Continuous-flow low-energy-consumption PCR (Polymerase Chain Reaction) chip

Abstract

The invention discloses a continuous flow low energy consumption PCR chip, which comprises a chip base, a chip body and a thermal resistance adjusting layer, a heater is arranged on the chip base, the heater is divided into a plurality of temperature zones, the temperatures of the plurality of temperature zones are different, the thermal resistance adjusting layer is located above the heater, a flow channel is arranged in the chip body, the flow channel is located above the thermal resistance adjusting layer, and the flow channel is located above the thermal resistance adjusting layer. Heat generated by the heater is transmitted to the chip body through the thermal resistance adjusting layer. The fluid flows in the flow channel, the fluid located in different areas of the flow channel obtains different heat to achieve temperature zoning of the fluid, the structure is reasonable, the thermal resistance adjusting layer is matched with the different temperature areas, the temperature of the fluid in the flow channel can be continuously changed in the continuous flowing process, and the temperature zoning effect of the fluid is improved. Therefore, a plurality of processes of denaturation, annealing and extension circulation processes can be completed by PCR under different temperature conditions.

Description

Technical Field

[0001] The invention relates to the technical field of PCR, and in particular to a continuous-flow low-energy-consumption PCR chip. Background Art

[0002] Polymerase chain reaction (PCR) is a technique for amplifying specific DNA or RNA fragments in an in vitro environment, and is commonly used in the fields of bacteria and virus detection, population research, and life detection. PCR usually requires repeated denaturation, annealing, and extension cycles at 95°C, 55°C, and 72°C.

[0003] In order to meet the requirement of raising and lowering the temperature between three temperature conditions during the cycle, existing PCR reaction devices usually use Peltier for bidirectional heating and cooling; some devices use heaters for unidirectional heating and cooling with radiators; some devices use multiple resistance heaters arranged in different heating areas to achieve temperature control through fluid circulation; and some devices use a single resistance heater to heat special-shaped parts to obtain different temperature areas, and achieve temperature control by wrapping pipes around the special-shaped parts for fluid circulation.

[0004] When conducting unattended detection in special environments, new requirements are put forward for energy consumption, space, heat dissipation conditions and stability. For the Peltier-type temperature control solution, although it can achieve rapid heating and cooling cycles, the Peltier needs to be used with a radiator during the cooling process, which requires additional space, and the heating and cooling processes will generate additional energy consumption. The use conditions also limit the form of the radiator. For the case of using a heater to heat up in one direction and a radiator to cool down, the cooling rate is slower than the Peltier solution, which reduces the energy consumption during the cooling process, but other problems are not significantly improved. For the solution of using multiple resistance heaters to generate different temperature control areas, the energy consumption during the heating and cooling cycle is reduced, but multi-channel temperature measurement and temperature control are required, and additional driving and conversion elements are added. For the solution of using a heater to heat the special-shaped part to generate a temperature gradient and then wrapping the pipe around the special-shaped part, the consistency problem of multiple contact surfaces will make it difficult to ensure temperature accuracy, lack of stability and large longitudinal space occupation. In response to the above problems, a solution is proposed below. Summary of the invention

[0005] The purpose of the present invention is to provide a continuous flow low-energy consumption PCR chip, which controls the temperature of different areas of the continuous flow PCR microfluidic chip by adjusting the heat generated by the heating element and the heat transfer resistance, so that the heat conduction in different areas matches the required heat.

[0006] The above technical objectives of the present invention are achieved through the following technical solutions: A continuous-flow low-energy consumption PCR chip, comprising a chip base, a chip body and a thermal resistance adjustment layer. A heater is provided on the chip base. The heater is divided into several temperature zones with different temperatures. The thermal resistance adjustment layer is located above the heater. A flow channel is provided below the chip body, and the flow channel is located above the thermal resistance adjustment layer. The heat generated by the heater is transferred to the chip body through the thermal resistance adjustment layer. There is fluid in the flow channel, and the fluid flows in the flow channel. The fluid in different regions of the flow channel obtains different amounts of heat to achieve heating of the chip body.

[0007] Preferably, the several temperature zones on the heater are respectively a denaturation zone, an annealing zone and an extension zone. The denaturation zone, the annealing zone and the extension zone are arranged in sequence from left to right. The section width of the denaturation zone is d1, the section width of the extension zone is d2, and the section width of the annealing zone is d3. d1 < d3 < d2, and the interval widths of the three regions are set according to the time required for the three stages of denaturation, annealing and extension.

[0008] Preferably, the flow channel is horizontally arranged above the thermal resistance adjustment layer, and the fluid will sequentially pass above the denaturation zone, the annealing zone and the extension zone during one flow.

[0009] Preferably, the heater is one of copper thin film, aluminum thin film or conductive polymer material.

[0010] Preferably, the material of the chip base is one of glass, quartz, polymer, aluminum alloy, stainless steel.

[0011] Preferably, the thermal resistance adjustment layer includes a conduction layer and a heat insulation layer. The conduction layer is located above the heat insulation layer. The thickness of the heat insulation layer above the denaturation zone is h1, the thickness of the heat insulation layer above the annealing zone is h2, and the thickness of the heat insulation layer above the extension zone is h3. h2 > h3 > h1.

[0012] The beneficial effects of the present invention are as follows: 1. The present invention proposes a scheme for controlling the temperatures of different regions of a continuous-flow PCR microfluidic chip by adjusting the heat generation amount of the heating element and the heat transfer resistance, so that the heat conduction in different regions matches the required heat, greatly reducing the attached cooling and measurement and control components, and solving the problems of PCR energy consumption, space occupation and heat dissipation under special conditions; 2. The continuous-flow PCR chip combined with the thermal resistance adjustment layer and the adjustable heating element has low energy consumption, little environmental impact, low requirements for the use environment and saves space; 3. The chip-type design is flexible and stable during use. Description of the Drawings

[0013] Figure 1It is a structural schematic diagram of an embodiment; Figure 2 is a schematic structural diagram of a heater in an embodiment; Figure 3 It is a schematic diagram of the structure of the embodiment for showing the temperature area of ​​the heater; Figure 4 Schematic diagram of fluid temperature distribution in the embodiment; Figure 5 Schematic diagram of fluid temperature and duration in an embodiment.

[0014] Figure numerals: 1. chip base; 2. chip body; 3. thermal resistance adjustment layer; 31. conduction layer; 32. thermal insulation layer; 4. heater; 41. denaturation zone; 42. annealing zone; 43. extension zone; 5. flow channel. DETAILED DESCRIPTION

[0015] The following is only a preferred embodiment of the present invention, and the protection scope is not limited to this embodiment. All technical solutions under the concept of the present invention should belong to the protection scope of the present invention. The same parts are represented by the same reference numerals.

[0016] like Figures 1 to 5 As shown, a continuous flow low energy consumption PCR chip comprises a chip base 1, a chip body 2 and a thermal resistance adjustment layer 3. The chip base 1 is made of one of glass, quartz, polymer, aluminum alloy and stainless steel, and is stable and not easily damaged.

[0017] A heater 4 is provided on the upper end surface of the chip base 1. The material of the heater 4 can be a metal film such as copper or aluminum, or a conductive polymer, etc., and it will generate heat when powered on. The heater 4 is divided into several temperature zones, and the zone width d of the heater 4 in different temperature zones is different. By adjusting the zone width d, different temperature zones can have different heating powers, so that different temperature zones can reach different temperatures at the same time.

[0018] The several temperature zones on the heater 4 are respectively a denaturation zone 41, an annealing zone 42 and an extension zone 43. The heater 4 is made of a copper film, an aluminum film or a conductive polymer material. The denaturation zone 41, the annealing zone 42 and the extension zone 43 are arranged in sequence from left to right. The section width of the denaturation zone 41 is d1, the section width of the annealing zone 42 is d2, and the section width of the extension zone 43 is d3. In this design, d1 is d2. The thermal resistance regulating layer 3 is located above the heater 4, and the thermal resistance regulating layer 3 includes a conductive layer 31 and a heat insulating layer 32. The conductive layer 31 can be made of copper, stainless steel or other materials with good thermal conductivity. The conductive layer 31 needs to be partially connected to the external environment to obtain a larger temperature gradient. The conductive layers are not connected to each other to prevent the temperatures corresponding to each temperature zone from being mixed in the conductive layer 31. The heat insulating layer 32 is made of ceramics, plastics, foamed cement and other materials with low thermal conductivity.

[0019] The thermal resistance adjustment layer 3 is also divided into corresponding adjustment zones according to different temperature zones of the heater 4. Different temperature zones on the heater 4 heat the adjustment zones of the thermal resistance adjustment layer 3 respectively. The thickness of the thermal insulation layer 32 used in different adjustment zones is also different. The thickness of the thermal insulation layer 32 located above the denaturation zone 41 is h1, the thickness of the thermal insulation layer 32 located above the annealing zone 42 is h2, and the thickness of the thermal insulation layer 32 located above the extension zone 43 is h3. h2>h3>h1. The specific thickness setting needs to match the heater and the material. In this design, h1:h2:h3 is 0.1:2:1.5.

[0020] The conductive layer 31 is located above the heat insulating layer 32, and the heat insulating layer 32 can smoothly transfer the heat generated by the heater 4 to the conductive layer 31, so that the temperature on the conductive layer 31 is stable and will not change significantly. The heater 4 and the thermal resistance adjustment layer 3 work together so that the conductive layer 31 in contact with the flow channel 5 has different temperatures in different adjustment zones, and the temperature in each adjustment zone can reach the target temperature. In this design, the target temperature of each adjustment zone is: the temperature of the position corresponding to the denaturation zone 41 is 95°C; the temperature of the position corresponding to the annealing zone 42 is 55°C, and the temperature of the position corresponding to the extension zone 43 is 72°C.

[0021] A flow channel 5 is provided below the chip body 2 . The flow channel 5 is located above the thermal resistance adjustment layer 3 and is in contact with the conductive layer 31 . The heat on the conductive layer 31 is transferred to the flow channel 5 and heats the fluid in the flow channel 5 .

[0022] The direction in which the three temperature zones are arranged is set to be horizontal, and the direction in which each temperature zone itself spans is set to be vertical. Then each pipe in the flow channel 5 is arranged horizontally, and several pipes are arranged in the vertical direction, and adjacent pipes are connected by arc pipes at the ends. When the fluid in the flow channel 5 flows from one end of a pipe to the other end, it will pass through the three temperature zones in turn and obtain different temperatures, so that the fluid can repeatedly provide different temperature conditions to the chip body 2. The fluid in the flow channel 5 flows along the flow channel 5, so that the chip body 2 is repeatedly placed under temperature conditions of 95°C, 55°C, and 72°C to promote the PCR reaction.

[0023] This design combines the use of a thermal resistance adjustment layer 3 and a continuous flow PCR chip with an adjustable heating element, which consumes less energy, has little impact on the environment, has low requirements for the use environment, and saves space. Under the premise of simple optimization, through simulation, it is found that only 3W of heating power is required to realize the PCR reaction process.

[0024] The specific embodiments described above further illustrate the technical problems, technical solutions and beneficial effects solved by the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A continuous flow low energy consumption PCR chip, comprising a chip base (1), a chip body (2) and a thermal resistance adjustment layer (3), characterized in that: The chip base (1) is provided with a heater (4), the heater (4) is divided into a plurality of temperature zones, the temperatures of the plurality of temperature zones are different, the thermal resistance adjustment layer (3) is located above the heater (4), a flow channel (5) is provided below the chip body (2), the flow channel (5) is located above the thermal resistance adjustment layer (3), the heat generated by the heater (4) is transferred to the chip body (2) through the thermal resistance adjustment layer (3), the temperature of the chip body (2) in different areas is different, there is a fluid in the flow channel (5), the fluid flows in the flow channel (5), the flow rate of the fluid in the flow channel is in the order of nl / s to ul / s, and the fluid in different areas of the flow channel (5) obtains different amounts of heat.

2. A continuous flow low energy consumption PCR chip according to claim 1, characterized in that: The plurality of temperature zones on the heater (4) are a denaturation zone (41), an annealing zone (42) and an extension zone (43), the denaturation zone (41), the annealing zone (42) and the extension zone (43) are arranged in sequence from left to right, the segment width of the denaturation zone (41) is d1, the segment width of the extension zone (43) is d2, the segment width of the annealing zone (42) is d3, and the segment width of d1 is d4. <d3<d2。 3. A continuous flow low energy consumption PCR chip according to claim 2, characterized in that: The flow channel (5) is arranged transversely above the thermal resistance adjustment layer (3), and a flow of the fluid will sequentially pass above the denaturation zone (41), the annealing zone (42), and the extension zone (43).

4. A continuous flow low energy consumption PCR chip according to claim 1, characterized in that: The heater (4) is one of a copper film, an aluminum film or a conductive polymer material.

5. The continuous flow low energy consumption PCR chip according to claim 1, characterized in that: The chip base (1) is made of one of glass, quartz, polymer, aluminum alloy and stainless steel.

6. A continuous flow low energy consumption PCR chip according to claim 1, characterized in that: The thermal resistance adjustment layer (3) comprises a conductive layer (31) and a heat insulating layer (32), wherein the conductive layer (31) is located on the heat insulating layer (32), the thickness of the heat insulating layer (32) located above the denaturation zone (41) is h1, the thickness of the heat insulating layer (32) located above the annealing zone (42) is h2, and the thickness of the heat insulating layer (32) located above the extension zone (43) is h3, wherein h2>h3>h1.

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