Microorganism extraction system

Abstract

The utility model belongs to the field of microorganism extraction equipment, and particularly relates to a microorganism extraction system. Comprising a microorganism extraction container, the microorganism extraction container contains a to-be-extracted solution and magnetic beads for adsorbing microorganisms, and the to-be-extracted solution contains the microorganisms to be extracted; the workbench is provided with a container placing part for placing the microorganism extraction container; the magnetic bead adsorption mechanism comprises a magnetic attraction assembly and a magnetic attraction state switching assembly used for changing the magnetic attraction state, and the magnetic attraction assembly has magnetism and is used for adsorbing magnetic beads located in the microorganism extraction container; and the controller is electrically connected with the magnetic attraction state switching assembly and is used for changing the state of the magnetic attraction assembly or changing the relative state of the magnetic attraction assembly and the microorganism extraction container. The operation is simple and convenient, the automatic operation of microorganism extraction is favorably realized, the difficulty of microorganism extraction is reduced, and the efficiency of microorganism extraction is improved.

Description

Technical Field

[0001] The utility model belongs to the field of microbial extraction equipment, in particular to a microbial extraction system. Background Art

[0002] Bloodstream infection (BSI) is a general term for sepsis and bacteremia. Septicemia is caused by the invasion of the bloodstream by various pathogenic microorganisms (bacteria or fungi) and toxins. Its primary clinical manifestations include a series of severe symptoms, including sudden chills, high fever, tachycardia, rapid breathing, rash, hepatosplenomegaly, and altered mental status. Severe cases can lead to shock, disseminated intravascular coagulation (DIC), and multiple organ failure. Bacteremia is defined as short-lived bacterial invasion of the bloodstream without clinically apparent toxic symptoms (e.g., vascular-related infection). In recent years, with the widespread development of invasive diagnostic and treatment techniques and the widespread use of broad-spectrum antibiotics and hormones, the incidence of BSI has been increasing annually. BSIs carry a high mortality rate, prolong hospitalizations, and increase hospital costs, resulting in significant complications. Consequently, the control of BSIs is receiving increasing attention.

[0003] Currently, blood culture is the gold standard for diagnosing bloodstream infections. It involves inoculating a freshly isolated blood sample onto a nutrient medium and, under certain conditions of temperature and humidity, allowing the growth and reproduction of nutrient-demanding bacteria to be identified and subsequently confirmed, thereby identifying the pathogen. It is commonly used for the etiological diagnosis of bacteremia, fungemia, sepsis, and septicemia.

[0004] The culture medium is usually placed in a culture bottle, and an automated blood culture instrument is used to enrich the culture bottle until it tests positive, which indicates a bloodstream infection.

[0005] In order to analyze the types of pathogens in the positive samples, the pathogens need to be extracted for identification.

[0006] Pathogens (microorganisms) are typically purified by streaking. This involves streaking a sample onto a culture medium, where the microorganisms will grow as colonies, allowing for the isolation of a single species. However, this method requires 24-48 hours of incubation. Considering the commute time of the examiner, the incubation period often takes up to three days. However, for patients with bloodstream infections, expedited identification is crucial.

[0007] There are also methods for rapidly extracting microorganisms through repeated centrifugation. However, these methods cannot achieve the same accuracy as streaking, and the multiple centrifugation and precipitation method is very complex and requires high precision, so it is not widely used. Furthermore, this method of extracting microorganisms is generally performed in a test tube, which has low extraction efficiency. There is a lack of a microbial extraction system to improve microbial extraction efficiency. Utility Model Content

[0008] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a microorganism extraction system for solving the problems of difficulty in microorganism extraction and low extraction efficiency in the prior art.

[0009] To achieve the above-mentioned and other related purposes, the present invention provides a microbial extraction system, comprising:

[0010] a microorganism extraction container configured to contain a liquid to be extracted and magnetic beads for adsorbing microorganisms, wherein the liquid to be extracted contains the microorganisms to be extracted;

[0011] A workbench is provided with a container placement portion, wherein the container placement portion is used to place the microorganism extraction container;

[0012] A magnetic bead adsorption mechanism is installed on the workbench, and the magnetic bead adsorption mechanism includes a magnetic attraction component and a magnetic attraction state switching component for changing the magnetic attraction state. The magnetic attraction component has magnetism and is used to adsorb the magnetic beads located in the microorganism extraction container;

[0013] A controller is electrically connected to the magnetic state switching component and is used to change the state of the magnetic component or change the relative state between the magnetic component and the microorganism extraction container.

[0014] It should be noted that the extracted microorganisms mentioned in this application refer to the extraction of fungi, bacteria or viruses, rather than the nucleic acids or other specific tissues therein.

[0015] The microbial extract can be a processed product of a positive blood culture sample, a processed product of a urine sample, or a processed product of an industrial sample requiring microbial testing. The processed product refers to a mixed solution obtained by pre-treating the sample, such as a mixed solution obtained by adding a buffer, a mixed solution obtained by centrifugation, or a mixed solution obtained by lysing blood cells.

[0016] Changing the state of the magnetic component mentioned here refers to changing the size or presence of magnetism, and changing the relative state of the magnetic component and the microorganism extraction container refers to changing the influence of the magnetism of the magnetic component on the magnetic beads in the microorganism extraction container, such as changing the relative position of the two, or adding a structure that can isolate magnetism between the two, as long as it can change the influence of magnetism on the magnetic beads.

[0017] The microorganism extraction container has multiple hole positions, and the magnetic bead adsorption mechanism is a transfer mechanism for adsorbing and transferring the magnetic beads between the hole positions; the transfer mechanism includes a magnetic attraction component and a magnetic attraction state switching component, and the magnetic attraction state switching component includes a driving component, and the driving component is used to drive the magnetic attraction component to move relative to the microorganism extraction container or to change the magnetism of the magnetic attraction component.

[0018] By using a drive assembly to drive the relative movement of the magnetic assembly and the microorganism extraction container, magnetic beads can be transferred between the various wells of the microorganism extraction container. Of course, the drive assembly can drive the movement of both the magnetic assembly and the microorganism extraction container, or both the magnetic assembly and the microorganism extraction container. In applications where the drive assembly changes the magnetism of the magnetic assembly, the magnetic assembly includes an electromagnet, and the drive assembly includes an electromagnet controller. The electromagnet controller controls the switching of the electromagnet between a magnetic state in which it has magnetism and a released state in which it loses magnetism. In addition to the electromagnet controller, the drive assembly can also include an electromagnet driver capable of driving the electromagnet to move. The electromagnet driver can also drive the electromagnet to move, thereby achieving magnetic bead transfer.

[0019] Optionally, the microorganism extraction container is a 96-well plate having 96 wells, a 48-well plate having 48 wells, or a 32-well plate having 32 wells.

[0020] Optionally, the microorganism extraction container has at least three wells, and magnetic beads, a washing solution, and an eluent are pre-embedded in at least three of the wells, respectively.

[0021] Optionally, the magnetic attraction assembly includes a magnetic rod and a magnetic rod sleeve, the magnetic rod sleeve is used to extend into the microorganism extraction container, the magnetic rod is built into the magnetic rod sleeve, and can move relative to the magnetic rod sleeve to adsorb and release magnetic beads located outside the magnetic rod sleeve with the magnetic rod sleeve.

[0022] Specifically, when it is necessary to adsorb magnetic beads, place the magnetic rod in the magnetic rod sleeve and extend it into the microorganism extraction container. The magnetic beads will be adsorbed to the outer wall of the magnetic rod sleeve. The simultaneous movement of the magnetic rod and the magnetic rod sleeve can drive the magnetic beads to move together. When it is necessary to release the magnetic beads, place the magnetic beads in the microorganism extraction container, and the magnetic rod moves and separates from the magnetic rod sleeve, and the magnetic beads will naturally separate from the magnetic rod sleeve. Another advantage of this form is that the magnetic rod can move alone in the microorganism extraction container to achieve mixing of the liquid to be extracted. It is understandable that this is only one form of implementation of the magnetic attraction component. Other forms of magnetic attraction components can also achieve magnetic bead transfer. For example, only a magnetic rod is used without a magnetic rod sleeve. The magnetic rod uses an electromagnet, which can also achieve the adsorption and release of magnetic beads by controlling the presence or absence of magnetism of the magnetic rod.

[0023] Optionally, the magnetic rod sleeve is installed on the workbench or placed on the microorganism extraction container.

[0024] The magnetic rod cover is placed on the microbial extraction container, so that the magnetic rod cover can be flexibly replaced. The magnetic rod cover can be replaced every time an extraction is performed, which can avoid contamination and eliminate the need for a cleaning step.

[0025] The driving assembly includes a first driving member, the output end of which is connected to the magnetic rod through a first movable bracket to drive the magnetic rod to move up and down;

[0026] The driving assembly further includes a second driving member, an output end of which is connected to the magnetic rod sleeve via a second movable bracket, and the first driving member is mounted on the second movable bracket, and the second driving member drives the first driving member and the second movable bracket to move synchronously up and down through the second movable bracket;

[0027] The top end of the magnetic rod sleeve is open, the first movable bracket is located above the second movable bracket, and the first driving member drives the magnetic rod to move up and down through the first movable bracket to extend into and out of the magnetic rod sleeve;

[0028] The driving assembly further includes a third driving member, which is connected to the second driving member to drive the second driving member and the second movable bracket to move in a horizontal direction.

[0029] Optionally, a rear limit block and a side limit block are provided on the workbench, and the rear limit block, the side limit block and the container placement portion cooperate to define a plurality of container placement positions distributed along the horizontal movement direction of the second movable bracket.

[0030] Optionally, there are multiple magnetic bars, and the multiple magnetic bars are arranged in multiple rows along the arrangement direction of the multiple container placement positions; the magnetic bar sleeves are arranged in a one-to-one correspondence with the magnetic bars.

[0031] Optionally, it further includes a liquid replacement mechanism for absorbing and discharging liquid to replace the liquid in the microorganism extraction container. The magnetic bead adsorption mechanism is located outside the microorganism extraction container and is used to adsorb the magnetic beads on the microorganism extraction container.

[0032] Optionally, the liquid exchange mechanism is a pipette needle, a pipette line or multiple pipette lines.

[0033] The liquid exchange mechanism can be a pipette needle or a pipette line, etc., which can automatically complete the liquid exchange. In the figure, a pipette line is used as an example. After the magnetic component absorbs the mixture of magnetic beads and microorganisms, the liquid is sucked away through the pipette line, leaving the mixture of magnetic beads and microorganisms. Then, washing liquid is added through the pipette line. Similarly, the washing liquid can be replaced with a new one or an elution liquid. Similarly, this function can also be achieved by using a pipette needle or other liquid exchange mechanism.

[0034] Optionally, the magnetic bead adsorption mechanism is an electromagnet, and the magnetic state switching component is an electromagnet controller.

[0035] The magnetism of the electromagnet is controlled by the electromagnet controller, so that the adsorption or release of the magnetic beads can be controlled.

[0036] Optionally, the magnetic state switching component includes a fifth driving member for driving the microorganism extraction container and / or a sixth driving member for driving the magnetic component to move. The magnetic state switching component changes the magnetic state by changing the relative position of the microorganism extraction container and the magnetic component.

[0037] Optionally, the fifth driving member is a linear motion mechanism or a rotational motion mechanism.

[0038] Optionally, the particle size of the magnetic beads is not greater than 3 μm; preferably, the particle size of the magnetic beads is 5 nm-1 μm.

[0039] Optionally, the magnetic beads are positively charged. Positively charged magnetic beads refer to the surface of the magnetic beads being positively charged, for example, the surface of the magnetic beads has positively charged groups, such as nitrogen-containing groups.

[0040] As described above, the microorganism extraction system of the present invention has at least the following beneficial effects: the container placement part and the magnetic bead adsorption mechanism are integrated on the workbench, and are electrically connected to the magnetic state switching component through the controller to change the state of the magnetic component or change the relative state of the magnetic component and the microorganism extraction container. The operation is simple and convenient, which is conducive to the realization of automated operation of microorganism extraction, reduces the difficulty of microorganism extraction and improves the efficiency of microorganism extraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1This is a schematic structural diagram of an embodiment of the microbial extraction system of the present invention;

[0042] Figure 2 for Figure 1 Schematic diagram of the internal structure of the microbial extraction system;

[0043] Figure 3 This is a schematic diagram of the operation of another embodiment of the microorganism extraction system of the present invention when the magnetic attraction component is an electromagnet;

[0044] Figure 4 This is a schematic diagram of the operation of another embodiment of the microorganism extraction system of the present invention when the magnetic state switching component is a linear motion mechanism;

[0045] Figure 5 This is a schematic diagram of the operation of another embodiment of the microorganism extraction system of the present invention when the magnetic state switching component is a rotary motion mechanism;

[0046] Figure 6 This is a simplified structural diagram of a microorganism extraction container of an embodiment of the microorganism extraction system of the present invention.

[0047] Part Number Description

[0048] Housing 1, workbench 2, rear limit block 21, side limit block 22, container placement position 23, magnetic suction assembly 3, magnetic rod 31, magnetic rod sleeve 32, first driving member 41, first movable bracket 42, first fixed bracket 43, second driving member 44, second movable bracket 45, second fixed bracket 46, third driving member 47, third fixed bracket 48, microorganism extraction container 5, washing liquid pipeline 6, fifth driving member 71, sixth driving member 72. DETAILED DESCRIPTION

[0049] The following describes the implementation of the present invention through specific embodiments. People familiar with this technology can easily understand other advantages and effects of the present invention from the content disclosed in this specification.

[0050] It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for understanding and reading by those familiar with this technology, and are not used to limit the conditions for implementation of the present invention. Therefore, they have no substantial technical significance. Any modification of the structure, change in the proportional relationship, or adjustment of the size should still fall within the scope of the technical content disclosed in the present invention without affecting the efficacy and purpose of the present invention. At the same time, terms such as "upper", "lower", "left", "right", "middle" and "one" quoted in this specification are only for the convenience of description and are not used to limit the scope of implementation of the present invention. Changes or adjustments in their relative relationships should also be considered as the scope of implementation of the present invention without substantially changing the technical content.

[0051] In order to solve the problems of traditional microbial extraction methods being too time-consuming, or manual extraction steps being too complicated and having high operational bottlenecks, this patent application provides a microbial extraction system for extracting microorganisms, comprising:

[0052] A microorganism extraction container, configured to contain a liquid to be extracted and magnetic beads for adsorbing microorganisms, wherein the liquid to be extracted contains the microorganisms to be extracted;

[0053] A workbench is provided with a container placement portion for placing a microorganism extraction container;

[0054] A magnetic bead adsorption mechanism is installed on the workbench and can be used to adsorb the magnetic beads. The magnetic bead adsorption mechanism includes a magnetic attraction component and a magnetic attraction state switching component for changing the magnetic attraction state. The magnetic attraction component has magnetism and is used to adsorb the magnetic beads located in the microorganism extraction container.

[0055] The controller is electrically connected to the magnetic state switching component and is used to change the state of the magnetic component or the relative state between the magnetic component and the microorganism extraction container.

[0056] The system works by placing a microbial extraction container containing a liquid containing microorganisms to be extracted and magnetic beads on a workbench. After the microorganisms in the liquid are adsorbed by the magnetic beads, a magnetic state switching component controls the magnetic component to adsorb the mixture of magnetic beads and microorganisms. The magnetic state switching component also controls the magnetic component to release the magnetic beads, thereby washing and eluting the mixture and obtaining a sample suitable for microbial identification.

[0057] The adsorption principle of magnetic beads is to use the specific adsorption of special magnetic beads or to use charged magnetic beads to adsorb bacteria.

[0058] Positively charged magnetic beads refer to beads with positive charges on their surface, for example, beads with positively charged groups on their surface or beads with positive charges by other means, such as nitrogen-containing groups. This embodiment uses amino magnetic beads and imidazole groups as examples.

[0059] This patent application primarily studies the use of charged magnetic beads to adsorb bacteria. While charged magnetic beads have a certain degree of universality and are more suitable for the application scenarios described in this patent application, specialized magnetic beads can also be used for specific adsorption to achieve targeted microbial extraction.

[0060] It should be noted that the extracted microorganisms mentioned in this application refer to the extraction of fungi, bacteria or viruses, rather than the nucleic acids or other specific tissues therein.

[0061] The microbial extract is a processed product of a positive blood culture sample, a processed product of a urine sample, or a processed product of an industrial sample requiring microbial testing. The processed product refers to a mixed solution obtained by pre-processing the sample, such as a mixed solution obtained by adding a buffer, a mixed solution obtained after centrifugation, or a mixed solution obtained by lysing blood cells.

[0062] Changing the state of the magnetic component mentioned here refers to changing the size or presence of magnetism, and changing the relative state of the magnetic component and the microorganism extraction container refers to changing the influence of the magnetism of the magnetic component on the magnetic beads in the microorganism extraction container, such as changing the relative position of the two, or adding a structure that can isolate magnetism between the two, as long as it can change the influence of magnetism on the magnetic beads.

[0063] In one optional embodiment, the magnetic beads have a particle size of no greater than 3 μm. Furthermore, the particle size can be between 5 nm and 1 μm. The particle size of the magnetic beads affects their ability to adsorb microorganisms. Using a particle size within an appropriate range can improve the adsorption efficiency of the magnetic beads.

[0064] In the above process, the microorganism extraction container can be of different types, and the magnetic attraction component and the magnetic attraction state switching component can also be of different structures, which will be described below through different embodiments.

[0065] Example 1

[0066] See also Figure 1 and Figure 2In this embodiment, a microorganism extraction system for extracting microorganisms includes a microorganism extraction container containing a liquid to be extracted and magnetic beads for adsorbing microorganisms; a workbench 2, a magnetic bead adsorption mechanism, and a controller; and a magnetic bead adsorption mechanism for selecting and transferring the magnetic bead. In addition to the aforementioned components, the microorganism extraction system may also include a housing 1. The workbench 2 and the transfer mechanism may be located inside the housing 1 to prevent external factors from affecting the working environment. The controller may be located inside or outside the housing 1. The workbench 2 is provided with a container placement portion for placing the microorganism extraction container. The microorganism extraction container has multiple wells. The transfer mechanism is mounted on the workbench 2 and is used to adsorb and transfer magnetic beads between the wells. The transfer mechanism includes a magnetic attraction component and a drive component (i.e., one embodiment of a magnetic state switching component). The magnetic attraction component is used to adsorb magnetic beads within the microorganism extraction container. The drive component is connected to the magnetic attraction component and is adapted to drive the magnetic attraction component and the microorganism extraction container to move relative to each other to adsorb and release the magnetic beads. The controller is electrically connected to the drive component to control the operation of the drive component to drive the magnetic attraction component to move relative to the microorganism extraction container.

[0067] Optionally, the magnetic assembly 3 includes a magnetic rod 31 and a magnetic rod sleeve 32. The magnetic rod sleeve 32 is used to extend into the microorganism extraction container. The magnetic rod 31 is built into the magnetic rod sleeve 32 and can move relative to the magnetic rod sleeve 32 to absorb and release magnetic beads located outside the magnetic rod sleeve 32. When the magnetic rod 31 is extended into the magnetic rod sleeve 32, the magnetic beads located outside the magnetic rod sleeve 32 are absorbed by the magnetic rod sleeve 32 under the action of the magnetic attraction of the magnetic rod 31; when the magnetic rod 31 is withdrawn from the magnetic rod sleeve 32, the magnetic attraction in the magnetic rod sleeve 32 disappears, and the magnetic beads absorbed on the outside of the magnetic rod sleeve 32 are released and separated from the magnetic rod sleeve 32.

[0068] Optionally, the magnetic rod cover 32 is installed on the workbench 2 to facilitate the transfer of the magnetic rod cover 32 and the magnetic rod 31 together. Especially when magnetic beads need to be transferred, the magnetic rod cover 32 installed on the workbench 2 is conducive to moving together with the magnetic rod 31.

[0069] Optionally, the magnetic rod sleeve 32 is placed on the microorganism extraction container, eliminating the need for additional support components inside the housing 1 to provide mounting support for the magnetic rod sleeve 32, which helps simplify the structure, especially when there is no need to transfer magnetic beads. At the same time, a disposable magnetic rod sleeve is provided for use with a disposable microorganism extraction container. When the microorganism extraction container is mounted on the container placement portion of the workbench, the magnetic rod sleeve is simply placed into the microorganism extraction container. The magnetic rod sleeve is placed on the microorganism extraction container, allowing the magnetic rod sleeve to be flexibly replaced. The magnetic rod sleeve can be replaced for each extraction, thus avoiding contamination and eliminating the need for a cleaning step.

[0070] Optionally, the microorganism extraction container is a container with multiple wells, and the multiple wells can respectively perform different process operations, wherein the processes include lysis / adsorption, washing and elution.

[0071] Optionally, the microorganism extraction container is pre-embedded with magnetic beads, washing solution and eluent in different wells during production (before use). Optionally, the microorganism extraction container has at least three wells, in which magnetic beads, washing solution and eluent are pre-embedded respectively.

[0072] See also Figure 6 The microbial extraction container can be a 96-well plate, with 96 wells arranged in 12 rows, with 8 wells in each row. It is understood that the microbial extraction container is not limited to 96-well plates and can also be a 48-well plate or a 32-well plate. The following uses a 96-well plate as an example for specific description. Of course, other microbial extraction containers can also be used as long as they can meet the transfer requirements of the magnetic beads and microbial mixture.

[0073] Continue to see Figure 6 The eight positions AH can target 8 or 16 samples at the same time. Position 1 is used to add samples, position 2 is pre-buried with magnetic beads, positions 3 / 4 are pre-buried with washing (cleaning) solution, and position 6 is pre-buried with elution solution.

[0074] Add the pre-treated blood culture strip extract to position 1, and then start the microbial extraction system to start working:

[0075] The magnetic rod 31 moves to position 2 to adsorb the magnetic beads and transfer them to position 1. After the microorganisms are adsorbed at position 1, the mixture of magnetic beads and microorganisms is transferred to the washing solution under the magnetic attraction of the magnetic rod 31. After washing is completed, the mixture is transferred to the elution solution through the magnetic rod 31 to elute the microbial components into the liquid. Generally, formic acid and acetonitrile are added to destroy the microorganisms. After the elution is completed, the magnetic beads are removed with the magnetic rod 31, leaving a solution for identification.

[0076] In the microorganism extraction system of the above embodiment, the controller controls the operation of the driving component to control the operating state of the magnetic attraction component, thereby adjusting the operating time of each process to improve the quality and efficiency of microorganism extraction.

[0077] See also Figure 1 and Figure 2 (The microorganism extraction container is not shown), in some optional embodiments, the driving assembly includes a first driving member 41, and the output end of the first driving member 41 is connected to the magnetic rod 31 through a first movable bracket 42 to drive the magnetic rod 31 to move up and down.

[0078] Optionally, the drive assembly further includes a second drive member 44, the output end of which is connected to the magnetic rod sleeve 32 via a second movable bracket 45. The first drive member 41 is mounted on the second movable bracket 45, and the second drive member 44 drives the first drive member 41 and the second movable bracket 45 to move synchronously up and down. Furthermore, the first drive member 41 is connected to the output end of the second drive member 44 via a first fixed bracket 43, and the second movable bracket 45 is also connected to the output end of the second drive member 44 via the first fixed bracket 43. This structural design enables the first movable bracket 42 to move independently relative to the second movable bracket 45. In other words, the magnetic rod 31 can move up and down relative to the magnetic rod sleeve 32, while the second drive member 44 can drive the first movable bracket 42 and the second movable bracket 45 to move synchronously up and down via the first fixed bracket 43. In other words, the second drive member 44 can drive the magnetic rod 31 and the magnetic rod sleeve 32 to move simultaneously. This embodiment is only one type of drive configuration. It is foreseeable that any method that can drive the magnetic rod 31 and the magnetic rod sleeve 32 to move independently can achieve the objectives of this patent application. Of course, the driving structure of the magnetic rod sleeve may not be provided, that is, the adsorption is completed by directly using the magnetic rod to contact the magnetic beads, but it is necessary to clean the magnetic rod after one operation.

[0079] Optionally, the top of the magnetic rod sleeve 32 is open, while the bottom of the magnetic rod sleeve 32 is closed. The first movable bracket 42 is positioned above the second movable bracket 45. The first driving member 41, through the first movable bracket 42, drives the magnetic rod 31 to move upward and downward, extending into and out of the magnetic rod sleeve 32. When the magnetic rod 31 is inserted into the magnetic rod sleeve 32, the magnetic attraction of the magnetic rod 31 causes the microbial magnetic beads to be attracted to the outer wall of the magnetic rod sleeve 32. The combined movement of the magnetic rod 31 and the magnetic rod sleeve 32 enables the transfer of the magnetic bead and microorganism mixture. When the magnetic rod 31 exits the magnetic rod sleeve 32, the magnetic attraction of the magnetic rod 31 disappears, the magnetic beads are separated from the magnetic rod sleeve 32, and the magnetic rod sleeve can also be removed from the microorganism extraction container. The upward and downward movement of the magnetic rod sleeve 32 enables the magnetic beads to be agitated. Agitation can be achieved either with the magnetic beads separated from the magnetic rod sleeve or with the magnetic beads attached to the magnetic rod sleeve.

[0080] Optionally, the drive assembly further includes a third drive member 47, which is connected to the second drive member 44 to drive the second drive member 44 and the second movable bracket 45 to move in the horizontal direction. The third drive member 47 is mounted on the workbench 2 via a third fixed bracket 48. Furthermore, the output end of the third drive member 47 is connected to the second drive member 44 via a second fixed bracket 46. Alternatively, the second drive member 44 is mounted on the second fixed bracket 46, and the second fixed bracket 46 is connected to the output end of the third drive member 47, so that the operation of the third drive member 47 can drive the first movable bracket 42 and the second movable bracket 45 to move synchronously, thereby achieving synchronous movement of the magnetic bar 31 and the magnetic bar sleeve 32.

[0081] Optionally, the first driving member 41 , the second driving member 44 and the third driving member 47 may be motors, cylinders, hydraulic cylinders or other driving members capable of providing power.

[0082] The microbial extraction system of the above embodiment has a simple structure and a compact layout. The first driving member 41, the second driving member 44 and the third driving member 47 cooperate with each other to realize the independent lifting and lowering of the magnetic rod 31, and the lifting and moving of the magnetic rod 31 and the magnetic rod sleeve 32 together, so as to adapt to the operation requirements of different processes.

[0083] See also Figure 1 and Figure 2 In some optional embodiments, the workbench 2 is provided with a rear limit block 21 and a side limit block 22. The rear limit block 21, the side limit block 22, and the container placement portion cooperate to define a plurality of container placement positions 23 distributed along the horizontal movement direction of the second movable bracket 45. The microorganism extraction container is placed on the container placement portion, and the rear limit block 21 and the side limit block 22 cooperate to achieve positioning of the microorganism extraction container, simplifying positioning and reducing the difficulty of the microorganism extraction operation.

[0084] Optionally, there are multiple magnetic rods 31, and the multiple magnetic rods 31 are arranged in multiple rows along the arrangement direction of the multiple container placement positions 23. In the application of a 96-well plate, 8 magnetic rods 31 are arranged in one row, corresponding to the eight well positions of AH respectively. Two rows of magnetic rods are set. When the first row of magnetic rods corresponds to position 1, the other row corresponds to position 7. In this way, extraction of 16 positions can be achieved simultaneously. The number of magnetic rod sleeves 32 is the same as that of magnetic rods 31 and they are arranged in a one-to-one correspondence. Multiple magnetic rod sleeves 32 and magnetic rods 31 can be used in conjunction with each other to simultaneously achieve the extraction operation of multiple microorganisms, which is beneficial to improving the efficiency of microbial extraction.

[0085] Specifically, in an optional embodiment, the microbial extraction process includes magnetic bead transfer, binding, wash 1, wash 2, elution, and termination. Six columns or rows of wells are selected to form a well group, with the six wells in the same well group being sequentially numbered 1, 2, 3, 4, 5, and 6. Multiple well groups can be used to perform microbial extraction simultaneously, which improves operational efficiency. The specific operations of each process are detailed in Table 1 below.

[0086] Table 1

[0087]

[0088] That is, first add the pre-treated positive sample to position 1, use the magnetic rod and magnetic rod cover to adsorb the magnetic beads in position 2 for 10 seconds, then transfer the magnetic beads to position 1 and mix for 0.5 minutes to allow the magnetic beads and microorganisms to fully combine; then adsorb the magnetic beads in position 2 for 10 seconds, transfer the magnetic beads to position 3 and mix for 0.3 minutes to complete the first wash; then adsorb the magnetic beads in position 3 for 10 seconds, transfer the magnetic beads to position 4 and mix for 0.3 minutes to complete the second wash; then adsorb the magnetic beads in position 4 for 10 seconds, transfer the magnetic beads to position 6 and mix for 0.5 minutes to complete the system; finally, adsorb the magnetic beads and transfer them to position 2. At this point, the solution in position 6 is the extracted solution for identification.

[0089] The mixing time refers to the time it takes for the magnetic rod sleeve 32 to vibrate up and down while inserted into the liquid in the well. The magnetic rod sleeve can vibrate up and down while still in the sleeve. Alternatively, the controller can control the first drive member 41 to rise and exit the magnetic rod sleeve 32, and then control the second drive member 44 to drive the magnetic rod sleeve 32 up and down, allowing the non-magnetic magnetic rod sleeve 32 to move up and down and vibrate the liquid in the well. The magnetic attraction time refers to the time it takes for the magnetic beads to be attracted to the magnetic rod sleeve and transferred to the target location.

[0090] Vibration allows the magnetic beads to be fully and evenly mixed with the liquid in the corresponding wells, ensuring the quality and efficiency of microbial extraction.

[0091] During this process, the controller controls the mixing time to less than 1 minute, achieving a thorough mixing effect, which facilitates rapid extraction of microorganisms and maintains their activity without compromising the adsorption effect. The magnetic attraction time only needs to be sufficient to complete the adsorption and transfer process of the magnetic beads. This embodiment only illustrates one way to set the magnetic attraction time. It can be seen that with the above settings, the extraction process can be completed in just 3 minutes.

[0092] In the microbial extraction system of this embodiment, the controller controls the operation of the driving component to adjust the operating state of the magnetic attraction component, thereby achieving adsorption, release and stirring of magnetic beads to adapt to the operational requirements of different processes in the microbial extraction operation, reducing the operational difficulty and facilitating improved quality and efficiency of the microbial extraction operation.

[0093] Example 2

[0094] See also Figure 3 、 Figure 4 and Figure 5 In this embodiment, a microbial extraction system includes:

[0095] A microorganism extraction container 5, wherein the microorganism extraction container contains a liquid to be extracted and magnetic beads for adsorbing microorganisms, and the liquid to be extracted contains the microorganisms to be extracted;

[0096] A workbench (not shown), on which a container placement portion is provided for placing a microorganism extraction container;

[0097] A magnetic bead adsorption mechanism is installed on the workbench and is used to adsorb magnetic beads;

[0098] The magnetic bead adsorption mechanism includes a magnetic attraction component and a magnetic attraction state switching component for changing the magnetic attraction state. The magnetic attraction component has magnetism and is used to adsorb magnetic beads located in the microorganism extraction container.

[0099] The controller is electrically connected to the magnetic state switching component and is used to change the state of the magnetic component or the relative state between the magnetic component and the microorganism extraction container.

[0100] The microbial extraction system also includes a liquid replacement mechanism, which is used to absorb and discharge liquid to replace the liquid in the microbial extraction container. The liquid replacement mechanism can be a liquid suction needle, a liquid suction pipeline or multiple liquid suction pipelines, and any structure that can automatically complete the liquid replacement can be used. Figure 3 Taking a liquid pipette as an example, after the magnetic component 3 absorbs the mixture of magnetic beads and microorganisms, the liquid is aspirated through the liquid pipette 6, leaving the mixture of magnetic beads and microorganisms. Washing liquid is then added through the liquid pipette 6. Similarly, the washing liquid can be replaced with a new one or an eluent. Similarly, this function can also be achieved using a liquid pipette needle or other liquid replacement mechanism.

[0101] For details, see Figure 3 The magnetic bead adsorption mechanism is located outside the microorganism extraction container 5 and is used to adsorb magnetic beads 51 on the microorganism extraction container 5. The magnetic attraction component 3 is an electromagnet, and the magnetic attraction state switching component is an electromagnet controller. The electromagnet controller controls the magnetism of the electromagnet to control the adsorption or release of the magnetic beads.

[0102] See also Figure 4 and Figure 5 The magnetic state switching component can be configured to include a fifth driving member 71 for driving the microorganism extraction container 5 and / or a sixth driving member 72 for driving the magnetic assembly 3 to move. The magnetic state switching component changes the magnetic state by changing the relative position of the microorganism extraction container 5 and the magnetic assembly 3. Figure 4 , the fifth driving member 71 is a linear motion mechanism; or further see Figure 5 , the fifth driving member 71 is a rotary motion mechanism.

[0103] As described above, this embodiment has a different microorganism extraction container from that of embodiment 1, and a different magnetic bead adsorption mechanism, but can also achieve liquid replacement by adsorbing magnetic beads. The other principles of the two are the same, and both can also use magnetic beads to extract microorganisms.

[0104] Example 3

[0105] In this embodiment, a microbial extraction system includes:

[0106] A microorganism extraction container, wherein the microorganism extraction container contains a liquid to be extracted and magnetic beads for adsorbing microorganisms, and the liquid to be extracted contains the microorganisms to be extracted;

[0107] A workbench is provided with a container placement portion, the container placement portion is used to place a microorganism extraction container, and the microorganism extraction container has multiple holes;

[0108] Magnetic bead adsorption mechanism and controller, the magnetic bead adsorption mechanism is installed on the workbench, and is used to adsorb and transfer magnetic beads between each well position. The magnetic bead adsorption mechanism includes a magnetic attraction component and a driving component. The driving component is used to change the magnetism of the magnetic attraction component (that is, another embodiment of the magnetic attraction state switching component). The controller is electrically connected to the driving component to control the operation of the driving component to drive the magnetic attraction component to move relative to the microorganism extraction container; wherein, the magnetic attraction component includes an electromagnet, and the driving component includes an electromagnet controller. In addition to the electromagnet controller, the driving component can also include an electromagnet driver. The controller is electrically connected to the electromagnet controller and the electromagnet driver to control the magnetism and movement of the electromagnet. The electromagnet controller can control the power on and off of the electromagnet to switch between a magnetic attraction state with magnetism and a release state with loss of magnetism. The electromagnet driver can drive the electromagnet to move to achieve magnetic bead transfer.

[0109] Example 4

[0110] This patent application discovered that the particle size and electrical properties of the magnetic beads would affect the extraction effect. To verify this solution, the experiment of this embodiment was conducted.

[0111] First, we list several ways to obtain magnetic beads to better understand this solution.

[0112] A. Synthesis of amino magnetic beads

[0113] (1) Synthesis of amino magnetic beads 1

[0114] 46 g of ferric chloride hexahydrate and 16 g of ferrous chloride tetrahydrate were dissolved in 400 g of water, followed by the addition of 120 g of ammonia water. The mixture was then heated to 70°C and allowed to react for 2 h. After cooling, the mixture was subjected to magnetic separation and washed with water until the pH of the clear solution was neutral. The magnetic particles were then dispersed in 400 g of water, 400 g of ethanol and 10 g of an aminosilane reagent [γ-aminopropyltriethoxysilane (brand A-1100, KH-550)] were added, and the mixture was allowed to react at 70°C for 5 h. After cooling, the mixture was subjected to magnetic separation and washed with ethanol three times and water three times to obtain amino magnetic beads 1. The particle size of the amino magnetic beads was determined to be approximately 80 nm by SEM.

[0115] (2) Synthesis of amino magnetic beads 2

[0116] 46 g of ferric chloride hexahydrate and 16 g of ferrous chloride tetrahydrate were dissolved in 400 g of water, followed by the addition of 120 g of ammonia water. The mixture was heated to 70°C and allowed to react for 2 h. After cooling, magnetic separation was performed and vacuum drying was performed to obtain magnetic particle powder. The magnetic particle powder was then dispersed in 400 g of isopropanol, followed by the addition of polyaminoalkyltrialkoxysilane (Y-5691) and 50 g of water. The mixture was allowed to react at 70°C for 5 h. After cooling, magnetic separation was performed and the mixture was washed three times with ethanol and three times with water to obtain amino magnetic beads 2. SEM analysis showed that the particle size of the amino magnetic beads was approximately 200 nm.

[0117] (3) Synthesis of amino magnetic beads 3

[0118] 6g of ferric chloride hexahydrate was dissolved in 30mL of ethylene glycol, and 0.6g of sodium acetate and 1g of sodium citrate were dissolved in 30mL of ethylene glycol. The mixture was placed in a 100mL hydrothermal reactor and reacted at 200°C for 15h. After completion of the reaction, magnetic separation was performed and the mixture was washed three times with ethanol and water to obtain magnetic particles. The magnetic particles were dispersed in 80mL of ethanol, followed by the addition of 20mL of water and 8mL of ammonia water, followed by the addition of 0.5g of tetraethyl orthosilicate, and the reaction was continued for 6h. After completion of the reaction, the mixture was washed three times with ethanol and water to obtain magnetic beads. The magnetic beads were dispersed in 40mL of ethanol, followed by the addition of 40mL of water and γ-aminopropyltrimethoxysilane, and the reaction was continued for 6h. After completion of the reaction, the mixture was washed three times with ethanol and water to obtain amino magnetic beads 3. The particle size of the amino magnetic beads was determined by SEM to be approximately 2μm.

[0119] (4) Synthesis of amino magnetic beads 4

[0120] 10 g of commercial carboxyl magnetic beads (Dynabeads TM5 g of polyethyleneimine was dissolved in 400 mL of the same MES buffer (4-morpholineethanesulfonic acid) (20 mM, pH = 5.5), the two were mixed, and then 2 g of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) (dissolved in 100 mL of MES) was added and reacted for 5 h. After completion of the reaction, the beads were magnetically separated and washed four times with water to obtain amino magnetic beads 4. The particle size of the amino magnetic beads was determined by SEM to be approximately 1 μm.

[0121] (5) Amino magnetic beads 5

[0122] Purchased from Nanjing Dongna Biotechnology Co., Ltd. (Mag9002), particle size 100 nm.

[0123] B. Synthesis of Imidazole-Based Magnetic Beads

[0124] Synthesis of imidazole-based magnetic beads

[0125] 1. Synthesis of magnetic core

[0126] 120 mL of ethylene glycol, 6 g of ferric chloride hexahydrate, and 9.72 g of anhydrous sodium acetate were mixed and dissolved by ultrasonication. The mixture was placed in a 200° oven and reacted for 12 h. The mixture was then magnetically separated, washed with ethanol three times, washed with water three times, and dried in vacuo to obtain a solid powder.

[0127] 2. Preparation of transition layer

[0128] Step 1: Disperse 10g of magnetic core in 400g of ethanol, add 100g of water and 20g of acetic acid, then add 5g of tetramethyl orthosilicate, and react at 50° for 10h; after the reaction is completed, perform magnetic separation, and wash with ethanol and water three times in sequence to obtain silica-coated magnetic particles;

[0129] Step 2: Disperse the magnetic particles prepared in step 1 in 400 g of ethanol, add 1 g of KH570, and react at 50° for 12 h. After the reaction is complete, perform magnetic separation and wash with ethanol and water three times in sequence to obtain KH570-modified magnetic particles.

[0130] 3. Preparation of hydrophobic layer

[0131] Step 3: The magnetic particles obtained in step 2 are dispersed in 800 g of PVA in DMSO (dimethyl sulfoxide) solution (1% wt), and then 0.5 g of azobisisobutyronitrile is added. After nitrogen replacement, the temperature is raised to 70°, and then a mixed solution of 5 g of N-vinylimidazole, 0.5 g of N,N'-methylenebisacrylamide and 20 g of DMSO is added dropwise. The mixture is added over 30 minutes and the reaction is continued for 20 hours. After cooling to room temperature, magnetic separation is performed and the mixture is washed three times with ethanol and water in sequence to obtain imidazole-coated magnetic particles.

[0132] C. Carboxyl magnetic beads

[0133] Carboxyl magnetic beads were purchased from Thermo Fisher Scientific (Dynabeads TM 65012), Suzhou Weidu Biotechnology Co., Ltd. (CMP0600CA), Xiamen PuriMag Biotechnology Co., Ltd. (PuriMag GS-COOH), and Suzhou Nanovitamin Life Science Co., Ltd. (DS415_MS500-COOH), which were named as Carboxyl Magnetic Beads 1, Carboxyl Magnetic Beads 2, Carboxyl Magnetic Beads 3, and Carboxyl Magnetic Beads 4, respectively.

[0134] It is worth noting that the various magnetic beads provided in this patent application can be obtained by purchase or preparation, and will not affect the results.

[0135] The experiment first verified the adsorption effect of positively charged magnetic beads. The following comparison test was conducted on the most common sample - blood culture sample, using different positively charged magnetic beads (amino magnetic beads, imidazole magnetic beads) and negatively charged magnetic beads (hydroxyl magnetic beads). The results of bacterial adsorption by different magnetic beads are shown in Table 2.

[0136] Table 2 Enrichment effect of different electromagnetic beads

[0137]

[0138]

[0139] As can be seen from Table 2, positively charged magnetic beads can extract most microorganisms, especially for blood culture positive samples, which is of great significance for clinical use.

[0140] Then, purchase or prepare amino magnetic beads with different particle sizes to test the particle size adsorption effect. The particle sizes of amino magnetic beads with different sizes include: 1nm, 5nm, 30nm, 150nm, 500nm, 1μm, and 3μm.

[0141] Among them, 1nm and 5nm were obtained using the synthesis method of amino magnetic beads 5 in the above magnetic bead preparation scheme, 30nm and 150nm were obtained using the synthesis method of amino magnetic beads 3, and 500nm were obtained using the synthesis method of amino magnetic beads 1. 1μm amino magnetic beads were purchased from Beijing Beike New Materials Technology Co., Ltd.; 3μm amino magnetic beads were purchased from Beijing Zhongke Keyou Technology Co., Ltd.

[0142] In the most common sample - blood culture sample, a comparative test was conducted using amino magnetic beads with different particle sizes. The results of bacterial adsorption by magnetic beads with different particle sizes are shown in Table 3:

[0143] Table 3 Enrichment effect of magnetic beads with different particle sizes

[0144]

[0145] As can be seen from Table 3, the magnetic beads have a good enrichment effect when the particle size is no more than 3 μm, and the magnetic beads have the best enrichment effect when the particle size is 5 nm-1 μm.

[0146] Throughout this specification, references to terms such as "this embodiment," "example," and "specific example" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0147] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Anyone skilled in the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by one of ordinary skill in the art without departing from the spirit and technical principles disclosed in the present invention are intended to be covered by the claims of the present invention.

Claims

1. A microbial extraction system, characterized in that: include: a microorganism extraction container configured to contain a liquid to be extracted and magnetic beads for adsorbing microorganisms, wherein the liquid to be extracted contains the microorganisms to be extracted; A workbench is provided with a container placement portion, wherein the container placement portion is used to place the microorganism extraction container; A magnetic bead adsorption mechanism is installed on the workbench, and the magnetic bead adsorption mechanism includes a magnetic attraction component and a magnetic attraction state switching component for changing the magnetic attraction state. The magnetic attraction component has magnetism and is used to adsorb the magnetic beads located in the microorganism extraction container; A controller is electrically connected to the magnetic state switching component and is used to change the state of the magnetic component or change the relative state between the magnetic component and the microorganism extraction container.

2. The microorganism extraction system according to claim 1, characterized in that The microorganism extraction container has multiple hole positions, and the magnetic bead adsorption mechanism is a transfer mechanism for adsorbing and transferring the magnetic beads between each of the hole positions; the transfer mechanism includes a magnetic attraction component and a magnetic attraction state switching component, and the magnetic attraction state switching component includes a driving component, and the driving component is used to drive the magnetic attraction component to move relative to the microorganism extraction container or to change the magnetism of the magnetic attraction component.

3. The microbial extraction system according to claim 2, characterized in that: The microorganism extraction container is a 96-well plate having 96 wells, a 48-well plate having 48 wells, or a 32-well plate having 32 wells.

4. The microorganism extraction system according to claim 2, characterized in that The microorganism extraction container has at least three holes, and magnetic beads, washing solution and eluent are pre-buried in at least three of the holes respectively.

5. The microorganism extraction system according to claim 2, 3 or 4, characterized in that: The magnetic attraction assembly includes a magnetic rod and a magnetic rod sleeve. The magnetic rod sleeve is used to extend into the microorganism extraction container. The magnetic rod is built into the magnetic rod sleeve and can move relative to the magnetic rod sleeve to adsorb and release magnetic beads located outside the magnetic rod sleeve.

6. The microorganism extraction system according to claim 5, characterized in that: The magnetic rod sleeve is installed on the workbench or placed on the microorganism extraction container.

7. The microorganism extraction system according to claim 5, characterized in that: The driving assembly includes a first driving member, the output end of which is connected to the magnetic rod through a first movable bracket to drive the magnetic rod to move up and down; The driving assembly further includes a second driving member, an output end of which is connected to the magnetic rod sleeve via a second movable bracket, and the first driving member is mounted on the second movable bracket, and the second driving member drives the first driving member and the second movable bracket to move synchronously up and down via the second movable bracket; The top end of the magnetic rod sleeve is open, the first movable bracket is located above the second movable bracket, and the first driving member drives the magnetic rod to move up and down through the first movable bracket to extend into and out of the magnetic rod sleeve; The driving assembly further includes a third driving member, which is connected to the second driving member to drive the second driving member and the second movable bracket to move in a horizontal direction.

8. The microorganism extraction system according to claim 7, characterized in that: The workbench is provided with a rear limit block and a side limit block, and the rear limit block, the side limit block and the container placement portion cooperate to define a plurality of container placement positions distributed along the horizontal movement direction of the second movable bracket.

9. The microorganism extraction system according to claim 8, characterized in that There are multiple magnetic bars, which are arranged in multiple rows along the arrangement direction of the multiple container placement positions; and the magnetic bar sleeves are arranged in a one-to-one correspondence with the magnetic bars.

10. The microorganism extraction system according to claim 1, characterized in that: It also includes a liquid replacement mechanism for sucking and discharging liquid to replace the liquid in the microorganism extraction container. The magnetic bead adsorption mechanism is located outside the microorganism extraction container and is used to adsorb the magnetic beads on the microorganism extraction container.

11. The microorganism extraction system according to claim 10, characterized in that: The liquid replacement mechanism is a liquid suction needle, a liquid suction pipeline or multiple liquid suction pipelines.

12. The microorganism extraction system according to claim 10 or 11, characterized in that: The magnetic bead adsorption mechanism is an electromagnet, and the magnetic state switching component is an electromagnet controller.

13. The microorganism extraction system according to claim 10 or 11, characterized in that: The magnetic state switching component includes a fifth driving member for driving the microorganism extraction container and / or a sixth driving member for driving the magnetic assembly to move. The magnetic state switching component changes the magnetic state by changing the relative position of the microorganism extraction container and the magnetic assembly.

14. The microorganism extraction system according to claim 13, characterized in that: The fifth driving member is a linear motion mechanism or a rotary motion mechanism.

15. The microorganism extraction system according to any one of claims 1-4, 6-11 or 14, characterized in that: The particle size of the magnetic beads is no greater than 3 μm.

16. The microorganism extraction system according to claim 15, characterized in that: The particle size of the magnetic beads is 5nm-1μm.

17. The microorganism extraction system according to any one of claims 1-4, 6-11 or 14, characterized in that: The magnetic beads are positively charged.

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