Sample pretreatment apparatus and sample pretreatment method

Abstract

The invention belongs to the technical field of sample detection, and discloses a sample pretreatment device and a sample pretreatment method. The sample pretreatment device comprises a sample positioning disc, an ultrasonic module and a driving part, the sample positioning disc is provided with a plurality of uniformly distributed accommodating grooves, the accommodating grooves are used for supporting test tubes, and the sample positioning disc is fixed on the outer shell; the ultrasonic module comprises an ultrasonic transducer and an ultrasonic vibration disc connected with the ultrasonic transducer, the ultrasonic vibration disc is in movable contact with the sample positioning disc, the ultrasonic vibration disc is provided with a plurality of limiting grooves, the multiple limiting grooves and the multiple containing grooves are arranged in a one-to-one correspondence mode, and the limiting grooves and the containing grooves define a containing space for clamping test tubes; the driving part is in transmission connection with the ultrasonic module and is configured to drive the ultrasonic module to ascend and descend. According to the sample pretreatment device, the structure is simplified, the processing and manufacturing cost is reduced, and the application range is wide. The sample pretreatment method provided by the invention is completed by using the sample pretreatment device.

Description

Technical Field

[0001] The present invention relates to the technical field of sample detection, and in particular to a sample pre-processing device and a sample pre-processing method. Background Art

[0002] In the fields of scientific research, biochemical analysis, inspection and quarantine, in order to obtain the required nucleic acids (DNA, RNA), proteins and other target molecules, the early processing of samples, especially the fragmentation of tissues and cells, is a very important and critical step.

[0003] There are two methods for biological sample disruption: chemical methods and mechanical methods. The chemical method targets the cell membrane or cell wall of the sample to be processed. Relevant enzymes or chemicals need to be added during the processing. The processing effect is better for specific samples, but the application is limited, and the residue of chemical reagents may affect downstream experiments. Mechanical methods include mortar grinding, bead milling, homogenizer disruption, high temperature disruption, freezing disruption, ultrasonic disruption, etc. Mechanical methods mainly disrupt tissues by shearing solids or liquids, destroying cell membranes or cell walls, and causing target molecules to overflow. Compared with chemical methods, mechanical disruption does not introduce chemical substances that may interfere with downstream experiments, but mechanical methods such as mortar grinding, bead milling, and homogenizer disruption cannot handle small volumes of samples, and are prone to loss of operating samples during the processing process, and cross contamination occurs, while high temperature or freezing disruption often takes a long time. Traditional ultrasonic disruption methods mainly include probe ultrasound and water bath ultrasound. Among them, probe ultrasound may cause cross contamination due to the direct contact of the probe with the sample. Water-soluble ultrasound requires a water bath environment, because the sample to be processed must be placed in a water tank, the transmission time is slow and the processing efficiency is low. Using these traditional methods, the yield of target molecules is low and the purity is poor, which has a great impact on the stability and repeatability of downstream experiments.

[0004] In view of the problems of cross-contamination and uneven sample crushing in traditional mechanical methods, the emergence of non-contact ultrasonic methods has better avoided the above problems. Non-contact ultrasonic methods can effectively improve experimental efficiency, save precious samples, reduce reagent costs, and obtain more accurate and reliable experimental data. There are two ways to implement non-contact ultrasound. One is that the ultrasonic probe contacts the sample tube and transmits ultrasound to the sample through the sample tube; the other is that the ultrasound is transmitted through the liquid and focuses the ultrasound on the sample. In existing non-contact ultrasonic devices, spring adjustment is required to control the contact pressure, and the structure is relatively complex. In particular, in order to realize multi-channel sample processing, multiple ultrasonic modules are required, which increases the cost of instrument production. In addition, it is difficult to balance and guarantee the uniformity and one-time processing of multiple samples. Summary of the invention

[0005] The purpose of the present invention is to provide a sample pretreatment device and a sample pretreatment method, aiming to solve the problems of complex structure, high manufacturing cost and complicated treatment process of existing treatment devices. The sample pretreatment device has a simple structure, low processing cost and simple treatment process. It can be applied to various sample pretreatments such as biological sample crushing, liquefaction of viscous samples, crushing of cells or cell walls, dissolution and dispersion of particles or powders or fragmentation of nucleic acids, so as to achieve comprehensive processing and crushing from tissues to cells, thereby effectively releasing various target molecules, ensuring the accuracy of information in subsequent analysis, and has a wide range of applications.

[0006] To achieve this object, the present invention adopts the following technical solutions:

[0007] A sample pre-processing device, comprising:

[0008] A sample positioning plate is provided with a plurality of evenly distributed receiving grooves, the receiving grooves are used to support test tubes, and the sample positioning plate is fixed to the external shell;

[0009] An ultrasonic module, comprising an ultrasonic transducer and an ultrasonic vibration disk connected to the ultrasonic transducer, wherein the ultrasonic transducer is used to generate ultrasonic vibration, the ultrasonic vibration disk is in active contact with the sample positioning disk, the ultrasonic vibration disk is provided with a plurality of limiting grooves, the plurality of limiting grooves and the plurality of containing grooves are arranged in a one-to-one correspondence, and the limiting grooves and the containing grooves are surrounded to form a storage space for clamping the test tube;

[0010] A driving member is transmission-connected to the ultrasonic module, and the driving member is configured to drive the ultrasonic module to move up and down so that the limiting groove and the containing groove clamp the test tube or release the test tube.

[0011] In some possible implementations, a guide hole is provided at the center of the sample positioning plate, the hole wall of the guide hole is recessed with the accommodating groove, the accommodating groove is inclined to support the test tube, and the ultrasonic vibration plate is movably disposed in the guide hole.

[0012] In some possible embodiments, the ultrasonic vibration disk includes a column and a conical structure that are connected to each other, the column is connected to the ultrasonic transducer, the small end of the conical structure is connected to the column, and the outer peripheral side of the large end of the conical structure is provided with the limiting groove, and the ultrasonic vibration disk can pass through the guide hole from top to bottom so that the limiting groove is pressed against the side wall of the test tube.

[0013] In some possible implementations, the limiting groove is configured as an arc-shaped groove.

[0014] In some possible implementations, the ultrasonic vibration disk is configured as a circular structure, a square structure, a trapezoidal structure, or a triangular structure.

[0015] In some possible implementations, the test tube is made of plastic, glass or metal.

[0016] In some possible implementations, the number of the limiting grooves is 8-40.

[0017] In some possible implementations, the accommodating groove includes an arc-shaped abutting groove and a hemispherical supporting groove that are interconnected, the bottom of the test tube fits with the hemispherical supporting groove, and the side wall of the test tube fits with the arc-shaped abutting groove;

[0018] And / or, a plurality of evenly distributed elastic members are arranged on the groove wall of the arc-shaped abutting groove.

[0019] The present invention also provides a sample pre-processing method, which is performed using a sample pre-processing device as described in any of the above schemes, and the sample pre-processing method comprises:

[0020] placing the test tube containing the sample to be processed in the holding tank;

[0021] adjusting the operating parameters of the ultrasonic transducer;

[0022] Turning on the driving member, adjusting the relative positions of the ultrasonic vibration disk and the sample positioning disk so that the test tube is clamped in the receiving space, and then the ultrasonic transducer performs ultrasonic treatment on the sample in the test tube;

[0023] After the ultrasonic treatment is completed, the test tube is removed.

[0024] In some possible implementations, the sample pre-treatment method satisfies at least one of the following conditions:

[0025] (1) The frequency of the ultrasonic transducer outputting ultrasonic waves is 20kHz-100kHz;

[0026] (2) The total working time of the ultrasonic transducer and the ultrasonic on time are both 1s-2000s;

[0027] (3) Ultrasonic off time 0s-1999s;

[0028] (4) The ultrasonic amplitude is 1%-100%;

[0029] (5) Duty cycle is 1%-100%;

[0030] (6) The pulse repetition frequency is 1Hz-1000Hz.

[0031] Beneficial effects of the present invention: When the sample pretreatment device provided by the present invention is used, the driving member drives the ultrasonic module to descend, so that the test tube is clamped in the storage space formed by the limiting groove and the receiving groove. The ultrasonic vibration generated by the ultrasonic transducer acts on the sample in the test tube via the ultrasonic vibration disk and the test tube to perform ultrasonic treatment on the sample. After the ultrasonic treatment is completed, the driving member drives the ultrasonic module to rise, and the limiting groove and the receiving groove release the test tube, so that the test tube can be taken away. Compared with the prior art, the structure is simplified and the processing and manufacturing cost is reduced. The sample pretreatment device can be applied to various sample pretreatments such as biological sample crushing, viscous sample liquefaction, cell or cell wall crushing, particle or powder dissolution and dispersion or nucleic acid fragmentation, so as to achieve comprehensive processing and crushing from tissue to cell, so that various target molecules can be effectively released, ensuring the accuracy of information in subsequent analysis. It has a wide range of applications, and the ultrasonic treatment process is simple and efficient.

[0032] The present invention also provides a sample pre-processing method, which is completed using the above-mentioned sample pre-processing device. The sample pre-processing method has a simple process and a short time, and effectively improves the efficiency of sample pre-processing. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 is a side view of a sample pre-treatment device according to an embodiment of the present invention;

[0034] Figure 2 yes Figure 1 Sectional view at AA in the middle;

[0035] Figure 3 is a top view of a sample pre-treatment device according to an embodiment of the present invention;

[0036] Figure 4 is a cross-sectional view of a sample pre-treatment device according to another embodiment of the present invention;

[0037] Figure 5 is a top view of a sample pre-treatment device according to another embodiment of the present invention;

[0038] Figure 6 It is a fluorescence quantitative qPCR test result diagram after the throat swab sample is processed by the sample pre-treatment device with 8 containing slots provided in an embodiment of the present invention (from left to right in each bar graph, the total working time of the device is 0s (control), 30s, 60s, 90s and 120s respectively);

[0039] Figure 7It is a comparison chart of the fluorescence quantitative qPCR detection results after the mixed solution of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-) is treated by the sample pretreatment device based on 8 containing tanks provided in an embodiment of the present invention in continuous and pulse working modes and without ultrasound (from left to right in each bar graph, the device processes in the following ways: none (control), pulse and continuous (time is 120s));

[0040] Figure 8 It is a sample pretreatment device based on 24 holding tanks and other treatment methods (grinding, heating and chemical lysis) provided in an embodiment of the present invention, and a comparison chart of the Ct value of the fluorescent quantitative qPCR detection results of the mixed solution of fungi and cocci after treatment compared with the control (original solution) (the treatment methods from left to right in each bar graph are ultrasound, grinding, thermal lysis and chemical lysis, respectively).

[0041] In the figure:

[0042] 100, sample positioning plate; 110, receiving groove; 200, test tube; 300, ultrasonic module; 310, ultrasonic transducer; 320, ultrasonic vibration plate; 321, column; 322, conical structure; 3221, limiting groove. DETAILED DESCRIPTION

[0043] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It is to be understood that the specific embodiments described herein are only used to explain the present invention, rather than to limit the present invention. It should also be noted that, for ease of description, only parts related to the present invention, rather than all structures, are shown in the accompanying drawings.

[0044] In the description of the present invention, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0045] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

[0046] In the description of this embodiment, the terms "upper", "lower", "right", etc., directions or positional relationships are based on the directions or positional relationships shown in the drawings, and are only for the convenience of description and simplification of operation, rather than indicating or implying that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are only used to distinguish in the description and have no special meaning.

[0047] The present embodiment provides a sample pretreatment device, which aims to solve the problems of complex structure, high manufacturing cost and complicated processing flow of existing processing devices. The sample pretreatment device has a simple structure and low processing cost, and can be applied to various sample pretreatments such as biological sample crushing, liquefaction of viscous samples, crushing of cells or cell walls, dissolution and dispersion of particles or powders, or fragmentation of nucleic acids, to achieve comprehensive processing and crushing from tissues to cells, thereby effectively releasing various target molecules, ensuring the accuracy of information in subsequent analysis, and has a wide range of applications.

[0048] like Figures 1 to 5 As shown, the sample pre-processing device includes a sample positioning plate 100, an ultrasonic module 300 and a driving member. The sample positioning plate 100 is provided with a plurality of evenly distributed receiving grooves 110, the receiving grooves 110 are used to support the test tube 200, and the sample positioning plate 100 is fixed to the external shell; the ultrasonic module 300 includes an ultrasonic transducer 310 and an ultrasonic vibration plate 320 connected to the ultrasonic transducer 310, the ultrasonic transducer 310 is used to generate ultrasonic vibration, the ultrasonic vibration plate 320 and the sample positioning plate 100 are in active contact, the ultrasonic vibration plate 320 is provided with a plurality of limiting grooves 3221, the plurality of limiting grooves 3221 and the plurality of receiving grooves 110 are arranged one by one, and the limiting grooves 3221 and the receiving grooves 110 can enclose and form a storage space for clamping the test tube 200; the driving member is transmission-connected to the ultrasonic module 300, and the driving member is configured to drive the ultrasonic module 300 to rise and fall, so that the limiting grooves 3221 and the receiving grooves 110 clamp the test tube 200 or release the test tube 200.

[0049] When the sample pretreatment device is used, the driving member drives the ultrasonic module 300 to descend, so that the test tube 200 is clamped in the storage space formed by the limiting groove 3221 and the receiving groove 110, and the ultrasonic vibration generated by the ultrasonic transducer 310 acts on the sample in the test tube 200 via the ultrasonic vibration disk 320 and the test tube 200 to perform ultrasonic treatment on the sample. After the ultrasonic treatment is completed, the driving member drives the ultrasonic module 300 to rise, and the limiting groove 3221 and the receiving groove 110 release the test tube 200, so that the test tube 200 can be taken away. Compared with the prior art, the structure is simplified, the processing and manufacturing cost is reduced, and the processing flow is simple. The sample pretreatment device can be applied to various sample pretreatments such as the crushing of biological samples, the liquefaction of viscous samples, the crushing of cells or cell walls, the dissolution and dispersion of particles or powders, or the fragmentation of nucleic acids, so as to achieve comprehensive processing and crushing from tissues to cells, so that various target molecules can be effectively released, and the accuracy of information in subsequent analysis is guaranteed. The application range is wide, and the ultrasonic treatment process is simple and efficient.

[0050] Optionally, a guide hole is provided at the center of the sample positioning plate 100, and the ultrasonic vibration plate 320 is movably arranged in the guide hole. The hole wall of the guide hole is recessed with a receiving groove 110, and the receiving groove 110 is inclined to support the test tube 200 and prevent the test tube 200 from falling. Preferably, the receiving groove 110 includes an arc-shaped abutment groove and a hemispherical support groove that are interconnected, the bottom of the test tube 200 is fitted with the hemispherical support groove, and the side wall of the test tube 200 is fitted with the arc-shaped abutment groove to ensure that the test tube 200 is supported stably and reliably. Specifically, the inclination angle and size of the arc-shaped abutment groove and the size of the hemispherical support groove can be set according to the size of the test tube 200 to ensure that the test tube 200 will not fall. In order to improve the stability of the test tube 200 during ultrasonic treatment, the groove wall of the arc-shaped abutment groove can be provided with a plurality of evenly distributed elastic members, such as springs or rubber pads, to slow down vibration.

[0051] In this embodiment, the ultrasonic vibration disk 320 includes a column 321 and a conical structure 322 connected to each other, the column 321 is connected to the ultrasonic transducer 310, the small end of the conical structure 322 is connected to the column 321, and the outer peripheral side of the large end of the conical structure 322 is provided with a limiting groove 3221, and the ultrasonic vibration disk 320 can pass through the guide hole from top to bottom, so that the limiting groove 3221 is closely fitted to the side wall of the test tube 200. By providing the conical structure 322, the conical structure 322 can better cooperate with the inclined receiving groove 110 to clamp the test tube 200. Preferably, the limiting groove 3221 is set as an arc groove to cooperate with the side wall of the test tube 200.

[0052] In this embodiment, the sample pre-processing device further includes an ultrasonic controller, which is electrically connected to the ultrasonic transducer 310 . The ultrasonic controller is used to control and adjust the working parameters of the ultrasonic transducer 310 .

[0053] In some other embodiments, the ultrasonic vibration disk 320 can be configured as a circular structure, a square structure (such as a square or a rectangle), a trapezoidal structure or a triangular structure, which can be configured as required.

[0054] Optionally, the test tube 200 is made of plastic, glass or metal. In actual implementation, the specific material of the test tube 200 can be set as needed.

[0055] Optionally, the number of the limiting slots 3221 is 8 to 40. In other embodiments, the number of the limiting slots 3221 can be 1, 2, 3, 4, 6, 8, 10, 12, 24, 32, 48, 96, etc., which can be set as needed.

[0056] This embodiment also provides a sample pre-processing method, which is performed using the above-mentioned sample pre-processing device. The sample pre-processing method includes:

[0057] S1, placing the test tube 200 containing the sample to be processed in the receiving tank 110;

[0058] S2, adjusting the working parameters of the ultrasonic transducer 310;

[0059] S3, turning on the driving member, adjusting the relative positions of the ultrasonic vibration plate 320 and the sample positioning plate 100, so that the test tube 200 is clamped in the storage space, and then the ultrasonic transducer 310 performs ultrasonic treatment on the sample in the test tube 200;

[0060] S4. After the ultrasonic treatment is completed, the test tube 200 is removed.

[0061] The sample pretreatment method is completed using the above-mentioned sample pretreatment device. The sample pretreatment method has a simple process and a short time, and effectively improves the efficiency of sample pretreatment.

[0062] Optionally, when the sample is ultrasonically treated using the sample pretreatment method, at least one of the following conditions is met:

[0063] (1) The frequency of ultrasonic wave output by ultrasonic transducer 310 is 20kHz-100kHz;

[0064] For example, the frequency of the ultrasonic wave may be 20 kHz, 28 kHz, 34 kHz, 38 kHz, 42 kHz or 60 kHz, 70 kHz, 80 kHz, 90 kHz or 100 kHz, and may be set as required.

[0065] (2) The total working time of the ultrasonic transducer 310 and the ultrasonic on time are both 1s-2000s;

[0066] (3) Ultrasonic off time 0s-1999s;

[0067] (4) The ultrasonic amplitude is 1%-100%;

[0068] (5) Duty cycle is 1%-100%;

[0069] (6) The pulse repetition frequency is 1Hz-1000Hz.

[0070] Hereinafter, the present invention will be further described based on specific examples.

[0071] Example 1

[0072] The pharyngeal swab is processed by the sample pre-processing device, wherein 8 receiving slots 110 are provided, and the specific operation process includes:

[0073] (1) Take throat swab samples provided by 4 volunteers and place them in a test tube with 200 μL of preservation solution. After standing for a certain period of time, take 100 μL of the solution from each volunteer, mark it, and store it for later use, and record it as a control;

[0074] (2) Place the test tube 200 on the sample positioning plate 100, and set the ultrasonic working parameters: total working time 30 seconds, ultrasonic on 10 seconds, off 5 seconds, ultrasonic amplitude 80%;

[0075] (3) The motor drives the ultrasonic module 300 downward, so that the limiting groove 3221 of the ultrasonic vibration disk 320 is in contact with the side wall of the test tube 200, and the operation starts;

[0076] (4) After the work is completed, the ultrasonic vibration plate 320 and the sample positioning plate 100 are automatically released, and 100 μL of solution is taken from each plate, marked and saved for later use, which is recorded as 30 seconds;

[0077] (5) Repeat the above steps (2) to (4) and record the total working time as 60s, 90s and 120s respectively;

[0078] (6) Using the control and 30s, 60s, 90s, and 120s solutions as templates, perform fluorescence quantitative qPCR experiments;

[0079] (7) After the qPCR experiment, record the Ct value of each amplification of each sample and plot it. Figure 6 shown.

[0080] Depend on Figure 6 It can be seen that the use of the above sample pre-treatment device has a significant treatment effect, and a good treatment effect is achieved in a very short time (within 60 seconds), which is very efficient.

[0081] Example 2

[0082] The sample pretreatment device is used to treat the mixed solution of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-) in different working modes, wherein 8 containing tanks 110 are provided, and the specific operation process includes:

[0083] (1) Prepare a mixed solution of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-), let it stand for a certain period of time, take out 100 μL of the solution, mark it, save it for later use, and record it as a control;

[0084] (2) Take 2 mL of each of the above solutions and place them in test tube 200A and test tube 200B respectively;

[0085] (3) Place the test tube 200A on the sample positioning plate 100 and set the ultrasonic working parameters: total working time 120 s, ultrasonic on 10 s, off 5 s, ultrasonic amplitude 100%;

[0086] (4) The motor drives the ultrasonic module 300 downward, so that the limiting groove 3221 of the ultrasonic vibration disk 320 is in contact with the side wall of the test tube 200, and the operation starts;

[0087] (5) After the work is completed, the ultrasonic vibration plate 320 and the sample positioning plate 100 are automatically released, and 100 μL of the solution is taken, marked and saved for later use, which is recorded as pulse ultrasound;

[0088] (6) Place the test tube 200B on the sample positioning plate 100, and set the ultrasonic working parameters: total working time 120 s, ultrasonic off 0 s, ultrasonic amplitude 100%, duty cycle 40%, pulse repetition frequency 40 Hz;

[0089] (7) Repeat the above steps (4) and (5), which is recorded as continuous ultrasound;

[0090] (8) Using the control, pulsed ultrasound and continuous ultrasound solutions as templates, a fluorescence quantitative qPCR experiment was performed;

[0091] (9) After the qPCR experiment, record the Ct value of each amplification of each sample and plot it. Figure 7 shown.

[0092] from Figure 7 It can be seen that the use of the above sample pretreatment device has a significant treatment effect, and the continuous ultrasound working mode is better than the pulse ultrasound.

[0093] Example 3

[0094] The sample pre-treatment device and other treatment methods (grinding, heating and chemical lysis) are used to treat the fungus and cocci mixed solution respectively, wherein 8 containing tanks 110 are provided, and the specific operation process includes:

[0095] (1) Prepare a mixed solution of fungi and cocci, let it stand for a certain period of time, take 100 μL of the solution, mark it, save it for later use, and record it as a control;

[0096] (2) Take 2 mL of the above solution and place it in test tubes 200A, B, C and D respectively;

[0097] (3) Place the test tube 200A on the sample positioning plate 100 and set the ultrasonic working parameters: total working time 120 s, continuous working mode, ultrasonic amplitude 100%;

[0098] (4) The motor drives the ultrasonic module 300 downward, so that the limiting groove 3221 of the ultrasonic vibration disk 320 is in contact with the side wall of the test tube 200, and the operation starts;

[0099] (5) After the work is completed, the ultrasonic vibration plate 320 and the sample positioning plate 100 are automatically released and taken away for storage;

[0100] (6) Test tubes 200B, C, and D are ground, heated, and chemically lysed in sequence according to the instructions. The samples in test tubes 200A, B, C, and D are transferred to a magnetic bead-based fully automatic nucleic acid extractor, and nucleic acid is extracted and purified according to the operating instructions. The purified nucleic acids are recorded as ultrasonic, grinding, thermal lysis, and chemical lysis, respectively;

[0101] (7) Using the control and the above solutions as templates, perform fluorescence quantitative qPCR experiments;

[0102] (8) After the qPCR experiment, record the Ct value of each sample and calculate the Ct value difference, such as Figure 8 shown.

[0103] from Figure 8 It can be seen that the pre-treatment effect of using the above sample pre-treatment device is the best.

[0104] Example 4

[0105] The sample pretreatment device is used to treat the mixed solution of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-), respectively. There are 24 holding tanks 110, and the specific operation process includes:

[0106] (1) Prepare a mixed solution of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-), let it stand for a certain period of time, and then take 2 mL of the solution and place it in test tubes 200#1, #2, #3, ..., #12 respectively;

[0107] (2) Place the test tubes 200#1-#12 symmetrically on the sample positioning plate 100, and set the ultrasonic working parameters: continuous working mode, total working time 120s, ultrasonic amplitude 100%;

[0108] (3) The motor drives the ultrasonic module 300 downward, so that the limiting groove 3221 of the ultrasonic vibration disk 320 is in contact with the side wall of the test tube 200, and the operation starts;

[0109] (4) After the work is completed, the ultrasonic vibration plate 320 and the sample positioning plate 100 are automatically released, and 100 μL of solution is taken from each plate, marked with numbers, and stored for later use;

[0110] (5) Conducting fluorescence quantitative qPCR experiments using the above solutions as templates;

[0111] (6) After the qPCR experiment, the Ct value of each amplification of each sample was recorded and tabulated as shown in Table 1.

[0112] It can be seen from Table 1 below that the use of the above sample pre-treatment device has good processing consistency.

[0113] Table 1 Treatment results of the 24-channel sample pretreatment device for mixed solutions of fungi, Gram-positive bacteria (G+) and Gram-negative bacteria (G-)

[0114]

[0115] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, various obvious changes, readjustments and substitutions can be made without departing from the protection scope of the present invention. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A sample pre-processing device, characterized in that: include: A sample positioning plate (100) is provided with a plurality of evenly distributed receiving grooves (110), wherein the receiving grooves (110) are used to support test tubes (200), and the sample positioning plate (100) is fixed to an external housing; An ultrasonic module (300) comprises an ultrasonic transducer (310) and an ultrasonic vibration disk (320) connected to the ultrasonic transducer (310), wherein the ultrasonic transducer (310) is used to generate ultrasonic vibrations, the ultrasonic vibration disk (320) is in active contact with the sample positioning disk (100), the ultrasonic vibration disk (320) is provided with a plurality of limiting grooves (3221), the plurality of limiting grooves (3221) and the plurality of containing grooves (110) are arranged in a one-to-one correspondence, and the limiting grooves (3221) and the containing grooves (110) can be combined to form a storage space for clamping the test tube (200); A driving member is drivingly connected to the ultrasonic module (300), and the driving member is configured to be able to drive the ultrasonic module (300) to rise and fall, so that the limiting groove (3221) and the containing groove (110) clamp the test tube (200) or release the test tube (200).

2. The sample pre-treatment device according to claim 1, characterized in that: A guide hole is provided at the center of the sample positioning plate (100), the hole wall of the guide hole is recessed with the receiving groove (110), the receiving groove (110) is inclined to support the test tube (200), and the ultrasonic vibration plate (320) is movably inserted into the guide hole.

3. The sample pre-processing device according to claim 2, characterized in that: The ultrasonic vibration disk (320) comprises a column (321) and a conical structure (322) which are connected to each other. The column (321) is connected to the ultrasonic transducer (310). The small end of the conical structure (322) is connected to the column (321). The outer peripheral side of the large end of the conical structure (322) is provided with the limiting groove (3221). The ultrasonic vibration disk (320) can pass through the guide hole from top to bottom, so that the limiting groove (3221) is pressed against the side wall of the test tube (200).

4. The sample pre-processing device according to claim 3, characterized in that: The limiting groove (3221) is configured as an arc-shaped groove.

5. The sample pre-processing device according to claim 1, characterized in that: The ultrasonic vibration disk (320) is configured as a circular structure, a square structure, a trapezoidal structure or a triangular structure.

6. The sample pre-treatment device according to any one of claims 1 to 5, characterized in that: The test tube (200) is made of plastic, glass or metal.

7. The sample pre-processing device according to any one of claims 1 to 5, characterized in that: The number of the limiting grooves (3221) is set to be 8-40.

8. The sample pre-treatment device according to any one of claims 1 to 5, characterized in that: The containing groove (110) comprises an arc-shaped abutting groove and a hemispherical supporting groove which are connected to each other, the bottom of the test tube (200) is fitted with the hemispherical supporting groove, and the side wall of the test tube (200) is fitted with the arc-shaped abutting groove; And / or, a plurality of evenly distributed elastic members are arranged on the groove wall of the arc-shaped abutting groove.

9. A sample pre-treatment method, characterized in that: The sample pre-processing method is performed using the sample pre-processing device according to any one of claims 1 to 8, wherein the sample pre-processing method comprises: Placing the test tube (200) containing the sample to be processed in the containing tank (110); adjusting the operating parameters of the ultrasonic transducer (310); The driving member is turned on, and the relative positions of the ultrasonic vibration disk (320) and the sample positioning disk (100) are adjusted so that the test tube (200) is clamped in the storage space, and then the ultrasonic transducer (310) performs ultrasonic treatment on the sample in the test tube (200); After the ultrasonic treatment is completed, the test tube (200) is removed.

10. The sample pre-treatment method according to claim 9, characterized in that: Satisfy at least one of the following conditions: (1) The frequency of the ultrasonic wave output by the ultrasonic transducer (310) is 20kHz-100kHz; (2) The total working time of the ultrasonic transducer (310) and the ultrasonic on time are both 1s-2000s; (3) Ultrasonic off time 0s-1999s; (4) The ultrasonic amplitude is 1%-100%; (5) Duty cycle is 1%-100%; (6) The pulse repetition frequency is 1Hz-1000Hz.

Images