MICROFLUIDIC CHIP ASSEMBLY FOR RAPIDLY PERFORMING DIGITAL POLYMERASE CHAIN REACTION (PCR), AND USE THEREOF

Abstract

A microfluidic chip assembly for rapidly performing digital polymerase chain reaction (PCR) and use thereof are provided. The microfluidic chip assembly includes at least one microfluidic chip, a heat sink arranged below the microfluidic chip, a heater arranged above the microfluidic chip, a semiconductor cooler arranged between the heat sink and the microfluidic chip, and a heat-conducting plate arranged above the semiconductor cooler. A thin film layer is bonded at the bottom of the microfluidic chip, and the thin film layer abuts against the heat-conducting plate.

Claims

1. A microfluidic chip assembly for rapidly performing digital polymerase chain reaction (PCR), comprising: at least one microfluidic chip, a heat sink arranged below the at least one microfluidic chip, a semiconductor cooler arranged between the heat sink and the at least one microfluidic chip, and a heat-conducting plate arranged above the semiconductor cooler; wherein the at least one microfluidic chip abuts against the heat-conducting plate.

2. The microfluidic chip assembly according to claim 1, wherein a heat insulation layer is wrapped outside each of the heat-conducting plate and the semiconductor cooler.

3. The microfluidic chip assembly according to claim 2, wherein each of the at least one microfluidic chip comprises a chip body; an oil well, a sample well and a waste liquid well are protrudingly provided on the chip body; the chip body is provided with a sealing cover; through holes matched with the oil well, the sample well and the waste liquid well are formed on the sealing cover; and a sealing filter is provided inside the through holes.

4. The microfluidic chip assembly according to claim 3, wherein a heater is provided above the microfluidic chip, and a pneumatic press plate is provided above the heater.

5. The microfluidic chip assembly according to claim 3, wherein a thin film layer is bonded at a bottom of the microfluidic chip, and the thin film layer at the bottom of the microfluidic chip has a thickness of 10 μm to 500 μm.

6. The microfluidic chip assembly according to claim 1, wherein two or more microfluidic chips are arranged in parallel, and the two or more microfluidic chips are provided on a chip tray.

7. The microfluidic chip assembly according to claim 1, wherein the heat-conducting plate is circumferentially provided with a flexible heat insulation pad.

8. An integrated device with the microfluidic chip assembly according to claim 1, comprising a rack, wherein a sliding rail is provided on the rack, and the microfluidic chip assembly is provided on the sliding rail; a driving motor is provided on the rack, and the driving motor is configured to drive the microfluidic chip assembly to move on the sliding rail; a driving mechanism is provided above the microfluidic chip assembly, and the driving mechanism is configured to drive the pneumatic press plate to move; and a temperature control system and a pneumatic control system are provided on a side of the microfluidic chip assembly and are electrically connected to the microfluidic chip assembly.

9. The integrated device according to claim 8, wherein the rack is further provided with an industrial personal computer (IPC), wherein the IPC is electrically connected to the temperature control system and the pneumatic control system.

10. A reaction method of an integrated device with a microfluidic chip assembly for rapidly performing digital polymerase chain reaction (PCR), comprising the following steps: S1: driving the microfluidic chip assembly by a driving motor to move out of the integrated device until a chip tray is exposed outside the integrated device; S2: loading an oil phase reagent and a water phase reagent respectively in an oil well and a sample well of a microfluidic chip through a pipettor; S3: sealing the oil phase reagent and the water phase reagent in the microfluidic chip in S2 using a sealing cover; S4: driving the microfluidic chip assembly by the driving motor to move into the integrated device until the microfluidic chip assembly is exactly below a pneumatic press plate; S5: driving the pneumatic press plate and a heater by a driving mechanism to move in a direction of the microfluidic chip until the heater is tightly attached to an upper surface of the microfluidic chip and a bottom of the microfluidic chip abuts against a heat-conducting plate; S6: controlling a temperature control system and a pneumatic control system by an industrial personal computer (IPC) to respectively control a temperature and an air pressure of the microfluidic chip assembly to complete a thermal cycling process of PCR; and S7: driving, by the driving motor, the microfluidic chip assembly to move out of the integrated device, wherein the microfluidic chip assembly completes a PCR process in S6, and taking out the microfluidic chip to complete an entire process.

11. The microfluidic chip assembly according to claim 6, wherein the heat-conducting plate is circumferentially provided with a flexible heat insulation pad.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0029] FIG. 1 is a schematic diagram illustrating a structural relationship between the microfluidic chip and the thin film layer according to the present invention.

[0030] FIG. 2 is a schematic diagram illustrating a structure of the microfluidic chip with a sealing cover according to the present invention.

[0031] FIG. 3 is a schematic diagram illustrating a structure of the microfluidic chip assembly according to the present invention.

[0032] FIG. 4 is a schematic diagram of the microfluidic chip assembly according to the present invention.

[0033] FIG. 5 is a schematic diagram illustrating a structure of the microfluidic chip assembly in a specific example according to the present invention.

[0034] FIG. 6 shows a device provided with the microfluidic chip assembly according to the present invention.

[0035] FIG. 7 shows a heating rate curve of the heat-conducting plate during a practical application according to the present invention.

[0036] FIG. 8 shows a heating rate curve of a sample in the microfluidic chip during a practical application according to the present invention.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] The present invention discloses a microfluidic chip assembly for rapidly performing digital PCR, and a device provided with the microfluidic chip assembly, which will be described in detail below with reference to FIG. 1 to FIG. 6.

[0038] The device including a microfluidic chip assembly for rapidly performing digital PCR includes a rack 56, and the rack 56 is provided with a sliding rail 55. A guide post 51 is provided at an outer side of the sliding rail, a driving plate 91 is perpendicularly installed on the guide post 51, and a driving mechanism 92 is provided in the driving plate 91. The driving mechanism 92 can be a driving motor, which is not limited here.

[0039] The microfluidic chip assembly is provided on the sliding rail 55, and a driving motor (not shown in the figures) is provided on the rack 56 and configured to drive the microfluidic chip assembly to move on the sliding rail 55. The microfluidic chip assembly is arranged below the driving plate 91. In order to well realize automatic control, a temperature control system 52 and a pneumatic control system 53 are provided on a side of the microfluidic chip assembly and are electrically connected to the microfluidic chip assembly. The rack 56 may be further provided with an IPC 54 electrically connected to the temperature control system 52 and the pneumatic control system 53.

[0040] The microfluidic chip assembly includes at least one microfluidic chip 11, a heat sink 5 arranged below the microfluidic chip 11, and a heater 8 arranged above the microfluidic chip 11, where a pneumatic press plate 9 is arranged above the heater 8, and outlets connected to an oil well, a sample well and a waste liquid well on the microfluidic chip are formed on the pneumatic press plate 9. The pneumatic press plate 9 is connected to the driving mechanism 92, and the driving mechanism 92 can work to drive the pneumatic press plate 9 and the heater 8 to move in a direction of the guide post.

[0041] The heater is provided with a metal block, which can well transmit heat to the microfluidic chip 11, and thus an area of the metal block is set according to a heating area required by the microfluidic chip. In order to ensure the heating effect, the area of the metal block is 5% to 100% larger than the heating area required by the microfluidic chip.

[0042] The microfluidic chip 11 includes a chip body 1, protruded wells 2 are provided on the chip body 1, including an oil well, a sample well and a waste liquid well. The chip body is provided with a sealing cover 4, through holes 41 matched with the oil well, the sample well and the waste liquid well are formed on the sealing cover 4, and a sealing filter is provided inside the through holes 41, which can protect generated microdroplets from being contaminated externally and can also effectively increase the airtightness of the pneumatic structure. Since the microfluidic chip only needs to handle a small amount of sample, a thin film layer 3 is bonded with a bottom of the chip body 1 to form a seal with the chip body 1 for flow channels in the microfluidic chip. The microfluidic chip has a small size. The thin film layer 3 has a thickness only of 10 μm to 500 μm, and the thin film layer 3 and the chip body 1 have a total thickness only of 1 mm to 5 mm.

[0043] A semiconductor cooler 7 is provided between the heat sink 5 and the microfluidic chip 11, a heat-conducting plate 71 is provided above the semiconductor cooler 7, and the thin film layer 3 abuts against the heat-conducting plate 71. For effective heat conduction, a design idea of an area of the heat-conducting plate 71 is the same as that of the heater, which is generally 5% to 100% larger than the heating area required by the microfluidic chip 11.

[0044] In order to avoid heat loss, a heat insulation layer 72 may be wrapped outside each of the heat-conducting plate 71 and the semiconductor cooler 7.

[0045] The microfluidic chip assembly of the present invention is not only suitable for a single chip structure, but also can meet the requirements of high throughput. That is, when other structures are arranged in the same way, the number of microfluidic chips can be changed to adapt new detection conditions, for example, by arranging the microfluidic chips in parallel on the chip tray. In an example of the present invention, 4 microfluidic chips are arranged in parallel, as shown in FIG. 5. In order to further reduce the gap between materials and increase the thermal conductivity, the heat-conducting plate 71 is circumferentially provided with a flexible heat insulation pad 73. The pneumatic press plate and the heater can be an integrated structure.

[0046] The present invention also discloses a reaction method of an integrated device with a microfluidic chip assembly for rapidly performing digital PCR, including the following steps.

[0047] I. Actual Loading of Microfluidic Chips

[0048] S1. The microfluidic chip assembly is driven by the driving motor to move out of the integrated device along the sliding rail 55 until the chip tray is exposed outside the integrated device, and then a microfluidic chip is loaded on the chip tray.

[0049] S2. An oil phase reagent and a water phase reagent are loaded respectively in an oil well and a sample well of a microfluidic chip 11 through a pipettor.

[0050] S3. The reagents in the microfluidic chip in S2 are sealed using the sealing cover 4.

[0051] II. Microdroplet Generation and PCR

[0052] S4. The microfluidic chip assembly is driven by the driving motor to move into the integrated device until the microfluidic chip assembly is exactly below the pneumatic press plate 9.

[0053] S5. The pneumatic press plate 9 and the heater 8 are driven by the driving mechanism 92 to move in a direction of the microfluidic chip until the heater 8 is tightly attached to the upper surface of the microfluidic chip and the bottom of the microfluidic chip abuts against the heat-conducting plate 71.

[0054] S6. The temperature control system and the pneumatic control system are controlled by an IPC to respectively control an air pressure and a temperature of the microfluidic chip assembly to complete a thermal cycling process of PCR The pneumatic control system applies a high-precision air pressure to the sample well and the oil well of the microfluidic chip, such that the water phase reagent and the oil phase reagent enter the microfluidic chip through microfluidic channels to form a stable and uniform microdroplet emulsion, and then the emulsion enters a PCR chamber through a microfluidic channel.

[0055] The temperature control system achieves the temperature control, and protects the droplets in combination with the pneumatic control system to make a PCR process completed.

[0056] III. Process Completion

[0057] S7. The microfluidic chip assembly that completes a PCR process in S6 is driven by the driving motor to move out of the integrated device, and the microfluidic chip is taken out to complete the entire process.

[0058] The heat-conducting plate in the structure of the present invention has a high heating rate. As shown in FIG. 7, in the temperature range of 58° C. to 98° C., an average heating rate can reach 14.6 C/s and an average cooling rate can reach 8.8 C/s within the interval of 20% to 80% of the temperature difference. As shown in FIG. 8, heating and cooling rates of a sample inside the microfluidic chip all exceed 5 C/s, which far exceed the heating and cooling rates in the prior art.

[0059] Apparently, there are many specific implementations of the present invention, which are not listed here. All technical solutions formed from equivalent replacements or equivalent transformations should fall within the protection scope of the present invention.