CN115651824A - PCR reaction system and method - Google Patents
PCR reaction system and method Download PDFInfo
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- CN115651824A CN115651824A CN202210732160.XA CN202210732160A CN115651824A CN 115651824 A CN115651824 A CN 115651824A CN 202210732160 A CN202210732160 A CN 202210732160A CN 115651824 A CN115651824 A CN 115651824A
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Abstract
The invention provides a PCR reaction system and a method, wherein the PCR reaction system comprises a bottom plate and a PCR reaction chip, and the PCR reaction chip is provided with a reagent cavity; further comprising: the heat conducting component is internally provided with a fluid inlet, a plurality of fluid channels and a fluid outlet which are sequentially communicated, the inlets are sequentially communicated, and the plurality of fluid channels are arranged in parallel; the PCR chip is positioned in the enclosed area of the annular component; the heating unit is arranged in the area; the fixing plate is provided with a bulge, the bulge is clamped in the area, and the fixing plate, the heating unit, the PCR reaction chip, the heat conducting part and the bottom plate are sequentially arranged. The invention has the advantages of simple structure, uniform temperature and the like.
Description
Technical Field
The present invention relates to PCR reactions, and more particularly to PCR reaction systems and methods.
Background
Polymerase Chain Reaction (PCR) is a molecular biology technique used to perform rapid amplification of specific DNAs in vitro. The PCR equipment has the characteristics of high sensitivity, strong specificity, simple operation and the like, can be used for basic research such as gene separation, cloning, nucleic acid sequence analysis and the like, and can also be used for disease diagnosis or any place with DNA and RNA, so that the PCR equipment is widely applied and rapidly developed in the fields of life science research, biochemical analysis, clinical diagnosis, forensic identification, epidemic situation rapid inspection and the like.
The PCR process is generally composed of three basic reaction steps of high-temperature denaturation, low-temperature annealing and moderate-temperature extension, which are circularly repeated. Therefore, the temperature control system is extremely important for the PCR reaction result, and directly determines the performance of the PCR equipment. The heating module temperature rise and fall speed and the temperature uniformity are important evaluation indexes for measuring the excellence of the temperature control system of the PCR equipment. Most of PCR equipment in the market at present adopts a semiconductor refrigeration piece as a heat source, and utilizes a PID algorithm to realize accurate circulating temperature control on a temperature control system, and the following problems are generally existed:
1. the heating module has limited temperature rising and falling speed;
2. the heating module may have an edge effect, i.e. the intermediate temperature is higher than the edge temperature.
Disclosure of Invention
In order to overcome the defects in the prior art scheme, the invention provides a PCR reaction system.
The purpose of the invention is realized by the following technical scheme:
the PCR reaction system comprises a bottom plate and a PCR reaction chip, wherein the PCR reaction chip is provided with a reagent cavity; the PCR reaction system further comprises:
the heat conduction component is internally provided with a fluid inlet, a plurality of fluid channels and a fluid outlet which are sequentially communicated, and the fluid channels are arranged in parallel;
a ring-shaped member, the PCR chip being located in a surrounding region of the ring-shaped member;
a heating unit disposed within the region;
the fixing plate is provided with a protrusion, the protrusion is clamped in the area, and the fixing plate, the heating unit, the PCR reaction chip, the heat conducting component and the bottom plate are sequentially arranged.
The invention also aims to provide a PCR reaction method, and the aim is realized by the following technical scheme:
the PCR reaction method comprises the following steps:
the fluid enters the heat-conducting component and sequentially passes through the fluid inlet, the multiple fluid channels and the fluid outlet, and the multiple fluid channels are arranged in parallel;
the reagent cavity at one side of the PCR reaction chip contacting with the heat conducting component is heated to a set temperature, and at the same time, the other side of the PCR reaction chip contacting with the heat conducting component is heated to the set temperature by the heating unit;
the PCR chip and the heating unit are respectively located in the enclosed region of the annular component, the fixing plate, the heating unit, the PCR reaction chip, the heat conducting component and the bottom plate are sequentially arranged, the fixing plate is provided with a protrusion, and the protrusion is clamped into the region.
Compared with the prior art, the invention has the beneficial effects that:
1, the PCR reaction time is short;
the PCR reaction of the flat plate type microfluidic chip can quickly realize the cyclic change of temperature, thereby greatly reducing the whole PCR reaction time;
2. the temperature consistency is good;
the design of the multi-stage flow channels ensures that the length and the flow resistance of the flow channels are consistent, the flow of the heat conducting liquid in each flow channel can be ensured to be consistent, and the heating unit heats the other side of the PCR reaction chip, so that the temperature consistency of the samples corresponding to each flow channel is improved;
3. miniaturization;
the internal heat-conducting liquid is recycled, so that the situation that the whole system is too large due to too much heat-conducting liquid is avoided, and the miniaturization of equipment is facilitated.
Drawings
The disclosure of the present invention will become more readily understood with reference to the accompanying drawings. As is readily understood by those skilled in the art: these drawings are only for illustrating the technical solutions of the present invention and are not intended to limit the scope of the present invention. In the figure:
FIG. 1 is a schematic structural view of a PCR reaction system according to an embodiment of the present invention;
fig. 2 is a schematic view of a fluid channel structure according to an embodiment of the present invention.
Detailed Description
Fig. 1-2 and the following description depict alternative embodiments of the invention to teach those skilled in the art how to make and use the invention. Some conventional aspects have been simplified or omitted for the purpose of explaining the technical solution of the present invention. Those skilled in the art will appreciate variations or substitutions from these embodiments that will be within the scope of the invention. Those skilled in the art will appreciate that the features described below can be combined in various ways to form multiple variations of the invention. Thus, the present invention is not limited to the following alternative embodiments, but is only limited by the claims and their equivalents.
Example 1:
FIG. 1 is a schematic view showing a schematic configuration of a PCR reaction system according to an embodiment of the present invention, which comprises, as shown in FIG. 1:
the kit comprises a base plate 2 and a PCR reaction chip 7, wherein the PCR reaction chip 7 is provided with a reagent cavity 71;
a heat-conducting member 1, as shown in fig. 2, the heat-conducting member 1 having a fluid inlet 5, a plurality of fluid channels and a fluid outlet 11 connected in sequence, the plurality of fluid channels being arranged in parallel;
a ring member 3, the PCR chip 7 being located in a region surrounded by the ring member 3;
a heating unit disposed within the region;
the fixing plate 4 is provided with a protrusion, the protrusion is clamped in the area, and the fixing plate 4, the heating unit, the PCR reaction chip 7, the heat conducting component 1 and the bottom plate 2 are sequentially arranged.
In order to fix the heat conduction member, further, one side of the base plate 2 has a groove in which the heat conduction member 1 is located.
In order to prevent the temperature of the PCR reaction chip from being inconsistent due to heat loss, the heating unit further includes:
the heating film, the heat transfer component and the flat PCR reaction chip are arranged in sequence.
In order to prevent heat loss, further, the PCR reaction system further includes:
a first annular seal disposed between the heat-conducting member and a wall of the recess;
a second annular seal disposed between the annular member and the stationary plate and surrounding a side opening of the area.
In order to maintain temperature consistency, further, as shown in fig. 2, the fluid channel includes:
a first set of flow channels comprising at least two stages of flow channels, such as a first stage flow channel 13, a second stage flow channel 14 and a third stage flow channel 15, such that the number of flow channels increases exponentially, the fluid inlet 5 communicating with the outlet ends of the first set of flow channels;
the second group of flow passages 16 are arranged in parallel and are respectively communicated with the outlet of the last stage of the first group of flow passages, and a plurality of parallel partition pieces 17 are arranged in each flow passage of the second group of flow passages 16; the projection of the reagent chambers 71 onto the heat-conducting member falls in the area of the second set of flow channels 16
And the third group of flow passages comprises at least two stages of flow passages, so that the number of the flow passages is decreased exponentially, the inlet ends of the first group of flow passages are communicated with the second group of flow passages, and the outlet ends of the first group of flow passages are communicated with the fluid outlet 11.
To circulate the heating fluid, further, the PCR reaction system further includes:
a first switching module for selectively communicating the fluid outlet with any one of the fluid supply units;
the fluid supply units comprise heating modules, containers and pumps, wherein the heating modules, the containers and the pumps are sequentially passed by fluid, and the temperature control module is used for controlling the heating modules so that the temperature of the fluid in the containers reaches a set value;
a second switching module for selectively communicating the fluid inlet with the pump of either fluid supply unit.
The PCR reaction method of the embodiment of the invention comprises the following steps:
the fluid enters the heat-conducting component and sequentially passes through the fluid inlet, the multiple fluid channels and the fluid outlet, and the multiple fluid channels are arranged in parallel;
the reagent cavity at one side of the PCR reaction chip contacting with the heat conducting component is heated to a set temperature, and at the same time, the other side of the PCR reaction chip contacting with the heat conducting component is heated to the set temperature by the heating unit;
the PCR chip and the heating unit are respectively located in the enclosed region of the annular component, the fixing plate, the heating unit, the PCR reaction chip, the heat conducting component and the bottom plate are sequentially arranged, the fixing plate is provided with a protrusion, and the protrusion is clamped into the region.
In order to ensure the consistency of the temperature of the fluid in the channel, furthermore, the mode of the fluid passing through the fluid channel is as follows:
fluid entering the fluid inlet sequentially passes through a first group of flow channels, a second group of flow channels, a third group of flow channels and a fluid outlet, wherein the first group of flow channels comprises at least two stages of flow channels, so that the number of the flow channels is exponentially increased, and the fluid inlet is communicated with the outlet end of the third group of flow channels; the second group of flow passages are arranged in parallel and are respectively communicated with the outlet of the last stage of the first group of flow passages, and a plurality of parallel-arranged isolating pieces are arranged in each flow passage in the second group of flow passages; the third group of flow passages comprises at least two stages of flow passages, so that the number of the flow passages is exponentially decreased, the inlet ends of the first group of flow passages are communicated with the second group of flow passages, and the outlet ends of the first group of flow passages are communicated with the fluid outlet;
the flow rate of the fluid in the second group of flow channels is the same, and the projection of the reagent cavity on the heat conducting part falls in the area of the second group of flow channels.
In order to fix the heat-conducting member and the PCR reaction chip, further, one side of the base plate has a groove, and the heat-conducting member is positioned in the groove.
In order to recycle the heating fluid, the fluid is discharged from the fluid outlet, passes through the switching module, selectively enters the fluid supply unit with the same set temperature, is heated to the set temperature, is sent into the container, and is finally sent to the fluid inlet after passing through the pump.
Example 2:
an application example of the PCR reaction system and method according to embodiment 1 of the present invention.
In the present application example, as shown in fig. 1, the bottom plate 2 is rectangular and has a rectangular groove on the upper side; the annular member 3 encloses a rectangular region; the heat conducting component 1 is in a rectangular flat plate shape and is arranged in the rectangular groove, and a first annular sealing element 6 is arranged between the lower side of the heat conducting component and the bottom wall of the groove; the PCR reaction chip 7 is in a rectangular flat plate shape, the lower side of the PCR reaction chip is provided with a reagent cavity 71, and the PCR reaction chip is arranged in the rectangular area; the heating unit includes a heating film 9 and a rectangular heat transfer member 8 disposed in the rectangular region; the fixing plate 4 is in a rectangular flat plate shape, the lower side of the fixing plate is provided with quadrangular-prism-shaped bulges, the bulges are clamped in the rectangular area, the upper side of the annular component 3 is provided with a second annular sealing element 10 which surrounds an opening at the upper end of the rectangular area, and the fixing plate 4, the heating film 9, the heat transfer component 8, the PCR reaction chip 7, the heat conducting component 1 and the bottom plate 2 are sequentially arranged;
as shown in fig. 2, the heat-conducting member 1 has therein a fluid inlet 5, a plurality of fluid channels arranged in parallel, and a fluid outlet 11 which are communicated in this order; the fluid channel includes:
the first set of flow channels comprises at least two stages of flow channels, such as 2 first stage flow channels 13, 4 second stage flow channels 14 and 8 third stage flow channels 15, so that the number of flow channels increases exponentially, and the fluid inlet 5 is communicated with the outlet ends of the first set of flow channels;
the 8 second-group flow passages 16 are arranged in parallel and are respectively communicated with the outlet of the last stage of the first-group flow passages, and a plurality of parallel partition pieces 17 are arranged in each flow passage of the second-group flow passages 16; the projection of the reagent chamber 71 onto the heat-conducting member falls within the area of the second set of flow channels 16
A third group of flow passages, which includes at least two stages of flow passages, such that the number of flow passages decreases exponentially, for example, 8, 4, and 2 tertiary flow passages, respectively, and the inlet ends of the first group of flow passages are communicated with the second group of flow passages, and the outlet ends are communicated with the fluid outlet 11;
the first switching module adopts a four-way valve and is used for enabling the fluid outlet to be selectively communicated with any one fluid supply unit;
a plurality of fluid supply units, such as three fluid supply units, including a heating module, a container and a pump through which fluid passes in sequence, and a temperature control module for controlling the heating module so that the temperature of the fluid in the container reaches a set value, such as 60 degrees, 72 degrees and 95 degrees, respectively;
the second switching module employs a four-way valve for selectively communicating the fluid inlet with the pump of any one of the fluid supply units.
The PCR reaction method of the embodiment of the present invention, that is, the working method of the PCR reaction system of the embodiment, is:
the liquid enters the heat conducting part 1 and sequentially passes through the fluid inlet 5, the plurality of fluid channels and the fluid outlet 11, and the plurality of fluid channels are arranged in parallel, namely the liquid sequentially flows through the first group of flow channels, the second group of flow channels and the third group of flow channels; the liquid flow rates in the 8 second groups of flow channels are respectively as follows: v1=0.042m/s, v2=0.040m/s, v3=0.039m/s, v4=0.041m/s, v5=0.041m/s, v6=0.039m/s, v7=0.040m/s, and v8=0.042m/s, and it can be seen that the liquid flow rates are substantially uniform, and the uniformity of the temperature of the PCR chip is improved;
the reagent chamber 71 contacting one side of the PCR reaction chip 7 of the heat conductive member 1 is heated to a set temperature, for example, 95 degrees, and at the same time, the other side contacting the PCR reaction chip 7 is heated to a set temperature by the heating unit;
after being discharged from the fluid outlet 11, the fluid passes through the first switching module, selectively enters the fluid supply unit with the same set temperature, is heated to the set temperature and is sent into the container, and finally is sent to the fluid inlet after passing through the pump and the second switching module, so that the circulation of the liquid is realized.
Claims (10)
- The PCR reaction system comprises a bottom plate and a PCR reaction chip, wherein the PCR reaction chip is provided with a reagent cavity; characterized in that, the PCR reaction system further comprises:the heat conduction component is internally provided with a fluid inlet, a plurality of fluid channels and a fluid outlet which are sequentially communicated, and the fluid channels are arranged in parallel;a ring-shaped member, the PCR chip being located in a surrounding region of the ring-shaped member;a heating unit disposed within the region;the fixing plate is provided with a protrusion, the protrusion is clamped in the area, and the fixing plate, the heating unit, the PCR reaction chip, the heat conducting component and the bottom plate are sequentially arranged.
- 2. The PCR reaction system of claim 1, wherein one side of the base plate has a groove, and the heat-conducting member is disposed in the groove.
- 3. The PCR reaction system of claim 1, wherein the heating unit comprises:the heating film, the heat transfer component and the flat PCR reaction chip are arranged in sequence.
- 4. The PCR reaction system of claim 1, further comprising:a first annular seal disposed between the heat conductive member and a wall of the groove;a second annular seal disposed between the annular member and the stationary plate and surrounding a side opening of the area.
- 5. The PCR reaction system of claim 1, wherein the fluid channel comprises:a first set of flow passages including at least two stages of flow passages such that the number of flow passages increases exponentially, the fluid inlet communicating with the outlet ends of the first set of flow passages;the second group of flow passages are arranged in parallel and are respectively communicated with the outlets of the last stage of the first group of flow passages, and a plurality of parallel-arranged isolating pieces are arranged in each flow passage of the second group of flow passages; the projection of the reagent chamber on the heat-conducting member falls within the area of the second set of flow channelsAnd the third group of flow passages comprises at least two stages of flow passages, so that the number of the flow passages is decreased exponentially, the inlet ends of the first group of flow passages are communicated with the second group of flow passages, and the outlet ends of the first group of flow passages are communicated with the fluid outlet.
- 6. The PCR reaction system of claim 1, further comprising:a first switching module for selectively communicating the fluid outlet with any one of the fluid supply units;the fluid supply units comprise heating modules, containers and pumps, wherein the heating modules, the containers and the pumps are sequentially passed by fluid, and the temperature control module is used for controlling the heating modules so that the temperature of the fluid in the containers reaches a set value;a second switching module for selectively communicating the fluid inlet with the pump of either fluid supply unit.
- A PCR reaction method, the PCR reaction method comprising:the fluid enters the heat-conducting component and sequentially passes through the fluid inlet, the multiple fluid channels and the fluid outlet, and the multiple fluid channels are arranged in parallel;the reagent cavity at one side of the PCR reaction chip contacting with the heat conducting component is heated to a set temperature, and at the same time, the other side of the PCR reaction chip contacting with the heat conducting component is heated to the set temperature by the heating unit;the PCR chip and the heating unit are respectively located in the enclosed region of the annular component, the fixing plate, the heating unit, the PCR reaction chip, the heat conducting component and the bottom plate are sequentially arranged, the fixing plate is provided with a protrusion, and the protrusion is clamped into the region.
- 8. The PCR reaction method according to claim 7, wherein the fluid passes through the fluid channel by:fluid entering the fluid inlet sequentially passes through a first group of flow channels, a second group of flow channels, a third group of flow channels and a fluid outlet, wherein the first group of flow channels comprises at least two stages of flow channels, so that the number of the flow channels is exponentially increased, and the fluid inlet is communicated with the outlet end of the third group of flow channels; the second group of flow passages are arranged in parallel and are respectively communicated with the outlet of the last stage of the first group of flow passages, and a plurality of parallel-arranged isolating pieces are arranged in each flow passage in the second group of flow passages; the third group of flow channels comprises at least two stages of flow channels, the number of the flow channels is reduced in an exponential manner, the inlet ends of the first group of flow channels are communicated with the second group of flow channels, and the outlet ends of the first group of flow channels are communicated with the fluid outlet;the flow velocity of the fluid in the second group of flow channels is the same, and the projection of the reagent cavity on the heat conducting component falls in the area of the second group of flow channels.
- 9. The PCR reaction method of claim 7, wherein the bottom plate has a groove on one side thereof, and the heat-conducting member is disposed in the groove.
- 10. The PCR reaction method of claim 7, wherein the fluid is discharged from the fluid outlet, passes through the switching module, and then selectively enters the fluid supply unit having the same temperature as the set temperature, and the fluid is heated to the set temperature and then is sent into the container, and finally is sent to the fluid inlet after being pumped.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210732160.XA CN115651824A (en) | 2022-06-27 | 2022-06-27 | PCR reaction system and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210732160.XA CN115651824A (en) | 2022-06-27 | 2022-06-27 | PCR reaction system and method |
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| Publication Number | Publication Date |
|---|---|
| CN115651824A true CN115651824A (en) | 2023-01-31 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210732160.XA Pending CN115651824A (en) | 2022-06-27 | 2022-06-27 | PCR reaction system and method |
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| CN (1) | CN115651824A (en) |
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- 2022-06-27 CN CN202210732160.XA patent/CN115651824A/en active Pending
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