controlled by the heat supply block 131 at 4°C to pass through, but closes to block the heat transfer medium 132 temperature-controlled by the heat supply block 131 at other temperatures, and the valve located in the position corresponding to the second holder opens to allow the heat transfer medium 132 temperature-controlled by the heat supply block 131 at 60°C to pass through, but closes to block the heat transfer medium 132 temperature-controlled by the heat supply block 131 at other temperatures.

[0085] First Embodiment

[0086] The PCR instrument's transport module has 24 holders, and depending on the operation of the transport module, 24 samples can be sequentially placed in the holders to initiate the reaction. The operating mode is intermittent, and the holders stop at 24 stations, remaining at each station for 1 minute and 45 seconds. The heat supply module is air-cooled and consists of a fan that functions as an output unit and three heat supply blocks. The gas output from the fan acts as a heat transfer medium. The output unit, heat supply blocks, and holders are connected by pipelines to allow the flow of the heat transfer medium. Valves are placed on the pipelines between each heat supply block and the corresponding holder to control the contact between the holder and the gas. The gas output from the fan flows into three different channels and reaches each of the three heat supply blocks. The gas, heated by each heat supply block, then flows into 24 different channels and reaches each of the 24 holders. In heat supply mode, the three heat supply blocks require heat supply temperatures of 94°C, 60°C, and 72°C, respectively, and heat supply times of 20 seconds, 60 seconds, and 25 seconds, respectively. The heat supply mode is the same for each holder. The total heat supply time in heat supply mode is 1 minute and 45 seconds, synchronized with the dwell time in operating mode.

[0087] Second Embodiment

[0088] The PCR instrument's transport module has 48 holders, and depending on the operation of the transport module, 48 samples can be sequentially placed in the holders to initiate the reaction. The transport module operates in a continuous mode. Therefore, the transport module operates continuously so that the holders do not stop at any station. Each holder is connected to the heat supply module pipeline for 1 minute. The heat supply module is air-cooled and consists of a fan that functions as an output unit and three heat supply blocks. The gas output from the fan functions as a heat transfer medium. The output unit, heat supply blocks, and holders are connected by pipelines, allowing the heat transfer medium to flow. Valves are placed on the pipelines between each heat supply block and the corresponding holder to control the contact between the holder and the gas. The gas output from the fan flows into three different channels and reaches each of the three heat supply blocks. The gas, heated by each heat supply block, then flows into 48 different channels and reaches each of the 48 holders. In heat supply mode, the three heat supply blocks require heat supply temperatures of 95°C, 65°C, and 77°C, respectively, and heat supply times of 25 seconds, 55 seconds, and 20 seconds, respectively. The heat supply mode is the same for each holder. The total heat supply time in heat supply mode is 1 minute and 40 seconds, synchronized with the connection time with each holder in operating mode.

[0089] Third Embodiment

[0090] The PCR instrument's transport module has 60 holders, and depending on the operation of the transport module, 60 samples can be sequentially placed in the holders to initiate the reaction. The operating mode is intermittent, and the holders can stop at 60 stations, remaining at each station for 1 minute. The heat supply module is liquid-cooled and comprises three reservoirs that function as output units and three heat supply blocks. The liquid output from the reservoirs functions as a heat transfer medium. The output units, heat supply blocks, and holders are connected by pipelines, allowing the heat transfer medium to flow. Valves are placed on the pipelines between each heat supply block and the corresponding holder to control contact between the holder and the liquid. The liquid output from the reservoirs flows into three different channels, each reaching one of the three heat supply blocks. The liquid, heated by each heat supply block, then flows into 60 different channels, each reaching one of the 60 holders. In heat supply mode, the three heat supply blocks require heat supply temperatures of 92°C, 55°C, and 67°C, respectively, and heat supply times of 15 seconds, 30 seconds, and 15 seconds, respectively. The heat supply module for each holder is the same. The heat supply time in heat supply mode is synchronized with the dwell time in operating mode, totaling 1 minute.

[0091] Fourth Embodiment

[0092] The PCR instrument's transport module has 60 holders, and depending on the operation of the transport module, 60 samples can be sequentially placed in the holders to initiate the reaction. The operating mode is intermittent, and the holders stop at 60 stations, remaining at each station for 1 minute. The heat supply module is liquid-cooled and consists of three reservoirs that function as output units and three heat supply blocks. The liquid output from the reservoirs functions as a heat transfer medium. The output units, heat supply blocks, and holders are connected by pipelines, allowing the heat transfer medium to flow. Valves are placed on the pipelines between each heat supply block and its corresponding holder to control contact between the holder and the liquid. The liquid output from the reservoirs flows into four different channels and reaches each of the four heat supply blocks. The liquid, heated by each heat supply block, then flows into 60 different channels and reaches each of the 60 holders. In heat supply mode, the four heat supply blocks require heat supply temperatures of 95°C, 55°C, 72°C, and 4°C respectively. Holders 1 through 10 require a heat supply temperature of 95°C and a heat supply time of 1 minute to perform a hot-start step (hot-start PCR) in which polymerase is activated in the presence of high heat. Holders 11 through 55 require heat supply temperatures of 95°C, 55°C, and 72°C, and heat supply times of 15 seconds, 30 seconds, and 15 seconds, respectively. Each holder can complete 1 PCR cycle, for a total of 45 cycles. 56 Holder 60 requires a heat supply temperature of 4°C and a heat supply time of 1 minute. Holder 60 in this embodiment has a total of three different heat supply modes. The heat supply time in each heat supply mode is synchronized with the shutdown time in the operating mode, totaling 1 minute.

[0093] Therefore, the PCR instrument of this disclosure achieves the objective of loading a small number of samples into the PCR instrument and initiating the reaction as quickly as possible. Unlike conventional batch instruments, the cyclic PCR instrument provided by this disclosure allows holders to move between different stations as a result of the operation of the transport module, and while the holders are moving between stations, the samples in the reaction tubes placed on the holders can undergo PCR. The arriving samples are placed in the holders idling in the PCR instrument and only need to wait for the duration of one PCR cycle before the reaction begins.

[0094] This disclosure provides a transport module for sharing a heat supply block, heat transfer medium, and / or output section in order to save energy, reduce the required equipment volume, and save space.

[0095] The operation of the transport module allows the holders to move synchronously. Therefore, the time spent by the reaction tube for one PCR cycle is the same, which not only facilitates quality control but also makes it easier to record the number of completed cycles. Although the total time required for the reaction tube to perform one PCR cycle is the same, the mode of each step in the PCR cycle can be freely adjusted and controlled, thus meeting the diverse requirements of different samples.

[0096] The above description is illustrative and not limiting. Equivalent modifications or changes made by those skilled in the art to an embodiment preferred by the art without departing from the spirit of this disclosure shall be deemed to be included in the claims of this disclosure.

Claims

1. It is a PCR device, A transport module, The bench and, A loop-shaped rail that allows the bench to move along the rail, A holder, which is positioned on the bench and allows at least one reaction tube to be detachably positioned in the holder, and A transport module including, A plurality of stations arranged along the rail, each of which is provided to perform at least one PCR cycle while at least one of the reaction tubes is stopped at each of the plurality of stations, A heat supply module including at least one heat supply block for adjusting the temperature of at least one reaction tube, The system includes a control module for controlling the operating mode of the transport module and the heat supply mode of the heat supply module, The heat supply module has at least one heat supply mode, The heat supply module adjusts the temperature of a first portion of the holder at a first time point in a first heat supply mode, and adjusts the temperature of a second portion of the holder in a second heat supply mode, wherein the first heat supply mode and the second heat supply mode differ in at least one of the heat supply temperature and heat supply time. PCR device.

2. The number of holders is 1 to 100. The PCR apparatus according to claim 1.

3. The heat supply module further comprises a heat transfer medium and an output unit. The PCR apparatus according to claim 1.

4. The output unit stores and / or outputs the heat transfer medium, the heat supply block adjusts the temperature of the heat transfer medium, and the output unit is connected to the heat supply block by a pipeline to enable the flow of the heat transfer medium. The PCR apparatus according to claim 3.

5. The heat transfer medium is a gas or a liquid. The PCR apparatus according to claim 3.

6. Some of the aforementioned heat supply blocks share the same output section. The PCR apparatus according to claim 3.

7. The heat supply module further comprises a valve for controlling contact between the heat transfer medium and the at least one reaction tube. The PCR apparatus according to claim 3.

8. The operating mode of the transport module includes the number of stations on which the first holder stops, and the stop time during which the first holder remains at each of the stations until the first holder returns to its initial position at the end of one cycle of operation of the transport module. The PCR apparatus according to claim 1.

9. The number of stations at which the at least one holder stops is 1 to 100. The PCR apparatus according to claim 1.

10. The stop time during which at least one holder remains at each of the stations is 1 to 300 seconds. The PCR apparatus according to claim 8.

11. The at least one holder stops for the same amount of time at each of the stations. The PCR apparatus according to claim 1.

12. The heat supply mode includes a heat supply temperature and a heat supply time. The PCR apparatus according to claim 1.

13. The heat supply temperatures are sequentially 92-96°C, 45-70°C, and 67-77°C during the denaturation, annealing, and expansion phases of one PCR cycle, respectively. The PCR apparatus according to claim 12.

14. The total heat supply time for the denaturation, annealing, and expansion phases in one PCR cycle is 1 to 300 seconds. The PCR apparatus according to claim 12.

15. The bench further comprises a drive module for supplying the power necessary to drive the bench. The PCR apparatus according to claim 1.

16. The system further comprises an inspection module for inspecting the product content in at least one of the reaction tubes. The PCR apparatus according to claim 1.

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