A nucleic acid amplification chip and a nucleic acid amplification method thereof

Abstract

The present application belongs to the field of medical technology, and relates to a nucleic acid amplification chip, including a chip body and an air outlet valve, wherein the air outlet valve is installed on the chip body; the chip body is provided with a sample injection structure, a flow control channel and at least one nucleic acid amplification structure, wherein the sample injection structure is connected to the liquid inlet end of the nucleic acid amplification structure, the flow control channel is connected to the liquid outlet end, and the end of the flow control channel away from the nucleic acid amplification structure is connected to the air outlet valve; the nucleic acid amplification structure includes a dissolution chamber, a flow stop channel and an amplification chamber connected in sequence from top to bottom, wherein one end of the flow stop channel is connected to the dissolution chamber, and the other end is connected to the amplification chamber after at least one bend. The present application also relates to a nucleic acid amplification method for a nucleic acid amplification chip. The technical solution provided by the present application can realize the control of fluid flow in the chip without the need for additional equipment, is simple to operate, has a low risk of liquid crosstalk, and can accurately and efficiently realize nucleic acid amplification of the sample liquid.

Description

Technical Field

[0001] The present application relates to the field of medical technology, and more specifically, to a nucleic acid amplification chip and a nucleic acid amplification method thereof. Background Art

[0002] The microfluidic platform is a device that can miniaturize and integrate sample preparation, nucleic acid amplification and nucleic acid detection in the process of nucleic acid detection.

[0003] At present, microfluidic technology faces the following problems in the portability, popularization and household use of nucleic acid amplification: 1. Additional equipment is required to control the flow of fluid in the nucleic acid amplification chip; 2. The fluid flows fast in the nucleic acid amplification chip, the residence time is short, and the lyophilized reagent cannot be fully dissolved in the fluid; therefore, the nucleic acid amplification chip cannot be portable and the quality of the nucleic acid amplification reaction is poor. Summary of the invention

[0004] The technical problem to be solved by the embodiments of the present application is that the existing nucleic acid amplification chip has poor portability and poor quality of nucleic acid amplification reaction.

[0005] In order to solve the above technical problems, the present application provides a nucleic acid amplification chip, which adopts the following technical solution:

[0006] A nucleic acid amplification chip comprises a chip body and an air outlet valve, wherein the air outlet valve is mounted on the chip body;

[0007] The chip body is provided with a sample injection structure, a flow control channel and at least one nucleic acid amplification structure, the sample injection structure is connected to the liquid inlet end of the nucleic acid amplification structure, the flow control channel is connected to the liquid outlet end, and one end of the flow control channel away from the nucleic acid amplification structure is connected to the gas outlet valve;

[0008] The nucleic acid amplification structure comprises a dissolution chamber, a flow-stopping channel and an amplification chamber which are sequentially connected from top to bottom, wherein one end of the flow-stopping channel is connected to the dissolution chamber, and the other end is connected to the amplification chamber after being bent at least once.

[0009] Furthermore, the flow-stopping channel includes a capillary drive section and a gravity drive section, the capillary drive section is connected to the dissolution chamber, and the gravity drive section is connected to the amplification chamber, wherein the diameter of the capillary drive section is less than or equal to the diameter of the gravity drive section;

[0010] A first angle is formed between the capillary driving section and the horizontal plane, and the first angle is 5° to 45°;

[0011] A second angle is formed between the capillary drive section and the gravity drive section, and the second angle is 90° to 135°.

[0012] Furthermore, the flow-stopping channel also includes a gravity buffer section, which is vertically connected between the dissolution chamber and the capillary drive section, wherein the diameter of the capillary drive section is smaller than or equal to the diameter of the gravity buffer section.

[0013] Furthermore, the capillary drive section has a diameter of 0.25 mm to 1.5 mm; and / or

[0014] The depth of the capillary drive section is 0.2 mm to 1.5 mm; and / or

[0015] The diameter of the gravity-driven section is 0.5 mm to 3 mm; and / or

[0016] The depth of the gravity driven section is 0.2 mm to 1.5 mm; and / or

[0017] The diameter of the gravity buffer section is 0.5 mm to 3 mm; and / or

[0018] The depth of the gravity buffer section is 0.2 mm to 1.5 mm; and / or

[0019] The length of the gravity buffer section is 0 to 3 mm.

[0020] Furthermore, the air outlet valve is a tube-breaking handle integrally formed on the chip body, and the flow control channel is connected to the outside by breaking the tube-breaking handle off from the chip body.

[0021] Furthermore, the nucleic acid amplification structure further includes an overflow channel and a waste liquid chamber, the waste liquid chamber is connected to the amplification chamber through the overflow channel, and the flow control channel is connected to the top of the waste liquid chamber;

[0022] The sum of the volume of the waste liquid chamber and the volume of the amplification chamber is greater than the volume of the lysis chamber.

[0023] Furthermore, the dissolution chamber is provided with an air hole, and the chip body is also provided with an exhaust flow channel, the dissolution chamber is connected to the exhaust flow channel through the air hole, and the exhaust flow channel is connected to the outside;

[0024] The air vent is provided with a hydrophobic air-permeable member.

[0025] Further, a first optical element position is provided above the amplification chamber; and / or a second optical element position is provided below the amplification chamber;

[0026] The first optical element position and the second optical element position are used for placing optical components.

[0027] In order to solve the above technical problems, the present application also provides a nucleic acid amplification method of a nucleic acid amplification chip, which adopts the following technical solution:

[0028] A nucleic acid amplification method for a nucleic acid amplification chip, applied to the nucleic acid amplification chip as described above, comprises the following steps:

[0029] Providing a sample liquid, injecting the sample liquid into the injection structure, the sample liquid enters the dissolution chamber through the injection structure, and under the capillary action and gas pressure of the stop flow channel, the sample liquid stays in the dissolution chamber to fully dissolve the freeze-dried ball to form a liquid to be reacted;

[0030] Open the gas outlet valve, and the liquid to be reacted breaks through the capillary action of the flow-stopping channel under the action of gravity and enters the amplification chamber until all the liquid to be reacted enters the amplification chamber;

[0031] The sample injection structure and the gas outlet valve are sealed, and the liquid to be reacted undergoes a nucleic acid amplification reaction in the amplification chamber.

[0032] Furthermore, a first optical element position is provided above the amplification chamber; a second optical element position is provided below the amplification chamber; the first optical element position and the second optical element position are used to place optical components;

[0033] After the step of performing a nucleic acid amplification reaction on the liquid to be reacted in the amplification chamber, the following steps are also included:

[0034] The optical components are started to detect the reaction liquid in the amplification chamber to obtain nucleic acid amplification data.

[0035] Compared with the prior art, the embodiments of the present application have the following beneficial effects:

[0036] The nucleic acid amplification chip provided by the present application realizes the control of the air pressure balance in the nucleic acid amplification structure by controlling the opening and closing of the switch valve through the sequentially connected sample injection structure, nucleic acid amplification structure and flow control channel, and combined with the setting of the air outlet valve connected to the end of the flow control channel, thereby achieving the effect of controlling the flow of internal fluid without additional equipment; at the same time, combined with the setting of the flow-stop channel in the nucleic acid amplification structure, the dissolution chamber is limited, so that the sample liquid can stay in the dissolution chamber to fully dissolve the freeze-dried ball, thereby improving the quality of the subsequent nucleic acid amplification reaction. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the scheme of the present application, a brief introduction is given below to the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0038] Figure 1 is a schematic structural diagram of a nucleic acid amplification chip according to an embodiment of the present application;

[0039] Figure 2 is a schematic structural diagram of the nucleic acid amplification chip in another direction of an embodiment of the present application;

[0040] Figure 3 It is a schematic diagram of the structure of the chip body and the air outlet valve of the embodiment of the present application;

[0041] Figure 4 is a front view of the chip body and the gas outlet valve of the embodiment of the present application;

[0042] Figure 5 About Figure 4 A schematic diagram of the structure enlargement at A;

[0043] Figure 6 It is a flow chart of the nucleic acid amplification method of the nucleic acid amplification chip according to an embodiment of the present application.

[0044] Reference numerals:

[0045] 1. Chip body; 11. Sample injection structure; 111. Sample injection port; 112. Sample injection channel; 113. Liquid separation channel; 12. Nucleic acid amplification structure; 121. Dissolution chamber; 1211. Air vent; 1212. Hydrophobic air permeable member; 122. Stop flow channel; 1221. Capillary drive section; 1222. Gravity drive section; 1223; Gravity buffer section; 123. Amplification chamber; 1231. First optical element position; 1232. Second optical element position; 124. Overflow channel; 125. Waste liquid chamber; 126. Heat conducting member; 13. Flow control channel; 2. Exhaust valve; 3. Sealing member. DETAILED DESCRIPTION

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by technicians in the technical field of this application; the terms used in the specification of the application herein are only for the purpose of describing specific embodiments and are not intended to limit this application; the terms "including" and "having" and any variations thereof in the specification and claims of this application and the above-mentioned drawings are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of this application or the above-mentioned drawings are used to distinguish different objects, not to describe a specific order.

[0047] Reference to "embodiments" herein means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present application. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0048] See also Figure 1 , Figure 2 As shown, an embodiment of the present application provides a nucleic acid amplification chip, including a chip body 1 and an exhaust valve 2, wherein the exhaust valve 2 is mounted on the chip body 1. In this embodiment, the exhaust valve 2 is configured as a tube breaking handle integrally formed on the chip body 1.

[0049] In some embodiments, the chip body 1 is provided with a sample injection structure 11, a flow control channel 13 and at least one nucleic acid amplification structure 12, the sample injection structure 11 is connected to the liquid inlet end of the nucleic acid amplification structure 12, the flow control channel 13 is connected to the liquid outlet end, and the end of the flow control channel 13 away from the nucleic acid amplification structure 12 is connected to the gas outlet valve 2. In this embodiment, the gas outlet valve 2 is a tube breaking handle, and the tube breaking handle closes the end of the flow control channel 13. By breaking the tube breaking handle off the chip body 1, the flow control channel 13 is connected to the outside world.

[0050] The nucleic acid amplification structure 12 includes a dissolution chamber 121, a flow-stopping channel 122 and an amplification chamber 123 which are sequentially connected from top to bottom, wherein one end of the flow-stopping channel 122 is connected to the dissolution chamber 121, and the other end is connected to the amplification chamber 123 after being bent at least once.

[0051] When the outlet valve 2 is closed, the sample liquid is injected into the chip body 1, and the sample liquid enters the dissolution chamber 121 through the injection structure 11. Under the capillary action and gas pressure of the stop flow channel 122, the sample liquid stays in the dissolution chamber to fully dissolve the freeze-dried ball to form a liquid to be reacted;

[0052] When the gas outlet valve 2 is opened, the amplification chamber 123 is connected to the outside through the flow control channel 13, and the reaction liquid flows downward under the action of gravity, breaking through the capillary action of the flow-stopping channel 122, and the reaction liquid all enters the amplification chamber 123 and undergoes nucleic acid amplification reaction.

[0053] The nucleic acid amplification chip provided in the embodiment of the present application realizes the control of the air pressure balance in the nucleic acid amplification structure 12 by controlling the opening and closing of the switch valve through the arrangement of the sample injection structure 11, the nucleic acid amplification structure 12 and the flow control channel 13 connected in sequence, and in combination with the arrangement of the air outlet valve 2 connected to the end of the flow control channel 13, thereby achieving the effect of controlling the flow of the internal fluid without the need for additional equipment; at the same time, in combination with the arrangement of the flow stop channel 122 in the nucleic acid amplification structure 12, the dissolution chamber 121 is limited in flow, so that the sample liquid can stay in the dissolution chamber 121 to fully dissolve the freeze-dried ball, thereby improving the quality of the subsequent nucleic acid amplification reaction.

[0054] The following description will be made by taking the chip body 1 having four nucleic acid amplification structures 12 as an example. Figure 1 , Figure 3 , Figure 4 shown.

[0055] See also Figure 1 to Figure 4 As shown, in some embodiments, the injection structure 11 includes an injection port 111, an injection channel 112 and at least one liquid separation channel 113, the injection port 111 is connected to the injection channel 112, the injection channel 112 is connected to the liquid separation channel 113, the number of the liquid separation channels 113 is consistent with the number of the nucleic acid amplification structures 12. In this embodiment, the number of the liquid separation channels 113 is four, and each of the liquid separation channels 113 is connected to a corresponding nucleic acid amplification structure 12.

[0056] In this embodiment, the liquid separation channels 113 are distributed in an even number of tree-like shapes and are expanded layer by layer to form four liquid outlets respectively connected to the corresponding nucleic acid amplification structures 12 .

[0057] In other embodiments, four liquid separation channels 113 may be arranged in parallel, and the liquid inlets of the four liquid separation channels 113 are connected to the sample injection channel 112 .

[0058] See also Figures 2 to 4 As shown, in some embodiments, the dissolution chamber 121 is provided with an air hole 1211, and the chip body 1 is also provided with an exhaust channel (not shown in the figure), the dissolution chamber 121 is connected to the exhaust channel through the air hole 1211, and the exhaust channel is connected to the outside. In this embodiment, since the dissolution chamber 121 is provided with an air hole 1211, when the sample liquid enters the dissolution chamber 121 through the liquid inlet structure, the gas in the dissolution chamber 121 is discharged from the air hole 1211 until all the sample liquid enters the dissolution chamber 121, and the freeze-dried ball is fully dissolved to form a liquid to be reacted.

[0059] In some embodiments, a hydrophobic breathable member 1212 is provided on the air hole 1211. In this embodiment, the hydrophobic breathable member 1212 may be a hydrophobic breathable membrane. The hydrophobic breathable membrane completely covers the air hole 1211 by means of adhesive to achieve exhaust and prevent leakage of sample liquid.

[0060] In other embodiments, the hydrophobic breathable member 1212 may also be a hydrophobic breathable filler, and the hydrophobic breathable filler is interference-fitted with the breathable hole 1211 to achieve exhaust and prevent leakage of the sample liquid.

[0061] In other embodiments, the hydrophobic breathable member 1212 and the breathable hole 1211 may be welded together by ultrasonic welding or laser welding.

[0062] See also Figure 1 , Figure 3 , Figure 4 , Figure 5 As shown, in some embodiments, the flow-stopping channel 122 is bent at least once, and the flow-stopping channel 122 forms an angle at the bending point.

[0063] The embodiment of the present application designs the flow-stopping channel 122 to be bent at least once, so that when the sample liquid enters the flow-stopping channel 122 , the component velocity of the sample liquid in the downward direction can be reduced, thereby achieving a flow-stopping effect on the sample liquid.

[0064] In this embodiment, the stop flow channel 122 is bent once to be in a “<” shape, and the stop flow channel 122 includes a capillary drive section 1221 and a gravity drive section 1222. The capillary drive section 1221 is connected to the dissolution chamber 121, and the gravity drive section 1222 is connected to the amplification chamber 123.

[0065] See also Figure 5 As shown, in this embodiment, a first angle is formed between the capillary drive segment 1221 and the horizontal plane, and the first angle is 5° to 45°. Specifically, the first angle can be set to any one of 5°, 10°, 15°, 20°, 25°, 30°, 35°, 40°, 45° or a range formed between any two values.

[0066] In this embodiment, a second angle is formed between the capillary drive section 1221 and the gravity drive section 1222, and the second angle is 90° to 135°. Specifically, the first angle can be set to any one of 90°, 95°, 100°, 105°, 110°, 115°, 120°, 125°, 130°, and 135°, or a range formed between any two values.

[0067] The embodiment of the present application uses the capillary driving section 1221 to be tilted so that the capillary driving section 1221 can buffer the sample liquid at the sample injection structure 11. At the same time, combined with the capillary effect of the capillary driving section 1221 itself and the closed setting of the nucleic acid amplification structure 12, it can prevent the sample liquid from flowing downward, so that the sample liquid can fully react and dissolve with the freeze-dried ball in the dissolution chamber 121. In addition, by setting the angle between the capillary driving section 1221 and the gravity driving section 1222, it is ensured that the gravity driving section 1222 is tilted downward or vertically set, so that after the outlet valve 2 is opened, the reaction liquid can flow downward under the action of gravity and enter the amplification chamber 123.

[0068] Please continue reading Figure 5 As shown, in some embodiments, the flow-stopping channel 122 further includes a gravity buffer section 1223 , and the gravity buffer section 1223 is vertically connected between the dissolution chamber 121 and the capillary driving section 1221 .

[0069] In the embodiment of the present application, the gravity buffer section 1223 is provided to buffer the sample liquid, thereby reducing the downward flow speed of the sample liquid, so that the sample liquid can fully react and dissolve with the freeze-dried ball in the dissolution chamber 121 .

[0070] In some embodiments, the diameter of the capillary driving section 1221 is smaller than or equal to the diameter of the gravity driving section 1222 ; the diameter of the capillary driving section 1221 is smaller than or equal to the diameter of the gravity buffer section 1223 .

[0071] The embodiment of the present application reduces the volume of the sample liquid entering the capillary driving section 1221 by limiting the tube diameter, thereby further reducing the downward flow speed of the sample liquid, so that the sample liquid can fully react and dissolve with the freeze-dried ball in the dissolution chamber 121.

[0072] In this embodiment, the volume of the sample liquid in each section of the flow channel in the flow-stopping flow channel 122 is also related to the depth. The depth mentioned below refers to the depth of the depression of each section of the flow channel in the flow-stopping flow channel 122 on the plane of the chip body 1 .

[0073] In some embodiments, the diameter of the capillary driving section 1221 is 0.25 mm to 1.5 mm. Specifically, the diameter of the capillary driving section 1221 can be set to any value of 0.25 mm, 0.5 mm, 1.0 mm, 1.5 mm or a range formed between any two values.

[0074] In some embodiments, the depth of the capillary driving section 1221 is 0.2 mm to 1.5 mm. Specifically, the depth of the capillary driving section 1221 can be set to any one of 0.2 mm, 0.5 mm, 1.0 mm, and 1.5 mm or a range formed between any two values.

[0075] In some embodiments, the diameter of the gravity-driven section 1222 is 0.5 mm to 3 mm. Specifically, the diameter of the gravity-driven section 1222 can be set to any value of 0.5 mm, 1.0 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm or a range formed between any two values.

[0076] In some embodiments, the depth of the gravity-driven section 1222 is 0.2 mm to 1.5 mm. Specifically, the depth of the gravity-driven section 1222 can be set to any one of 0.2 mm, 0.5 mm, 1.0 mm, and 1.5 mm, or a range formed between any two values.

[0077] In some embodiments, the diameter of the gravity buffer section 1223 is 0.5 mm to 3 mm. Specifically, the diameter of the gravity buffer section 1223 can be set to any value of 0.5 mm, 1.0 mm, 1.5 mm, 2.0 mm, 2.5 mm, 3 mm or a range formed between any two values.

[0078] In some embodiments, the depth of the gravity buffer section 1223 is 0.2 mm to 1.5 mm. Specifically, the depth of the gravity buffer section 1223 can be set to any value of 0.2 mm, 0.5 mm, 1.0 mm, 1.5 mm or a range formed between any two values.

[0079] The embodiment of the present application controls the sample liquid capacity of the capillary driving section 1221 and the gravity buffer section 1223 by limiting the length and depth of the capillary driving section 1221, and limiting the length and depth of the gravity buffer section 1223. By limiting the sample liquid capacity of the capillary driving section 1221, the downward flow speed of the sample liquid is further reduced when the end of the flow control channel 13 is closed, so that the sample liquid can fully react and dissolve with the freeze-dried ball in the dissolution chamber 121.

[0080] In some embodiments, the length of the gravity buffer section 1223 is 0 to 3 mm. Specifically, in the present embodiment, when the flow-stop channel 122 is bent once to form the capillary drive section 1221 and the gravity drive section 1222, the length of the gravity buffer section 1223 is 0 mm. In other embodiments, when the flow-stop channel 122 is bent twice to form the gravity buffer section 1223, the capillary drive section 1221 and the gravity drive section 1222, the length of the gravity buffer section 1223 can be set to any one of 0.5 mm, 1 mm, 1.5 mm, 2 mm, 3 mm or a range formed between any two values.

[0081] The embodiment of the present application buffers the sample liquid by adjusting the length of the gravity buffer section. The longer the length of the gravity buffer section is, the better the effect of limiting the downward flow speed of the sample liquid is, so that the sample liquid can fully react and dissolve with the freeze-dried ball in the dissolution chamber 121.

[0082] See also Figure 3 , Figure 4 As shown, in some embodiments, the thickness of the amplification chamber 123 is less than or equal to 2 mm. Preferably, the thickness of the amplification chamber 123 is less than or equal to 1 mm.

[0083] The embodiment of the present application limits the thickness of the amplification chamber 123 so that it is convenient for the staff to perform nucleic acid amplification detection after the nucleic acid amplification reaction is completed.

[0084] Please continue reading Figure 3 , Figure 4 As shown, the nucleic acid amplification structure 12 further includes an overflow channel 124 and a waste liquid chamber 125, the waste liquid chamber 125 is connected to the amplification chamber 123 through the overflow channel 124, and the flow control channel 13 is connected to the top of the waste liquid chamber 125;

[0085] The sum of the volume of the waste liquid chamber 125 and the volume of the amplification chamber 123 is greater than the volume of the lysis chamber 121 .

[0086] The embodiment of the present application collects the reaction liquid overflowing the amplification chamber 123 by disposing the overflow channel 124 and the waste liquid chamber 125 to prevent the sample liquid from flowing back to the amplification chamber 123 after some splashed sample liquid enters the flow control channel 13 .

[0087] See also Figure 1 , Figure 2 As shown, in some embodiments, the nucleic acid amplification structure 12 includes a heat conductor 126, which is installed on the chip body 1 and corresponds to the position of the amplification chamber 123, and is used to control the reaction temperature in the amplification chamber 123, so that the nucleic acid amplification reaction can be carried out at a suitable temperature, thereby improving the quality of the nucleic acid amplification reaction.

[0088] See also Figure 3 As shown, in some embodiments, a first optical element position 1231 is provided below the amplification chamber 123; and / or a second optical element position 1232 is provided above the amplification chamber 123; the first optical element position 1231 and the second optical element position 1232 are used to place optical components.

[0089] In this embodiment, a first optical element position 1231 is provided directly above the amplification chamber 123, and a second optical element position 1232 is provided directly below the amplification chamber 123, wherein the optical component that can be placed in the first optical element position 1231 can be a light reflecting element or structure, and the optical component that can be placed in the second optical element position 1232 can be a light source or an optical sensor.

[0090] When the detection method is fluorescence-localized PCR, a light reflecting element is placed at the first optical element position 1231 directly above the amplification chamber 123, and an optical sensor is placed at the second optical element position 1232 directly below the amplification chamber 123. Through the above arrangement, the amount of fluorescence that the optical sensor can accept can be increased, thereby improving the quality of detection.

[0091] When the detection method is colorimetry, the first optical element directly above the amplification chamber 123 has no optical components, and the second optical element position 1232 directly below the amplification chamber 123 is used to place a light source. Through the above arrangement, the brightness of the solution during detection can be increased, thereby improving the quality of detection.

[0092] See also Figure 1 , Figure 2 As shown, in some embodiments, the nucleic acid amplification chip further includes a seal 3 , which covers the chip body 1 and the gas outlet valve 2 and is used to seal the sample injection structure, the nucleic acid amplification structure 12 and the flow control channel 13 .

[0093] In this embodiment, the seal 3 can be made of high-grade synthetic glue, which is resistant to strong acids and alkalis, has a strong sealing effect on the sample injection structure, the nucleic acid amplification structure 12 and the flow control channel 13, has no backing glue and residue during the bonding process, has good biocompatibility, and will not react with reagents or samples.

[0094] In other embodiments, the sealing member 3 may also be a plastic thin plate, and the plastic thin plate is sealed and fixed on the chip body 1 by ultrasonic welding, laser welding, hot pressing welding or the like.

[0095] See also Figure 6 As shown, based on the above-mentioned nucleic acid amplification chip, an embodiment of the present application also provides a nucleic acid amplification method for a nucleic acid amplification chip. In this embodiment, before injecting the sample liquid, it is necessary to ensure that the nucleic acid amplification chip is placed upright with the front side facing forward to ensure that the sample liquid can flow from top to bottom into the dissolution chamber under the action of gravity. At the same time, it is also necessary to ensure that the tube breaking handle closes the flow control channel to ensure that the sample liquid will not break through the capillary action of the stop flow channel and flow further downward before it fully reacts with the freeze-dried ball.

[0096] In some embodiments, the nucleic acid amplification method comprises the following steps:

[0097] Step S100, providing a sample liquid, injecting the sample liquid into an injection structure, and the sample liquid enters a dissolution chamber through the injection structure. Under the capillary action of the stop flow channel and the gas pressure, the sample liquid stays in the dissolution chamber to fully dissolve the freeze-dried ball to form a liquid to be reacted.

[0098] In this embodiment, the sample liquid is added to the injection channel through the injection port, and then flows along the four liquid separation channels to the dissolution chambers in the corresponding nucleic acid decomposition structure under the action of gravity; the capillary action of the capillary driving section of the stop flow channel, the gas pressure and the setting of the inclined flow channel can slow down or even limit the downward flow of the sample liquid, so that the sample liquid stays in the dissolution chamber to fully dissolve the freeze-dried balls to form a liquid to be reacted.

[0099] Step S200, opening the gas outlet valve, the liquid to be reacted breaks through the capillary action of the flow-stopping channel under the action of gravity and enters the amplification chamber until all the liquid to be reacted enters the amplification chamber.

[0100] In this embodiment, the outlet valve is a tube breaking handle, which is broken to connect the flow control channel to the outside world. The reaction liquid breaks through the slowing effect of capillary action under the action of gravity and enters the amplification chamber until all the reaction liquid enters the amplification chamber.

[0101] In this embodiment, the nucleic acid amplification structure also includes an overflow channel and a waste liquid chamber. After all the reaction liquid enters the amplification chamber, the excess reaction liquid flows along the overflow channel to the waste liquid chamber to collect the reaction liquid overflowing the amplification chamber, thereby preventing part of the splashed sample liquid from entering the flow control channel and causing the sample liquid to flow back into the amplification chamber.

[0102] Step S300, the sample injection structure and the gas outlet valve are closed, and the liquid to be reacted undergoes a nucleic acid amplification reaction in the amplification chamber.

[0103] The nucleic acid amplification method provided in the embodiment of the present application controls the opening and closing of the switch valve by executing corresponding steps in the above-mentioned nucleic acid amplification chip, thereby controlling the air pressure balance in the nucleic acid amplification structure, thereby achieving the effect of controlling the internal fluid flow without additional equipment; at the same time, combined with the setting of the stop flow channel in the nucleic acid amplification structure, the dissolution chamber is limited, so that the sample liquid can stay in the dissolution chamber to fully dissolve the freeze-dried ball, thereby improving the quality of the subsequent nucleic acid amplification reaction.

[0104] In some embodiments, a first optical element position is disposed above the amplification chamber; a second optical element position is disposed below the amplification chamber; and the first optical element position and the second optical element position are used to place optical components.

[0105] In step S300, after the reaction liquid undergoes nucleic acid amplification reaction in the amplification chamber, the following steps are further included:

[0106] The optical components are started to detect the reaction liquid in the amplification chamber to obtain nucleic acid amplification data.

[0107] In this embodiment, when the detection method is fluorescence positioning PCR, a light reflecting element is placed at the first optical element position directly above the amplification chamber, and an optical sensor is placed at the second optical element position directly below the amplification chamber. Through the above settings, the amount of fluorescence that the optical sensor can accept can be increased, thereby improving the quality of detection.

[0108] In other embodiments, when the detection method is colorimetry, the first optical element directly above the amplification chamber has no optical components, and the second optical element directly below the amplification chamber is placed with a light source. Through the above arrangement, the brightness of the solution during detection can be increased, thereby improving the quality of detection.

[0109] Obviously, the embodiments described above are only some embodiments of the present application, rather than all embodiments. The preferred embodiments of the present application are given in the accompanying drawings, but they do not limit the patent scope of the present application. The present application can be implemented in many different forms. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive. Although the present application is described in detail with reference to the aforementioned embodiments, for those skilled in the art, it is still possible to modify the technical solutions recorded in the aforementioned specific implementation methods, or to perform equivalent replacement of some of the technical features therein. Any equivalent structure made using the contents of the specification and drawings of this application, directly or indirectly used in other related technical fields, is similarly within the scope of patent protection of this application.

Claims

1. A nucleic acid amplification chip, characterized in that: It includes a chip body and an air outlet valve, wherein the air outlet valve is installed on the chip body; The chip body is provided with a sample injection structure, a flow control channel and at least one nucleic acid amplification structure, the sample injection structure is connected to the liquid inlet end of the nucleic acid amplification structure, the flow control channel is connected to the liquid outlet end, and one end of the flow control channel away from the nucleic acid amplification structure is connected to the gas outlet valve; The nucleic acid amplification structure comprises a dissolution chamber, a flow-stopping channel and an amplification chamber which are sequentially connected from top to bottom, wherein one end of the flow-stopping channel is connected to the dissolution chamber, and the other end is connected to the amplification chamber after being bent at least once.

2. The nucleic acid amplification chip according to claim 1, characterized in that: The flow-stopping channel includes a capillary drive section and a gravity drive section, wherein the capillary drive section is connected to the dissolution chamber, and the gravity drive section is connected to the amplification chamber, wherein the diameter of the capillary drive section is smaller than or equal to the diameter of the gravity drive section; A first angle is formed between the capillary driving section and the horizontal plane, and the first angle is 5° to 45°; A second angle is formed between the capillary drive section and the gravity drive section, and the second angle is 90° to 135°.

3. The nucleic acid amplification chip according to claim 2, characterized in that: The flow-stopping channel further includes a gravity buffer section, which is vertically connected between the dissolution chamber and the capillary driving section, wherein the diameter of the capillary driving section is smaller than or equal to the diameter of the gravity buffer section.

4. The nucleic acid amplification chip according to claim 3, characterized in that: The diameter of the capillary drive section is 0.25 mm to 1.5 mm; and / or The depth of the capillary drive section is 0.2 mm to 1.5 mm; and / or The diameter of the gravity-driven section is 0.5 mm to 3 mm; and / or The depth of the gravity driven section is 0.2 mm to 1.5 mm; and / or The diameter of the gravity buffer section is 0.5 mm to 3 mm; and / or The depth of the gravity buffer section is 0.2 mm to 1.5 mm; and / or The length of the gravity buffer section is 0 to 3 mm.

5. The nucleic acid amplification chip according to claim 1, characterized in that: The air outlet valve is a tube-breaking handle integrally formed on the chip body. By breaking the tube-breaking handle off from the chip body, the flow control channel is connected to the outside.

6. The nucleic acid amplification chip according to claim 1, characterized in that: The nucleic acid amplification structure further includes an overflow channel and a waste liquid chamber, wherein the waste liquid chamber is connected to the amplification chamber via the overflow channel, and the flow control channel is connected to the top of the waste liquid chamber; The sum of the volume of the waste liquid chamber and the volume of the amplification chamber is greater than the volume of the lysis chamber.

7. The nucleic acid amplification chip according to claim 1, characterized in that: The dissolution chamber is provided with an air hole, and the chip body is also provided with an exhaust flow channel. The dissolution chamber is connected to the exhaust flow channel through the air hole, and the exhaust flow channel is connected to the outside world; The air vent is provided with a hydrophobic air-permeable member.

8. The nucleic acid amplification chip according to claim 1, characterized in that: A first optical element position is disposed above the amplification chamber; and / or a second optical element position is disposed below the amplification chamber; The first optical element position and the second optical element position are used for placing optical components.

9. A nucleic acid amplification method for a nucleic acid amplification chip, applied to the nucleic acid amplification chip according to any one of claims 1 to 8, characterized in that: The following steps are involved: Providing a sample liquid, injecting the sample liquid into a sample injection structure, the sample liquid enters a dissolution chamber through the sample injection structure, and under the capillary action of a flow-stopping channel and gas pressure, the sample liquid stays in the dissolution chamber to fully dissolve the freeze-dried ball to form a liquid to be reacted; Open the gas outlet valve, and the liquid to be reacted breaks through the capillary action of the flow-stopping channel under the action of gravity and enters the amplification chamber until all the liquid to be reacted enters the amplification chamber; The sample injection structure and the gas outlet valve are sealed, and the liquid to be reacted undergoes a nucleic acid amplification reaction in the amplification chamber.

10. The nucleic acid amplification method of the nucleic acid amplification chip according to claim 9, characterized in that: A first optical element position is provided above the amplification chamber; a second optical element position is provided below the amplification chamber; the first optical element position and the second optical element position are used to place optical components; After the step of performing a nucleic acid amplification reaction on the liquid to be reacted in the amplification chamber, the following steps are also included: The optical components are started to detect the reaction liquid in the amplification chamber to obtain nucleic acid amplification data.

Images