Integrated nucleic acid detection chip

Abstract

The utility model provides an integrated nucleic acid detection chip which comprises a cracking cavity, a first cavity, a first cleaning cavity, a second cavity, a reaction cavity, a sample adding port, an elution liquid adding port and a first cleaning liquid adding port which are sequentially communicated from top to bottom, the sample adding port is communicated with the cracking cavity, the first cleaning liquid adding port is communicated with the first cleaning cavity, and the second cavity is communicated with the second cleaning cavity. The elution liquid adding opening is communicated with the reaction cavity; the micro-fluidic chip further comprises a storage cavity, a first filling port, a second filling port, a third filling port, a storage port and a storage cover; the storage cover covers the storage port of the storage cavity; the first cavity is filled with a first isolator; and the second cavity is filled with a second isolator. The storage cavity is located above the reaction cavity and communicated with the reaction cavity, and the storage cavity and the reaction cavity are separated through a third isolator; the first filling port is communicated with the first cavity; the second filling port is communicated with the second cavity; and the third filling port is communicated with the storage cavity and the reaction cavity. The chip is convenient to operate, good in sealing effect and small in size.

Description

Technical Field

[0001] The utility model relates to the technical field of microfluidics, in particular to an integrated nucleic acid detection chip. Background Art

[0002] Nucleic acid analysis and testing in a laboratory environment has problems such as long testing time and the need for professional operation, which makes it difficult to meet the needs of on-site, rapid and portable testing of additives. Microfluidic chip technology developed in recent years can not only greatly shorten the testing time, but also has the advantages of large detection throughput, high degree of automation, and can be used by non-professionals. It is expected to be widely used in the field of nucleic acid analysis and testing.

[0003] The complete nucleic acid extraction and detection process generally requires four nucleic acid extraction steps, including lysis, binding, washing, and elution, plus subsequent nucleic acid molecular hybridization, polymerase chain reaction (PCR), and biochip detection steps. As for the transfer of active ingredients in each step, the existing technology mostly uses manual transfer, which is not only cumbersome, time-consuming and labor-intensive, but also difficult to fully and efficiently transfer samples, and manual operation can easily lead to unstable results.

[0004] An existing microfluidic chip for biological sample processing includes multiple reagent storage areas, a first functional area, a waste liquid area, and a microchannel switch. The reagent storage area is used to store reagents for processing biological samples, the first functional area is used to process biological samples using the reagents stored in the reagent storage area, and reagent microchannels are respectively arranged between each reagent storage area and the sample area; the microchannel switch is used to switch the opening and closing of each reagent microchannel, and the waste liquid area is used to receive waste liquid discharged from the first functional area.

[0005] However, in this solution, after each reaction with a reagent, the waste liquid needs to be discharged to the waste liquid area before it can be mixed with the next reaction reagent. The operation is complicated, and the large number of cavities leads to a large chip volume, a complex structure, and a high production cost. In addition, the sealing effect of the mechanical valve is poor. Summary of the invention

[0006] In order to solve the problems existing in the above-mentioned prior art, the purpose of the utility model is to provide an integrated nucleic acid detection chip with simple structure, convenient operation, good sealing effect and small size.

[0007] In order to achieve the above-mentioned purpose, the integrated nucleic acid detection chip provided by the utility model includes a lysis chamber, a first cavity, a first cleaning chamber, a second cavity, and a reaction chamber which are connected in sequence from top to bottom; the microfluidic chip also includes a sample addition port, an elution liquid addition port and a first cleaning liquid addition port, the sample addition port is connected to the lysis chamber, the first cleaning liquid addition port is connected to the first cleaning chamber, and the elution liquid addition port is connected to the reaction chamber; the microfluidic chip also includes a storage chamber, a first filling port, a second filling port, a third filling port, a storage port and a storage cover, the storage port is connected to the storage chamber, and the storage cover is closed at the storage port; the first cavity is filled with a first insulating material; the second cavity is filled with a second insulating material; the storage chamber is connected to the reaction chamber, the storage chamber is located above the reaction chamber, and the storage chamber and the reaction chamber are separated by a third insulating material; the first insulating material, the second insulating material and the third insulating material are all hydrophobic insulating materials; the first filling port is connected to the first cavity; the second filling port is connected to the second cavity; the third filling port is located between the storage chamber and the reaction chamber, and the third filling port is connected to the storage chamber and the reaction chamber respectively.

[0008] As can be seen from the above, the microfluidic chip provided by the utility model includes a lysis chamber, a cleaning chamber, a reaction chamber and a storage chamber, and does not need to be provided with an elution chamber and a waste liquid chamber, etc., and has a simple structure, which greatly reduces the volume of the chip. The provided storage chamber and the storage port connected to the storage chamber can facilitate the user to add reaction reagents according to different detection requirements, and the provided hydrophobic first and second isolation objects can effectively prevent the reaction reagents between different chambers from flowing and interfering with each other during transportation and storage, and the sealing effect is good.

[0009] A further solution is that a first positioning blind hole for assisting the positioning of the ultrasonic device is provided on the microfluidic chip, and the first positioning blind hole is provided corresponding to the lysis chamber; and / or a second positioning blind hole for assisting the positioning of the ultrasonic device is also provided on the microfluidic chip, and the second positioning blind hole is provided corresponding to the first cleaning chamber.

[0010] It can be seen from the above that, by setting the first positioning blind hole and the second positioning blind hole, it is convenient for the ultrasonic device of the amplification detection equipment to be quickly adjusted to the corresponding position and perform ultrasonic mixing operations.

[0011] A further solution is that the microfluidic chip also includes a second cleaning chamber, a second cleaning liquid filling port, a fourth filling port and a third cavity, the third cavity is located between the first cleaning chamber and the second cleaning chamber, and the second cleaning chamber is located between the third cavity and the second cavity; the second cleaning liquid filling port is connected to the second cleaning chamber; the fourth filling port is connected to the third cavity; and the third cavity is filled with a hydrophobic fourth insulator.

[0012] It can be seen from the above that the cleaning effect can be effectively improved by providing the first cleaning chamber and the second cleaning chamber.

[0013] A further solution is that a third positioning blind hole for assisting the positioning of the ultrasonic device is also provided on the microfluidic chip, and the third positioning blind hole is provided corresponding to the second cleaning chamber.

[0014] A further solution is that the second cavity includes a first cavity and a second cavity which are connected in sequence from top to bottom, and along the thickness direction of the microfluidic chip, the thickness of the second cavity is less than the thickness of the first cavity, and the second cavity is located between the first cavity and the reaction cavity; the second insulator includes silicone oil, and the first cavity and the second cavity are both filled with silicone oil; or, the second insulator includes paraffin and silicone oil, the second cavity is filled with paraffin, and the first cavity is filled with silicone oil; the second filling port is connected to the first cavity.

[0015] As can be seen from the above, when the silicone oil flows downward from the first cavity to the second cavity, the thickness of the first cavity is greater than that of the second cavity, and the channel becomes narrower. When the silicone oil passes through, the viscosity of the channel wall to the liquid increases, making it difficult for the silicone oil to flow, thereby preventing the silicone oil from entering the reaction chamber during the PCR reaction.

[0016] A further solution is that the first cavity includes a third cavity and a fourth cavity which are connected in sequence from top to bottom, and along the thickness direction of the microfluidic chip, the thickness of the third cavity is greater than the thickness of the fourth cavity; the number of first filling ports is two, one first filling port is connected to the third cavity, and the other first filling port is connected to the fourth cavity; the first insulator includes paraffin and silicone oil, the third cavity is filled with paraffin, and the fourth cavity is filled with silicone oil.

[0017] In summary, the integrated nucleic acid detection chip provided by the utility model has a simple structure, is easy to operate, has a good sealing effect, and is small in size. It can effectively reduce the risk of bubbles generated when filling reagents and is convenient for observation. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a structural diagram of an embodiment of the integrated nucleic acid detection chip of the utility model.

[0019] Figure 2 This is a diagram showing the internal structure of the back shell of the integrated nucleic acid detection chip embodiment of the present utility model.

[0020] Figure 3 It is a cross-sectional view of an embodiment of the integrated nucleic acid detection chip of the utility model. DETAILED DESCRIPTION

[0021] See also Figures 1 to 3The integrated nucleic acid detection chip provided in this embodiment includes a lysis chamber 1, a first cavity 2, a first cleaning chamber 3, a third cavity 4, a second cleaning chamber 5, a second cavity 6, a reaction chamber 7, a storage chamber 8, a storage port 9, a first filling port 100, a first filling port 101, a second filling port 11, a third filling port 12, a fourth filling port 13, an elution filling port 30, a first cleaning filling port 31, a second cleaning filling port 32 and a sample filling port 14. The sample filling port 14 is connected to the lysis chamber 1, the elution filling port 30 is connected to the reaction chamber 7, the first cleaning filling port 31 is connected to the first cleaning chamber 3 and is arranged near the bottom of the second cleaning chamber 3, and the second cleaning filling port 32 is connected to the second cleaning chamber 5 and is arranged near the bottom of the second cleaning chamber 5. The fourth filling port 13 is connected to the third cavity 4 and is arranged near the bottom of the third cavity 4. The third filling port 12 is located between the storage chamber 8 and the reaction chamber 7 and is connected to the storage chamber 8 and the reaction chamber 7. The sample adding port 14 is covered with a sample adding cover 15 .

[0022] The cracking chamber 1, the first cavity 2, the first cleaning chamber 3, the third cavity 4, the second cleaning chamber 5, the second cavity 6 and the reaction chamber 7 are connected in sequence from top to bottom. The storage chamber 8 is located above the reaction chamber 7 and is connected to the reaction chamber 7. The storage chamber 8 and the reaction chamber 7 are filled with a third insulator (not shown in the figure) for spacing. The storage chamber 8 is connected to the storage port 9, and the storage cover 19 covers the storage port 9. The user can open the storage cover 19 to add the required reaction reagents to the storage port 9, or the reaction reagents are pre-stored in the storage chamber 8 in advance. The first cavity 2 and the second cavity 6 are respectively filled with a first insulator (not shown in the figure) and a second insulator (not shown in the figure). The third cavity 4 is filled with a fourth insulator (not shown in the figure). The first insulator, the second insulator, the third insulator and the fourth insulator are all hydrophobic.

[0023] The microfluidic chip is provided with a first positioning blind hole 16, a second positioning blind hole 17 and a third positioning blind hole 18 for assisting the positioning of the ultrasonic device. The first positioning blind hole 16 is arranged corresponding to the lysis chamber 1, the second positioning blind hole 17 is arranged corresponding to the first cleaning chamber 3, and the third positioning blind hole 18 is arranged corresponding to the second cleaning chamber 5.

[0024] The first cavity 2 includes a third cavity 20 and a fourth cavity 21 which are connected in sequence from top to bottom. Along the thickness direction of the microfluidic chip, the thickness of the third cavity 20 is greater than that of the fourth cavity 21. The third cavity 20 is connected to the lysis cavity 1, and the fourth cavity 21 is connected to the first cleaning cavity 3. The fourth cavity 21 is located between the third cavity 20 and the first cleaning cavity 3. The first filling port 100 is connected to the third cavity 20 and is arranged near the bottom of the third cavity 20. The first filling port 101 is connected to the fourth cavity 21 and is arranged near the bottom of the fourth cavity 21.

[0025] The second cavity 6 includes a first cavity 60 and a second cavity 61 which are connected in sequence from top to bottom. The first cavity 60 is connected to the second cleaning cavity 5, and the second cavity 61 is connected to the reaction cavity 7. The second cavity 61 is located between the first cavity 60 and the reaction cavity 7. Along the thickness direction of the microfluidic chip, the thickness of the second cavity 61 is less than the thickness of the first cavity 60. The second filling port 110 is connected to the first cavity 60 and is arranged near the bottom of the first cavity 60. The elution liquid filling port 30 is connected to the second cavity 61 and is arranged near the bottom of the second cavity 61.

[0026] This embodiment also provides a method for adding reagents to the microfluidic chip, which further includes the following steps:

[0027] S1: Add eluent to the eluent adding port 30 until the eluent fills the reaction chamber 7;

[0028] S2: Add a third insulating material into the third filling port 12, wherein the third insulating material includes paraffin wax, so as to separate the reaction chamber 7 from the storage chamber 8;

[0029] S3: The second insulator includes silicone oil, and silicone oil is added to the elution liquid filling port 30 until the silicone oil fills the second cavity 61, and then silicone oil is added to the second filling port 11 until the silicone oil fills the first cavity 60, and then the second cleaning liquid is added to the second cleaning liquid filling port 32 until the second cleaning liquid fills the second cleaning cavity 5, and finally silicone oil is added to the fourth filling port 13 until the silicone oil fills the third cavity 4;

[0030] S4: Add the first cleaning liquid into the first cleaning liquid adding port 31 until the first cleaning liquid fills the first cleaning chamber 3;

[0031] S5: Add some silicone oil into one first filling port 101, and then add paraffin into another first filling port 100 until the paraffin fills the third cavity 20.

[0032] Optionally, the second insulator includes paraffin and silicone oil,

[0033] Step S3 includes: adding paraffin to the elution liquid filling port 30 until the paraffin fills the second cavity 61; then adding silicone oil to the second filling port 11 until the silicone oil fills the first cavity 60, separating the second cleaning cavity 5 and the reaction cavity 7, then adding the second cleaning liquid to the second cleaning liquid filling port 32 until the second cleaning liquid fills the second cleaning cavity 5, and finally adding silicone oil to the fourth filling port 13 until the silicone oil fills the third cavity 4.

[0034] When the microfluidic chip is used, the sample loading cover 15 is opened, and after adding the magnetic bead reagent and the sample to be tested to the sample loading port 14, the lysis reagent is added. After the sample loading is completed, the sample loading cover 15 is covered, and the microfluidic chip is placed vertically into the detection device in the manner that the lysis chamber 1 is on the upper reaction chamber 7 and the lower reaction chamber 7. The ultrasonic mechanism in the device closely contacts the first positioning blind hole 16 and scatters the magnetic beads. After the sample to be tested in the lysis chamber 1 is fully lysed, the magnetic beads are adsorbed and gathered by the magnetic rod device in the detection device. The detection device heats the third cavity 20 so that the paraffin melts into a liquid phase, and the magnetic rod device drives the gathered magnetic beads to enter the first cleaning chamber 3 through the paraffin and silicone oil in the first cavity 2 in turn. The ultrasonic mechanism closely contacts the second positioning blind hole 17 and scatters the magnetic beads in the first cleaning chamber 3. After the sample to be tested is fully washed in the first cleaning chamber 3. The magnetic rod device adsorbs and gathers the magnetic beads, and the magnetic rod device controls the gathered magnetic beads to pass through the silicone oil in the third cavity 4 and enter the second cleaning chamber 5. The ultrasonic mechanism closely contacts the third positioning blind hole 18 and scatters the magnetic beads. After the sample to be tested is fully washed, the magnetic rod device absorbs and gathers the magnetic beads. The magnetic rod device controls the gathered magnetic beads to pass through the silicone oil in the second cavity 6 and enter the reaction cavity 7 in sequence. The magnetic beads and samples in the reaction chamber 7 are fully heated and mixed with the elution liquid through the magnetic rod device and the heating device of the detection equipment, so that the magnetic beads are fully eluted and the purified nucleic acid is released. Then, the third insulation between the storage chamber 8 and the reaction chamber 7 is heated by the heating device of the detection equipment to melt it into a liquid phase. The magnetic rod device drags the dried reaction reagents (including magnetic beads and other magnetic carriers) in the storage chamber 8 downward into the reaction chamber 7, and repeatedly shakes the reaction reagents up and down to make them fully contact with the liquid in the reaction chamber 7 and redissolve. After standing for a period of time, the magnetic carrier is brought back to the storage chamber 8 by the magnetic rod device, and waits for the third insulation to solidify. The magnetic rod device continues to adsorb the magnetic carrier until the PCR reaction is completed to ensure that the magnetic carrier will not fall naturally due to gravity during the reaction process. Finally, the temperature control module and the heating device of the detection equipment perform a PCR reaction on the reaction chamber 7, and read the fluorescence data to complete the nucleic acid extraction and detection process.

[0035] In summary, the integrated nucleic acid detection chip of the utility model has a simple structure, is easy to operate, has a good sealing effect, and is small in size.

[0036] Finally, it should be emphasized that the above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various changes and modifications. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An integrated nucleic acid detection chip, characterized in that: It includes a cracking chamber, a first cavity, a first cleaning chamber, a second cavity, and a reaction chamber which are connected in sequence from top to bottom; The integrated nucleic acid detection chip further includes a sample addition port, an elution liquid addition port and a first cleaning liquid addition port, wherein the sample addition port is connected to the lysis chamber, the first cleaning liquid addition port is connected to the first cleaning chamber, and the elution liquid addition port is connected to the reaction chamber; The integrated nucleic acid detection chip further includes a storage cavity, a first filling port, a second filling port, a third filling port, a storage port and a storage cover, wherein the storage port is connected to the storage cavity, and the storage cover is covered at the storage port; The first cavity is filled with a first insulating material; The second cavity is filled with a second insulating material; The storage chamber is communicated with the reaction chamber, the storage chamber is located above the reaction chamber, and the storage chamber and the reaction chamber are separated by a third insulating member; The first insulator, the second insulator and the third insulator are all hydrophobic insulators; The first filling port is in communication with the first cavity; The second filling port is in communication with the second cavity; The third filling port is located between the storage chamber and the reaction chamber, and the third filling port is communicated with the storage chamber and the reaction chamber respectively.

2. The integrated nucleic acid detection chip according to claim 1, characterized in that: The integrated nucleic acid detection chip is provided with a first positioning blind hole for assisting the positioning of the ultrasonic device, and the first positioning blind hole is arranged corresponding to the lysis chamber; And / or the integrated nucleic acid detection chip is also provided with a second positioning blind hole for assisting the positioning of the ultrasonic device, and the second positioning blind hole is arranged corresponding to the first cleaning chamber.

3. The integrated nucleic acid detection chip according to claim 2, characterized in that: The integrated nucleic acid detection chip further includes a second cleaning chamber, a second cleaning liquid filling port, a fourth filling port and a third cavity, wherein the third cavity is located between the first cleaning chamber and the second cleaning chamber, and the second cleaning chamber is located between the third cavity and the second cavity; The second cleaning liquid filling port is in communication with the second cleaning chamber; The fourth filling port is in communication with the third cavity; The third cavity is filled with a fourth hydrophobic insulator.

4. The integrated nucleic acid detection chip according to claim 3, characterized in that: The integrated nucleic acid detection chip is also provided with a third positioning blind hole for assisting the positioning of the ultrasonic device, and the third positioning blind hole is arranged corresponding to the second cleaning chamber.

5. The integrated nucleic acid detection chip according to any one of claims 1 to 4, characterized in that: The second cavity comprises a first cavity and a second cavity which are sequentially connected from top to bottom, and along the thickness direction of the integrated nucleic acid detection chip, the thickness of the second cavity is less than the thickness of the first cavity, and the second cavity is located between the first cavity and the reaction chamber; The second insulating material includes silicone oil, and the first cavity and the second cavity are both filled with silicone oil; Alternatively, the second insulating material includes paraffin and silicone oil, the second cavity is filled with paraffin, and the first cavity is filled with silicone oil; The second filling port is in communication with the first cavity.

6. The integrated nucleic acid detection chip according to claim 5, characterized in that: The first cavity comprises a third cavity and a fourth cavity which are sequentially connected from top to bottom, and along the thickness direction of the integrated nucleic acid detection chip, the thickness of the third cavity is greater than the thickness of the fourth cavity; There are two first filling ports, one of which is connected to the third cavity, and the other of which is connected to the fourth cavity; The first insulating material includes paraffin and silicone oil, the third cavity is filled with paraffin, and the fourth cavity is filled with silicone oil.

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