Nucleic acid cleaving device and control method thereof

Abstract

The application provides a nucleic acid cleavage device and a control method thereof, wherein the nucleic acid cleavage device comprises a device main body and a vibrating needle body, the vibrating needle body is detachably connected with the device main body, an ultrasonic unit is arranged in the device main body, an ultrasonic head of the ultrasonic unit is vibrationally coupled with the vibrating needle body, and the vibrating needle body has a cleavage working state of being at least partially inserted into a nucleic acid reaction liquid in a reaction liquid container. The ultrasonic unit directly conducts the action on the vibrating needle body to generate high-frequency vibration and then directly conducts the action on the nucleic acid reaction liquid, so that strong vibration and micro-flow effect are caused in the nucleic acid reaction liquid, the shearing force of the cleavage liquid on the cell membrane is enhanced, and the cleavage is more uniform. Since the ultrasonic wave can be directly conducted to the liquid in which the vibrating needle body is inserted and is not absorbed and scattered by a container base body with a large size, the utilization efficiency of the ultrasonic wave energy is significantly improved, and the cleavage time is shorter.

Description

Technical Field

[0001] This invention belongs to the field of molecular diagnostic technology, specifically relating to a nucleic acid lysis device and its control method. Background Technology

[0002] Nucleic acid testing is an important means of modern medical diagnosis. Its basic process includes sample collection, cell lysis, nucleic acid extraction, nucleic acid purification, amplification and detection. Among these steps, cell lysis is the key step for nucleic acid release, and the lysis efficiency directly affects the sensitivity and accuracy of subsequent detection.

[0003] Traditional cell lysis methods mainly include two categories: chemical lysis and physical lysis.

[0004] Chemical lysis uses surfactants, proteases, and other components in the lysis buffer to disrupt the cell membrane and cell wall structure, releasing nucleic acids into the solution. This method is relatively simple to operate, but the lysis rate is slow, especially for difficult-to-lyse samples such as FFPE samples and fungi, where the lysis time often takes more than 30 minutes.

[0005] Physical lysis methods mainly include mechanical grinding, ultrasonic lysis, and thermal lysis. Ultrasonic lysis utilizes the cavitation effect generated by ultrasound in liquids to create a local high temperature and high pressure environment, effectively breaking down cell structures and releasing nucleic acids. Ultrasonic lysis has the advantages of high lysis efficiency and wide applicability. However, traditional ultrasound equipment is large in size and consumes a lot of energy, making it difficult to apply to POCT (Point-of-Care Testing) scenarios.

[0006] Specifically, existing ultrasonic lysis techniques mainly suffer from the following technical problems: 1. Difficulty in ultrasonic integration: Ultrasonic pyrolysis requires high-frequency vibration to provide stable energy to form a cavitation effect. However, micro piezoelectric transducers have problems such as low power density and poor energy coupling efficiency, and cannot form a uniform cavitation field in the microcavity. 2. Low energy utilization efficiency: In traditional ultrasonic pyrolysis devices, ultrasonic energy is easily absorbed and scattered by the container substrate material, resulting in local overheating, uneven pyrolysis effect, and low energy utilization efficiency.

[0007] 3. Cost and compatibility: Ultrasonic pyrolysis has a significant cost disadvantage. It requires the integration of core components such as miniature piezoelectric transducers into the cartridge, and its material and process costs are 3-5 times that of traditional chemical pyrolysis cartridges. Furthermore, it is incompatible with existing POCT testing instruments. Summary of the Invention

[0008] Therefore, the technical problem to be solved by the present invention is to provide a nucleic acid lysis device and its control method, which effectively overcomes the shortcomings of the prior art, which uses an ultrasonic device outside the reaction liquid container to indirectly sonicate the reaction liquid, resulting in low energy utilization efficiency and uneven lysis effect, and uses an ultrasonic device inside the reaction liquid container, which has high cost or poor lysis effect due to the use of a micro ultrasonic device, making it unsuitable for POCT detection scenarios.

[0009] To address the aforementioned problems, the present invention provides a nucleic acid lysis device, comprising a device body and a vibrating needle, wherein the vibrating needle is detachably connected to the device body, an ultrasonic unit is disposed within the device body, the ultrasonic head of the ultrasonic unit is vibratingly coupled to the vibrating needle, and the vibrating needle is in a lysis working state in which it is at least partially inserted into the nucleic acid reaction solution within the reaction solution container.

[0010] In some embodiments, the main body of the device is further provided with a suction unit, the vibrating needle is a hollow needle, the first end of the hollow needle is detachably connected to the suction interface of the suction unit, and when the vibrating needle is in the lysis working state, the second end of the hollow needle is below the surface of the nucleic acid reaction solution.

[0011] In some embodiments, the ultrasonic head of the ultrasonic unit is coupled to the suction interface.

[0012] In some embodiments, the inner diameter of the hollow needle body is d, 0.1mm≤d≤10mm; and / or, the wall thickness of the hollow needle body is δ, 0.5mm≤δ≤5mm; and / or, the length of the hollow needle body is L, 10mm≤L≤200mm.

[0013] In some embodiments, the hollow needle body comprises a metal matrix.

[0014] In some embodiments, the hollow needle body further includes a biocompatible outer layer, and the portion of the metal matrix in contact with the nucleic acid reaction solution when the vibrating needle body is in the lysis working state is a first portion, and the biocompatible outer layer at least covers the outer wall surface of the first portion.

[0015] In some embodiments, the material of the metal substrate is at least one of aluminum alloy, stainless steel, and iron; and / or, the biocompatible outer layer is at least one of PP coating and metal oxide layer.

[0016] In some embodiments, a temperature control unit is also provided inside the main body of the device, and the temperature control unit is heat-exchange connected with the vibrating needle body.

[0017] The present invention also provides a control method for the above-mentioned nucleic acid lysis device, comprising the following steps: Place the reaction solution container in a preset position so that the second end of the vibrating needle is below the surface of the nucleic acid reaction solution in the reaction solution container; Upon receiving a nucleic acid lysis command, the ultrasonic unit is controlled to operate.

[0018] In some embodiments, when a suction unit is included and the vibrating needle body is a hollow needle body, after receiving the nucleic acid lysis command, the suction unit is also controlled to operate so that the nucleic acid reaction solution is repeatedly sucked in and expelled from the hollow cavity of the hollow needle body. And / or, When a temperature control unit is included, after receiving a nucleic acid lysis command, the temperature control unit is also controlled to operate so that the temperature inside the hollow cavity of the hollow needle is at the set temperature.

[0019] This invention provides a nucleic acid lysis device and its control method. An ultrasonic unit within the device body transmits ultrasonic energy to a vibrating needle coupled to it. Simultaneously, the vibrating needle is in a lysis state inserted into the nucleic acid reaction solution. When the ultrasonic unit operates, it directly transmits high-frequency vibrations to the vibrating needle, which in turn directly transmits these vibrations to the nucleic acid reaction solution, inducing strong vibrations and microfluidic effects. This enhances the shear force of the lysis solution on the cell membrane (nucleic acid cell membrane), resulting in more uniform lysis. Because the ultrasonic waves can be directly transmitted to the liquid into which the vibrating needle is inserted, they are no longer absorbed or scattered by the larger container substrate, significantly improving the utilization efficiency of ultrasonic energy and shortening the lysis time. The ultrasonic unit is integrated within the device body, eliminating the need for integration into the corresponding reaction solution container (disposable consumable). This ensures energy utilization efficiency while significantly reducing consumable costs, better meeting the low-cost requirements for reaction solution containers in POCT scenarios. Attached Figure Description

[0020] Figure 1 This is a schematic diagram of the structure of a nucleic acid lysis device according to an embodiment of the present invention; Figure 2 This is a schematic diagram of a nucleic acid lysis device according to another embodiment of the present invention. The vibrating needle in the figure is in the lysis working state. The radial solid arrow in the figure indicates ultrasonic vibration, and the axial dashed arrow indicates the direction of nucleic acid reaction liquid aspiration and expulsion.

[0021] The reference numerals in the attached figures are as follows: 1. Vibrating needle body; 2. Ultrasonic unit; 3. Aspiration unit; 31. Aspiration interface; 4. Temperature control unit; 100. Reaction liquid container; 101. Nucleic acid reaction solution. Detailed Implementation

[0022] See Figures 1 to 2As shown in the figure, according to an embodiment of the present invention, a nucleic acid lysis device is provided, including a device body (not shown) and a vibrating needle 1. The vibrating needle 1 is detachably connected to the device body. An ultrasonic unit 2 is disposed within the device body. The ultrasonic head (not shown) of the ultrasonic unit 2 is vibratingly coupled to the vibrating needle 1. The vibrating needle 1 is in a lysis working state where it is at least partially inserted into the nucleic acid reaction solution 101 within the reaction solution container 100. The aforementioned vibrating coupling connection refers to the connection where, during the operation of the ultrasonic unit 2, ultrasound can be directly or efficiently indirectly (e.g., by applying an ultrasonic coupling agent between the ultrasonic head and the vibrating needle 1) to allow ultrasound to act efficiently on the vibrating needle 1. In practical applications, various specific implementation methods can be adopted, but these specific implementation methods should ensure that the proportion of the portion of the ultrasonic energy generated by the ultrasonic unit 2 that acts on the vibrating needle 1 to produce vibration is too low in the total ultrasonic energy generated. The aforementioned ultrasonic unit 2 can be a commercially available component, and the specific operating frequency, maximum power, and specific structural type are selected according to the application conditions.

[0023] In this technical solution, the ultrasonic unit 2 configured inside the main body of the device can transmit ultrasonic energy to the vibrating needle 1 coupled to it. At the same time, the vibrating needle 1 is in a lysis working state inserted into the nucleic acid reaction solution 101. Thus, when the ultrasonic unit 2 is running, the ultrasonic unit 2 will directly transmit and act on the vibrating needle 1 to generate high-frequency vibration, which will then directly transmit and act on the nucleic acid reaction solution 101, thereby inducing strong vibration and microfluidic effect in the nucleic acid reaction solution 101, enhancing the shear force of the lysis solution on the cell membrane (nucleic acid cell membrane), and making the lysis more uniform. Since the ultrasonic waves can be directly transmitted and act on the liquid into which the vibrating needle 1 is inserted, they are no longer absorbed or scattered by the large container substrate, significantly improving the utilization efficiency of ultrasonic energy and shortening the lysis time. The ultrasonic unit 2 is configured inside the main body of the device and does not need to be integrated into the corresponding reaction solution container (disposable consumable). While ensuring energy utilization efficiency, it also significantly reduces the cost of consumable use and can better meet the low-cost requirements of the reaction solution container 100 in POCT scenarios.

[0024] It is particularly important to emphasize that the vibrating needle body 1 in this invention is a cantilever structure with one end connected to the ultrasonic head of the ultrasonic unit 2 in actual application. This cantilever structure has a better vibration transmission effect on the nucleic acid reaction solution 101 under the enable of ultrasonic vibration.

[0025] In some embodiments, the ultrasonic unit 2 operates at a frequency of 10-100kHz, preferably 20-50kHz; and has an ultrasonic power of 1-100W, preferably 10-50W. In one specific embodiment, the ultrasonic unit 2 uses a commercially available piezoelectric ceramic transducer with an operating frequency of 40kHz and a maximum power of 50W.

[0026] See details Figure 1 As shown, the ultrasonic unit 2 applies ultrasonic waves to the vibrating needle body 1. The ultrasonic waves include radial and axial components, with the radial component being dominant. The ultrasonic unit 2 completes the transmission of ultrasonic waves by being in close contact with the vibrating needle body 1, without passing through the substrate of the larger reaction liquid container 100.

[0027] In some embodiments, the main body of the device is further provided with a suction unit 3, and the vibrating needle 1 is a hollow needle (which can also be called a capillary needle). The first end of the hollow needle is detachably connected to the suction interface 31 of the suction unit 3. When the vibrating needle 1 is in the lysis working state, the second end of the hollow needle is below the surface of the nucleic acid reaction solution 101. In a specific embodiment, the hollow cavity of the aforementioned hollow needle is a central through hole, which extends from the first end of the hollow needle to its second end.

[0028] In this technical solution, a suction unit 3 is further configured inside the main body of the device. At this time, the vibrating needle body 1 is designed as a hollow needle body, and its first end is detachably connected to the suction interface 31 of the suction unit 3. The nucleic acid reaction solution can be further suctioned into its hollow cavity by the suction unit 3. In this way, the nucleic acid reaction solution can be further formed into a stronger ultrasonic field in the vibrating needle body 1, the energy is utilized more concentratedly, and the lysis is more complete and uniform.

[0029] It is understandable that during the lysis of nucleic acids, the operation of the aforementioned suction unit 3 enables the reaction solution to be repeatedly drawn in and expelled within the vibrating needle body 1, thereby achieving efficient, uniform, and complete lysis of all nucleic acid reaction solutions 101 within the entire reaction solution container 100.

[0030] The aforementioned suction unit 3 can specifically adopt a commercially available miniature plunger pump. Other types of pneumatic suction components can be selected according to the actual operational accuracy requirements. In a specific embodiment, the flow control accuracy is 0.1 μL.

[0031] In some embodiments, the ultrasonic head of the ultrasonic unit 2 is coupled to the suction port 31, that is, the ultrasonic head of the ultrasonic unit 2 acts on the vibrating needle body 1 through the suction port 31.

[0032] In this technical solution, the ultrasonic head of the ultrasonic unit 2 is coupled to the suction interface 31 to achieve ultrasonic transmission of the vibrating needle 1, which makes the device structure more compact and more convenient in actual operation. It is understood that the aforementioned vibrating needle 1 is used as a consumable in actual use, and the corresponding vibrating needle 1 needs to be replaced when cracking different reaction solutions. At this time, since the ultrasonic head of the ultrasonic unit 2 is vibratingly coupled to the suction interface 31, when replacing the vibrating needle 1, it is only necessary to ensure the reliability of the connection between the vibrating needle 1 and the suction interface 31, without considering whether the coupling connection between the vibrating needle 1 and the ultrasonic head of the ultrasonic unit 2 is reliable.

[0033] In some embodiments, the first end of the aforementioned vibrating needle body 1 and the suction interface 31 can be detachably connected by means of threaded connection, snap-fit ​​connection or interference fit connection.

[0034] In some embodiments, the inner diameter of the hollow needle body is d, 0.1mm≤d≤10mm, preferably 1mm≤d≤4mm; and / or, the wall thickness of the hollow needle body is δ, 0.5mm≤δ≤5mm, preferably 0.5mm≤δ≤1mm; and / or, the length of the hollow needle body is L, 10mm≤L≤200mm, preferably 20mm≤L≤50mm.

[0035] In some embodiments, the hollow needle body includes a metal matrix, and the material of the metal matrix can be at least one of aluminum alloy, stainless steel, and iron. Using a metal matrix can ensure the high efficiency of ultrasonic wave transmission and meet the structural reliability requirements under high-frequency vibration.

[0036] In some embodiments, the hollow needle body further includes a biocompatible outer layer. When the vibrating needle body 1 is in the lysis working state, the part of the metal substrate that contacts the nucleic acid reaction solution 101 is the first part. The biocompatible outer layer at least covers the outer wall surface of the first part. The biocompatible outer layer is, for example, at least one of PP coating and metal oxide layer. The metal of the aforementioned metal oxide layer can be consistent with the specific material of the aforementioned metal substrate.

[0037] In this technical solution, by setting a biocompatible outer layer on the outer wall of the metal substrate, it is possible to effectively prevent the vibrating needle 1 from damaging or adsorbing enzymes, nucleic acids, etc. in the reaction solution.

[0038] In some embodiments, the main body of the device is further provided with a temperature control unit 4, which is heat-exchange connected to the vibrating needle body 1. The aforementioned heat exchange connection means that the temperature control unit 4 can regulate the temperature of the vibrating needle body 1 and the nucleic acid reaction solution in its cavity to ensure that the temperature is at a preset value (i.e., the set temperature).

[0039] In this technical solution, the heat exchange connection between the temperature control unit 4 and the vibrating needle 1 configured in the main body of the device achieves a more direct and efficient temperature control of the reaction liquid, which can further improve the nucleic acid lysis rate. At the same time, compared with the traditional indirect temperature control method on the outer wall (side wall or bottom wall) of the reaction liquid container, the heat of the temperature control unit 4 is directly conducted to the vibrating needle 1 of the metal substrate with higher thermal conductivity, which can significantly improve the heat utilization efficiency. It should be noted that since the volume of the vibrating needle 1 of the metal substrate is much smaller than that of the reaction liquid container, the cost of consumables can be significantly reduced while ensuring efficient temperature control.

[0040] In addition, it is worth emphasizing that the aforementioned temperature control unit 4 can generate heat to heat the vibrating needle 1 when the temperature is insufficient, and can also actively control the temperature to absorb (i.e. balance) the waste heat caused by ultrasound when the temperature is high, so as to avoid the reaction solution temperature being too high and causing the protease to become inactive.

[0041] It is understood that the aforementioned temperature control unit 4 includes a heat source component (not shown in the figure) and a heat conduction component (not shown in the figure). The heat source component is used to generate a preset amount of heat under the action of an electric current, and the heat conduction component is used to conduct the preset amount of heat to the target location. In some embodiments, the aforementioned heat conduction component is a heat conduction ring (not shown in the figure). The aforementioned heat conduction ring is designed as an open and closed structure (e.g., two symmetrically arranged semicircular rings). When it is in the open state (the two semicircular rings are far apart from each other and form a large gap in the middle), it does not contact the vibrating needle body 1, so as to facilitate the convenient replacement and connection of the vibrating needle body 1 and the aforementioned suction interface 31. When it is in the closed state (the two semicircular rings are close to each other and form a ring around the vibrating needle body 1 between them), it rings the outer wall surface of the vibrating needle body 1 to achieve efficient heat conduction.

[0042] See details Figure 1 As shown, in some embodiments, the aforementioned temperature control unit 4 is disposed on the side of the vibrating needle body 1 near the reaction liquid container 100, and is arranged at intervals with the aforementioned ultrasonic unit 2 and suction unit 3 in the axial direction of the vibrating needle body 1. Under the premise of realizing reaction liquid suction, ultrasonic vibration and temperature control, the various module components are reasonably arranged, the structure is simple and compact, and the functional modules have high compatibility.

[0043] The aforementioned heat source component uses a semiconductor thermoelectric cooler (TEC) or a thin-film heating element to achieve rapid heating and cooling control of the hollow vibrating needle body 1. For example, a commercially available Peltier semiconductor cooling chip can be used to achieve rapid heating and cooling. In terms of the specific specifications, the requirements for heating and cooling speed should be ensured. In some embodiments, the temperature control unit 4 should have the ability to achieve a heating and cooling speed of the vibrating needle body 1 greater than 10°C / s.

[0044] According to an embodiment of the present invention, a control method for the above-described nucleic acid lysis apparatus is also provided, comprising the following steps: Place the reaction solution container 100 in a preset position so that the second end of the vibrating needle body 1 (i.e., the bottom end in the use state) is below the surface of the nucleic acid reaction solution 101 in the reaction solution container 100. Upon receiving a nucleic acid lysis command, the ultrasonic unit 2 is controlled to operate.

[0045] In this technical solution, the ultrasonic unit 2 directly conducts high-frequency vibrations onto the vibrating needle 1, which in turn directly conducts these vibrations onto the nucleic acid reaction solution 101. This induces strong vibrations and microfluidic effects within the nucleic acid reaction solution 101, enhancing the shear force of the lysis buffer on the cell membrane (nucleic acid cell membrane), resulting in more uniform lysis. Because the ultrasonic waves can be directly conducted into the liquid into which the vibrating needle 1 is inserted, they are no longer absorbed or scattered by the larger container substrate, significantly improving the utilization efficiency of ultrasonic energy and shortening the lysis time. The ultrasonic unit 2 is configured within the main body of the device, eliminating the need for integration into the corresponding reaction solution container (disposable consumable). This ensures energy utilization efficiency while significantly reducing consumable costs, better meeting the low-cost requirements of the reaction solution container 100 in POCT scenarios. In some embodiments, when the aspiration unit 3 is included and the vibrating needle body 1 is a hollow needle body, after receiving the nucleic acid lysis command, the aspiration unit 3 is also controlled to operate so that the nucleic acid reaction solution 101 is repeatedly aspirated and expelled from the hollow cavity of the hollow needle body. The aspiration unit 3 is used to repeatedly aspirate the nucleic acid reaction solution into its hollow cavity. The reciprocating motion of the liquid flow in the hollow cavity promotes the full mixing of the sample and the lysis agent and enhances the lysis effect of ultrasound. When the temperature control unit 4 is included, after receiving the nucleic acid lysis command, the temperature control unit 4 is also controlled to operate so that the temperature inside the hollow cavity of the hollow needle body is at the set temperature. The heat exchange connection between the temperature control unit 4 and the vibrating needle body 1 achieves a more direct and efficient temperature regulation of the reaction solution, which can further improve the nucleic acid lysis speed.

[0046] Experiments have shown that traditional chemical pyrolysis takes 10-30 minutes, while this invention uses ultrasonic-enhanced pyrolysis, reducing the pyrolysis time to less than 1 minute, increasing the speed by more than 10 times.

[0047] The nucleic acid lysis device using the aforementioned technical solution of the present invention has a faster lysis speed, higher energy utilization efficiency, simpler structure, lower consumable cost, and high compatibility with detection platforms. It is especially suitable for POCT detection scenarios. By adjusting the ultrasound parameters and temperature, it can be applied to the lysis processing of various sample types such as swab samples, blood samples, and FFPE samples.

[0048] The technical solution of the present invention will be further described below with reference to specific embodiments: Example 1: Nucleic acid lysis of influenza virus swab samples 1. Device Configuration The nucleic acid lysis device used in this embodiment is configured as follows:

[0049] 2. Ligation reagent The lysis reagent used in this embodiment has the following composition: Lysis buffer: containing 4M guanidine hydrochloride, 50mM Tris-HCl (pH 8.0), 20mM EDTA, 1% Triton X-100, 0.5% SDS, and 10mM DTT; Proteinase K: concentration 20 mg / mL; Magnetic beads: Silicon-based magnetic beads, 1μm in size, 10mg / mL in concentration.

[0050] 3. Operating Procedures Step 1: Sample aspiration (5 seconds) A throat swab sample tube containing influenza virus is placed in the device, and a capillary needle is inserted into the sample tube under the control of the pneumatic unit to aspirate a total of 200 μL of sample in two separate applications.

[0051] Step 2: Pyrolysis treatment (60 seconds) The capillary needle was used to transfer the lysis buffer into a lysis well containing 500 μL of lysis buffer and 20 μL of proteinase K. The sonication and temperature control units were activated (target temperature 65°C) to rapidly lyse cells under the combined action of ultrasound and heating. A cyclic aspiration-ejection mode was adopted—after the capillary needle aspirated the lysis buffer, it was sonicated inside the tube, while the pneumatic unit drove the liquid column to reciprocate. The ultrasonic vibration generated a strong cavitation effect and convection inside the tube, accelerating the lysis process.

[0052] Step 3: Nucleic acid capture (10 seconds) After lysis, 50 μL of the magnetic bead suspension was aspirated into a capillary needle and mixed with the lysis buffer. The magnetic beads bound to nucleic acids were captured by an external magnetization device, the liquid was discharged, and the magnetic beads remained in the tube.

[0053] Step 4: Cleaning and purification (60 seconds) The capillary needle was sequentially transferred to four cleaning wells, each containing 300 μL of 80% ethanol cleaning solution. In each well, the following steps were performed: (1) aspiration of cleaning solution; (2) ultrasonic cleaning for 10 seconds with an ultrasonic source (30W power); (3) drainage of cleaning solution, with magnetic beads retained.

[0054] Step 5: Nucleic acid elution (15 seconds) The purified magnetic beads were transferred to 50 μL of elution buffer (10 mM Tris-HCl, pH 8.0), heated to 70 °C and held for 10 seconds to release the nucleic acid template. After demagnetization, the nucleic acid solution was aspirated for subsequent qPCR detection.

[0055] 4. RT-qPCR detection The following RT-qPCR reagents and conditions were used for detection: RT-qPCR reaction system (25μL): 12.5μL of 2× RT-qPCR One-step Mix, 2.5μL of influenza A virus H1N1 primer and probe mixture, 5μL of nucleic acid template, and 5μL of nuclease-free water; RT-qPCR reaction conditions: 50℃ / 2min; 95℃ / 30s; then 40 cycles (95℃ / 5s, 60℃ / 10s).

[0056] 5. Experimental Results Using the method of this embodiment, the total time from sample aspiration to obtaining RT-qPCR detection results is less than 18 minutes. The Ct value for RT-qPCR detection of influenza A virus H1N1 is 28.5, indicating high nucleic acid extraction efficiency. Compared with the traditional chemical lysis method (lysis time 300 seconds, Ct value 30.6), the method of this invention shortens the lysis time by 80% while releasing a higher amount of nucleic acid.

[0057] Example 2: Experiment on Optimization of Ultrasonic Intervention Time To determine the optimal sonication lysis time, influenza virus swab samples were used as a model, and different sonication time groups (10s, 20s, 30s, 40s, 50s, 60s, 90s, 120s, 180s) were set up. Other conditions were the same as in Example 1, and the nucleic acid release efficiency of each group was compared (expressed as qPCR Ct value).

[0058] The experimental results are shown in the table below:

[0059] The results showed that from 10 to 50 seconds, the longer the ultrasonic time, the smaller the Ct value, indicating that the ultrasonic lysis release was higher; from 60 to 180 seconds, the Ct value did not decrease significantly with increasing ultrasonic time, indicating that it was close to saturation. Considering both lysis efficiency and time cost, the recommended ultrasonic lysis time is 60-90 seconds.

[0060] Example 3: Comparison of lysis effects with and without ultrasound Using influenza virus swab samples as a model, the lysis effects of sonication versus no sonication were compared under the same lysis reagent conditions. The experiment was divided into two groups: Experimental group: Sonication for 30 seconds (power 50W, frequency 50kHz), temperature 65℃; Control group: No ultrasound was added, and the samples were placed at 65°C for different times (60s, 120s, 180s, 240s, 300s).

[0061] The experimental results are shown in the table below:

[0062] The results showed that the Ct values ​​of the samples subjected to sonication for 30 seconds and those subjected to 300 seconds without sonication were similar, indicating that sonication could significantly shorten the nucleic acid extraction time and increase the lysis rate by about 10 times.

[0063] It will be readily understood by those skilled in the art that the aforementioned advantageous methods can be freely combined and superimposed without conflict.

[0064] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention. The above are merely preferred embodiments of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the protection scope of the present invention.

Claims

1. A nucleic acid lysis device, characterized in that, The device includes a main body and a vibrating needle (1). The vibrating needle (1) is detachably connected to the main body. An ultrasonic unit (2) is disposed inside the main body. The ultrasonic head of the ultrasonic unit (2) is vibratingly coupled to the vibrating needle (1). The vibrating needle (1) is in a lysis working state where it is at least partially inserted into the nucleic acid reaction solution (101) inside the reaction solution container (100).

2. The nucleic acid lysis apparatus according to claim 1, characterized in that, The device body is also equipped with a suction unit (3). The vibrating needle body (1) is a hollow needle body. The first end of the hollow needle body is detachably connected to the suction interface (31) of the suction unit (3). When the vibrating needle body (1) is in the lysis working state, the second end of the hollow needle body is below the surface of the nucleic acid reaction solution (101).

3. The nucleic acid lysis apparatus according to claim 2, characterized in that, The ultrasonic head of the ultrasonic unit (2) is coupled to the suction port (31).

4. The nucleic acid lysis apparatus according to claim 2, characterized in that, The inner diameter of the hollow needle body is d, 0.1mm≤d≤10mm; and / or, the wall thickness of the hollow needle body is δ, 0.5mm≤δ≤5mm; and / or, the length of the hollow needle body is L, 10mm≤L≤200mm.

5. The nucleic acid lysis apparatus according to claim 2, characterized in that, The hollow needle body includes a metal matrix.

6. The nucleic acid lysis apparatus according to claim 6, characterized in that, The hollow needle body also includes a biocompatible outer layer. When the vibrating needle body (1) is in the lysis working state, the part in contact with the metal matrix and the nucleic acid reaction solution (101) is the first part, and the biocompatible outer layer at least covers the outer wall surface of the first part.

7. The nucleic acid lysis apparatus according to claim 6, characterized in that, The material of the metal substrate is at least one of aluminum alloy, stainless steel, and iron; and / or the biocompatible outer layer is at least one of PP coating and metal oxide layer.

8. The nucleic acid lysis apparatus according to claim 1, characterized in that, The device body is also equipped with a temperature control unit (4), which is heat exchanged with the vibrating needle body (1).

9. A method for controlling a nucleic acid lysis apparatus according to any one of claims 1 to 8, characterized in that, Includes the following steps: Place the reaction solution container (100) in a preset position so that the second end of the vibrating needle (1) is below the surface of the nucleic acid reaction solution (101) in the reaction solution container (100); Upon receiving the nucleic acid lysis command, the ultrasonic unit (2) is controlled to operate.

10. The control method according to claim 9, characterized in that, When the suction unit (3) is included and the vibrating needle body (1) is a hollow needle body, after obtaining the nucleic acid lysis command, the suction unit (3) is also controlled to run so that the nucleic acid reaction solution (101) is repeatedly sucked in and expelled from the hollow cavity of the hollow needle body; And / or, When the temperature control unit (4) is included, after receiving the nucleic acid lysis command, the temperature control unit (4) is also controlled to operate so that the temperature inside the hollow cavity of the hollow needle is at the set temperature.

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