A system for separating and extracting a microbiota

Abstract

The utility model discloses a kind of flora separation extraction systems, including stirring filter device, circulating filter device, waste liquid waste gas treatment device, collecting device and liquid supplementing air supplementing device, stirring filter device, circulating filter device, collecting device are sequentially connected, stirring filter device, circulating filter device are respectively connected with waste liquid waste gas treatment device, liquid supplementing air supplementing device;Stirring filter device is used to stir and filter sample to obtain primary filtrate;Circulating filter device is used to circulate and filter primary filtrate to obtain target bacteria suspension, circulating filter device includes with the bacteria liquid bottle and microfiltration device that stirring filter device communicates, and for the second weighing device for weighing target bacteria suspension, microfiltration device communicates with bacteria liquid bottle;Liquid supplementing air supplementing device communicates with bacteria liquid bottle, microfiltration device communicates with waste liquid waste gas treatment device, and liquid supplementing air supplementing device is used to give stirring filter device and bacteria liquid bottle supplement buffer solution and gas.

Description

Technical Field

[0001] This utility model relates to the field of intestinal flora collection, and in particular to a flora separation and extraction system. Background Technology

[0002] Fecal microbiota transplantation (FMT) is a treatment method that involves transplanting the gut microbiota (intestinal flora) from the feces of healthy individuals into the body of patients, allowing the patients to obtain the gut microbiota of healthy individuals, thereby regulating their imbalanced gut microbiota and rebuilding a normally functioning gut microecological system. The most important issue in FMT is how to isolate and preserve fecal microbiota without affecting the function of the microbiota.

[0003] A prior art patent, such as Chinese Patent Publication No. CN114177684B, discloses a stirring and filtering device for bacterial colony separation, comprising a stirring tank and a filter tank located below the stirring tank. The stirring tank has a drain outlet at its bottom, and an openable sealing cap below the drain outlet. The stirring tank has an inlet, and the filter tank has an outlet at its bottom. The filter tank contains a filtering device and a stirring anti-clogging device. The filtering device includes an upper filter screen frame that completely encloses the drain outlet. An upper filter screen is horizontally arranged within the upper filter screen frame. A side filter frame is provided at the lower end, and multiple layers of side filter screens are vertically arranged on the side filter frame. A lower filter screen is provided at the bottom of the inner surface of the side filter frame, and the diameter of the lower filter screen is the same as the inner diameter of the side filter frame. The stirring and anti-clogging device includes a rotatable rotating rod, and an upper stirring brush is provided at the top of the rotating rod. The upper stirring brush is located above the upper filter screen, and the lower end of the upper stirring brush can contact the surface of the upper filter screen. A side stirring brush is provided on the rotating rod. The side stirring brush is located inside the hollow side filter frame, and the side stirring brush can contact the surface of the innermost side filter screen.

[0004] Although the stirring and filtering device for bacterial separation disclosed in the aforementioned patent is equipped with a stirring anti-clogging device, it lacks a recovery device. As unfilterable particles accumulate, the filtering device will eventually become clogged. To recover the unfilterable particles, the entire filtering device needs to be disassembled and cleaned, which is not very practical. Furthermore, the aforementioned device lacks activity protection measures during the bacterial extraction process, which will negatively affect the activity of the extracted bacterial population. Utility Model Content

[0005] To address the problems existing in the prior art, this utility model discloses a microbial community isolation and extraction system.

[0006] A microbial community isolation and extraction system includes a stirring filtration device, a circulating filtration device, a waste liquid and waste gas treatment device, a collection device, and a liquid and gas replenishment device. The stirring filtration device, the circulating filtration device, and the collection device are connected in sequence. The stirring filtration device and the circulating filtration device are respectively connected to the waste liquid and waste gas treatment device and the liquid and gas replenishment device.

[0007] The stirring and filtering device is used to stir and filter the sample to obtain a primary filtrate;

[0008] The circulating filtration device is used to circulate and filter the primary filtrate to obtain the target bacterial suspension. The circulating filtration device includes a bacterial liquid bottle and a microfiltration device connected to the stirring filtration device, as well as a second weighing device for weighing the target bacterial suspension. The microfiltration device is connected to the bacterial liquid bottle, and the liquid replenishment and gas replenishment device is connected to the bacterial liquid bottle. The microfiltration device is also connected to the waste liquid and waste gas treatment device.

[0009] The liquid and gas replenishment device is used to replenish buffer solution and gas to the stirring and filtering device and the bacterial culture bottle.

[0010] Specifically, the sample is first-stage filtered using a stirring filtration device to obtain primary filtrate. Then, a circulating filtration device is used to circulate the primary filtrate. The specific process of circulating filtration is as follows: gas is passed into the bacterial culture bottle to maintain its environment (e.g., nitrogen is passed in if an anaerobic environment is required). The primary filtrate enters the bacterial culture bottle. When the primary filtrate reaches a certain weight, it is passed through a microfiltration device for filtration and concentration. The concentrate is then returned to the bacterial culture bottle, and a buffer solution is added to the bacterial culture bottle. The mixture of buffer solution and concentrate is then passed through the microfiltration device again for filtration and concentration, and then returned to the bacterial culture bottle. This cycle is repeated until the target bacterial suspension meets the requirements. Finally, the target bacterial suspension in the bacterial culture bottle is placed into a collection bag. The process of circulating filtration of the primary filtrate in the circulating filtration device is actually the process of washing the target bacterial suspension. Washing the target bacterial suspension can further and effectively ensure the safety of the bacterial culture.

[0011] In addition, both liquids and gases flow along the pipeline. In order to ensure that the liquids and gases flow smoothly in the system in the specified direction, various valves and power devices (such as pumps) can be installed in the system according to the specific situation.

[0012] Preferably, the stirring and filtering device includes a stirring and filtering tank, which includes a detachably connected tank body and a tank cover, and also includes a stirring device;

[0013] The tank is equipped with a filter device, which forms a stirring zone. The filter device is separated from the side wall of the tank and forms a liquid storage zone between the filter device and the side wall of the tank.

[0014] The distance between the bottom of the tank corresponding to the stirring zone and the tank cover decreases towards the center. The bottom of the tank corresponding to the liquid storage zone is provided with a liquid outlet, which connects the liquid storage zone and the bacterial liquid bottle.

[0015] Specifically, the stirring and filtering device needs to stir and filter the sample. After the sample is stirred and filtered, there will be residue deposited at the bottom of the tank corresponding to the stirring zone. The residue will reduce the collection rate of bacteria. Therefore, in order to prevent the residue from being deposited in the middle of the bottom of the tank, it is appropriate to set the bottom of the tank corresponding to the stirring zone as the highest structure in the middle.

[0016] Preferably, the bottom of the tank corresponding to the liquid storage area is provided with a highest point and a lowest point, the highest point and the lowest point are connected in a streamline shape, and the liquid outlet is located at the lowest point.

[0017] Specifically, in this structure, the filtrate after the first stage flows out of the tank through the lowest point of the tank. This structure can ensure that there is basically no filtrate residue in the tank, and can also prevent sedimentation at the bottom of the tank corresponding to the storage area by utilizing the flow of filtrate and gravity.

[0018] Preferably, the stirring device includes a stirring shaft with a stirring paddle and a motor. The stirring shaft passes through the bottom of the tank and is connected to the motor located outside the tank. The stirring shaft is sealed to the bottom of the tank. The highest point of the bottom of the tank corresponding to the stirring zone falls on the central axis of the stirring shaft.

[0019] Specifically, the highest point at the bottom of the container falls on the central axis of the stirring shaft, meaning the stirring shaft is positioned at the center of the bottom of the container. This structural arrangement allows the stirring shaft to more evenly mix the sample (mixed in the buffer solution).

[0020] Preferably, the stirring and filtering device is further provided with a first weighing device, which is used to measure the weight of the sample inside the tank.

[0021] Specifically, the weight of the sample inside the tank is obtained by reading the value of the first weighing device, and then buffer solution is added into the tank according to the weight of the sample and a preset ratio.

[0022] Preferably, the waste liquid and waste gas treatment device includes a waste liquid bag connected to a microfiltration device; the waste liquid and waste gas treatment device includes a deodorization device connected to a stirring filter and / or the waste liquid bag.

[0023] Specifically, samples used for bacterial isolation may have a strong odor. In order to protect the health of operators and maintain a good working environment, it is reasonable to install a deodorization device connected to the stirring and filtering device. Microfiltration devices are used to circulate and concentrate the primary filtrate. Waste liquid will inevitably be generated in this process. Therefore, it is necessary to install a waste liquid bag connected to the microfiltration device.

[0024] Preferably, it also includes a sampling port through which samples can be taken from the bacterial culture bottle;

[0025] A pressure monitoring device, a flow regulating pump, and a pressure regulating valve are provided between the bacterial culture bottle and the microfiltration device. The pressure monitoring device is used to monitor the pressure of the liquid flowing from the bacterial culture bottle to the microfiltration device or from the microfiltration device to the bacterial culture bottle in real time. The flow regulating pump is used to regulate the flow rate of the liquid flowing from the bacterial culture bottle to the microfiltration device. The pressure regulating valve is used to regulate the pressure of the liquid flowing from the microfiltration device to the bacterial culture bottle.

[0026] Specifically, when the pressure monitoring device detects that the liquid pressure is too high, there is a risk of pipe bursting. At this time, the liquid pressure is reduced by adjusting the pressure regulating valve. When the pressure regulating valve has reached the pressure regulation limit but the liquid pressure is still too high, the liquid flow rate is reduced by adjusting the flow regulating pump to achieve the effect of pressure reduction. When the flow regulating pump has brought the liquid to the target flow rate, but the pressure monitoring device detects that the liquid pressure value is too low, the pressure on the liquid is increased by adjusting the pressure regulating valve.

[0027] Preferably, the collection device includes a cryopreservation solution replenishment device and a collection bag equipped with a mixing device. The collection bag is connected to the cryopreservation solution replenishment device and the bacterial suspension bottle. The mixing device is used to mix the cryopreservation solution and the target bacterial suspension in the collection bag.

[0028] Preferably, the microfiltration device is a tangential flow device, which is a hollow fiber column or a membrane package.

[0029] Preferably, the stirring filter, circulating filter, collecting device, and liquid replenishment and gas replenishment device are not connected to the environment outside the microbial community separation system.

[0030] Specifically, the structure of the microbial community isolation system, which is not connected to the external environment, makes it easier to maintain the internal environment of the microbial community isolation system. Maintaining the internal environment of the microbial community extraction system according to the characteristics of the target microbial community helps to maintain the activity and stability of the target microbial community.

[0031] Compared with the prior art, the advantages of this utility model are:

[0032] By setting up a stirring filtration device and a circulating filtration device, the sample was subjected to two-stage filtration, including circulating filtration. At the same time, a waste liquid and waste gas treatment device, a liquid replenishment and gas replenishment device, and a collection device were set up to achieve a high degree of automation in the separation of bacterial communities, including environmental maintenance and waste liquid and waste gas treatment. In addition, the circulating filtration device also achieved the purpose of washing the target bacterial suspension, effectively ensuring the safety of the target bacterial suspension. Attached Figure Description

[0033] Figure 1 This is a schematic diagram of the microbial community separation system provided by this utility model;

[0034] Figure 2 A schematic diagram of the structure of the stirring filter tank provided by this utility model;

[0035] Figure 3 This is a cross-sectional schematic diagram of the stirring filter tank provided by this utility model. Detailed Implementation

[0036] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0037] like Figure 1 As shown, the microbial community isolation and extraction system includes a stirring filter device 10, a circulating filter device, a waste liquid and waste gas treatment device, a collection device, and a liquid and gas replenishment device 50. The stirring filter device 10, the circulating filter device, and the collection device are connected in sequence. The stirring filter device 10 and the circulating filter device are respectively connected to the waste liquid and waste gas treatment device and the liquid and gas replenishment device 50.

[0038] The stirring and filtering device 10 is used to stir and filter the sample to obtain the primary filtrate;

[0039] The circulating filtration device is used to circulate and filter the primary filtrate to obtain the target bacterial suspension. The circulating filtration device includes a bacterial liquid bottle 22 and a microfiltration device 21 connected to the stirring filtration device 10, as well as a second weighing device 62 for weighing the target bacterial suspension. The microfiltration device 21 is connected to the bacterial liquid bottle 22, the liquid replenishment and gas replenishment device 50 is connected to the bacterial liquid bottle 22, and the microfiltration device 21 is connected to the waste liquid and waste gas treatment device.

[0040] The liquid and gas replenishment device 50 is used to replenish buffer solution and gas to the stirring and filtering device 10 and the bacterial culture bottle 22.

[0041] The sample is first-stage filtered using a stirring filter device 10 to obtain a primary filtrate. Then, a circulating filter device is used to circulate the primary filtrate. The specific process of circulating filtration is as follows: gas is introduced into the bacterial culture bottle 22 to maintain its environment (e.g., nitrogen is introduced if an anaerobic environment is required). The primary filtrate enters the bacterial culture bottle 22. When the primary filtrate reaches a certain weight, it is passed through a microfiltration device 21 for filtration and concentration. The concentrate is then returned to the bacterial culture bottle 22. Simultaneously, buffer solution is added to the bacterial culture bottle 22. The mixture of buffer solution and concentrate is then passed through the microfiltration device 21 again for filtration and concentration, and then returned to the bacterial culture bottle 22. This cycle is repeated until the target bacterial suspension meets the requirements. Finally, the target bacterial suspension in the bacterial culture bottle 22 is placed into a collection bag. The process of circulating filtration of the primary filtrate in the circulating filter device is essentially the washing process of the target bacterial suspension. Washing the target bacterial suspension further effectively ensures its safety.

[0042] In addition, both liquids and gases flow along the pipeline. In order to ensure that the liquids and gases flow smoothly in the system in the specified direction, various valves and power devices (such as pumps) can be installed in the system according to the specific situation.

[0043] like Figure 2-3 As shown, the stirring and filtering device 10 includes a stirring and filtering tank, which includes a tank body 11 and a tank cover 12 that are detachably connected, and also includes a stirring device 13.

[0044] A filter device 14 is installed inside the tank body 11. The filter device 14 surrounds and forms a stirring zone, and the filter device 14 is separated from the side wall of the tank body 11 and forms a liquid storage zone between the filter device 14 and the side wall of the tank body 11.

[0045] The distance between the bottom of the tank 11 corresponding to the stirring zone and the tank cover 12 decreases towards the center. The bottom of the tank 11 corresponding to the liquid storage zone is provided with a liquid outlet 15, which connects the liquid storage zone and the bacterial liquid bottle 22.

[0046] The stirring and filtering device 10 needs to stir and filter the sample. After the sample is stirred and filtered, there will be residue deposited at the bottom of the tank 11 corresponding to the stirring zone. The residue will reduce the collection rate of the microbial community. Therefore, in order to prevent the residue from being deposited in the middle of the bottom of the tank 11, it is appropriate to set the bottom of the tank 11 corresponding to the stirring zone to be the highest in the middle.

[0047] The tank 11 corresponding to the liquid storage area has a highest point and a lowest point at its bottom, and the highest point and the lowest point are connected in a streamlined manner. The liquid outlet 15 is located at the lowest point.

[0048] In this structure, the primary filtrate eventually flows out of the tank 11 through the lowest point of the tank 11. This structure can ensure that there is basically no filtrate residue in the tank 11, and can also prevent sedimentation at the bottom of the tank 11 corresponding to the storage area by utilizing the flow of the filtrate and the effect of gravity.

[0049] The stirring device 13 includes a stirring shaft with a stirring paddle and a motor. The stirring shaft passes through the bottom of the tank 11 and is connected to the motor located outside the tank 11. The stirring shaft is sealed to the bottom of the tank 11. The highest point of the bottom of the tank 11 corresponding to the stirring zone falls on the central axis of the stirring shaft.

[0050] The highest point of the bottom of the container 11 falls on the central axis of the stirring shaft, that is, the stirring shaft is located at the center of the bottom of the container 11. This position structure makes the stirring shaft more uniform in stirring the sample (mixed in the buffer solution).

[0051] The stirring and filtering device 10 is also equipped with a first weighing device 61, which is used to measure the weight of the sample inside the tank 11.

[0052] The weight of the sample in the tank 11 is obtained by reading the value of the first weighing device 61, and then buffer solution is added into the tank 11 according to the sample weight and a preset ratio.

[0053] The waste liquid and waste gas treatment device includes a waste liquid bag 31, which is connected to a microfiltration device 21.

[0054] The waste liquid and waste gas treatment device includes a deodorization device 32, which is connected to a stirring and filtering device 10 and / or a waste liquid bag 31.

[0055] Samples used for bacterial isolation may have a strong odor. In order to protect the health of operators and maintain a good working environment, it is reasonable to set up a deodorization device 32 connected to the stirring and filtering device 10. The microfiltration device 21 is used to circulate and concentrate the primary filtrate. Waste liquid will inevitably be generated in this process. Therefore, it is necessary to set up a waste liquid bag 31 connected to the microfiltration device 21.

[0056] It also includes a sampling port through which samples can be taken from the bacterial culture bottle 22;

[0057] A pressure monitoring device 71, a flow regulating pump 72, and a pressure regulating valve 73 are provided between the bacterial culture bottle 22 and the microfiltration device 21. The pressure monitoring device 71 is used to monitor the pressure of the liquid flowing from the bacterial culture bottle 22 to the microfiltration device 21 or from the microfiltration device 21 to the bacterial culture bottle 22 in real time. The flow regulating pump 72 is used to regulate the flow rate of the liquid flowing from the bacterial culture bottle 22 to the microfiltration device 21. The pressure regulating valve 73 is used to regulate the pressure of the liquid flowing from the microfiltration device 21 to the bacterial culture bottle 22.

[0058] When the pressure monitoring device 71 detects that the liquid pressure is too high, there is a risk of pipe bursting. At this time, the liquid pressure is reduced by adjusting the pressure regulating valve 73. When the pressure regulating valve 73 has reached the pressure regulation limit but the liquid pressure is still too high, the liquid flow rate is reduced by adjusting the flow regulating pump 72 to achieve the effect of pressure reduction. When the flow regulating pump 72 has brought the liquid to the target flow rate, but the pressure monitoring device 71 detects that the liquid pressure value is too low, the pressure on the liquid is increased by adjusting the pressure regulating valve 73.

[0059] The collection device includes a cryopreservation liquid replenishment device 41 and a collection bag 43 equipped with a mixing device 42. The collection bag 43 is connected to the cryopreservation liquid replenishment device 41 and the bacterial culture bottle 22. The mixing device 42 is used to mix the cryopreservation liquid and the target bacterial suspension in the collection bag 43.

[0060] The microfiltration device 21 is a tangential flow device, which is a hollow fiber column or a membrane package.

[0061] The stirring filter device 10, the circulating filter device, the collection device, and the liquid replenishment and gas replenishment device 50 are all not connected to the environment outside the microbial community separation system.

[0062] The structure of the microbial community isolation system, which is not connected to the external environment, makes it easier to maintain the internal environment of the microbial community isolation system. Maintaining the internal environment of the microbial community extraction system according to the characteristics of the target microbial community helps to maintain the activity and stability of the target microbial community.

Claims

1. A microbial community isolation and extraction system, comprising a stirring and filtering device, a circulating and filtering device, a waste liquid and waste gas treatment device, a collection device, and a liquid and gas replenishment device, characterized in that, The stirring filter, the circulating filter, and the collecting device are connected in sequence. The stirring filter and the circulating filter are respectively connected to the waste liquid and waste gas treatment device and the liquid replenishment and gas replenishment device. The stirring and filtering device is used to stir and filter the sample to obtain a primary filtrate; The circulating filtration device is used to circulate and filter the primary filtrate to obtain the target bacterial suspension. The circulating filtration device includes a bacterial liquid bottle and a microfiltration device connected to the stirring filtration device, as well as a second weighing device for weighing the target bacterial suspension. The microfiltration device is connected to the bacterial liquid bottle, and the liquid replenishment and gas replenishment device is connected to the bacterial liquid bottle. The microfiltration device is also connected to the waste liquid and waste gas treatment device. The liquid and gas replenishment device is used to replenish buffer solution and gas to the stirring and filtering device and the bacterial culture bottle.

2. The microbial community isolation and extraction system according to claim 1, characterized in that, The stirring and filtering device includes a stirring and filtering tank, which includes a detachably connected tank body and a tank cover, and also includes a stirring device. The tank is equipped with a filter device, which forms a stirring zone. The filter device is separated from the side wall of the tank and forms a liquid storage zone between the filter device and the side wall of the tank. The distance between the bottom of the tank corresponding to the stirring zone and the tank cover decreases towards the center. The bottom of the tank corresponding to the liquid storage zone is provided with a liquid outlet, which connects the liquid storage zone and the bacterial liquid bottle.

3. The microbial community isolation and extraction system according to claim 2, characterized in that, The tank bottom corresponding to the liquid storage area has a highest point and a lowest point, which are connected in a streamlined manner, and the liquid outlet is located at the lowest point.

4. The microbial community isolation and extraction system according to claim 2, characterized in that, The stirring device includes a stirring shaft with a stirring paddle and a motor. The stirring shaft passes through the bottom of the tank and is connected to the motor located outside the tank. The stirring shaft is sealed to the bottom of the tank. The highest point of the bottom of the tank corresponding to the stirring zone falls on the central axis of the stirring shaft.

5. The microbial community isolation and extraction system according to claim 2, characterized in that, The stirring and filtering device is also equipped with a first weighing device, which is used to measure the weight of the sample inside the tank.

6. The microbial community isolation and extraction system according to claim 1, characterized in that, The waste liquid and waste gas treatment device includes a waste liquid bag connected to a microfiltration device; the waste liquid and waste gas treatment device includes a deodorization device connected to a stirring filter and / or the waste liquid bag.

7. The microbial community isolation and extraction system according to claim 1, characterized in that, It also includes a sampling port through which samples can be taken from the bacterial culture bottle; A pressure monitoring device, a flow regulating pump, and a pressure regulating valve are provided between the bacterial culture bottle and the microfiltration device. The pressure monitoring device is used to monitor the pressure of the liquid flowing from the bacterial culture bottle to the microfiltration device or from the microfiltration device to the bacterial culture bottle in real time. The flow regulating pump is used to regulate the flow rate of the liquid flowing from the bacterial culture bottle to the microfiltration device. The pressure regulating valve is used to regulate the pressure of the liquid flowing from the microfiltration device to the bacterial culture bottle.

8. The microbial community isolation and extraction system according to claim 1, characterized in that, The collection device includes a cryopreservation liquid replenishment device and a collection bag equipped with a mixing device. The collection bag is connected to the cryopreservation liquid replenishment device and the bacterial suspension bottle. The mixing device is used to mix the cryopreservation liquid and the target bacterial suspension in the collection bag.

9. The microbial community isolation and extraction system according to claim 1, characterized in that, The microfiltration device is a tangential flow device, which is a hollow fiber column or membrane package.

10. The microbial community isolation and extraction system according to claim 1, characterized in that, The stirring and filtering device, the circulating and filtering device, the collecting device, and the liquid and gas replenishment device are all not connected to the environment outside the microbial community separation system.

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