Separation and concentration device for intestinal flora

Abstract

The utility model discloses a separation and concentration device for intestinal flora, which belongs to the technical field of biological extraction and comprises a flora extraction device and a negative pressure drainage apparatus, the flora extraction device is sequentially connected with a plurality of filters from top to bottom, and every two adjacent filters are detachably connected. The mesh size of the screen of each stage of filter is gradually reduced from the uppermost stage to the lower stage, the lowermost stage of filter is used for enriching and concentrating the intestinal flora suspension separated by the previous stage of filter, and the negative pressure drainage device is connected with the lowermost stage of filter and is used for collecting liquid generated in the enriching and concentrating process of the lowermost stage of filter; according to the separation and concentration device, the homogenizing device, the multi-stage filtering device and the enrichment and concentration device are integrated into a whole, the flora separation and concentration process is simplified, and the extraction efficiency of intestinal flora is improved.

Description

Technical Field

[0001] The utility model relates to the field of intestinal flora technical equipment, in particular to an intestinal flora separation and concentration device. Background Art

[0002] The human intestinal flora is a group of bacteria in the digestive tract, mainly in the intestines. It is a huge and extremely active micro-ecological system that changes dynamically with the health of the human body. A typical human intestinal flora can contain hundreds of microbial species, and each species contains multiple strains with different functions. Among them, Bifidobacterium and Bacteroides are the main species, followed by Enterobacter, Enterococcus, Lactobacillus and many other groups that have not yet been isolated or are difficult to culture in vitro.

[0003] There have been reports that the imbalance of human intestinal flora is associated with the occurrence and development of almost all diseases, including metabolic diseases, intestinal diseases, behavioral and cognitive disorders, autoimmune diseases, cancer, etc. Therefore, studying the characteristics of intestinal flora is of great significance for understanding the health status of the human body, but there are still methodological challenges in isolation, culture, genomic analysis and functional analysis research. In particular, in recent years, it has been discovered that the "fecal microbiota transplantation" (FMT) treatment strategy, which transplants functional flora from fecal samples of healthy donors or in vitro modified autologous feces into the recipient's gastrointestinal tract, can reconstruct new intestinal flora in the recipient's gastrointestinal tract, and has a significant therapeutic effect on various intractable intestinal diseases and even aging, obesity and mental diseases.

[0004] It is particularly important to establish a large-scale human intestinal flora separation method, which is a prerequisite for conducting such scientific research and clinical applications. However, the bacterial preparation preparation devices for fecal microbiota transplantation reported in the literature are usually based on conventional laboratory agitators, centrifuges and filtration equipment, and the entire technical process is completed manually by technicians. This type of technology has a low degree of automation, is difficult to standardize, and is time-consuming. The composition and activity of the microbiota are affected to a certain extent, and the manual operation process has a strong odor. There are also equipment that facilitates peristaltic pumps or other power devices to connect the above steps with latex pipes for step-by-step operation. This technology has improved in automation and standardization compared to manual operation, but a large number of pipe connections and pumps make the overall size of the device bulky, and the whole process is time-consuming, the bacterial activity and yield are greatly affected, and there are many disposable consumables and the preparation cost is high. In summary, it is of great significance and application value to design and prepare a device and method that can be used to quickly and efficiently separate and concentrate large sample sizes of human fecal samples so as to freeze-dry them into dry powder to preserve activity. Utility Model Content

[0005] In view of the problems existing in the prior art, the utility model provides a device for separating and concentrating intestinal flora, which can realize rapid and efficient extraction of flora concentrate.

[0006] The utility model is realized by the following technical solutions:

[0007] A device for separating and concentrating intestinal flora, comprising a flora extraction device and a negative pressure drainer, wherein the flora extraction device comprises a plurality of filters connected in sequence from top to bottom;

[0008] The filter includes a filter body and a filter cavity formed therein. A screen is provided at the inner bottom of the filter body. A stirring device is provided inside the filter body. The bottom of the upper filter is detachably connected to the upper end of the lower filter. The mesh size of the screen of each level of filter gradually decreases from the uppermost level to the lowermost level. The lowermost filter is used to enrich and concentrate the intestinal flora suspension separated by the uppermost filter. The negative pressure drainer is connected to the lowermost filter to collect the liquid generated during the enrichment and concentration process of the lowermost filter.

[0009] Preferably, the stirring device of the lower filter is fixedly connected to the bottom of the upper filter.

[0010] Preferably, the filter further comprises a connector, the screen is laid in the connector, the lower end of the filter body is connected to the connector and the screen is press-fitted into the connector.

[0011] Preferably, a raised support platform is provided in the connector, the edge of the screen is arranged on the support platform, and the lower end of the filter body is connected to the connector and sealed and crimped with the screen.

[0012] Preferably, the stirring device comprises a bracket, a motor and a stirring mechanism;

[0013] The bracket of the lower filter is connected to the support platform of the upper filter, the motor is fixed on the bracket, and the output shaft of the motor extends downward and is connected to the stirring mechanism.

[0014] Preferably, the stirring head of the stirring device of the uppermost filter is a multi-petal blunt-edged knife;

[0015] The stirring head of the stirring device of the lowest filter is a flexible stirring head, and the lower end of the flexible stirring head is in contact with the screen;

[0016] The stirring heads of the multiple filters between the uppermost filter and the lowermost filter are hard stirring heads, and the lower ends of the hard stirring heads are in contact with the screen.

[0017] Preferably, the screen of the bottom filter is a filter membrane or a filter screen provided with sieve holes, and the diameter of the sieve holes is smaller than the particle size of the bacterial strain.

[0018] Preferably, a base is provided at the lower end of the lowermost filter, a liquid collecting chamber is formed between the base and the screen of the lowermost filter, and the negative pressure drainer is connected to the liquid collecting chamber through the base.

[0019] Preferably, the pressure drainage device comprises a primary drainage bottle, a secondary drainage bottle and a negative pressure pump;

[0020] The inlet of the first-level drainage bottle is connected to the lowest-level filter through a pipeline, a certain amount of deodorizing liquid for removing odor is poured into the second-level drainage bottle, the outlet of the first-level drainage bottle is connected to the second-level drainage bottle through a pipeline, and the outlet of the pipeline extends into the deodorizing liquid, and the outlet of the second-level drainage bottle is connected to a negative pressure pump.

[0021] A method for extracting intestinal flora using an intestinal flora separation and concentration device comprises the following steps:

[0022] Step 1: Load the sample and solution into the top filter, stir the sample and solution under constant pressure to form a homogenate, filter the homogenate under negative pressure, and the filtered homogenate enters the lower filter;

[0023] Step 2: Under constant pressure, the filters at each level sequentially stir the homogenate leaking from the upper filter, and filter the homogenate step by step under negative pressure, so that the filtered homogenate leaks to the next filter, until the homogenate forms an intestinal flora suspension and enters the lowest filter;

[0024] Step 3: Under negative pressure, the lowest filter separates the liquid in the intestinal flora suspension to obtain an intestinal flora concentrate.

[0025] Compared with the prior art, the utility model has the following beneficial technical effects:

[0026] The utility model provides a device for separating and concentrating intestinal flora, wherein a plurality of filters are connected in sequence to form the flora extraction device, the lowest filter is connected to a negative pressure drainage device, the negative pressure drainage device applies negative pressure to each level of filtration step by step as a filtration power for homogenized slurry, the mesh openings of the filter screens of each level are gradually reduced, the mesh opening diameter of the lowest filter is smaller than the particle size of the bacterial species, the highest filter is used as a homogenizer to stir the sample and the solution to form a homogenized slurry, the middle multi-stage filter is used as a filtering device to filter impurities of different particle sizes in the homogenized slurry step by step The lowest filter is used as an enrichment and concentrator to separate the liquid in the intestinal flora suspension to achieve the extraction of intestinal flora; during the stirring process of each level of filters, the negative pressure drainage device stops working, and during the filtration process, the negative pressure drainage device works to apply negative pressure to each level of filters, so that a pressure difference is generated on both sides of the screen, and under the action of the pressure difference, the upper homogenate leaks to the next level; the flora extraction device integrates the homogenization device, the multi-stage filtration device and the enrichment and concentration device into a whole, which simplifies the process of flora separation and concentration and improves the extraction efficiency of intestinal flora. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a schematic diagram of the structure of the separation and concentration device of the utility model;

[0028] Figure 2 This is the internal structure diagram of the separation and concentration device of the utility model;

[0029] Figure 3 It is a structural schematic diagram of the separation device of the utility model;

[0030] Figure 4 This is the internal structure diagram of the separation device of the utility model;

[0031] Figure 5 This is a schematic diagram of the internal structure of the primary filter in the separation device of the utility model;

[0032] Figure 6 This is a schematic diagram of the internal structure of the two-stage filter of the utility model;

[0033] Figure a is a cross-sectional view of the secondary filter, and Figure b is a schematic diagram of the structure of the bracket;

[0034] Figure 7 This is a schematic diagram of the internal structure of the three-stage filter of the utility model;

[0035] Figure 8 This is a schematic diagram of the internal structure of the four-stage filter of the utility model;

[0036] Fig. 9 This is a structural diagram of the five-stage filter of the utility model;

[0037] Figure a is a cross-sectional view of the five-stage filter, and Figure b is a schematic diagram of the screen installation.

[0038] In the figure: 1. bacterial flora extraction device; 2. primary drainage bottle; 3. secondary drainage bottle; 4. deodorizing liquid; 5. motor; 6. cover plate; 7. filter body; 8. connector; 9. base; 10. primary filter chamber; 11. secondary filter chamber; 12. tertiary filter chamber; 13. quaternary filter chamber; 14. fifth filter chamber; 15. liquid collecting chamber; 16. liquid injection hole; 17. output shaft; 18. multi-petal metal blunt blade stirring head; 19. support platform; 20. bracket; 21. first screen; 22. sealing ring; 23. waterproof adhesive layer; 24. plastic long-handled stirring head; 25. second screen; 26. third screen; 27. fourth screen; 28. flexible stirring head; 29. ​​filter membrane; 30. fixing frame. DETAILED DESCRIPTION

[0039] The present invention will be further described in detail below in conjunction with the accompanying drawings, which are intended to explain the present invention rather than to limit it.

[0040] See also Figure 1-9 , a device for separating and concentrating intestinal flora, comprising a flora extraction device 1 and a negative pressure drain;

[0041] The flora extraction device 1 includes a plurality of filters connected in sequence from top to bottom, the filter includes a filter body 7 and a filter cavity formed therein, a screen is provided at the inner bottom of the filter body 7, a stirring device is provided inside the filter body, the bottom of the upper filter is detachably connected to the upper end of the lower filter, the mesh size of the screen of each level of filter gradually decreases from the uppermost level to the lowermost level, a filter membrane 29 is provided in the lowermost filter for enriching and concentrating the intestinal flora suspension separated in the upper level, and a negative pressure drainer is connected to the lowermost filter for collecting the liquid generated during the enrichment and concentration process of the lowermost filter.

[0042] The bacterial flora extraction device comprises a plurality of filters with screens at the bottom, and a stirring device is loaded in each filter, the uppermost filter is also used as a homogenizing chamber, the stool sample and the dilution solution are injected into the uppermost filter, the stirring device inside the uppermost filter rotates at a high speed, so that the stool sample and the dilution solution are fully mixed to form a homogenized liquid, a negative pressure is applied to each filter by a negative pressure drainer, a pressure difference is generated on both sides of the filter screen, the homogenized liquid leaks down to the lower filter through the uppermost screen, and the aperture of the screen separated at each level is gradually reduced, so that impurities of different sizes in the homogenized liquid are filtered, and an intestinal flora suspension is formed in the lowermost filter after multi-stage separation, the lowermost filter enriches and concentrates the intestinal flora suspension through a filter membrane, and finally the liquid generated in the enrichment and concentration process is collected by the negative pressure drainer. The utility model integrates a homogenizing device, a multi-stage filtering device and an enrichment and concentration device into a whole, simplifies the bacterial flora separation and concentration process, and improves the extraction efficiency of the intestinal flora.

[0043] The filter includes a filter body 7 and a connector 8. A support platform 19 for supporting the screen is formed on the inner wall of the connector 8 and is located at the center of the height of the connector 8. The edge of the screen is fixedly pressed on the support platform. The lower end of the filter body 7 is connected to the connector by threads. The lower end of the filter body 7 is pressed onto the top of the screen through a sealing ring 22, that is, the screen is pressed between the lower end of the filter body 7 and the support platform 19. The lower end of the connector 8 of the upper filter is connected to the upper end of the filter body of the lower filter by threads.

[0044] In this embodiment, the filter body 7 and the connector 8 are both hollow cylindrical structures. The lower end of the filter body 7 is connected to the inner wall of the connector through a thread. The support platform 19 is a circular ring arranged on the inner wall of the connector, and the edge of the screen overlaps the top of the support platform.

[0045] The stirring device of the lower filter is connected to the bottom of the upper filter. When the two adjacent filters are disassembled, the two filters are separated. At the same time, the stirring device of the lower filter is synchronously separated from the corresponding filter body, solving the problem of secondary disassembly of the stirring device.

[0046] The stirring device includes a bracket 20, a motor 5 and a stirring mechanism. The bracket 20 is installed on the support platform of the upper filter and is located at the bottom of the screen. The motor 5 is fixedly connected to the center of the bracket. The output shaft 17 of the motor extends downward. The stirring mechanism is arranged at the lower end of the output shaft. The motor drives the stirring mechanism to rotate to achieve stirring of the homogenized slurry.

[0047] The stirring mechanism in the top filter is a multi-petal metal blunt blade stirring wheel 18, and multiple blunt blades are evenly distributed around the circumference and fixed at the lower end of the output shaft. The rotation of the multi-petal metal blunt blade can separate the food residue and bacteria in the fecal sample without crushing the food residue, thereby preventing the food residue particles from becoming smaller and flowing into the lower filter.

[0048] The stirring mechanism in the lowest-level filter is a flexible stirring head 28, which includes a T-shaped fixing rod, the top center of the fixing rod is fixedly connected to the lower end of the motor output shaft, and the lower end of the T-shaped fixing rod is fixedly connected to a high-temperature resistant foamed silicone rotor. The lower end of the foamed silicone rotor is in contact with the filter membrane. The foamed silicone rotor close to the surface of the filter membrane will not scratch or wear the filter membrane, and at the same time can push the intestinal bacteria trapped on the upper surface of the filter membrane so that it does not block the holes in the filter membrane to ensure that physiological saline and other small molecular impurities can be effectively filtered out.

[0049] The stirring mechanism of the multiple filters in the middle is a hard plastic long-handled stirring head 24, which is parallel and close to the screen at the bottom of the filter cavity, and can push the food residues and other impurities trapped on the upper surface of the screen at the bottom of each filter cavity so that they do not block the screen holes.

[0050] The bracket includes an outer ring and an inner ring arranged concentrically, and the outer ring and the inner ring are connected by a plurality of connecting arms, and the plurality of connecting arms are evenly distributed around the circumference, and the upper end of the motor is fixedly connected to the inner ring. Since the homogenate in the filter is liquid, to avoid damage to the motor, the outside of the motor is coated with a waterproof adhesive layer 23 to block the homogenate, thereby increasing the service life of the motor.

[0051] See also Figure 5 A cover plate 6 is provided on the top filter, a motor is fixed on the top of the cover plate, an output shaft of the motor passes through the cover plate and extends into the filter body, an injection hole 16 is provided on the cover plate 6 for injecting physiological saline to form a homogenous slurry through stirring the fecal sample.

[0052] A base 9 is provided at the lower end of the connector of the lowest-level filter, and the base is threadedly connected to the connector of the lowest-level filter, and a liquid collecting chamber 15 is formed between the base and the filter membrane. The negative pressure drainer is connected to the base and communicates with the liquid collecting chamber 15. A fixing frame 30 is provided on the support platform of the connector of the lowest-level filter, and the filter membrane is laid on the fixing frame, and the edge of the filter membrane is pressed onto the bottom of the filter body.

[0053] In this embodiment, the top filter serves as a homogenizer, and the multiple filters between the top filter and the bottom filter serve as a multi-stage filtering device. The top filter serves as an enrichment concentrator, and the motor speeds in the filters at each stage are independently controlled. The screen in the homogenizer is used to filter large-sized food residues in the homogenate. The filtered homogenate enters the multi-stage filtering device. The apertures of the multiple screens in the multi-stage filtering device gradually decrease, and are used to filter and separate smaller-sized food residues or other particulate impurities step by step to obtain an intestinal flora suspension and enter the enrichment concentrator through the screen. The enrichment concentrator separates the liquid in the intestinal flora suspension through the filter membrane 29 to achieve the concentration of the intestinal flora suspension, and forms an intestinal flora concentrate in the enrichment concentrator. The separated liquid is recovered and processed through a negative pressure drain.

[0054] The negative pressure drainage device comprises a primary drainage bottle 2, a secondary drainage bottle 3 and a negative pressure pump which are connected in series.

[0055] The inlet of the primary drainage bottle 2 is connected to the base through a pipeline, a certain amount of deodorizing liquid 4 for removing odor is poured into the secondary drainage bottle 3, the outlet of the primary drainage bottle 2 is connected to the secondary drainage bottle 3 through a pipeline, and the outlet of the pipeline extends into the deodorizing liquid, and the outlet of the secondary drainage bottle 3 is connected to a negative pressure pump.

[0056] The deodorant liquid 4 includes, by mass percentage, 0.1% surfactant, 10% alkali solution and the rest water, the alkali solution contains 0.01% fragrance agent, the alkali solution can react with organic matter such as indoles in feces to eliminate odor, the surfactant forms a large amount of foam with the pipeline input gas, increases the combination of alkali solution with internal odor molecules, promotes the reaction efficiency of alkali solution with organic matter such as indoles, and the fragrance agent further eliminates the odor of the gas discharged from the final outlet of the negative pressure pump. The deodorant is poured into the secondary drainage bottle, which effectively eliminates the odor generated during the separation and concentration of the bacterial flora, and increases the friendliness of the device to the environment and operators.

[0057] Example 1

[0058] See also Figure 1-2 A device for separating and concentrating intestinal flora comprises a flora extraction device 1 and a negative pressure drainer; the flora extraction device 1 comprises five filters connected in sequence from top to bottom, the top filter serves as a homogenizer, the middle three-stage filter serves as a filtering device, and the bottom filter serves as an enrichment and concentration device; the negative pressure drainer comprises a primary drainage bottle 2, a secondary drainage bottle 3 and a negative pressure pump.

[0059] See also Figure 3 and 4The lowest filter is connected to the inlet of the primary drainage bottle 2 through a hose, the secondary drainage bottle 3 is filled with 1 / 2 of the deodorant 4 of the infusion solvent, the outlet of the primary drainage bottle 2 is connected to the inlet of the secondary drainage bottle through a hose, and the outlet of the hose extends into the deodorant.

[0060] For ease of description, the five-stage filter is respectively a primary filter, a secondary filter, a tertiary filter, a quaternary filter and a fifth-stage filter from top to bottom. The structures of the five-stage filters are basically the same, and the difference lies in the aperture of the sieve holes and the stirring mechanism.

[0061] Figure 5 It is a primary filter, namely a homogenizer, which comprises a motor 5 , a cover plate 6 , a filter body 7 , a connector 8 , a stirring mechanism and a first screen 21 .

[0062] The filter as a whole is a cylindrical structure, the filter body 7 is a stainless steel hollow cylinder, the connector 8 is a circular ring structure, a support platform 19 protruding toward the center of the circle is provided in the middle of the inner wall of the connector, an internal thread is provided on the inner wall of the connector, and is located on both sides of the support platform, the edge of the first screen 21 is overlapped on the support platform, a bracket 20 for the secondary filter is also provided on the support platform, and is located at the bottom of the first screen, and a sealing ring of soft rubber material is also provided on the top of the first screen, an external thread matching the internal thread is provided at the lower end of the outer wall of the filter body 7, the filter body 7 is detachably connected to the connector through threads, and a primary filter chamber 10 is formed in the filter body 7, the cover plate 6 is connected to the upper end of the connector through threads, the motor is fixed at the top center of the cover plate 6, the output shaft 17 of the motor passes through the cover plate and extends into the primary filter chamber 10, and an injection hole 16 is provided on the cover plate for injecting physiological saline into the primary filter chamber.

[0063] The stirring mechanism comprises a multi-petal metal blunt blade stirring wheel 18, and a motor provides power for the multi-petal metal blunt blade stirring wheel to rotate at a high speed, so as to quickly homogenize the physiological saline and the fecal sample, and separate the food residue and the bacterial cells in the baked fecal sample, but does not crush the food residue.

[0064] The aperture of the first screen is 1 mm, and is used to filter out larger food residues and allow the remaining homogenate to flow down into the secondary filter.

[0065] See also Figure 6-8 , respectively a two-stage filter, a three-stage filter and a four-stage filter. The structures of the three filters are the same, but the difference lies in the mesh diameter of the screen. The mesh diameters of the three filters decrease successively. The following only takes the two-stage filter as an example to explain its structure in detail.

[0066] See again Figure 6The secondary filter includes a motor 5, a cover plate 6, a filter body 7, a connector 8, a stirring mechanism, a second screen 21 and a bracket 20 for fixing the motor.

[0067] The bracket is installed on the support platform of the first-level filter, the motor is fixed at the center of the bracket, the lower end of the motor output shaft is connected to the stirring mechanism, the filter body 7 is connected to the connector through threads, and the second screen 21 is pressed onto the support platform of the connecting machine to form a secondary filter chamber 11 in the filter body 7.

[0068] The stirring mechanism of the secondary filter is a hard plastic long-handled stirring head 24, the top center of the plastic long-handled stirring head is fixedly connected to the output shaft, the two ends of the plastic long-handled stirring head extend toward the edge of the secondary filter cavity, and the lower end of the plastic long-handled stirring head contacts the surface of the second screen, which can push the food residues and other impurities trapped on the upper surface of the second screen to avoid clogging of the mesh.

[0069] The mesh size of the second screen is 500 microns, which is used to retain larger-sized food and other impurities with a particle size of more than 500 microns in the secondary filtration cavity, while particles with a particle size of less than 500 microns enter the tertiary cavity with the saline.

[0070] See also Figure 7 , which is a three-stage filter, a three-stage filter chamber 12 is formed inside the filter, and the mesh diameter of the third screen 26 in the filter is 150 microns. The third screen 26 retains smaller-sized food and other impurities with a particle size of more than 150 microns in the three-stage filter chamber, while particles with a particle size of less than 150 microns enter the four-stage chamber with the saline.

[0071] See also Figure 8 , is a four-stage filter, a four-stage filter chamber 13 is formed inside the filter, the mesh diameter of the fourth screen 27 in the filter is 50 microns, the fourth screen 27 retains small-sized food and other impurities with a particle size of more than 50 microns in the four-stage filter chamber, and at the same time, particles with a particle size of less than 50 microns, that is, various groups of intestinal bacteria, enter the five-stage chamber with the physiological saline to form an intestinal flora suspension.

[0072] See also Fig. 9 , is a five-stage filter, i.e., an enrichment and concentration device, whose main function is to separate the physiological saline in the intestinal flora suspension to form an intestinal flora concentrate.

[0073] The five-stage filter differs from the four-stage filter in the screen and the stirring mechanism, and a base 9 is provided at the bottom of the connector.

[0074] The screen in the five-stage filter is a filter membrane 29. A fixing frame 30 is provided on the support platform of the filter. The filter membrane 29 is laid on the fixing frame. The lower end of the filter body is pressed onto the edge of the filter membrane through a sealing ring. The lower end of the connector is connected to the base 9 through a thread, and a liquid collecting chamber 15 is formed between the base and the filter membrane. The first-stage drainer is connected to the base through a hose, so that the liquid in the liquid collecting chamber enters the drainage bottle under the action of negative pressure.

[0075] The lower end of the motor output shaft is connected to a flexible stirring head 28. The flexible stirring head 28 is close to the foamed silicone rotor on the surface of the filter membrane and will not scratch or wear the filter membrane. At the same time, it can push the intestinal bacteria trapped on the upper surface of the filter membrane so that they do not block the holes in the filter membrane to ensure that physiological saline and other small molecule impurities can be effectively filtered out, and the filtered liquid and small molecules enter the liquid collecting chamber 15.

[0076] In another embodiment, the mesh in the five-stage filter is a mesh with a pore size of 0.22 microns, which retains various groups of intestinal bacteria with a particle size of more than 0.22 microns in the five-stage filter cavity 14, while small molecular impurities with a particle size of less than 0.22 microns enter the waste liquid bottle with the physiological saline, and all bacteria eventually form a thick liquid in the five-stage filter cavity to form an intestinal flora concentrate.

[0077] Example 2

[0078] A method for extracting intestinal flora concentrate using the intestinal flora separation and concentration device comprises the following steps:

[0079] Step 1: Place a stool sample in the primary filter and inject saline.

[0080] When the negative pressure pump is turned off, open the cover of the primary filter and put a stool sample, such as 100g, in the primary filter chamber. Then inject 1000ml of normal saline through the injection hole of the cover and turn on the homogenization drive motor for high-speed homogenization. During the homogenization process, the negative pressure pump is always in the closed state. There is no pressure difference on both sides of the first screen. Therefore, the homogenate in the primary filter chamber does not leak or leaks a small amount, and most of the homogenate will not pass through the first screen into the secondary filter.

[0081] After the separation and homogenization operation is completed, turn on the bottom negative pressure pump. At this time, a pressure difference is generated above and below the bottom screen of the first filter chamber, and the homogenized liquid begins to leak into the secondary filter chamber of the secondary filter. The first screen retains large-sized food and other impurities with a particle size of more than 1 mm in the first filter chamber, while particles with a particle size of less than 1 mm enter the secondary filter chamber with the physiological saline.

[0082] Step 2: The secondary filter stirs and filters the homogenate flowing into the primary filter, and the filtered homogenate enters the tertiary filter.

[0083] The secondary filter stirs and filters the homogenate, and the second screen with a pore size of 500 microns filters the homogenate. Under negative pressure, a pressure difference is formed above and below the second screen, and the homogenate in the secondary filter chamber begins to leak down to the tertiary filter chamber. The second screen retains larger food and other impurities with a particle size of more than 500 microns in the secondary chamber. At the same time, particles with a particle size of less than 500 microns enter the tertiary filter chamber of the tertiary filter with the physiological saline.

[0084] Step 3: The third-stage filter stirs and filters the homogenate flowing into the second-stage filter, and the filtered homogenate enters the fourth-stage filter.

[0085] The tertiary filter stirs and filters the homogenate, and the third screen with a pore size of 150 microns filters the homogenate. Under negative pressure, a pressure difference is formed above and below the third screen, and the homogenate in the tertiary filter chamber begins to leak down to the fourth filter chamber. The third screen retains larger food and other impurities with a particle size of more than 150 microns in the secondary chamber. At the same time, particles with a particle size of less than 150 microns enter the fourth filter chamber of the fourth filter with the saline.

[0086] Step 4: The fourth-stage filter stirs and filters the homogenate flowing into the third-stage filter, and the filtered homogenate enters the fifth-stage filter.

[0087] The four-stage filter stirs and filters the homogenate, and the fourth screen with a pore size of 50 microns filters the homogenate. Under negative pressure, a pressure difference is formed above and below the fourth screen, and the homogenate in the four-stage filter chamber begins to leak down to the five-stage filter chamber. The fourth screen retains larger-sized food and other impurities with a particle size of more than 50 microns in the secondary chamber. At the same time, particles with a particle size of less than 50 microns (various groups of intestinal bacteria) enter the five-stage filter chamber of the five-stage filter with the physiological saline, that is, the intestinal flora suspension enters the five-stage filter chamber.

[0088] Step 5: The five-stage filter is used to enrich and concentrate the intestinal flora suspension to obtain an intestinal flora concentrate.

[0089] The four-stage filter stirs and filters the intestinal flora suspension. Under negative pressure, a pressure difference is formed on both sides of the 0.22-micron screen. The liquid in the intestinal flora suspension leaks down through the filter membrane into the liquid collection chamber and is sucked into the drainage bottle under the action of negative pressure. The screen retains various intestinal bacteria with a particle size of more than 0.22 microns in the five-stage chamber. At the same time, small molecular impurities with a particle size of less than 0.22 microns enter the liquid collection chamber with the physiological saline until the intestinal flora suspension forms a thick liquid to obtain an intestinal flora concentrate.

[0090] Step 6: The liquid produced by the five-stage filter enters the negative pressure drainer under the action of negative pressure for deodorization.

[0091] The liquid produced by the five-stage filter enters the primary drainage bottle 2 under the action of negative pressure, and the liquid in the primary drainage bottle enters the deodorant in the secondary drainage bottle under the action of negative pressure.

[0092] The 10% alkali solution in the deodorant can react with indoles and other organic substances in feces to eliminate odor. The 0.1% surfactant contained in the deodorant forms a large amount of foam after the gas in the conduit below the liquid surface overflows, increasing the combination of the alkali solution with the odor molecules in the conduit and promoting the reaction efficiency of the alkali solution with indoles and other organic substances. The 0.01% fragrance agent contained in the deodorant can further eliminate the odor of the gas discharged from the final outlet of the negative pressure pump.

[0093] Example 3

[0094] A method for preparing intestinal flora dry powder comprises the following steps:

[0095] The intestinal flora concentrate is cryogenically frozen at below -80°C until the intestinal flora concentrate forms a solid structure, and then the solid intestinal flora concentrate is vacuum freeze-dried until the solid intestinal flora concentrate is dry, and the dried solid intestinal flora concentrate is made into powder to obtain intestinal flora dry powder.

[0096] Specifically, the five-stage filter containing the intestinal flora concentrate is pre-frozen in a -80°C low-temperature refrigerator overnight until the intestinal flora concentrate forms a solid structure.

[0097] Turn on the compressor of the vacuum freeze dryer. After 30 minutes, the temperature of the cold hydrazine drops below -50 degrees Celsius. Place the pre-frozen chamber and the concentrated bacterial liquid in the sample placement chamber of the vacuum freeze dryer, cover with a transparent sealing cover, turn on the vacuum oil pump, and check the airtightness. Start timing when the pressure in the vacuum chamber drops below 100 Pa. Turn off the machine overnight or 24 hours later to check the status until the bacterial liquid is freeze-dried, grind into powder, put into capsules or glass bottles, and store in a refrigerator at 4 degrees Celsius.

[0098] Intestinal flora dry powder activity test

[0099] After being stored for different periods of time, the intestinal flora dry powder was taken out, and a certain weight or a capsule of intestinal flora dry powder was weighed under sterile conditions. The dry powder was resuspended in 1 ml of liquid LB culture medium that had been sterilized by moist heat to prepare a bacterial suspension. 50 μl was transferred to a plate loaded with a fixed LB culture medium layer using a pipette, spread evenly, waited to dry, and inverted in an incubator at 37 degrees Celsius overnight to observe the colony formation.

[0100] The utility model provides an intestinal flora separation and concentration device, which is composed of multiple stages of independent filters connected in series, and two adjacent filters are detachably sealed to form an integral flora extraction device, in which a stool sample and physiological saline are stirred and mixed in a primary filter to form a homogenate and filtered once, and under a negative pressure state, the homogenate is filtered step by step through three filters using screens of different apertures until an intestinal flora suspension is formed, and finally the intestinal flora suspension is concentrated through the lowest filter, the device integrates the existing homogenizer, filter and enrichment concentrator into a flora extraction device, and each filter is detachably connected, which greatly reduces the number of extraction devices and simplifies the extraction process of intestinal flora concentrate, solves the problems of manual sample transfer or multi-stage pipeline connection and multi-stage pump suction continuous operation in the existing conventional technology of homogenization, multi-stage filtration, enrichment and concentration independent instrument operation, so that the functional modules of homogenization, multi-stage filtration, enrichment and concentration are integrated into one device, so that the operation is simpler, less consumables are used, and the flora separation and concentration efficiency is higher.

[0101] Then, the motor in each filter is fixed to the bottom of the filter of the previous level through a bracket. When the two adjacent levels of filters are separated, the stirring device is separated at the same time, and a waterproof rubber layer is coated on the outside of the motor to increase the service life of the motor. The motors of each level of filters are independently controlled, and the stirring speed is controlled according to the state of the slurry in each filter chamber to avoid damage to the homogenous slurry.

[0102] Secondly, different structures of stirring devices are used in the filters at each level. The first-level filter uses a multi-petal metal blunt knife stirring head to quickly homogenize the fecal sample of saline, so that the food residue and bacteria in the baked fecal sample are separated, but the food residue is not crushed; the second-level filter, the third-level filter and the fourth-level filter use a hard stirring head, and the bottom surface of the stirring head is in contact with the screen, which can push the food residue and other impurities trapped on the upper surface of the screen at the bottom of each chamber so that they do not block the screen holes; the fifth-level filter uses a flexible stirring head, and the foamed soft silicone rotor close to the surface of the filter membrane will not scratch and wear the filter membrane, and at the same time can push the trapped intestinal bacteria so that they do not block the holes of the filter membrane to ensure that saline and other small molecular impurities can be effectively filtered out. Each motor controls the stirring head to rotate at a corresponding speed to achieve sample homogenization, accelerate the graded filtration and enrichment concentration of the sample, and solve the problems of inefficiency and cumbersomeness of existing devices and operating methods.

[0103] In addition, the utility model adopts negative pressure technology to drain each filter in the bacterial flora extraction device. The negative pressure pump applies a continuous negative pressure difference to each filter chamber from bottom to top, so that the slurry in each stage of the filter leaks down step by step to the filter of the next stage, thereby realizing step-by-step filtration of the slurry, avoiding the need for independent peristaltic pumps or centrifugal force or vacuum negative pressure and other driving forces for each step in conventional operation and the device. The device is simpler and more efficient.

[0104] Finally, a negative pressure drainer is set at the lower end of the bacterial community extraction device. The two drainage bottles connected in series collect the liquid under the action of negative pressure, and then neutralize the odorous gas in the liquid, effectively eliminating the odor generated during the bacterial community separation and concentration process, and increasing the friendliness of the device to the environment and operators.

[0105] The above content is only for explaining the technical idea of ​​the utility model and cannot be used to limit the protection scope of the utility model. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the utility model shall fall within the protection scope of the claims of the utility model.

Claims

1. A device for separating and concentrating intestinal flora, characterized in that: It comprises a bacterial flora extraction device (1) and a negative pressure drainer, wherein the bacterial flora extraction device (1) comprises a plurality of filters connected in sequence from top to bottom; The filter comprises a filter body (7) and a filter cavity formed inside the filter body, a screen is provided at the inner bottom of the filter body (7), a stirring device is provided inside the filter body, the bottom of the upper filter is detachably connected to the upper end of the lower filter, the mesh size of the screen of each filter level is gradually reduced from the uppermost level to the lowermost level, the lowermost filter is used to enrich and concentrate the intestinal flora suspension separated by the uppermost filter, and the negative pressure drainer is connected to the lowermost filter to collect the liquid generated during the enrichment and concentration process of the lowermost filter.

2. The intestinal flora separation and concentration device according to claim 1, characterized in that: The stirring device of the lower filter is fixedly connected to the bottom of the upper filter.

3. The intestinal flora separation and concentration device according to claim 1, characterized in that: The filter further comprises a connector (8), the screen is laid in the connector, the lower end of the filter body (7) is connected to the connector (8) and the screen is pressed into the connector.

4. The intestinal flora separation and concentration device according to claim 3, characterized in that: A raised support platform is arranged in the connector, the edge of the screen is arranged on the support platform, and the lower end of the filter body (7) is connected to the connector and sealed and crimped with the screen.

5. The intestinal flora separation and concentration device according to claim 4, characterized in that: The stirring device comprises a bracket, a motor and a stirring mechanism; The bracket of the lower filter is connected to the support platform of the upper filter, the motor is fixed on the bracket, and the output shaft of the motor extends downward and is connected to the stirring mechanism.

6. The intestinal flora separation and concentration device according to claim 3, characterized in that: The two ends of the connector are provided with threads, and the lower end of the upper filter and the upper end of the lower filter are connected to the two ends of the filter respectively.

7. The intestinal flora separation and concentration device according to claim 1, characterized in that: The stirring head of the stirring device of the top filter is a multi-petal blunt-edged knife; The stirring head of the stirring device of the lowest filter is a flexible stirring head, and the lower end of the flexible stirring head is in contact with the screen; The stirring heads of the multiple filters between the uppermost filter and the lowermost filter are hard stirring heads, and the lower ends of the hard stirring heads are in contact with the screen.

8. The intestinal flora separation and concentration device according to claim 1, characterized in that: The screen of the bottom filter is a filter membrane or a filter screen with sieve holes, and the diameter of the sieve holes is smaller than the particle size of the bacterial strain.

9. The intestinal flora separation and concentration device according to claim 1, characterized in that: A base is provided at the lower end of the bottom filter, a liquid collecting chamber (15) is formed between the base and the screen of the bottom filter, and the negative pressure drainer is connected to the liquid collecting chamber (15) through the base.

10. The intestinal flora separation and concentration device according to claim 1, characterized in that: The pressure drainage device comprises a primary drainage bottle (2), a secondary drainage bottle (3) and a negative pressure pump; The inlet of the first-stage drainage bottle (2) is connected to the lowest-stage filter via a pipeline, a certain amount of deodorizing liquid (4) for removing odor is poured into the second-stage drainage bottle (3), the outlet of the first-stage drainage bottle (2) is connected to the second-stage drainage bottle (3) via a pipeline, and the outlet of the pipeline extends into the deodorizing liquid, and the outlet of the second-stage drainage bottle (3) is connected to a negative pressure pump.

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