CN119464044A

CN119464044A - A continuous flow low energy consumption PCR chip

Info

Publication number: CN119464044A
Application number: CN202411508129.3A
Authority: CN
Inventors: 李欣欣
Original assignee: Shaoxing Research Institute Of Zhejiang University
Current assignee: Shaoxing Research Institute Of Zhejiang University
Priority date: 2024-10-28
Filing date: 2024-10-28
Publication date: 2025-02-18

Abstract

The present invention discloses a continuous flow low energy consumption PCR chip, comprising a chip base, a chip body and a thermal resistance adjustment layer. A heater is arranged on the chip base, and the heater is divided into a plurality of temperature zones, and the temperatures of the plurality of temperature zones are different. The thermal resistance adjustment layer is located above the heater, and a flow channel is arranged inside 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. The fluid flows in the flow channel, and the fluids in different areas of the flow channel obtain different amounts of heat to realize temperature zoning of the fluid. The present invention has a reasonable structure, and the thermal resistance adjustment layer cooperates with different temperature zones, so that the fluid in the flow channel can continuously change its own temperature during the continuous flow process, so that the PCR can complete multiple processes of denaturation, annealing and extension cycles under different temperature conditions.

Description

Continuous flowing low-energy consumption PCR chip

Technical Field

The invention relates to the technical field of PCR, in particular to a continuous flow low-energy consumption PCR chip.

Background

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, virus detection, population research, life detection, and the like. PCR generally requires repeated denaturation, annealing, extension cycles at temperatures of 95 ℃, 55 ℃, 72 ℃.

For the requirements of temperature rise and fall between three temperature conditions during circulation, the conventional PCR reaction device generally uses Peltier to carry out bidirectional temperature rise and fall, some devices use heaters for unidirectional temperature rise and cooling together with radiators, some devices use a plurality of resistance heaters to be arranged in different heating areas and realize temperature control in a fluid circulation flow mode, and other devices use a single resistance heater to heat a special-shaped piece to obtain different temperature areas and realize temperature control in a fluid circulation flow mode by winding a pipeline on the special-shaped piece.

When the unmanned detection is carried out in a special environment, new requirements on energy consumption, space, heat dissipation conditions and stability are provided. For the temperature control scheme of the peltier form, although rapid temperature rising and falling circulation can be realized, the peltier can be used only by matching with a radiator in the refrigeration process, extra space is required to occupy, extra energy consumption is generated in the temperature rising process and the temperature falling process, and the use condition also limits the radiator form. For the situation that the heater is used for unidirectional heating and matched with the radiator for cooling, the cooling rate is slower than that of the Peltier scheme, so that the energy consumption in the cooling process is reduced, but other problems are not obviously improved. For the scheme of using a plurality of resistance heaters to generate different temperature control areas, the energy consumption in the temperature rise and fall circulation process is reduced, but multiple temperature measurement and temperature control are needed, and additional driving and conversion elements are added. For the scheme that the heater is used for heating the special-shaped piece to generate a temperature gradient and then the pipeline is wound on the special-shaped piece, the consistency problem of a plurality of contact surfaces can lead to temperature accuracy being difficult to guarantee, stability is poor and longitudinal space occupation is large. In view of the above problems, a solution is proposed below.

Disclosure of Invention

The invention aims to provide a continuous flow low-energy consumption PCR chip, and the scheme of controlling the temperatures of different areas of the continuous flow PCR microfluidic chip by adjusting the heating value and the heat transfer resistance of a heating element is adopted, so that the heat conduction of the different areas is matched with the required heat.

The technical aim of the invention is realized by the following technical scheme:

the utility model provides a low energy consumption PCR chip of continuous flow, includes chip base, chip body and thermal resistance adjustment layer, be equipped with the heater on the chip base, the heater divide into a plurality of temperature areas, and is a plurality of the temperature of temperature area is different, the thermal resistance adjustment layer is located the top of heater, the below of chip body is equipped with the runner, the runner is located the top of thermal resistance adjustment layer, the heat that the heater produced is passed through the thermal resistance adjustment layer and is transmitted to on the chip body, there is fluid in the runner, fluid flows in the runner, be located fluid in the different regions of runner obtains different heat to realize the heating to the chip body.

Preferably, the plurality of temperature areas on the heater are respectively a denaturation area, an annealing area and an extension area, the denaturation area, the annealing area and the extension area are sequentially arranged from left to right, the section width of the denaturation area is d1, the section width of the extension area is d2, the section width of the annealing area is d3, d1 is less than d3 and less than d2, and the three areas are provided with interval widths according to the time required by the three stages of denaturation, annealing and extension.

Preferably, the flow channel is arranged transversely above the thermal resistance adjusting layer, and the primary flow of the fluid passes through the upper parts of the denaturation zone, the annealing zone and the extension zone in sequence.

Preferably, the heater is one of a copper film, an aluminum film or a conductive polymer material.

Preferably, the chip base is made of one of glass, quartz, polymer, aluminum alloy and stainless steel.

Preferably, the thermal resistance adjusting layer comprises a conductive layer and a heat insulating layer, the conductive layer is located on the heat insulating layer, the thickness of the heat insulating layer located above the denaturation region is h1, the thickness of the heat insulating layer located above the annealing region is h2, the thickness of the heat insulating layer located above the extension region is h3, and h2> h3> h1.

The beneficial effects of the invention are as follows:

1. The invention provides a scheme for controlling the temperatures of different areas of a continuous flow PCR microfluidic chip by adjusting the heating value of a heating element and the heat transfer resistance, so that the heat conduction of the different areas is matched with the required heat, the attached cooling and measurement and control components are greatly reduced, and the problems of PCR energy consumption, space occupation and heat dissipation under special conditions are solved;

2. The continuous flow PCR chip combining the thermal resistance adjusting layer and the adjustable heating body has the advantages of low energy consumption, small environmental impact, low requirement on the use environment and space saving;

3. the chip type design is flexible and stable in the use process.

Drawings

FIG. 1 is a schematic diagram of an embodiment;

FIG. 2 is a schematic view of a heater according to an embodiment;

FIG. 3 is a schematic diagram of an embodiment for showing a heater temperature region;

FIG. 4 is a schematic illustration of an example fluid temperature profile;

FIG. 5 is a schematic diagram of example fluid temperature and duration.

Reference numeral 1, chip base, 2, chip body, 3, thermal resistance regulating layer, 31, conductive layer, 32, heat insulating layer, 4, heater, 41, denaturation zone, 42, annealing zone, 43, extension zone, 5, runner.

Detailed Description

The following description is only of the preferred embodiments of the present invention, and the scope of the present invention should not be limited to the examples, but should be construed as falling within the scope of the present invention. Wherein like parts are designated by like reference numerals.

As shown in fig. 1 to 5, a continuous flow low power PCR chip includes 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 has stable property and is not easy to damage.

The upper end surface of the chip base 1 is provided with a heater 4, and the material of the heater 4 can be a metal film such as copper, aluminum and the like, or can be a material such as conductive polymer and the like, and heat can be generated after the power is electrified. The heater 4 is divided into a plurality of temperature areas, the section widths d of the heater 4 at different temperature areas are different, and different heating powers of different temperature areas can be achieved by adjusting the section widths d, so that different temperature areas can reach different temperatures at the same time.

The temperature zones on the heater 4 are a denaturation zone 41, an annealing zone 42 and an extension zone 43, respectively, and the heater 4 is one 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, the section width of the extension zone 43 is d3, and d1 in the present design

The thermal resistance adjusting layer 3 is located above the heater 4, and the thermal resistance adjusting layer 3 includes a conductive layer 31 and a heat insulating layer 32. The material of the conductive layer 31 may be copper, stainless steel or other materials with better heat conduction performance, and the conductive layer 31 needs to be partially communicated with the external environment to obtain a larger temperature gradient, and the conductive layers are not connected, so that the corresponding temperatures of the temperature areas are prevented from being mixed in the conductive layer 31. The material of the heat insulating layer 32 is a material with a low heat conductivity coefficient, such as ceramic, plastic, foamed cement, etc.

The thermal resistance adjusting layer 3 is also divided into corresponding adjusting areas according to different temperature areas of the heater 4, the adjusting areas of the thermal resistance adjusting layer 3 are respectively heated by the different temperature areas of the heater 4, the thickness of the heat insulating layer 32 used by the different adjusting areas is different, the thickness of the heat insulating layer 32 above the denaturation area 41 is h1, the thickness of the heat insulating layer 32 above the annealing area 42 is h2, the thickness of the heat insulating layer 32 above the extension area 43 is h3, h2> h3> h1, the thickness is specifically set to be matched with the heater and the materials, and the thickness of the heat insulating layer 32 above the extension area 43 is h1:h2:h3 to be 0.1:2:1.5.

The conducting layer 31 is located above the heat insulating layer 32, and the heat insulating layer 32 can stably send the heat generated by the heater 4 to the conducting layer 31, so that the temperature on the conducting layer 31 is stable and cannot be changed greatly. The heater 4 and the thermal resistance adjusting layer 3 cooperate such that the conductive layer 31 in contact with the flow passage 5 has different temperatures in different adjusting regions, and the temperature in each adjusting region can reach the target temperature. In this design, the target temperature of each conditioning zone is 95℃for the location corresponding to denaturation zone 41, 55℃for the location corresponding to annealing zone 42, and 72℃for the location corresponding to extension zone 43.

The runner 5 is arranged below the chip body 2, the runner 5 is arranged above the thermal resistance adjusting layer 3 and is in contact with the conducting layer 31, and heat on the conducting layer 31 can be transferred to the runner 5 and heat fluid in the runner 5.

The arrangement direction of the three temperature zones is set to be transverse, and the crossing direction of each temperature zone is set to be longitudinal. Each pipeline in the flow channel 5 is transversely arranged, a plurality of pipelines are longitudinally arranged, and adjacent pipelines are connected through arc-shaped pipes at the end parts. When the fluid in the flow channel 5 flows from one end of one pipeline to the other end, the fluid sequentially passes through the three temperature areas to obtain different temperatures, so that the fluid can repeatedly provide different temperature conditions for 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 subjected to the temperature conditions of 95 ℃, 55 ℃ and 72 ℃ to promote the PCR reaction.

The design combines the continuous flow PCR chip using the thermal resistance adjusting layer 3 and the adjustable heating body, has less energy consumption, small influence on environment, low requirement on the use environment and space saving. On the premise of simple optimization, the PCR reaction process can be realized by simulating and obtaining the heating power of 3W.

The technical problems, technical solutions and advantageous effects solved by the present invention have been further described in detail in the above-described embodiments, and it should be understood that the above-described embodiments are only illustrative of the present invention and are not intended to limit the present invention, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the scope of protection of the present invention.

Claims

1. The utility model provides a low energy consumption PCR chip of continuous flow, includes chip base (1), chip body (2) and thermal resistance adjustment layer (3), its characterized in that, be equipped with heater (4) on chip base (1), heater (4) divide into a plurality of warm district, and is a plurality of the temperature in warm district is different, thermal resistance adjustment layer (3) are located the top of heater (4), the below of chip body (2) is equipped with runner (5), runner (5) are located the top of thermal resistance adjustment layer (3), the heat that heater (4) produced is passed through thermal resistance adjustment layer (3) and is transmitted on chip body (2), chip body (2) are different in different district temperature, have fluid in runner (5), fluid flows in runner (5), fluid flow in the runner is the flow nl/s to ul/s order of magnitude, is located fluid in runner (5) different district obtains different heat.

2. The continuous flow low energy consumption PCR chip as claimed in claim 1, wherein the plurality of temperature zones on the heater (4) are respectively 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 sequentially arranged from left to right, the section width of the denaturation zone (41) is d1, the section width of the extension zone (43) is d2, the section width of the annealing zone (42) is d3, and d1< d3< d2.

3. A continuous flow low energy PCR chip according to claim 2, wherein the flow channel (5) is arranged laterally over the thermal resistance adjustment layer (3), and the primary flow of fluid passes over the denaturation zone (41), the annealing zone (42) and the extension zone (43) in sequence.

4. The continuous flow low energy PCR chip as claimed in claim 1, wherein 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 as claimed in claim 1, wherein said chip base (1) is made of one of glass, quartz, polymer, aluminum alloy, and stainless steel.

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