Solution sample processing device, apparatus and system and use thereof

Abstract

The present invention provides a device for processing a solution sample, which has a cavity for containing a solution and an ultra-high frequency bulk acoustic wave resonator arranged relatively in the cavity. The present invention also provides an apparatus for purifying a target substance in a solution sample and a system for detecting a target substance in a solution sample having the device. The present invention also provides a method for processing, purifying or detecting a target biological substance in a biological sample, such as nucleic acid or protein of a cell or virus, by using the device, the equipment and the system.

Description

[0001] This application claims priority to Chinese patent application No. 201910794754.1, filed on August 23, 2019, entitled “Solution sample processing device, apparatus and system and use thereof”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to the fields of cell and molecular biology and medical devices. Specifically, the present invention relates to a device, an apparatus and a system for processing a solution sample using relatively arranged ultrahigh frequency bulk acoustic wave resonators, and a method for separating and / or detecting a target biological substance in a biological sample, such as nucleic acid or protein of a cell or virus, using the device, an apparatus and a system. Background Art

[0003] The purification and detection of biomolecules, such as small molecules such as metabolites, as well as proteins and nucleic acids, are core issues of widespread concern in many basic biological mechanism research, medicine and disease diagnosis, and medical instrument development. The enrichment and mixing of these micron-sized organisms in small volume solutions has always been a hot research direction in the field of biochemistry, especially in applications such as nucleic acid extraction and solid phase synthesis. Efficient solid-liquid mixing of samples in milliliters or even hundreds of microliters and below is a prerequisite for effective biochemical reactions.

[0004] In the prior art, various physical fields are used to process the enrichment and mixing of solutions containing biomolecules. The existing related technologies include driving methods that apply mechanical energy or electrical energy. However, these technologies are limited by the disadvantages of high energy and low flux, and require a very large field strength distribution to be generated locally, which has great limitations on molecular manipulation. At the same time, the specificity of directly using physical fields to manipulate molecules is poor.

[0005] In addition, the portability of detection equipment is a major trend in the development of modern scientific instruments. Portable biosensor devices have a wider range of applications, such as home health testing, real-time detection of human health indicators, and diagnosis in remote areas with scarce medical resources. Generally speaking, portable sensors have the characteristics of small size, simple operation, fast and reliable, and relatively low price. Accordingly, only biosensor technologies or methods that meet the above characteristics are expected to be developed into portable biosensors first.

[0006] Therefore, there is an urgent need for a more effective and convenient method and system for enriching and mixing solution samples. Summary of the invention

[0007] In order to improve the solid-liquid mixing efficiency and further improve the biochemical reaction efficiency, the present invention provides a device for performing three-dimensional spatial particle operations on samples in a solution using relatively arranged ultra-high frequency bulk acoustic wave resonators, equipment and a system containing the device, as well as a method for mixing and enriching solution samples containing biological molecules and the like using the device, equipment or system. By adjusting the input power, the sample mixing function can be efficiently achieved, and the conversion between sample particle enrichment and mixing can be conveniently achieved.

[0008] Specifically, the present invention provides a device for processing a solution sample, which has a cavity for accommodating the solution and an ultra-high frequency bulk acoustic wave resonator relatively arranged in the cavity, wherein the ultra-high frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz to generate a vortex in the solution.

[0009] In the method of the present invention, the ultra-high frequency bulk acoustic wave resonator can generate high-frequency (about 0.5-50GHz, for example, not less than 1.0GHz) vibrations, and induce bulk acoustic waves of corresponding frequencies in the solution. In one aspect of the present invention, the ultra-high frequency bulk acoustic wave resonator is a thin film bulk acoustic wave resonator (FBAR) or a solid-state assembly type resonator (SMR), preferably a solid-state assembly type resonator. In another aspect of the present invention, the ultra-high frequency bulk acoustic wave resonator is an acoustic wave resonator in a thickness stretching vibration mode, and a piezoelectric material thin film layer is grown in a vertical direction, and vibrations are excited by coupling a vertical electric field through a d33 piezoelectric coefficient. The ultra-high frequency bulk acoustic wave resonator used in the present invention can generate a localized acoustic flow at the interface between the device and the liquid without the help of a coupling medium or structure. The bulk acoustic wave resonator of the present invention includes an acoustic wave reflection layer, a bottom electrode layer, a piezoelectric layer and a top electrode layer arranged in sequence from bottom to top. The overlapping area of ​​the bottom electrode layer, the piezoelectric layer, the top electrode layer and the acoustic wave reflection layer constitutes a bulk acoustic wave generation area. The top surface of the UHF BAW resonator can be arranged on the wall of the cavity or other devices in the cavity, generating a BAW propagating perpendicular to the top of the resonator to the opposite side. The thickness of the piezoelectric layer of the UHF BAW resonator of the present invention ranges from about 1nm to 2um.

[0010] In this patent, the relative arrangement refers to the arrangement position and manner of the corresponding two or more BAW resonators in the cavity so that the BAW waves emitted by each of them propagate in opposite directions. For example, the two BAW resonators are respectively arranged at the bottom and top of the cavity, and the sound wave emission directions are opposite. In the device of the present invention, the relatively arranged ultra-high frequency BAW resonators include at least one group of BAW resonators with opposite sound wave emission directions, and each group of BAW resonators with opposite sound wave emission directions may include two BAW resonators with opposite sound wave emission directions, or may be multiple ultra-high frequency BAW resonators, for example, there are one or more BAW resonators in one sound wave propagation direction, and the corresponding BAW resonators arranged in the opposite direction may also be one or more.

[0011] In one aspect of the present invention, the distance between the relatively arranged ultrahigh frequency bulk acoustic wave resonators is about 0.1-5cm, preferably about 0.5-2cm, for example, about 0.8-1cm. In another aspect of the present invention, the distance between the relatively arranged ultrahigh frequency bulk acoustic wave resonators is about 0.1-2.0cm, preferably about 0.2-1.0cm, for example, about 0.2-0.8cm. In this article, the distance refers to the distance between each bulk acoustic wave resonator and its corresponding bulk acoustic wave resonator arranged in the opposite direction. For example, when the relatively arranged bulk acoustic wave resonators are two bulk acoustic wave resonators respectively arranged at the bottom and top of the cavity, and the sound wave emission directions are opposite to each other, the distance refers to the distance between the sound wave emission surfaces of the two bulk acoustic wave resonators. In the device of the present invention, the acoustic jet generated by the bulk acoustic wave emitted by the bulk acoustic wave resonator in the solution causes a vortex of the solution, and the vortex influence range generated by the relatively arranged bulk acoustic wave resonators can reach the position of the corresponding bulk acoustic wave resonator arranged in the opposite direction.

[0012] In one aspect of the present invention, the positions of the relatively arranged UHF BAW resonators are adjustable. For example, one of the relatively arranged BAW resonators is fixed at the bottom of the cavity, and the other is connected to a device that can move up and down and is arranged at the top.

[0013] In one aspect of the present invention, the area of ​​the bulk acoustic wave generating region of the ultra-high frequency bulk acoustic wave resonator is about 1000-50000 μm 2 , preferably about 5000-20000 μm 2 .

[0014] In one aspect of the present invention, the device has a power regulating device that can independently regulate the power of the BAW generated by each of the relatively arranged UHF BAW resonators.

[0015] In one aspect of the present invention, the ultra-high frequency bulk acoustic wave resonator in the device generates bulk acoustic waves in an operating power range of about 100 mW or more, preferably 150-3000 mW, more preferably 200-2000 mW, for example about 400-800 mW.

[0016] In one aspect of the present invention, the ultra-high frequency bulk acoustic wave resonator in the device generates bulk acoustic waves in an operating power range of about 0.01-20 mW, preferably 0.1-10 mW, and more preferably 0.5-5 mW.

[0017] In one aspect of the present invention, the volume of the cavity in the device is about 100 μl-10 ml, preferably about 200 μl-5 ml, and more preferably about 500 μl-1 ml.

[0018] In one aspect of the present invention, the height of the cavity in the device is about 0.5-5 cm, preferably about 1-3 cm, such as about 1-2 cm.

[0019] In one aspect of the present invention, the solution sample is a biological sample (e.g., body fluid, blood, cell, virus, protein, lipid, carbohydrate, nucleic acid, aptamer, nucleoside). The solution may also contain magnetic or non-magnetic particles that can bind to the biological sample. Preferably, the size of the sample or particle is about 0.01 μm to about 100 μm, more preferably about 0.05 μm to about 20 μm, for example, about 0.5 μm to about 10 μm.

[0020] The present invention is based on the following unexpected findings by the inventors:

[0021] The UHF BAW resonator can induce eddies (or acoustic flow vortices) in the solution by emitting acoustic waves. The particles in the acoustic flow vortices are simultaneously affected by the acoustic radiation force and the drag force. When the UHF BAW resonators are arranged relatively and work at the same time, they have a chaotic convection effect, the vortex range is increased, and the particles in the blind area of ​​the cavity boundary also enter the 3D eddy space under the drag force.

[0022] When the ultra-high frequency bulk acoustic wave resonators are relatively arranged, but only one direction of the resonator is operated, a particle aggregation effect is generated: when the input power of the resonator is in a suitable range, for example, less than about 20 mW, the particles (such as particles with a size of 0.01 μm to about 100 μm) are mainly affected by the acoustic radiation force in the area close to the device, and will move from the outside to the inside along the vortex. When they move to the edge of the resonator, the central liquid is continuing to move from bottom to top. The particles will be pushed to the inner area deviating from the vortex trajectory due to the large horizontal component of the acoustic radiation force. When the particles are a certain distance away from the edge of the device, the acoustic radiation force decreases due to the distance, and the particles will be affected by the fluid drag force of the micro-vortex from top to bottom and move to the edge of the device again, and so on. When the particle size is small, the drag force is less affected, it is difficult to aggregate, and the aggregation time is longer.

[0023] In one aspect of the present invention, there is provided a device for purifying a target substance in a solution sample, which includes the device described above, having a cavity for containing the solution and an ultrahigh frequency bulk acoustic wave resonator relatively arranged in the cavity, wherein the ultrahigh frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz, thereby generating a vortex in the solution.

[0024] In the present invention, the sample may be a biological sample. The sample may be a culture fluid or a suspension thereof or a supernatant thereof, or a buffer containing a target substance, such as a culture fluid or a suspension thereof or a supernatant thereof of various viruses, bacteria or cells. The present invention is suitable for various body fluids of various animals such as humans, including blood, tissue fluid, extracellular fluid, lymph fluid, cerebrospinal fluid, aqueous humor, urine, sweat, etc. The method and device of the present invention are particularly suitable for samples with complex contents and / or samples with higher viscosity, such as whole blood and respiratory mucus, such as sputum or nasal mucus.

[0025] The target material of the sample processed by the method provided by the present invention can be a biological substance, generally a molecule with a specific structure, such as a peptide, a polypeptide, a protein, a lipoprotein, a glycoprotein, a nucleic acid (DNA, RNA, PNA, an aptomer) and a nucleic acid precursor (nucleoside and nucleotide), a polysaccharide, a lipid such as a lipid vesicle, etc. Among them, the protein may include a cytokine, a hormone, a vitamin, a surface receptor, a hapten, an antigen, an antibody, an enzyme, a growth factor, a recombinant protein, a toxin, and a fragment and combination thereof. The biological substance usually has a size of nanometer order or smaller, for example, less than 100 nm.

[0026] In one aspect of the present invention, the device includes a sample lysis module, in which a sample (e.g., cell, virus, etc.) is lysed by a lysing agent, enzyme, ultrasound, or physical grinding, so that the bio-small molecule target substance (e.g., nucleic acid) of the sample is released into a solution.

[0027] In one aspect of the present invention, the device comprises a target substance binding module. In the target substance binding module, magnetic particles or non-magnetic particles can be specifically contacted with the sample after the lysis step, thereby allowing the target substance (e.g., nucleic acid) in the sample solution to bind to the magnetic particles or non-magnetic particles.

[0028] In one aspect of the present invention, the device includes a target substance cleaning module. In the target substance cleaning module, the target substance (e.g., nucleic acid) in the sample solution can be washed with water or a buffer after binding to the magnetic particles or non-magnetic particles. The specifically bound nucleic acid remains bound to the magnetic particles or non-magnetic particles, and other unnecessary substances are washed.

[0029] In one aspect of the invention, the device comprises a target material elution module, in which, under suitable solvents and conditions, target materials (e.g., nucleic acids) bound to the magnetic or non-magnetic particles can be separated from the magnetic or non-magnetic particles and enter into a solution, while the magnetic or non-magnetic particles can be separated and removed by a magnetic field or other means.

[0030] In one aspect of the present invention, the device is configured such that one of the sample lysis module, target substance binding module, target substance cleaning module and target substance elution module is the aforementioned cavity having the relatively arranged ultrahigh frequency bulk acoustic wave resonator, or any combination thereof has the same cavity, and the cavity has the relatively arranged ultrahigh frequency bulk acoustic wave resonator. One of the advantages of the present invention is that in the method using the device, one or more of the steps of sample lysis, target substance binding, target substance cleaning and target substance elution can use the cavity having the relatively arranged ultrahigh frequency bulk acoustic wave resonator and utilize bulk acoustic waves to fully stir and mix the particles in the solution to improve the efficiency of sample lysis, target substance binding, target substance cleaning or target substance elution, especially the efficiency of target substance binding, target substance cleaning or target substance elution. Another advantage is that in the method and device of the present invention, one or more of the steps of sample lysis, target substance binding, target substance cleaning and target substance elution can be performed in the same chamber.

[0031] In one aspect of the present invention, the device is used to separate target substances (such as nucleic acids) in samples by magnetic bead method. The magnetic bead method uses magnetic beads that can specifically identify cells, proteins or nucleic acids. The magnetic beads can specifically bind to target molecules such as nucleic acids, and then separate active macromolecules from samples such as blood, animal tissues, food, pathogenic microorganisms, etc. under the action of a magnetic field. In another aspect of the present invention, the device has a magnetic device that can apply a magnetic field to control the magnetic material and / or the magnetic material that binds to the target substance (such as nucleic acid).

[0032] In another aspect of the present invention, a system for detecting a target substance in a solution sample is provided, comprising: a device or apparatus as described above, which has a cavity for containing a solution and an ultra-high frequency bulk acoustic wave resonator arranged relatively in the cavity, wherein the ultra-high frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz to generate a vortex in the solution;

[0033] and

[0034] A detection device that detects identifiable labels carried by target substances, including but not limited to fluorescence or other forms of luminescence (such as chemiluminescence, bioluminescence, radioluminescence, electroluminescence, electrochemiluminescence, mechanical luminescence, crystal luminescence, thermoluminescence, sonoluminescence, phosphorescence and photoluminescence, etc.), enzymatic reactions, radioactivity, etc.

[0035] In another aspect of the present invention, a system for detecting a target substance in a solution sample is provided, comprising: a device or apparatus as described above, which has a cavity for containing a solution and an ultra-high frequency bulk acoustic wave resonator arranged relatively in the cavity, wherein the ultra-high frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz to generate a vortex in the solution;

[0036] Nucleic acid amplification module; the nucleic acid amplification module is a module suitable for performing nucleic acid amplification in any manner, including but not limited to various constant temperature amplifications or PCRs, such as qPCR (fluorescence quantitative PCR), RT-PCR, hot start PCR, nested PCR, multiplex PCR, recovery condition PCR, dsRNA synthesis, COLD-PCR, digital PCR, etc.

[0037] and

[0038] A detection device that detects identifiable labels carried by target substances, including but not limited to fluorescence or other forms of luminescence (such as chemiluminescence, bioluminescence, radioluminescence, electroluminescence, electrochemiluminescence, mechanical luminescence, crystal luminescence, thermoluminescence, sonoluminescence, phosphorescence and photoluminescence, etc.), enzymatic reactions, radioactivity, etc.

[0039] The present invention provides a method for processing a sample in a solution, the method comprising: generating a bulk acoustic wave using relatively arranged ultra-high frequency bulk acoustic wave resonators in a cavity containing the solution, generating a vortex in the solution, and mixing the sample in the solution, wherein the ultra-high frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz.

[0040] In one aspect of the present invention, a method for processing a sample to be tested using the device or equipment or system described above is provided: the device or equipment has a cavity for containing a solution and an ultra-high frequency bulk acoustic wave resonator relatively arranged in the cavity, and the ultra-high frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of about 0.5-50 GHz to generate a vortex in the solution.

[0041] In one aspect of the present invention, the power of the BAW generated by each BAW resonator in the relatively arranged UHF BAW resonators can be independently adjusted. The power range of the BAW that generates a vortex in the solution to mix the sample in the solution is about 100 mW or more, preferably about 150-3000 mW, more preferably about 200-2000 mW, for example, about 400-800 mW. In the method of the present invention, the acoustic jet generated by the BAW resonator in the solution causes a vortex in the solution, and the vortex generated by the relatively arranged BAW resonators can reach the position of the corresponding BAW resonator arranged in the relative direction.

[0042] In one aspect of the present invention, each resonator in the relatively arranged ultrahigh frequency bulk acoustic wave resonators in the method generates bulk acoustic waves at the same time, and the generated vortex action range covers the position of the corresponding bulk acoustic wave resonator arranged in the opposite direction, and the vortex generated by the corresponding bulk acoustic wave resonator arranged in the opposite direction produces synergistic enhancement, so that the samples and / or particles in the entire solution are fully mixed and contacted.

[0043] In one aspect of the present invention, the method further comprises: one of the relatively arranged ultra-high frequency bulk acoustic wave resonators does not work, and the corresponding resonator arranged in the opposite direction works to generate bulk acoustic waves, so that the solution in the bulk acoustic wave region generates a vortex, and the sample in the solution or the target substance contained in the sample or the particles combined with the target substance gather in the vortex. In another aspect of the present invention, only one resonator in the relatively arranged ultra-high frequency bulk acoustic wave resonator in the direction of bulk acoustic wave emission works, and the power range of the bulk acoustic wave that generates a vortex in the solution to gather the sample / target substance / particles in the solution is about 0.01-20mW, preferably 0.1-10mW, and more preferably 0.5-5mW.

[0044] In one aspect of the present invention, the sample in the method is a biological sample (e.g., body fluid, blood, cell, virus, protein, lipid, carbohydrate, nucleic acid, aptamer, nucleoside). The method and device of the present invention are particularly suitable for samples with complex contents and / or samples with higher viscosity, such as whole blood and respiratory mucus, such as sputum or nasal mucus.

[0045] In one aspect of the present invention, the target substance in the method is a peptide, glycoprotein, lipoprotein, DNA, RNA, PNA, nucleotide or polysaccharide.

[0046] In one aspect of the invention, the solution in the method contains magnetic or non-magnetic particles that can bind to the sample / target substance. Preferably, the size of the sample or particle is about 0.01 μm to about 100 μm, more preferably about 0.05 μm to about 20 μm, such as about 0.5 μm to about 10 μm.

[0047] In one aspect of the present invention, the volume of the solution in the method is about 20 μl-1 ml, preferably about 50 μl-500 μl, and more preferably about 100 μl-200 μl. The method and device of the present invention perform well when processing smaller volumes, such as about 20 μl-1 ml, preferably about 50 μl-500 μl, and more preferably about 100 μl-200 μl of solution, especially when processing small volumes of samples with complex inclusions and / or samples with higher viscosity. In one aspect of the present invention, the distance between the relatively arranged ultrahigh frequency bulk acoustic wave resonators in the method is about 0.1-2.0 cm, preferably about 0.2-1.0 cm, for example about 0.2-0.8 cm.

[0048] In one aspect of the present invention, in the method, the volume of the chamber is about 100 μL-10 mL, preferably about 200 μL-5 mL, and more preferably about 500 μL-1 mL.

[0049] In one aspect of the present invention, in the method, the height of the cavity is about 0.5-5 cm, preferably about 1-3 cm, such as about 1-2 cm.

[0050] In one aspect of the present invention, the method is used to separate nucleic acids from a sample. In one aspect of the present invention, the method is used to separate nucleic acids from a sample by magnetic bead method.

[0051] In one aspect of the present invention, the method further comprises the step of amplifying the nucleic acid.

[0052] The present invention also provides a kit for the above method of treating a sample in a solution, the kit comprising:

[0053] A cavity containing a solution and an ultra-high frequency bulk acoustic wave resonator arranged relatively in the cavity, wherein the resonator can generate bulk acoustic waves, generate vortex in the solution, mix or enrich the sample in the solution, wherein the ultra-high frequency bulk acoustic wave resonator can generate bulk acoustic waves with a frequency of about 0.5-50 GHz.

[0054] In one aspect of the present invention, the kit includes magnetic or non-magnetic particles that can bind to a sample or a target substance (e.g., nucleic acid) in the sample. Preferably, the sample or particle has a size of about 0.01 μm to about 100 μm, preferably about 0.05 μm to about 20 μm, for example about 0.5 μm to about 10 μm.

[0055] In one aspect of the present invention, the kit is used to separate nucleic acids from a sample. In one aspect of the present invention, the kit contains reagents for separating nucleic acids from a sample by magnetic bead method, such as nucleic acid binding, nucleic acid cleaning or nucleic acid elution reagents.

[0056] In one aspect of the present invention, the kit further comprises reagents for nucleic acid amplification, for example, reagents for PCR such as amplification enzymes, primers, and the like. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0058] Figure 1 Schematic diagram and structural diagram of an exemplary solution sample mixing and enrichment device of the present invention.

[0059] Figure 2 Schematic diagram and experimental results of the ultra-high frequency bulk acoustic wave resonator in the exemplary device of the present invention generating bulk acoustic waves and acoustic flow vortices in the cavity, as well as the effect on particles in the solution.

[0060] Figure 3 A diagram showing experimental results of the generation of bulk acoustic waves and acoustic flow vortices in a cavity by an ultra-high frequency bulk acoustic wave resonator in an exemplary device of the present invention.

[0061] Figure 4 A diagram showing experimental results of the generation of bulk acoustic waves and acoustic flow vortices in a cavity by an ultra-high frequency bulk acoustic wave resonator in an exemplary device of the present invention.

[0062] Figure 5 A graph showing the experimental results of using the exemplary device of the present invention to extract adenovirus.

[0063] Figure 6 An exemplary device of the present invention is used to extract experimental results of coronavirus. DETAILED DESCRIPTION

[0064] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field based on the disclosure and teachings of the present invention belong to the scope of protection of the present invention.

[0065] Example 1 Solution sample mixing and enrichment device

[0066] This embodiment provides an exemplary solution sample mixing and enrichment device of the present invention. Figure 1 Schematic diagram and structural diagram of an exemplary solution sample mixing and enrichment device of the present invention.

[0067] in, Figure 1 (A) is a schematic diagram of the solution sample mixing and enrichment device. The device includes a cavity that can hold a solution, and an inlet and an outlet (not shown) that can have a solution. The wall of the cavity is a polytetrafluoroethylene material that is biocompatible with cells or biomacromolecules such as nucleic acids and proteins. Figure 1 Two UHF BAW resonators are respectively arranged at the bottom and top of the device shown, and the two UHF BAW resonators can generate UHF BAW in the solution in the cavity with the propagation direction being the opposite side of the cavity. The surface of the UHF BAW resonator can be pre-treated by soaking in bovine serum albumin solution to prevent nucleic acid adhesion.

[0068] The ultra-high frequency bulk acoustic wave resonator that can be used in the present invention can generate bulk acoustic waves with a frequency of about 0.5-50 GHz. The ultra-high frequency bulk acoustic wave resonator includes a thin film bulk acoustic wave resonator or a solid-state assembly type resonator, such as a thickness stretching vibration mode acoustic wave resonator.

[0069] Figure 1 (B) is a structural diagram of the ultra-high frequency bulk acoustic wave resonator device. The surface of the resonator is a pentagon with a side length of about 100 μm. The signal line on the EVB board is connected to the top electrode of the bulk acoustic wave device (the middle position of the three-pronged structure) through a gold wire, and the bottom electrode of the bulk acoustic wave device (the two sides of the three-pronged structure) is connected to the ground position (non-signal position) on the EVB board. Then, the EVB board is connected to the signal generator through an SMA adapter to realize the input of high-frequency signals.

[0070] An exemplary ultra-high frequency bulk acoustic wave resonator is a resonator having a Bragg acoustic reflection structure, Figure 1(C) is an internal structure diagram of the ultrahigh frequency bulk acoustic wave resonator, and its preparation method is as follows:

[0071] The chip is made step by step by performing chemical vapor deposition, metal sputtering, photolithography and other methods on silicon-based wafers. The specific methods are as follows:

[0072] 1. Use a piranha solution with a volume ratio of concentrated sulfuric acid to hydrogen peroxide of 3:1 to thoroughly clean the surface of the silicon wafer. This method can effectively remove organic and inorganic matter on the silicon wafer.

[0073] 2. On the cleaned silicon wafer, a layer of aluminum nitride film is formed by surface sputtering, and then a layer of silicon dioxide film is deposited by ion-enhanced chemical vapor deposition. Then, the same method is used to alternately deposit aluminum nitride film and silicon dioxide film to form a Bragg acoustic reflection structure in which aluminum nitride and silicon dioxide alternately overlap.

[0074] 3. On the Bragg reflection layer structure, a 600nm molybdenum film is sputtered as the bottom electrode. Then, standard photolithography techniques, including coating, exposure, and development, are used to photolithograph the molybdenum electrode film, and then etching is performed to form a bottom electrode with a target pattern.

[0075] 4. Sputter an aluminum nitride film on the molybdenum electrode as a piezoelectric layer. Use dry etching to define a pattern on the aluminum nitride film. In this exemplary embodiment, a pentagonal shape is used.

[0076] 5. Use negative photoresist to transfer the pattern on the mask, and then sputter a 50nm thick titanium-tungsten alloy layer, which can increase the adhesion of the gold electrode as an adhesion layer. Then use the evaporation method to grow a 300nm thick gold film on the upper electrode. Finally, use acetone to remove the gold film around the target pattern to form a gold electrode with the target pattern.

[0077] In this exemplary embodiment, the UHF resonator surface is pentagonal, with a side length of about 100 μm and an area of ​​about 25,000 μm. 2 .

[0078] The resonator device is connected to a network analyzer using a standard SMA interface, and the resonance peak is found by testing the spectrum. It can be measured that in this exemplary embodiment, the frequency of the bulk acoustic wave emitted by the ultra-high frequency resonator device in the cavity is 2.5 GHz.

[0079] Preparation of solution sample mixing and enrichment device

[0080] In one aspect of the present invention, the cavity in the device provided by the present invention is cylindrical, with a bottom diameter of about 0.2-2.0 cm, preferably 0.5-1.0 cm. The device of the present invention may also use cavities of other shapes. The cavity prepared in this embodiment is cylindrical, with a bottom diameter of about 0.6 cm. An ultra-high frequency bulk acoustic wave resonator provided according to the above preparation method is bonded and integrated with the bottom of the cavity, and another ultra-high frequency bulk acoustic wave resonator is connected to the EVB board through a tube shell adapter. The device surface is perpendicular to the EVB surface and can be inserted into the cavity from the top of the cavity. Since the bulk acoustic wave resonator at the top is separated from the cavity, the distance between the two devices can be adjusted according to the amount of sample to be processed. In the experiment, two ultra-high frequency bulk acoustic wave resonator devices are respectively arranged at both ends of the cavity, and can be adjusted in the range of 0.1 cm-2.0 cm according to the amount of sample to be processed. In one aspect of the present invention, the volume of the cavity in the device provided by the present invention is about 20 microliters-1 milliliter, preferably about 50 microliters-500 microliters, and more preferably about 100 microliters-200 microliters. In one aspect of the present invention, the device and method provided by the present invention are suitable for processing solutions of about 20 μl to 1 ml, preferably about 50 μl to 500 μl, and more preferably about 100 μl to 200 μl.

[0081] In an exemplary embodiment of the present invention, a device for generating a magnetic field may also be provided at the bottom of the cavity containing the solution of the device, such as a device for generating magnetism by electromagnetic action, including an electromagnet. The device for generating a magnetic field may generally be provided near (e.g., below) the UHF BAW resonator at the bottom. The generated magnetic field may be used to adsorb magnetic particles in the solution, such as magnetic beads (which may bind / adsorb target biomacromolecules such as nucleic acids or proteins), etc. In this embodiment, an electromagnet is provided below the UHF BAW resonator at the bottom of the cavity.

[0082] Example 2

[0083] In the method and device of the present invention, an ultra-high frequency bulk acoustic wave resonator emits an ultra-high frequency bulk acoustic wave in a solution in a cavity toward the opposite side, causing an acoustic flow vortex to appear in the solution; the forces acting on the particles in the solution include the fluid drag force (Stokes drag force) and the acoustic radiation force (acoustic radiation force) generated by the vortex. Figure 2 It is a schematic diagram and experimental result diagram of the generation of bulk acoustic waves and acoustic flow vortices in the cavity by the ultra-high frequency bulk acoustic wave resonator in the method and device provided by the present invention, and the effect on particles in the solution.

[0084] Figure 2 (A) is a schematic diagram of the generation of bulk acoustic waves and acoustic flow vortices in the cavity by the ultra-high frequency bulk acoustic wave resonator in the method and device provided by the present invention, and the effect on particles in the solution. Figure 2As shown in (A), the UHF BAW generates an acoustic flow vortex in the solution, and the particles in the solution are subject to fluid drag force (Fdrag) and acoustic radiation force (Frad). In the area close to the UHF BAW resonator, the particles are mainly affected by the acoustic radiation force and move from the outside to the inside along the vortex. When they move to the edge of the resonator, the particles are pushed to the inner area deviating from the vortex trajectory due to the large horizontal component of the acoustic radiation force. When the particles are a certain distance away from the edge of the device, the acoustic radiation force decreases due to the distance, and the particles are again affected by the fluid drag force of the micro-vortex from top to bottom and move to the edge of the device, and so on. When the particles in the solution are far away from the UHF BAW resonator, they exceed the range of the acoustic radiation force and are mainly affected by the fluid drag force and spread in space.

[0085] Figure 2 (B) Experimental results of the forces and motion states of particles in a solution when an ultra-high frequency bulk acoustic wave resonator generates bulk acoustic waves and acoustic flow vortices in a cavity in the exemplary methods and apparatus provided by the present invention. In the experiment, 200ul of polystyrene particles with a particle size of 5um (hereinafter referred to as PS particles or PS beads) were tested to observe their motion in the cavity of the device of the present invention. The motion of micron-scale particles can be observed under normal light. The velocity characterization of the flow field motion is accomplished by photographing the motion of the microspheres in the cavity under a high-speed microscope.

[0086] Figure 2 (B) The horizontal axis is time, and the vertical axis is the particle ratio (the ratio of particles to the total area of ​​the field of view within a specified range). The points on the curve in the figure are the particle ratio values ​​measured at different time points. Figure 2 As shown in (B), within the first second of the experiment (the inset is a photo of the device surface taken by a high-speed camera), when the input power of the UHF BAW resonator is lower than a certain power (for example, lower than about 20mW, preferably lower than 10mW, 5.4mW is used in the experiment in the figure), the center of the acoustic flow vortex is close to the device surface, and the PS spheres gather at the center of the vortex. When the power increases to a certain power (for example, greater than about 100mW, preferably greater than about 200mW, 200mW is used in the experiment in the figure), the center of the vortex in the cavity moves upward, and the acoustic flow effect is enhanced. At this time, the particles in the solution have moved away from the device and are beyond the range of the acoustic radiation force. They are mainly affected by the drag force of the fluid and quickly change from a focused state to a dispersed state in space. As shown Figure 2 As shown in (B), the small figure is a scatter diagram of the PS balls being scattered in space. The results show that within 1 second after the generation of the bulk acoustic wave, the PS balls can change from a focused state to a scattered state in space (i.e., being scattered).

[0087] Example 3

[0088] In the fields of biotechnology and clinical medicine, biological samples from which biomacromolecules such as nucleic acids or proteins need to be extracted can be samples with complex contents and high viscosity, such as whole blood samples and sputum samples. In other forms of applications that use acoustics to drive and stir solution samples, high power is usually required, which leads to high temperatures in the solution (especially small volume solutions), damage to the required cells or nucleic acids, or the need to add additional reagents, which affects subsequent reactions.

[0089] The inventors used the device and method of the present invention for generating bulk acoustic waves and swirls in a solution in a cavity using an ultra-high frequency bulk acoustic wave resonator to conduct experiments in a liquid with high viscosity. 0.5 μl of fluorescently labeled PS particles with a particle size of 30 μm (Shanghai Huizhi Biotechnology Co., Ltd., Cat. No: GF030C) was added to 100 μl of 20% ethylene glycol aqueous solution, and then added to the exemplary device described in Example 1, and first only the ultra-high frequency bulk acoustic wave resonator on one side (bottom) was turned on. Figure 3 Display the results of the experiment. Figure 3 (A) shows the movement speed of tracer particles at different working powers of the UHF BAW resonator observed under a high-speed fluorescence microscope. Figure 3 (B) shows the mixing efficiency. According to the method and definition described in RJ Shilton et al., Sens. Actuators B: Chem. 2011, 160, according to the formula

[0090]

[0091]

[0092] Where M is defined as the mixing efficiency (mixing index),

[0093] The mixing efficiency is calculated by grayscale processing of matlab software. The UHF BAW resonator is in the off state for the first 12 seconds, and after it is turned on (working power is 100mW), the mixing efficiency quickly reaches about 70% and then continues for another 12 seconds, and the mixing efficiency is basically stable at about 75%. Figure 3 (C) Repeat Figure 3(B) Experimental steps, after 12 seconds, the UHF BAW resonator is turned on, and the mixing efficiency that can be achieved at the power of 0mW, 100mW, 200mW, and 400mW at 12 seconds after turning on is tested respectively. As shown in the figure, about 80% can be achieved at 400mW. However, the experiment found that when the UHF BAW resonator on one side is turned on, the mixing efficiency will show a platform phenomenon when it reaches a certain level by adjusting the BAW power: it is difficult to significantly improve the mixing efficiency after reaching a certain height, thereby affecting the binding efficiency of the biomacromolecules (such as nucleic acids or proteins) in the sample with the particles (such as magnetic beads) used to bind and adsorb the target biomacromolecules.

[0094] Example 4

[0095] The inventors further observed the simultaneous activation of relatively arranged UHF BAW resonators and compared the activation of one side of the UHF BAW resonator with that of the other side. Figure 4 (A) is a UHF BAW resonator with the bottom open alone. At a power of 400 mW, the motion state of polystyrene microspheres in a vertical section perpendicular to the surface of the UHF BAW resonator was photographed. The double vortex caused by the BAW (the arrow in the figure shows the direction of liquid flow) and the mixing effect on the microspheres in the solution can be observed. Figure 4 (B) shows the motion state of polystyrene microspheres in a vertical section of the UHF BAW resonator surface, taken at 400mW power, with the top and bottom UHF BAW resonators turned on at the same time. Figure 4 The two vortices shown by the upper arrows in (B) are caused by the BAW resonator set at the bottom (e.g. Figure 4 (A) The vortex observed in Figure 4 (B) The two vortices indicated by the lower arrows are caused by the BAW resonator set at the top. It can be observed that the vortex motion caused by the upper and lower BAW resonators is opposite, so the vortex action volume increases, and can act on the resonator surface set on the opposite side, playing a role in cleaning the surface of each other, allowing more particles to move into the three-dimensional space, and achieving a stronger mixing effect on the particles in the solution. When there are biomacromolecules (such as nucleic acids or proteins) and particles that can bind to target biomacromolecules (such as magnetic beads) in the solution, higher mixing can improve their binding efficiency.

[0096] Figure 4 (C) shows the experimental results of the exemplary methods and devices provided by the present invention for nucleic acid extraction.

[0097] The surface of the ultra-high frequency bulk acoustic wave resonator and the inner wall of the cavity in the device prepared in the above-mentioned embodiment 1 were soaked with bovine serum albumin solution to prevent the adsorption of nucleic acid on the surface. Whole blood was collected from SD mice (provided by Tianjin Yishengyuan Biotechnology Co., Ltd.), and nucleic acids were extracted using the reagents (including magnetic beads, lysate, binding solution, washing solution, eluent, protease and its buffer) in the whole blood genomic DNA extraction kit (magnetic bead method) (Changzhou Jinmaige Biotechnology Co., Ltd., product number NA001-1). The main steps include:

[0098] 1. Add 200μl mouse blood sample, 200μl lysis buffer, and 10μl proteinase K to a 1.5ml nuclease-free centrifuge tube, mix thoroughly, and place in a 55℃ water bath for 10min. Invert and mix 3 times, 3-5 times each time. Then take out 20.5μL, 61.5μL, and 102.5μL of the lysis sample for nucleic acid extraction respectively;

[0099] 2. Add the three groups of samples after lysis into the cavity, and then add 15μl, 45μL, 75μL of binding solution and 10μl of magnetic beads respectively, open the bottom or both the top and bottom BAW resonators (input power 400mW) for 10min, and the nucleic acids bind to the magnetic beads;

[0100] 3. The bottom BAW resonator works at low power (input power is about 1-10mW) for 1 minute, combined with the electromagnetic suction outside the tube, so that all particles gather at the bottom and absorb the waste liquid;

[0101] 4. Use 500 μL of cleaning solution 1, open the bottom or both the top and bottom BAW resonators (input power 400 mW) to disperse the magnetic beads for 30 seconds, operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute, and combine with the electromagnetic suction outside the tube to make all the particles gather at the bottom and absorb the waste liquid;

[0102] 5. Use 500 μL of cleaning solution 2, open the bottom or both the top and bottom BAW resonators (input power 400 mW) to disperse the magnetic beads for 30 seconds, operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute, and combine with the electromagnetic suction outside the tube to make all the particles gather at the bottom and absorb the waste liquid;

[0103] 6. Repeat step 5;

[0104] 7. Ventilate and let stand, dry at room temperature for 10 minutes;

[0105] 8. 75°C elution buffer, 60 μL elution, 5 min at room temperature;

[0106] 9. Collect 60 μL of nucleic acid-containing eluate and use NanoDrop 2000 Spectrophotometer (Thermo Fisher, Waltham, MA) for nucleic acid quantitative detection.

[0107] The results are as follows Figure 4 As shown in (C), the effects of turning on the UHF BAW resonator on one side of the bottom alone and turning on the UHF BAW resonators on both sides at the same time are compared. When the BAW resonator power is 400mW, the nucleic acid extraction efficiency of the UHF BAW resonator on both sides is significantly improved from 50% of the UHF BAW resonator on the bottom side alone to about 80%. Without being limited by this theory, the inventors believe that the improvement in the extraction effect is due to the fact that the vortex motion caused by the upper and lower BAW resonators has a stronger mixing effect on the particles in the solution, thereby improving the binding efficiency of the nucleic acid and the magnetic beads and ultimately improving the nucleic acid extraction efficiency.

[0108] Example 5 Extraction and Detection of Adenovirus in Serum Samples

[0109] The device and method of the present invention were used to extract and detect adenovirus in serum. Adenovirus was purchased from (Hanheng Biotechnology (Shanghai) Co., Ltd., item number HB-AP2100001), and diluted in bovine serum (purchased from Hangzhou Tianhang Biotechnology Co., Ltd., item number 22012-8612) to prepare adenovirus with a content of 10 1 IU / mL,10 2 IU / mL,10 3 IU / mL,10 4 IU / mL,10 5 IU / mL,10 6 IU / mL gradient test solution.

[0110] The surface of the ultrahigh frequency bulk acoustic wave resonator and the inner wall of the cavity in the device prepared in the above-mentioned Example 1 are soaked with bovine serum albumin solution to prevent the adsorption of nucleic acid on the surface.

[0111] The viral RNA was extracted from the gradient test solution using the reagents (including magnetic beads, lysate, washing solution, elution solution, protease and its buffer) in the viral DNA / RNA extraction kit (magnetic bead method) (Changzhou Jinmaige Biotechnology Co., Ltd.). The main steps include:

[0112] 1. Add 50 μl of virus sample, 75 μl of lysis solution, 5 μl of proteinase K and 10 μl of magnetic beads into the chamber, and turn on the top and bottom BAW resonators (input power 400 mW) for 10 min to allow the nucleic acid to bind to the magnetic beads;

[0113] 2. The bottom BAW resonator works at low power (input power is about 1-10mW) for 1 minute, combined with the electromagnetic suction outside the tube, so that all particles gather at the bottom and absorb the waste liquid;

[0114] 3. Use 500 μL of cleaning solution 1, turn on the top and bottom BAW resonators (input power 400 mW) at the same time to disperse the magnetic beads for 30 seconds, and operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute. Combined with the electromagnetic suction outside the tube, all the particles are gathered at the bottom and the waste liquid is sucked away;

[0115] 4. Use 500 μL of cleaning solution 2, turn on the top and bottom BAW resonators (input power 400 mW) at the same time to disperse the magnetic beads for 30 seconds, and operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute. Combined with the electromagnetic suction outside the tube, all the particles are gathered at the bottom and the waste liquid is sucked away;

[0116] 5. Ventilate and let stand;

[0117] 6. 75°C elution buffer, 60 μL elution, 5 min at room temperature;

[0118] 7. Collect 60 μL of nucleic acid-containing eluate.

[0119] The virus extracted and purified by the method and device of the present invention was subjected to RT-PCR detection using TaKaRaTaq T M HotStart Version Kit (Takara BioInc. Japan, product number: R007Z). Forward primer 5'-TGCTGCCCGACAACCATC-3', reverse primer 5'-TCACGAACTCCAGCAGGACCA-3', probe 5'-TGA GCACCC AGT CCG CCC TGA GCA-3' (synthesized by Sangon Biotech (Shanghai) Co., Ltd.).

[0120] The results are as follows Figure 5 shown. Figure 5 (A) is the logarithm of virus amount-cycle number curve of RT-PCR. 1 IU / mL concentration, the intervals were basically the same, and the detection limit was 10 2 IU / mL concentration. Figure 6 (B) is a graph showing the fluorescence value-cycle number of RT-PCR. The results show that the device and method of the present invention can effectively extract low-concentration adenovirus from serum.

[0121] Example 6 Extraction and Detection of Coronavirus

[0122] The device and method of the present invention were used to extract and detect coronavirus samples, and compared with commercial virus extraction kits.

[0123] The coronavirus sample was FNV-2019-nCOV-abEN pseudovirus (Cat. No. FNV2001) purchased from Fuboao (Suzhou) Biotechnology Co., Ltd. The concentration of the sample stock solution was 10 8 copies / mL, which was diluted in PBS to obtain 10 6 Copies / mL, 10 5 Copies / mL, 10 4 Copies / mL, 10 3 copies / mL of gradient test solution.

[0124] The surface of the ultrahigh frequency bulk acoustic wave resonator and the inner wall of the cavity in the device prepared in the above-mentioned Example 1 are soaked with bovine serum albumin solution to prevent the adsorption of nucleic acid on the surface.

[0125] The viral RNA was extracted from the gradient test solution using the reagents (including magnetic beads, lysate, washing solution, elution solution, protease and its buffer) in the viral DNA / RNA extraction kit (magnetic bead method) (Changzhou Jinmaige Biotechnology Co., Ltd.). The main steps include:

[0126] 1. Add 100 μl of virus sample, 150 μl of lysis solution, 10 μl of proteinase K and 20 μl of magnetic beads into the chamber, and turn on the top and bottom BAW resonators (input power 400 mW) for 10 min to allow the nucleic acid to bind to the magnetic beads;

[0127] 2. The bottom BAW resonator works at low power (input power is about 1-10mW) for 1 minute, combined with the electromagnetic suction outside the tube, so that all particles gather at the bottom and absorb the waste liquid;

[0128] 3. Use 500 μL of cleaning solution 1, turn on the top and bottom BAW resonators (input power 400 mW) at the same time to disperse the magnetic beads for 30 seconds, and operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute. Combined with the electromagnetic suction outside the tube, all the particles are gathered at the bottom and the waste liquid is sucked away;

[0129] 4. Use 500 μL of cleaning solution 2, turn on the top and bottom BAW resonators (input power 400 mW) at the same time to disperse the magnetic beads for 30 seconds, and operate the bottom BAW resonator at low power (input power about 1-10 mW) for 1 minute. Combined with the electromagnetic suction outside the tube, all the particles are gathered at the bottom and the waste liquid is sucked away;

[0130] 5. Ventilate and let stand;

[0131] 6. 75°C elution buffer, 60 μL elution, 5 min at room temperature;

[0132] 7. Collect 60 μL of nucleic acid-containing eluate.

[0133] In addition, a viral DNA / RNA extraction kit (magnetic bead method) (Changzhou Jinmai Biotechnology Co., Ltd.) was used to extract nucleic acids from the prepared gradient test solution using the magnetic rod method according to the method provided in its instructions and in conjunction with a fully automatic nucleic acid extraction and purification instrument DOF-9848 (Changzhou Jinmai Biotechnology Co., Ltd.).

[0134] The viral nucleic acid extracted and purified by the method and apparatus of the present invention and the commercial nucleic acid extraction kit and method was subjected to RT-PCR detection, and TaKaRaTaq T M HotStart Version kit (Takara Bio Inc. Japan, product number: PR064A). Forward primer 5'-CCCTGTGGGTTTTACACTTAA-3', reverse primer 5'-ACGATTGTGCATCAGCTGA-3', probe 5'-CCGTCTGCGGTATGTGGAAAGGTTATGG-3' (synthesized by Sangon Biotech (Shanghai) Co., Ltd.).

[0135] The results are as follows Figure 6 shown. Figure 6 (A) is the logarithm of virus amount-cycle number curve of RT-PCR. Figure 6 (B) is a graph of fluorescence value-cycle number of RT-PCR. The results show that the device and method of the present invention can effectively extract low-concentration coronaviruses, and the extraction efficiency is higher than that of the instrument using the magnetic rod method.

[0136] Although the exemplary embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions and modifications may be made to these embodiments without departing from the spirit of the present invention and the scope of protection defined by the appended claims. For other examples, it should be readily understood by those skilled in the art that the order of the process steps may be changed while maintaining the scope of protection of the present invention.

[0137] In addition, the scope of application of the present invention is not limited to the processes, mechanisms, manufactures, material compositions, means, methods and steps of the specific embodiments described in the specification. From the disclosure of the present invention, it will be easily understood by those skilled in the art that the processes, mechanisms, manufactures, material compositions, means, methods or steps that currently exist or will be developed in the future, which perform substantially the same functions as the corresponding embodiments described in the present invention or obtain substantially the same results, can be applied according to the present invention. Therefore, the claims attached to the present invention are intended to include these processes, mechanisms, manufactures, material compositions, means, methods or steps within their scope of protection.

Claims

1. A device for processing a solution sample, characterized in that: The invention comprises a cavity for accommodating a solution and relatively arranged ultrahigh frequency bulk acoustic wave resonators in the cavity, wherein the ultrahigh frequency bulk acoustic wave resonators can generate bulk acoustic waves with a frequency of 0.5-50 GHz, and generate a vortex in the solution; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonators is reversed, and has a chaotic convection effect, so that the vortex action volume is increased, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, so as to clean the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side.

2. The device according to claim 1, characterized in that The distance between the relatively arranged ultrahigh frequency bulk acoustic wave resonators is 0.1-5 cm.

3. The device according to claim 1, characterized in that The area of ​​the BAW generation region of the UHF BAW resonator is 1000-50000 μm 2 .

4. The device according to claim 1, characterized in that It is provided with a power regulating device for independently regulating the power of the bulk acoustic wave generated by each of the relatively arranged ultra-high frequency bulk acoustic wave resonators.

5. The device according to claim 1, characterized in that The working power range of the ultra-high frequency bulk acoustic wave resonator to generate bulk acoustic waves includes 150mW-3000mW.

6. The device according to claim 1, characterized in that The working power range of the ultra-high frequency bulk acoustic wave resonator to generate bulk acoustic waves includes 0.01-20mW.

7. The device according to claim 1, characterized in that The ultra-high frequency bulk acoustic wave resonator is a thin film bulk acoustic wave resonator or a solid-state assembly type resonator.

8. The device according to claim 1, characterized in that The volume of the cavity is 100 μl-10 ml.

9. The device according to claim 1, characterized in that The height of the cavity is 0.5-5 cm.

10. The device according to claim 1, characterized in that The solution sample contains a biological sample and / or magnetic or non-magnetic particles that can bind to the biological sample, wherein the size of the biological sample or the particles is 0.01 μm to 100 μm.

11. A device for purifying a target substance in a solution sample, characterized in that: It includes: The device according to any one of claims 1 to 10, comprising a cavity for accommodating a solution and relatively arranged ultrahigh frequency bulk acoustic wave resonators in the cavity, wherein the ultrahigh frequency bulk acoustic wave resonators can generate bulk acoustic waves with a frequency of 0.5-50 GHz to generate a vortex in the solution; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonators is reversed, has a chaotic convection effect, increases the vortex action volume, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, thereby cleaning the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side.

12. The device according to claim 11, characterized in that The solution sample includes a biological sample.

13. The device according to claim 11, characterized in that The target substance is nucleic acid.

14. The device according to claim 11, characterized in that It includes a sample lysis module.

15. The device according to claim 11, characterized in that It includes a target substance binding module.

16. The device according to claim 11, characterized in that It includes a target material cleaning module.

17. The device according to claim 11, characterized in that It includes a target substance elution module.

18. The device according to claim 11, characterized in that It has a magnet device which controls the magnetic material and / or the magnetic material bound to a target substance by applying a magnetic field.

19. A system for detecting a target substance in a solution sample, characterized in that: It includes: The device according to any one of claims 1 to 10 or the equipment according to any one of claims 11 to 18, which has a cavity for accommodating a solution and relatively arranged ultrahigh frequency bulk acoustic wave resonators in the cavity, wherein the ultrahigh frequency bulk acoustic wave resonators can generate bulk acoustic waves with a frequency of 0.5-50 GHz to generate a vortex in the solution; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonators is reversed, has a chaotic convection effect, increases the vortex action volume, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, thereby cleaning the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side; and A detection device detects an identifiable label carried by a target substance, wherein the identifiable label includes fluorescence, chemiluminescence, bioluminescence, radioluminescence, electroluminescence, electrochemiluminescence, mechanical luminescence, crystal luminescence, thermoluminescence, sonoluminescence, phosphorescence, photoluminescence, enzymatic reaction, or radioactivity.

20. A system for amplifying and detecting a target substance in a solution sample, characterized in that: It includes: The device according to any one of claims 1 to 10 or the equipment according to any one of claims 11 to 18, which has a cavity for accommodating a solution and relatively arranged ultrahigh frequency bulk acoustic wave resonators in the cavity, wherein the ultrahigh frequency bulk acoustic wave resonators can generate bulk acoustic waves with a frequency of 0.5-50 GHz to generate a vortex in the solution; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonators is reversed, has a chaotic convection effect, increases the vortex action volume, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, thereby cleaning the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side; Nucleic acid amplification module; the nucleic acid amplification module is a module for performing nucleic acid amplification, including: isothermal amplification, qPCR, RT-PCR, hot start PCR, nested PCR, multiplex PCR, recovery condition PCR, dsRNA synthesis, COLD-PCR, or digital PCR; and A detection device detects identifiable labels carried by a target substance, wherein the identifiable labels include fluorescence, chemiluminescence, bioluminescence, radioluminescence, electroluminescence, electrochemiluminescence, mechanical luminescence, crystal luminescence, thermoluminescence, sonoluminescence, phosphorescence, photoluminescence, enzymatic reaction, or radioactivity.

21. A method for processing a sample in a solution, characterized in that: The method comprises: generating a bulk acoustic wave by using relatively arranged ultrahigh frequency bulk acoustic wave resonators in a cavity containing a solution, generating a vortex in the solution, and mixing a sample in the solution, wherein the ultrahigh frequency bulk acoustic wave resonator can generate a bulk acoustic wave with a frequency of 0.5-50 GHz; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonators is reversed, has a chaotic convection effect, increases the vortex action volume, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, thereby achieving cleaning of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side.

22. The method according to claim 21, characterized in that It also includes: one of the relatively arranged ultra-high frequency bulk acoustic wave resonators does not work, and the corresponding resonator arranged in the opposite direction works, generating bulk acoustic waves, so that the solution in the bulk acoustic wave area generates a vortex, and the sample in the solution or the target substance contained in the sample or the particles combined with the target substance gather in the vortex.

23. The method according to claim 21 or 22, characterized in that The sample is a biological sample; the biological sample includes: body fluid, cells, viruses, proteins, lipids, carbohydrates, nucleic acids, aptamers, or nucleosides.

24. The method of claim 22, wherein: The target substance is a peptide, glycoprotein, lipoprotein, DNA, RNA, PNA, nucleotide or polysaccharide.

25. The method according to claim 21 or 22, characterized in that The solution contains magnetic or non-magnetic particles that can bind to samples or target substances, wherein the size of the samples or particles is 0.01 μm to 100 μm.

26. The method according to claim 21 or 22, characterized in that The area of ​​the BAW generation region of the UHF BAW resonator is 1000-50000 μm 2 .

27. The method according to claim 21 or 22, characterized in that The volume of the cavity is 100 μl-10 ml.

28. The method according to claim 21 or 22, characterized in that The height of the cavity is 0.5-5 cm.

29. The method according to claim 21 or 22, characterized in that It is used to isolate nucleic acids from a sample.

30. The method of claim 28, wherein: It also includes the step of amplifying the nucleic acid.

31. A kit for use in the method for treating a sample in a solution according to any one of claims 21 to 30, characterized in that: The kit comprises: A cavity for accommodating a solution and an ultrahigh frequency bulk acoustic wave resonator arranged relatively in the cavity, wherein the resonator can generate bulk acoustic waves, generate a vortex in the solution, and mix or enrich the sample in the solution, wherein the ultrahigh frequency bulk acoustic wave resonator can generate bulk acoustic waves with a frequency of 0.5-50 GHz; wherein the vortex motion of the vortex generated by the relatively arranged ultrahigh frequency bulk acoustic wave resonator is reversed, has a chaotic convection effect, increases the vortex action volume, and the vortex influence range can reach the position of the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side, thereby cleaning the ultrahigh frequency bulk acoustic wave resonator surface on the opposite side.

32. The kit according to claim 31, wherein These include magnetic or non-magnetic particles that can bind to a sample or a target substance in the sample, wherein the size of the sample or the particle is 0.01 μm to 100 μm.

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