Intestinal bacterium directional separation system and method based on microbial antibody omics technology

Abstract

The invention discloses an intestinal bacteria directional separation system and method based on a microbial antibody omics technology, and relates to the technical field of biomedicine. The device comprises a negative pressure driving mechanism, a filtration and impurity removal device for performing filtration and impurity removal treatment on an excrement sample, a harmful bacterium adsorption assembly for removing harmful bacteria in a microorganism mixed solution, and a beneficial bacterium enrichment assembly for performing forward enrichment treatment on beneficial bacteria in the microorganism mixed solution, the beneficial bacterium enrichment assembly is used for enriching beneficial bacterium coupling bodies, the beneficial bacterium dissociation assembly is used for dissociating the beneficial bacterium coupling bodies, the beneficial bacterium collection bottle is used for collecting beneficial bacteria, and the negative pressure driving mechanism is used for enabling the beneficial bacteria in excrement samples to automatically enter the beneficial bacterium collection bottle after sequentially passing through the filtering and impurity removing device, the beneficial bacterium enrichment assembly and the beneficial bacterium dissociation assembly. According to the application, the target beneficial bacteria can be directly and selectively screened out from the excrement sample, and the requirements of precise transplantation of intestinal flora and precise treatment of specific related diseases of the intestinal tract in the future can be met.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and particularly relates to an intestinal bacteria directional separation system and method based on microbial antibodyomics technology. Background Art

[0002] At present, the existing intestinal microorganism separation and collection equipment mainly filters and removes impurities from feces to separate and collect intestinal microorganisms from feces. There are mainly the following two implementation methods: The first method is to use a multi-stage filtration device, and domestic patent reports such as CN105624027B, CN105624024B, CN108676704B, CN209052694U, and CN306895532S, etc.; The second method is to use nuclear pore membranes with different pore sizes, and domestic patent reports such as CN108949631A, CN108676704A, etc.

[0003] However, due to the high diversity and complexity of fecal samples themselves, the substances collected by the above two filtration and impurity removal methods are all intestinal microorganism mixtures (including various microorganisms such as beneficial bacteria, harmful bacteria, fungi, viruses, etc.), and they cannot directly and selectively screen out the target beneficial bacteria from fecal samples, making it difficult to meet the needs of future precise transplantation of intestinal flora and precise treatment of specific intestinal-related diseases. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an intestinal bacteria directional separation system and method based on microbial antibodyomics technology to directly and selectively screen out the target beneficial bacteria from fecal samples.

[0005] In the first aspect, to achieve the above purpose, the technical solution designed by the present invention is as follows:

[0006] An intestinal bacteria directional separation system based on microbial antibodyomics technology, which includes a negative pressure driving mechanism, a filtration and impurity removal device for filtering and removing impurities from fecal samples, a beneficial bacteria enrichment component for positively enriching beneficial bacteria in the microorganism mixture, a beneficial bacteria dissociation component for dissociating beneficial bacteria conjugates, and a beneficial bacteria collection bottle for collecting beneficial bacteria. The negative pressure driving mechanism is used to enable the beneficial bacteria in the fecal sample to automatically enter the beneficial bacteria collection bottle after passing through the filtration and impurity removal device, the beneficial bacteria enrichment component, and the beneficial bacteria dissociation component in sequence.

[0007] In the second aspect, to achieve the above purpose, the technical solution designed by the present invention is as follows:

[0008] An automatic filtering, enrichment and adsorption method for intestinal microorganisms, comprising the following steps: under the action of a negative pressure driving mechanism, a filtering and impurity removing device performs filtering and impurity removing treatment on a fecal sample to obtain a microbial mixed solution; under the action of the negative pressure driving mechanism, a beneficial bacteria enrichment component performs positive enrichment treatment on the beneficial bacteria in the microbial mixed solution to obtain a bacterial liquid mixed solution containing a beneficial bacteria coupling body, and the beneficial bacteria coupling body is formed by coupling beneficial bacteria and second antibody magnetic beads; under the action of the negative pressure driving mechanism, a beneficial bacteria dissociation component first dissociates the beneficial bacteria coupling body to obtain a beneficial bacteria solution containing second antibody magnetic beads and beneficial bacteria; the beneficial bacteria dissociation component then removes the second antibody magnetic beads in the beneficial bacteria solution to obtain a beneficial bacteria solution; under the action of the negative pressure driving mechanism, a beneficial bacteria collection bottle collects the beneficial bacteria solution.

[0009] The present application provides an intestinal bacteria directional separation system and method based on microbial antibody group technology, which can directly and selectively screen out target beneficial bacteria from fecal samples, and lay a solid foundation for future precise transplantation of intestinal flora and / or precise treatment of specific intestinal-related diseases. Brief Description of the Drawings

[0010] Figure 1 It is a schematic structural diagram of the intestinal bacteria directional separation system based on microbial antibody group technology described in the present application;

[0011] Figure 2 It is a schematic structural diagram of the filtering and impurity removing device;

[0012] Figure 3 It is a schematic structural diagram when the first filtering cover plate and the second filtering cover plate of the filtering and impurity removing box are opened;

[0013] Figure 4 For Figure 3 The front view without the first filtering cover plate and the second filtering cover plate;

[0014] Figure 5 For Figure 3 The rear view without the first filtering cover plate and the second filtering cover plate;

[0015] Figure 6 It is a schematic semi-sectional structural diagram of the primary filter or the secondary filter;

[0016] Figure 7 It is a schematic flow diagram of how the filtering and impurity removing device obtains a microbial mixed solution from a fecal sample;

[0017] Figure 8 It is a schematic structural diagram of the enrichment and adsorption device;

[0018] Figure 9 It is a schematic structural diagram of the enrichment and adsorption box without the first enrichment cover plate and the second enrichment cover plate;

[0019] Figure 10 is Figure 9 front view;

[0020] Figure 11 is Figure 9 rear view;

[0021] Figure 12 is a schematic flow chart of how the enrichment adsorption device obtains beneficial bacteria from the microbial mixture.

[0022] In the figure: 100, filtering and impurity removal device; 101, first printer; 102, first fan; 103, first buzzer; 104, first spotlight; 105, first lighting lamp; 106, first filter cover plate; 107, second filter cover plate; 108, filtering and impurity removal box; 111, first operation area; 112, first clean area; 113, first pollution area; 114, first backstage area; 121, first filter partition; 122, second filter partition; 123, first filter support plate; 124, second filter support plate; 125, third filter support plate; 126, fourth filter support plate; 130, shredding component; 131, sample tank; 132, shredding motor; 133, shredding coupling; 140, filtering component; 141, primary filter; 142, secondary filter; 150, dilution bottle; 160, first negative pressure component; 161, first filter peristaltic pump; 162, second filter peristaltic pump; 171, first purification bottle; 172, first negative pressure pump; 173, first buffer; 1731, first buffer bottle; 1732, second buffer bottle; 174, first filter; 175, first control valve; 1732, second control valve; 181, bacterial liquid collection rack; 182, bacterial liquid collection bottle; 191, first display controller; 192, first nitrogen cabinet; 193, first consumable cabinet;

[0023] 200. Enrichment adsorption device; 201. Second printer; 202. Second fan; 203. Second buzzer; 204. Second spotlight; 205. Second lighting lamp; 206. First enrichment cover plate; 207. Second enrichment cover plate; 208. Enrichment adsorption box; 210. Harmful bacteria adsorption component; 211. First antibody magnetic beads; 212. Harmful bacteria magnetic bead tank; 213. Harmful bacteria adsorption bottle; 214. Harmful bacteria coupling component; 2141. First stirring motor; 2142. First material port; 2143. First stirring shaft; 2144. First stirring blade; 215. Harmful bacteria adsorbing part; 2151. First electromagnetic induction coil; 2152. First coil controller; 220. Beneficial bacteria enrichment component; 221. Second antibody magnetic beads; 222. Beneficial bacteria magnetic bead tank; 223. Beneficial bacteria enrichment bottle; 224. Beneficial bacteria coupling component; 2241. Second stirring motor; 2242. Second material port; 2243. Second stirring shaft; 2244. Second stirring blade; 225. Beneficial bacteria enriching part; 2251. Second electromagnetic induction coil; 2252. Second coil controller; 226. Waste liquid collection bottle; 230. Beneficial bacteria dissociation component; 232. Beneficial bacteria dissociation tank; 233. Magnetic bead adsorption bottle; 234. Beneficial bacteria dissociation part; 2341. Third stirring motor; 2342. Third material port; 2343. Third stirring shaft; 2344. Third stirring blade; 235. Magnetic bead adsorbing part; 2351. Third electromagnetic induction coil; 2352. Third coil controller; 240. Beneficial bacteria collection bottle; 250. Second negative pressure component; 251. First enrichment peristaltic pump; 252. Second enrichment peristaltic pump; 253. Third enrichment peristaltic pump; 261. First enrichment partition board; 262. Second enrichment partition board; 263. Second operation area; 264. Second background area; 265. Second clean area; 266. Second pollution area; 271. Second purification bottle; 272. Second negative pressure pump; 273. Second buffer; 2731. Third buffer bottle; 2372. Fourth buffer bottle; 274. Second filter; 275. Third control valve; 276. Fourth control valve; 280. Second display controller; 291. Second nitrogen cabinet; 292. Second consumables cabinet. Detailed implementation manners

[0024] The present invention will be further described in detail below in conjunction with specific embodiments for the understanding of those skilled in the art.

[0025] The immunomagnetic bead enrichment and separation technology is a technique for separating target molecules or cells by utilizing the principle of specific binding between antigens and antibodies, which is a conventional method in this field. The immunomagnetic bead enrichment and separation technology uses magnetic bead microspheres composed of core metal particles prepared from iron tetroxide or iron oxide, a polymer material, and a functional ligand. The chemical groups on the surface of the microspheres can be covalently coupled with antibodies to form a stable and firm immunomagnetic bead-antibody conjugate. The antibody can specifically bind to the target protein or the antigen on the cell surface through an immune reaction to form a microorganism-immunomagnetic bead-antibody complex. Iron oxide endows the magnetic beads with superparamagnetism. The magnetic bead microspheres are magnetized and acquire magnetism in a magnetic field environment and aggregate, while the magnetism disappears and the magnetic beads disperse when leaving the magnetic field environment. Under the action of an external magnetic field, the immunomagnetic beads can quickly aggregate the magnetic beads, facilitating the separation of the liquid.

[0026] Therefore, due to its advantages of high specificity, simple operation, and high purity, the immunomagnetic bead separation technology is currently often used in cell separation, immunoassay, molecular biology assay, microbiology assay, and other aspects.

[0027] The interaction between humans and microorganisms is extremely close. Soon after a newborn is born, a large number of external microorganisms begin to attach to its body surface and intestines. Most of these microorganisms adapt to the human body and eventually form a symbiotic relationship that lasts throughout life. Among these symbiotic microorganisms, about 100 trillion are colonized in the human intestine, covering various microorganisms such as bacteria, fungi, and viruses. The intestinal microbiota is the most complex and largest microecosystem in the human body and is known as the eighth organ of the human body. The genome of the intestinal microbiota contains approximately 5 million genes, about 150 times the number of human genes. Due to its huge quantity and rich variety characteristics, the intestinal microbiota is called the "second gene pool" of the human body.

[0028] The intestinal microbiota can produce short-chain fatty acids (SCFAs) by fermenting dietary fiber that is difficult for humans to digest, providing energy for colon epithelial cells, regulating the interaction between innate immunity and adaptive immunity, and the host's defense mechanism against intestinal pathogens. A normal and stable intestinal microbiota plays a key role in the body's defense against infection, maintaining the normal intestinal barrier, immunity, metabolism, nutrition, and maintaining internal environmental homeostasis through interactions with multiple systems such as the human immune, endocrine, and nervous systems.

[0029] Currently, the fecal microbiota transplantation (FMT) technology has gradually received extensive attention. In 2023, FMT was officially included in the "Notice on Issuing the Technical Specifications for National Medical Service Items" issued by the National Health Commission. As an innovative treatment method, the intestinal microbiota transplantation technology (FMT) can provide a new approach for the treatment of various diseases by reconstructing the intestinal microecological balance.

[0030] Fecal microbiota transplantation (FMT) originated from the works "Elbow Emergency Prescriptions" in the Eastern Jin Dynasty and "Plain Questions" in the Spring and Autumn and Warring States periods. Traditional Chinese medicine used human feces (recorded as Huanglong Decoction / Golden Juice) to treat intestinal diseases. In 1958, surgeon Eiseman used human fecal enema to treat patients with pseudomembranous colitis. In 2013, the American medical guidelines announced that human feces could be used for the treatment of Clostridium difficile.

[0031] Since then, intestinal microbiota transplantation technology has shown significant efficacy in the treatment of gastrointestinal diseases such as irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), recurrent Clostridium difficile infection (CDI), ulcerative colitis (UC), functional constipation (FC), etc. With the deepening of research and the continuous update of technical levels such as high-throughput sequencing, artificial intelligence big data analysis, and intelligent production systems, the number of its clinical application cases has also been increasing, showing broad application prospects.

[0032] Currently, for medical staff, how to isolate intestinal microbiota (including intestinal flora) from feces is a huge challenge. How to achieve the isolation and collection of intestinal microbiota is of great significance for medical staff, which can lay a solid foundation for the precise transplantation of intestinal flora and the precise treatment of specific intestinal-related diseases.

[0033] Existing intestinal microbiota isolation and collection devices mainly filter and remove impurities from feces to isolate and collect intestinal microbiota. The fecal bacteria obtained by separation are only the flora (including various microorganisms such as beneficial bacteria, harmful bacteria, fungi, viruses, etc.) that have removed fecal residues and metabolites. It is impossible to selectively screen specific intestinal microbiota. It generally relies on algorithms to achieve a certain degree of matching, and the process adjustment of the flora separation and purification process is very limited. The effect of flora transplantation is severely restricted by the donor's own flora structure, and potential trace undetected harmful pathogens also bring huge clinical risks, making it difficult to meet the needs of precise treatment of intestinal flora diseases.

[0034] Such as Figure 1As shown in the figure, in order to more efficiently and selectively remove harmful bacteria and enrich beneficial bacteria in the fecal microbiota transplantation technology, and thus lay a solid foundation for the precise transplantation of intestinal bacteria in the future, the present invention provides an intestinal bacteria directional separation system based on microbial antibodyomics technology, which includes a negative pressure driving mechanism, a filtering and impurity removing device 100 for filtering and impurity removing treatment of fecal samples, a beneficial bacteria enrichment component 220 for positively enriching beneficial bacteria in the microbial mixture, a beneficial bacteria dissociation component 230 for dissociating beneficial bacteria conjugates, and a beneficial bacteria collection bottle 240 for collecting beneficial bacteria. The negative pressure driving mechanism is used to make the beneficial bacteria in the fecal sample automatically enter the beneficial bacteria collection bottle 240 after passing through the filtering and impurity removing device 100, the beneficial bacteria enrichment component 220 and the beneficial bacteria dissociation component 230 in sequence.

[0035] In this application, an antibody with the function of identifying harmful bacteria (such as Salmonella) is conjugated to magnetic beads to form the first antibody magnetic beads; at the same time, an antibody with the function of identifying beneficial bacteria (such as Bacteroides uniformis) is conjugated to magnetic beads to form the second antibody magnetic beads. The methods for obtaining the above first antibody magnetic beads and second antibody magnetic beads are prior arts and will not be elaborated here.

[0036] After preparing the above first antibody magnetic beads and second antibody magnetic beads, first process the fecal bacteria through the filtering and impurity removing device to obtain an intestinal microbial mixture (including various microorganisms such as beneficial bacteria, harmful bacteria, fungi, and viruses); then enrich the beneficial bacteria in the fecal bacteria through the beneficial bacteria enrichment component 220, and then achieve antigen-antibody dissociation through the beneficial bacteria dissociation component 230 to obtain the target beneficial bacteria from the fecal bacteria. Preferably, this application can also remove harmful bacteria in the fecal bacteria through the harmful bacteria adsorption component to improve the purity of the beneficial bacteria solution.

[0037] This application greatly improves the sorting efficiency of target probiotics, especially has a positive significance for the precise capture of low-abundance strains, and can make the separation and screening work of fecal bacteria for precise transplantation more efficient and specific, thereby promoting the improvement of production efficiency.

[0038] During actual work, the negative pressure driving mechanism includes a first negative pressure group 160 arranged in the filtering and impurity removing box 108 and a second negative pressure component 250 arranged in the enrichment and adsorption box 208. The first filtering support plate 123 and the second filtering support plate 124 can be collectively referred to as the filtering support plate. The filtering and impurity removing device 100 includes components such as the filtering support plate, a shredding component 130, and a filtering component 140. The harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, the beneficial bacteria dissociation component 230, and the beneficial bacteria collection bottle 240 can be collectively referred to as the enrichment and adsorption device 200.

[0039] Such as Figure 4 and Figure 5As shown in the figure, the first negative pressure assembly 160 includes a first filtered peristaltic pump 161 installed between the dilution bottle 150 and the sample tank 131, and a second filtered peristaltic pump 162 installed between the primary filter 141 and the secondary filter 142.

[0040] During actual operation, the first filtered peristaltic pump 161 can be installed in the first backstage area 114, and the second filtered peristaltic pump 162 can be installed in the first contamination area 113. Both the first filtered peristaltic pump 161 and the second filtered peristaltic pump 162 are electrically connected to the first display controller 191. The dilution bottle 150, the first filtered peristaltic pump 161, the sample tank 131, the primary filter 141, the second filtered peristaltic pump 162, the secondary filter 142, and the bacterial liquid collection bottle 182 are sequentially connected by pipelines.

[0041] As Figure 9 and Figure 10 shown in the figure, the second negative pressure assembly 250 includes a first enrichment peristaltic pump 251 installed between the harmful bacteria adsorption assembly 210 and the beneficial bacteria enrichment assembly 220, a second enrichment peristaltic pump 252 installed between the beneficial bacteria enrichment assembly 220 and the beneficial bacteria dissociation assembly 230, and a third enrichment peristaltic pump 253 installed between the beneficial bacteria dissociation assembly 230 and the beneficial bacteria collection bottle 240.

[0042] During actual operation, the first enrichment peristaltic pump 251, the second enrichment peristaltic pump 252, and the third enrichment peristaltic pump 253 can all be installed in the second contamination area 266 (as shown in Figure 9 and Figure 10 the figure). The first enrichment peristaltic pump 251, the second enrichment peristaltic pump 252, and the third enrichment peristaltic pump 253 are all electrically connected to the second display controller 280. The harmful bacteria magnetic bead tank 212, the first enrichment peristaltic pump 251, the harmful bacteria adsorption bottle 213, the beneficial bacteria magnetic bead tank 222, the second enrichment peristaltic pump 252, the beneficial bacteria enrichment bottle 223, the beneficial bacteria dissociation tank 232, the third enrichment peristaltic pump 253, the magnetic bead adsorption bottle 233, and the beneficial bacteria collection bottle 240 are sequentially connected by pipelines.

[0043] The filtering and impurity removing device 100 and the enrichment and adsorption device 200 of the present application can be used separately or combined. When the filtering and impurity removing device 100 is used alone, it can separate and collect intestinal microorganisms from fecal samples. When the enrichment and adsorption device 200 is used alone, it can enrich and collect beneficial bacteria from the microbial mixture. When the filtering and impurity removing device 100 and the enrichment and adsorption device 200 are used together, they can separate and collect beneficial bacteria from fecal samples.

[0044] During actual operation, to make the present application more convenient, cleaner, tidier and more hygienic, through holes can be provided on the first filter partition 121 and / or the second filter partition 122 to allow pipelines to pass through. This is a conventional means in the art and will not be elaborated here.

[0045] As Figure 3 shown, the filtering and impurity removing device 100 of the present application includes a filtering and impurity removing box 108, a first filter partition 121 and a second filter partition 122; the first filter partition 121 divides the interior of the filtering and impurity removing box 108 into a first operation area 111 and a first backstage area 114, and the second filter partition 122 divides the first operation area 111 into a first clean area 112 and a first pollution area 113.

[0046] The first operation area 111 is mainly used for placing components that need to be operated by operators and are frequently replaced, such as a primary filter 141, a secondary filter 142, a sample tank 131, a bacterial liquid collection bottle 182, a second filtered peristaltic pump 162, etc.; the first backstage area 114 is mainly used for placing components that do not need to be operated by operators, such as a first negative pressure pump 172, a first buffer bottle 1731, a second buffer bottle 1732, a first filtered peristaltic pump 161, etc.

[0047] The present application can avoid the problem that non-consumable parts (such as the first buffer bottle 1731 and the second buffer bottle 1732), the first pollution area 113 and the first clean area 112 are located on the same working surface. The staff only needs to frequently clean the first pollution area 113 and occasionally clean the first clean area 112 and the first backstage area 114, and the cleaning difficulty is relatively low and the maintenance difficulty is relatively small.

[0048] The front end face of the filtering and impurity removing box 108 of the present application forms a slope (60 - 85°, preferably 75°) to facilitate the staff to view the working conditions in the first pollution area 113 through the glass on the first filter cover 106, view the working conditions in the first clean area 112 through the glass on the second filter cover 107, and enhance the aesthetics of the present application.

[0049] Components such as a first printer 101, a first illuminating lamp 105, a first spotlight 104, a socket and a control cabinet are provided on the shell of the filtering and impurity removing box 108 to make the present application more convenient to use; further, a first fan 102 and a first buzzer 103 can be provided in the first backstage area 114. The first fan 102 can be used to reduce the odor concentration in the first pollution area 113, and the first buzzer 103 can be used for alarm reminder, thereby making the present application more convenient to use.

[0050] During actual operation, the above-mentioned first printer 101, first lighting lamp 105, first spotlight 104, socket, control cabinet, first fan 102 and first buzzer 103 are all conventional items in the art, and their installation methods and installation positions are all conventional means in the art, which will not be elaborated here.

[0051] In the first contaminated area 113, there is a shredding component 130 for shredding fecal samples and a filtering component 140 for filtering the mixture after shredding; in the first clean area 112, there is a dilution bottle 150; in the first backstage area 114, there is a negative pressure component 160 for sequentially delivering the water source in the dilution bottle 150 to the shredding component 130 and the filtering component 140.

[0052] Under the combined action of the above-mentioned shredding component 130, filtering component 140, dilution bottle 150 and first negative pressure component 160, the present application can automatically separate intestinal microorganisms (including various microorganisms such as beneficial bacteria, harmful bacteria, fungi, viruses, etc.) from fecal samples. When it is necessary to replace the sampling consumables (shredding component 130 and filtering component 140), people only need to open the first filter cover 106; when it is necessary to supplement or replace the diluent, the present application only needs to open the second filter cover 118; in this way, the first contaminated area 113, the first clean area 112 and the first backstage area 114 cooperate with each other and do not interfere with each other, which can avoid cross-contamination and thus make the present application more convenient to use.

[0053] The shredding component 130 includes a sample tank 131 and a shredding motor 132. A shredding shaft with shredding blades is arranged in the sample tank 131, and the output end of the shredding motor 132 is connected to the shredding shaft and drives the shredding shaft to rotate. The sample tank 131 is installed in the first contaminated area 113. When the shredding motor 132 drives the shredding shaft to rotate, the shredding shaft will drive the shredding blades to rotate, thereby realizing the shredding function of the sample.

[0054] The filtering component 140 includes a primary filter 141 for primary filtering of the mixture after shredding and a secondary filter 142 for secondary filtering of the mixture after shredding. The dilution bottle 150, the sample tank 131, the primary filter 141 and the secondary filter 142 are sequentially connected through pipelines (not shown in the figure). Under the combined action of the primary filter 141 and the secondary filter 142, the present application realizes the multi-stage filtering function of the mixture after shredding, so that the present application finally obtains an intestinal microorganism solution.

[0055] During actual operation, both the primary filter 141 and the secondary filter 142 include a barrel body and a barrel cover, as Figure 6As shown, there are multiple layers (such as six layers) of filter meshes arranged inside the barrel body, and spacers are arranged between adjacent filter meshes. The pore size of the filter meshes in the primary filter 141 and the secondary filter 142 is 40 - 500.

[0056] Preferably, a first filter support plate 123 is installed in the first pollution area 113, and a third filter support plate 125 which is installed on the first filter support plate 123 and used for placing the sample tank 131. The shredding motor 132 is installed below the first filter support plate 123. A shredding coupling 133 is arranged inside the third filter support plate 125. The output end of the shredding motor 132 is connected to the shredding shaft through the shredding coupling 133. In this way, when conducting experiments on different fecal samples, the staff only needs to replace the sample tank 131, making the application more convenient to use.

[0057] Furthermore, a second filter support plate 124 for placing the primary filter 141 and the secondary filter 142 is installed in the first pollution area 113. The first filter support plate 123 is located below the second filter support plate 124, and the first filter support plate 123 and the second filter support plate 124 are connected by a fourth filter support plate 126 installed at the bottom of the first pollution area 113. In this way, the structure of the application is more compact, and the arrangement of the sample tank 131, the primary filter 141 and the secondary filter 142 in the first pollution area 113 is more reasonable, making the application more convenient to use.

[0058] Even further, both the first filter support plate 123 and the second filter support plate 124 are horizontally arranged, the fourth filter support plate 126 is vertically arranged, the first filter support plate 123, the second filter support plate 124 and the fourth filter support plate 126 are all connected to the first partition plate 121, and the fourth filter support plate 126 is installed on the bottom plate of the first pollution area 113.

[0059] The application further includes a first purification bottle 171 and a first negative pressure pump 172. One end of the first negative pressure pump 172 is connected to the first purification bottle 171 and the shredding assembly 130 in sequence, and the other end is connected to the first buffer 173 and the first filter 174 in sequence. The above-mentioned first purification bottle 171 and first negative pressure pump 172 can remove the odor in the filter and impurity removal box 108, making the application cleaner, more hygienic and more convenient to use.

[0060] During actual operation, the first negative pressure pump 172 can be a negative pressure vacuum pump. One end of the first negative pressure pump 172 is connected to the first purification bottle 171 and the sample tank 131 through pipelines in sequence, and the other end is connected to the first buffer 173, the first filter 174 and the bacterial liquid collection bottle 182 through pipelines in sequence.

[0061] Preferably, as Figure 7As shown, the first buffer 173 includes a first buffer bottle 1731 and a second buffer bottle 1732 arranged in sequence along the gas flow direction. A first control valve 175 is provided between the first purification bottle 171 and the sample tank 131, and a second control valve 176 is provided between the first filter 174 and the bacterial liquid collection bottle 182. During actual operation, both the first buffer bottle 1731 and the second buffer bottle 1732 can be pneumatic diaphragm pump buffer bottles, and both the first control valve 175 and the second control valve 176 can be pneumatic control valves.

[0062] A sample collection rack 181 is also provided in the first contamination area 113. A plurality of bacterial liquid collection bottles 182 are installed on the sample collection rack 181, and the bacterial liquid collection bottles 182 are used to collect the mixed liquid after being processed by the filtration assembly 140.

[0063] The structure of the bacterial liquid collection bottle 182 is as Figure 3 shown. There can be 4 bacterial liquid collection bottles 182. The bacterial liquid collection bottle 182 includes a bottle body and a bottle cap. A scale is provided on the bottle body of the bacterial liquid collection bottle, and a liquid storage inlet and a liquid storage outlet are provided on the bottle cap of the bacterial liquid collection bottle. The bottle body and the bottle cap of the bacterial liquid collection bottle 182 are connected by threads.

[0064] During actual operation, the above-mentioned sample collection rack 181 and the bacterial liquid collection bottle 182 are non-essential structures of this application. People can collect the microbial mixed liquid through a large-volume liquid collection bottle (such as an Erlenmeyer flask).

[0065] Preferably, a plurality of bacterial liquid collection bottles 182 are arranged in series, that is, a plurality of bacterial liquid collection bottles 182 are sequentially connected to the discharge end of the secondary filter 142. After the previous bacterial liquid collection bottle 182 is filled with the microbial mixed liquid (referred to as bacterial liquid), the next bacterial liquid collection bottle 182 continues to be filled with the microbial mixed liquid until the bacterial liquid collection bottle 182 completes the collection of the bacterial liquid. Further, a liquid level sensor can also be provided on the sample collection rack 181 to make it more convenient for this application to collect the microbial mixed liquid.

[0066] A first display controller 191 is also provided on the outer shell of the first clean area 112. The shredding assembly 130, the filtration assembly 140, and the negative pressure assembly 160 are all electrically connected to the first display controller 191.

[0067] The first display controller 191 can be used to control the working states of components such as the shredding component 130, the filtering component 140, and the negative pressure component 160. The real-time working parameters of the shredding component 130, the filtering component 140, and the negative pressure component 160 can be directly displayed on the first display controller 191, enabling the staff to adjust the working parameters of the shredding component 130, the filtering component 140, and the negative pressure component 160 in real time according to the state of the fecal sample, thereby controlling the working states of the shredding component 130, the filtering component 140, and the negative pressure component 160 in real time.

[0068] A first nitrogen cabinet 192 for placing nitrogen and a first consumable cabinet 193 for placing consumables are provided at the lower end of the filtration and impurity removal box 108. The first nitrogen cabinet 192 is located between the filtration and impurity removal box 108 and the first consumable cabinet 193. Preferably, a nitrogen switch valve connected to a nitrogen cylinder and used to control the oxygen concentration in the filtration and impurity removal box 108 is provided on the filtration and impurity removal box 108.

[0069] Under the action of the above nitrogen cylinder, the present application can keep the filtration and impurity removal box 108 in a low-oxygen state, thereby increasing the survival rate of anaerobic bacteria in feces. Components such as an oxygen sensor can also be provided in the filtration and impurity removal box 108. The oxygen sensor can be used to detect the oxygen concentration in the filtration and impurity removal box 108. The first display controller can control the opening or closing of the nitrogen switch valve according to the oxygen concentration fed back by the oxygen sensor, thereby controlling the oxygen concentration in the filtration and impurity removal box 108.

[0070] As Figure 7 shown, the working steps of the filtration and impurity removal device of the present application are as follows:

[0071] S1. The water source (such as physiological saline) in the dilution bottle 150 first enters the sample tank 131 under the action of the first filtration peristaltic pump 161. At this time, the sample tank 131 contains a mixture of the water source (such as physiological saline) and the fecal sample, which is denoted as the first mixture.

[0072] S2. The shredding motor 132 below the sample tank 131 centrifugally shreds the first mixture to obtain a mixture after centrifugal pulverization of the fecal sample, which is denoted as the second mixture.

[0073] S3. Under the action of the second filtration peristaltic pump 162, the second mixture is sequentially processed by the primary filter 141, the second filtration peristaltic pump 162, and the secondary filter 142 to form an intestinal microorganism mixed solution (also called bacterial liquid), and enters the bacterial liquid collection bottle 182 (i.e., the bacterial liquid collection bottle 182).

[0074] At this time, the bacterial liquid collection bottle 182 mainly contains odor, intestinal microorganisms, and aqueous solution, etc. The intestinal microorganisms include various microorganisms such as beneficial bacteria, harmful bacteria, fungi, viruses, etc.; at this time, preferably, a part of the odor in the second mixture will directly enter the first purification bottle 171, and finally the waste gas is discharged from the first purification bottle 171;

[0075] S4. Under the action of the first negative pressure pump 172, a negative pressure is formed in the bacterial liquid collection bottle 182. The odor in the bacterial liquid collection bottle 182 passes through the gas first filter 174, the first buffer bottle 1731, the second buffer bottle 1732, and the vacuum pump in sequence under the action of the negative pressure and then enters the first purification bottle 171, and finally the waste gas is discharged from the first purification bottle 171;

[0076] In this step, both buffer bottles are pneumatic diaphragm buffer bottles. The two pneumatic diaphragm buffer bottles can realize the negative pressure buffering function, so as to achieve pressure equalization. Preferably, a negative pressure gauge can also be set in this application.

[0077] As Figure 8 shown, the enrichment adsorption device 200 of the present application includes an enrichment adsorption box 208; a harmful bacteria adsorption component 210 for removing harmful bacteria in the intestinal microorganism mixture, a beneficial bacteria enrichment component 220 for enriching beneficial bacteria in the microorganism mixture, a beneficial bacteria dissociation component 230 for dissociating the coupled beneficial bacteria and antibody magnetic beads, a beneficial bacteria collection bottle 240 for collecting beneficial bacteria, and a second negative pressure component 250 are provided on the enrichment adsorption box 208. The second negative pressure component 250 is used to make the intestinal microorganism mixture flow through the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230 in sequence and enter the beneficial bacteria collection bottle 240.

[0078] The present application realizes the functions of more efficiently selectively removing harmful bacteria and enriching beneficial bacteria in the intestinal microbiota transplantation technology. The present application greatly improves the sorting efficiency of target probiotics, especially for the precise capture of low-abundance strains, which can make the fecal bacteria separation and screening work for precise transplantation more efficient and specific, and thus promote the improvement of production efficiency. The present application can play a more important role in the field of intestinal microbiota transplantation, realizing the precise removal of harmful bacteria and the precise enrichment of beneficial bacteria flora, and it will become the innovation leading the standardized intestinal microbiota preparation technology.

[0079] The present application mainly conducts enrichment treatment on the microorganism mixture on the basis of the already collected microorganism mixture to remove harmful bacteria and enrich beneficial bacteria, so as to obtain a beneficial bacteria conjugate (second antibody magnetic bead 221 + beneficial bacteria); then the beneficial bacteria conjugate is dissociated to dissociate the beneficial bacteria and the antibody magnetic beads, so as to obtain beneficial bacteria. Specifically:

[0080] In this application, an antibody for identifying harmful bacteria is first conjugated to magnetic beads to obtain the first antibody magnetic beads 211 (which can also be called harmful bacteria antibody magnetic beads), and then reverse enrichment is carried out through a magnetic field to adsorb harmful bacteria in the intestinal microbe mixture. Then, according to the same principle, the remaining intestinal microbe mixture is used to enrich beneficial bacteria in fecal bacteria, and a dissociation enzyme is used to dissociate the coupled beneficial bacteria and the second antibody magnetic beads 221, and finally the target beneficial bacteria are obtained from fecal bacteria.

[0081] It should be noted that the harmful bacteria adsorption component 210 is an optimal solution in this application. The harmful bacteria adsorption component 210 can efficiently remove harmful bacteria in the microbe mixture, solve the problem of potential harmful bacteria remaining in traditional physical screening technologies, further improve the purity of the bacterial solution, and provide a safer and more reliable bacterial source for subsequent microbiota transplantation applications.

[0082] As Figure 9 shown, a first enrichment partition 261 and a second enrichment partition 262 are also provided in the enrichment adsorption box 208. The first enrichment partition 261 divides the interior of the enrichment adsorption box 208 into a second operation area 263 and a second background area 264. The second enrichment partition 262 divides the second operation area 263 into a second clean area 265 and a second pollution area 266. The harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230 are all located in the second pollution area 266. The beneficial bacteria collection bottle 240 is located in the second clean area 265, and the second negative pressure component 250 is installed on the first enrichment partition 261.

[0083] During actual operation, the second background area 264 is mainly used to place those components that do not require operation by the operator (and are often replaced), such as the second negative pressure pump 272, the third buffer bottle 2731, the fourth buffer bottle 2372, etc. The second operation area 263 is mainly used to place those components that require operation by the operator and are often replaced, such as the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, the beneficial bacteria dissociation component 230, and the beneficial bacteria collection bottle 240, etc.

[0084] After dividing the interior of the enrichment adsorption box 208 into a second operation area 263, a second backstage area 264, a second pollution area 266, and a second clean area 265, the present application can avoid cross-contamination, making the present application more convenient to use. When it is necessary to replace sampling consumables (such as the harmful bacteria magnetic bead tank 212, the beneficial bacteria magnetic bead tank 222, and the beneficial bacteria dissociation tank 232, etc.), people only need to open the first enrichment cover plate 206; when it is necessary to supplement or replace the beneficial bacteria collection bottle 240 or the waste liquid collection bottle 226, the present application only needs to open the second enrichment cover plate 207; in this way, the second pollution area 266, the second clean area 265, and the second backstage area 264 cooperate with each other and do not interfere with each other, which can avoid cross-contamination, making the present application cleaner and more convenient to use.

[0085] Such as Figure 8 And Figure 9 As shown, the present application forms a slope (60-85°, preferably 75°) on the front end face of the enrichment adsorption box 208, so as to facilitate the staff to view the working conditions of the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the magnetic bead dissociation collection component through the glass on the first enrichment cover plate 206, and increase the aesthetics of the present application.

[0086] A second display controller 280 is also provided on the outer shell of the second clean area 265, and the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230 are all electrically connected to the second display controller 280.

[0087] The second display controller 280 can be used to control the working states of components such as the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230, and the real-time working parameters of the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230 can be directly displayed on the second display controller 280, so that the staff can adjust the working states of the harmful bacteria adsorption component 210, the beneficial bacteria enrichment component 220, and the beneficial bacteria dissociation component 230 in real time.

[0088] The lower end of the enrichment adsorption box 208 is connected to a second nitrogen cabinet 291 and a second consumable cabinet 292. A nitrogen cylinder for filling nitrogen into the enrichment adsorption box 208 is provided in the second nitrogen cabinet 291, and a nitrogen switch valve connected to the nitrogen cylinder and used to control the oxygen concentration in the enrichment adsorption box 208 is provided on the enrichment adsorption box 208.

[0089] Under the action of the nitrogen gas cylinder, the present application can make the enrichment adsorption box 208 in a low-oxygen state, thereby improving the survival rate and activity of anaerobic bacteria in feces; components such as an oxygen sensor can also be arranged in the enrichment adsorption box 208. The oxygen sensor can be used to detect the oxygen concentration in the enrichment adsorption box 208, and the second display controller 280 can control the opening or closing of the nitrogen gas switch valve according to the oxygen concentration fed back by the oxygen sensor, so as to control the oxygen concentration in the enrichment adsorption box 208.

[0090] The present application ensures the activity and stability of the bacterial community to a greater extent during the separation process. By adding a low-oxygen device (nitrogen gas cylinder), it provides an anaerobic environment with a relatively low oxygen concentration, which is more beneficial to maintaining the abundance of beneficial bacteria and enhancing the activity of the bacterial community. The present application will play a more important role in the field of fecal microbiota transplantation, achieving the precise removal of harmful bacteria and the precise enrichment of beneficial bacteria communities, and it will lead the innovation of standardized fecal microbiota preparation technology.

[0091] One or more of a second printer 201, a second fan 202, a second buzzer 203, a second spotlight 204, and a second lighting lamp 205 are also arranged on the enrichment adsorption box 208. The second printer 201 can be used to print test records, the second fan 202 can be used to reduce the oxygen concentration in the second contamination area 266, the second buzzer 203 can be used for alarm prompts, the second lighting lamp 205 can be used to illuminate the second contamination area 266 and the second clean area 265, and the second spotlight 204 can be used to centrally reflect the working state of the second contamination area 266, so as to make the present application more convenient to use.

[0092] During actual work, the above-mentioned second printer 201, second lighting lamp 205, second spotlight 204, socket, control cabinet, second fan 202, and second buzzer 203 are all conventional items in the art, and their installation methods and installation positions are all conventional means in the art, which will not be elaborated here. Preferably, a plurality of through holes can be arranged on the first partition 261, the second partition 262, and the enrichment adsorption box 208 to allow pipelines to pass through, so as to make the present application more convenient to use.

[0093] As Figure 9 and Figure 10 shown, the harmful bacteria adsorption component 210 includes a first antibody magnetic bead 211, a harmful bacteria magnetic bead tank 212, and a harmful bacteria adsorption bottle 213. A harmful bacteria coupling member 214 for coupling harmful bacteria and the first antibody magnetic bead 211 together is arranged on the harmful bacteria magnetic bead tank 212, and a harmful bacteria adsorption member 215 for reversely enriching the harmful bacteria coupling body (that is, the coupled harmful bacteria and the first antibody magnetic bead 211) is arranged on the harmful bacteria adsorption bottle 213.

[0094] The harmful bacteria adsorption component 210 is located between the filtration and impurity removal device 100 and the beneficial bacteria enrichment component 220. The main function of the harmful bacteria adsorption component 210 is to remove the (extremely trace) harmful bacteria that may exist in the intestinal microorganism mixture, so as to reduce the competition and potential risks of harmful bacteria, thereby improving the safety of the bacterial liquid. Preferably, multiple harmful bacteria can be removed simultaneously in this step to further improve the safety of the bacterial liquid.

[0095] The harmful bacteria coupling part 214 includes a first stirring motor 2141 and a first material port 2142. The first material port 2142 is used to introduce the first antibody magnetic beads 211 into the harmful bacteria magnetic bead tank 212. The output end of the first stirring motor 2141 is connected with a first stirring shaft 2143, and a first stirring blade 2144 located in the harmful bacteria magnetic bead tank 212 is connected inside the first stirring shaft 2143. Under the combined action of the above-mentioned first stirring motor 2141 and the first stirring blade 2144, the first bacterial antibody magnetic beads and harmful bacteria will be coupled in the harmful bacteria magnetic bead tank 212, thereby obtaining harmful bacteria coupling bodies.

[0096] The harmful bacteria adsorption part 215 includes a first electromagnetic induction coil 2151 and a first coil controller 2152. The first electromagnetic induction coil 2151 is sleeved on the harmful bacteria adsorption bottle 213, and the first coil controller 2152 is electrically connected to the first electromagnetic induction coil 2151 to control the working state of the first electromagnetic induction coil 2151.

[0097] The first coil controller 2152 can make the first electromagnetic induction coil 2151 in a powered-on state or a disconnected state. When the first electromagnetic induction coil 2151 is in a powered-on state, the first electromagnetic induction coil 2151 will generate a magnetic field, thereby adsorbing the harmful bacteria coupling bodies, and further enabling the present application to achieve the purpose of removing harmful bacteria in intestinal microorganisms.

[0098] The beneficial bacteria enrichment component 220 includes a second antibody magnetic bead 221, a beneficial bacteria magnetic bead tank 222, a beneficial bacteria enrichment bottle 223 and a waste liquid collection bottle 226. A beneficial bacteria coupling part 224 for coupling beneficial bacteria and the second antibody magnetic bead 221 together is arranged on the harmful bacteria magnetic bead tank 212, and a beneficial bacteria enrichment part 225 for positively enriching the beneficial bacteria coupling bodies (that is, the beneficial bacteria and the second antibody magnetic bead 221 coupled together) is arranged on the beneficial bacteria enrichment bottle 223.

[0099] Furthermore, a waste liquid guiding port is arranged on the beneficial bacteria enrichment bottle 223, and the waste liquid collection bottle 226 is connected to the waste liquid guiding port to collect the waste liquid after positive enrichment treatment. In this way, the present application is more convenient to use.

[0100] The main function of the beneficial bacteria enrichment component 220 is to automatically enrich the (extremely trace) beneficial bacteria that may exist in the intestinal microorganism mixture. Preferably, multiple beneficial bacteria can be enriched simultaneously in this step to further improve the beneficial bacteria enrichment efficiency.

[0101] The beneficial bacteria coupling component 224 includes a second stirring motor 2241 and a second material port 2242. The second material port 2242 is used to introduce the second antibody magnetic beads 221 into the beneficial bacteria magnetic bead tank 222. The output end of the second stirring motor 2241 is connected to a second stirring shaft 2243, and a second stirring blade 2244 located in the beneficial bacteria magnetic bead tank 222 is connected inside the second stirring shaft 2243. Under the combined action of the above-mentioned second stirring motor 2241 and the second stirring blade 2244, the second antibody magnetic beads 221 and the beneficial bacteria will be coupled in the beneficial bacteria magnetic bead tank 222, thereby obtaining a beneficial bacteria coupling body.

[0102] The beneficial bacteria enrichment component 225 includes a second electromagnetic induction coil 2251 and a second coil controller 2252. The second electromagnetic induction coil 2251 is sleeved on the beneficial bacteria enrichment bottle 223, and the second coil controller 2252 is electrically connected to the second electromagnetic induction coil 2251 to control the working state of the second electromagnetic induction coil 2251.

[0103] The second coil controller 2252 can make the second electromagnetic induction coil 2251 in an energized state or a disconnected state. When the second electromagnetic induction coil 2251 is in an energized state, the second electromagnetic induction coil 2251 will generate a magnetic field, thereby adsorbing the beneficial bacteria coupling body, and further enabling the present application to achieve the purpose of enriching the beneficial bacteria in the intestinal microorganisms.

[0104] The beneficial bacteria dissociation component 230 includes a beneficial bacteria dissociation tank 232 and a magnetic bead adsorption bottle 233. A beneficial bacteria dissociation part 234 for dissociating the coupled beneficial bacteria and the second antibody magnetic beads 221 is provided on the beneficial bacteria dissociation tank 232, and a magnetic bead adsorption part 235 for adsorbing the second antibody magnetic beads 221 is provided on the magnetic bead adsorption bottle 233.

[0105] The main function of the beneficial bacteria dissociation component 230 is to dissociate the coupled beneficial bacteria and the second antibody magnetic beads 221 to facilitate the collection of beneficial bacteria.

[0106] The beneficial bacteria dissociation part 234 includes a third stirring motor 2341 and a third material port 2342. The third material port 2342 is used to introduce dissociation enzymes into the beneficial bacteria dissociation tank 232. The output end of the third stirring motor 2341 is connected with a third stirring shaft 2343, and a third stirring blade 2344 located in the beneficial bacteria dissociation tank 232 is connected inside the third stirring shaft 2343. Under the combined action of the above-mentioned third stirring motor 2341, third stirring blade 2344 and dissociation enzymes, the coupled beneficial bacteria and the second antibody magnetic beads 221 will dissociate, so as to achieve the purpose of collecting beneficial bacteria in this application.

[0107] The magnetic bead adsorption part 235 includes a third electromagnetic induction coil 2351 and a third coil controller 2352. The third electromagnetic induction coil 2351 is sleeved on the magnetic bead adsorption bottle 233, and the third coil controller 2352 is electrically connected to the third electromagnetic induction coil 2351 to control the working state of the third electromagnetic induction coil 2351.

[0108] The third coil controller 2352 can make the third electromagnetic induction coil 2351 in a powered-on state or a disconnected state. When the third electromagnetic induction coil 2351 is in a powered-on state, the third electromagnetic induction coil 2351 will generate a magnetic field, so as to adsorb the second antibody magnetic beads 221 and remove the second antibody magnetic beads 221 in the mixed liquid, so that this application can collect more pure beneficial bacteria.

[0109] During actual operation, the first electromagnetic induction coil 2151, the second electromagnetic induction coil 2251 and the third electromagnetic induction coil 2351 can be collectively referred to as electromagnetic induction coils. The first coil controller 2152, the second coil controller 2252 and the third coil controller 2352 can all be integrated on the display controller 280 and can be collectively referred to as coil controllers. Controlling the working state (powered-on state or disconnected state) of the electromagnetic induction coils connected to them by the coil controller is a conventional means in the art and will not be elaborated here. Preferably, the coil controller can also control the current intensity flowing through the electromagnetic induction coil to control the magnetic field intensity of the electromagnetic induction coil connected to it.

[0110] This application further includes a second purification bottle 271 and a second negative pressure pump 272. One end of the second negative pressure pump 272 is sequentially connected to the second purification bottle 271 and the harmful bacteria adsorption component 210, and the other end is sequentially connected to the second buffer 273, the second filter 274 and the beneficial bacteria collection bottle 240.

[0111] During actual operation, the second negative pressure pump 272 can be a negative pressure vacuum pump. One end of the second negative pressure pump 272 is connected to the second purification bottle 271 and the harmful bacteria magnetic bead tank 212 in sequence through a pipeline, and the other end is connected to the second buffer 273, the second filter 274 and the beneficial bacteria collection bottle 240 in sequence through a pipeline.

[0112] The second buffer 273 includes a third buffer bottle 2731 and a fourth buffer bottle 2372 arranged in sequence along the gas flow direction. A third control valve 275 is arranged between the second purification bottle 271 and the second purification bottle 271, and a fourth control valve 276 is arranged between the second filter 274 and the beneficial bacteria collection bottle 240. During actual operation, both the third buffer bottle 2731 and the fourth buffer bottle 2372 can adopt pneumatic diaphragm pump buffer bottles, and both the third control valve 275 and the fourth control valve 276 can adopt pneumatic control valves.

[0113] The entire experimental process of this application takes about 2.5 hours. Among them, the first antibody magnetic beads and the second antibody magnetic beads are both prepared in advance. Refer to Figure 12 , the working steps of the enrichment and adsorption device of this application include:

[0114] K1. Send the microbial mixture through a pipeline into the harmful bacteria magnetic bead tank 212. The first antibody magnetic beads 211 enter the harmful bacteria magnetic bead tank 212 from the first material port 2142. The first stirring motor 2141 drives the first stirring blade 2144 connected thereto to rotate and mix for 10 minutes, and then stand and incubate at room temperature for 40 minutes, so as to couple the harmful bacteria antibody magnetic beads with the harmful bacteria to obtain harmful bacteria coupling bodies;

[0115] This application couples the harmful bacteria magnetic beads that have been surface carboxyl-activated with the corresponding antibody of the harmful bacteria to obtain the first antibody magnetic beads 211. The first antibody magnetic beads 211 are prepared before being sent to the harmful bacteria magnetic bead tank 212 to avoid affecting the harmful bacteria coupling bodies obtained in this application. Taking Salmonella as an example, this application further illustrates how to adsorb harmful bacteria in this application to make the technical solution of the present invention easier to understand:

[0116] K11. Surface carboxyl activation of harmful bacteria magnetic beads

[0117] Take 1000 mL of Mag COOH sterile magnetic bead suspension (product number 70113-5, Suzhou Beaver Biotechnology) (magnetic bead concentration is 5 mg / mL) in a super clean bench into a 5000 mL sterile centrifuge bottle, magnetically separate to remove the supernatant, and perform magnetic separation washing 2 times with 2000 mL of MEST solution (100 mM MES, pH 5.0, 0.05% Tween 20), and then remove the supernatant; quickly add 1000 mL of freshly prepared EDC solution (10 mg / mL, using the above MEST solution as a dispersant) and 1000 mL of NHS (10 mg / mL, using the above MEST solution as a dispersant) solution into the centrifuge tube containing the magnetic beads, vortex and mix to fully suspend the magnetic beads, and activate at room temperature for 30 min.

[0118] Transfer the magnetic bead suspension to a new sterile centrifuge tube at 100 mL per tube. During this period, keep the magnetic beads in a suspended state (you can use a vertical mixer to invert and mix). After the above steps, the carboxyl groups on the surface of the magnetic beads have been activated and can be covalently coupled with bioligands carrying primary amino groups. (The activated state should not be stored for a long time. It is recommended to perform the coupling immediately).

[0119] K12. Covalent coupling of magnetic beads and harmful bacteria antibodies

[0120] Replace the monoclonal antibody buffer solution of Salmonella with 15 mM MES buffer solution (pH = 6.0), and dilute the antibody with MES buffer to 5 mg / mL. Take 4 mL of the antibody and mix it well with 100 mL (diameter 10 μm) of the above-activated carboxyl magnetic beads. React at room temperature for 2 h, and keep the magnetic beads in a suspended state during the coupling (you can use a vertical mixer to invert and mix). Perform magnetic separation, suck out the supernatant and simultaneously detect the remaining antibody content in the supernatant, calculate the amount and concentration of the antibody coupled to the magnetic beads, wash the magnetic beads 2 to 3 more times, and resuspend them with physiological saline to obtain the monoclonal antibody magnetic beads of Salmonella. The monoclonal antibody magnetic beads of Salmonella can accurately enrich Salmonella.

[0121] During actual work, the staff can also replace Salmonella with other harmful bacteria, such as Shigella and Clostridium difficile. At the same time, when performing the above step K12, this application can simultaneously place multiple harmful bacteria antibody magnetic beads (i.e., the first antibody magnetic beads 211) into the harmful bacteria magnetic bead tank 212 to simultaneously remove multiple harmful bacteria in the intestinal microorganism mixture.

[0122] Furthermore, to improve work efficiency, the harmful bacteria in this application can be one or more of Salmonella, Shigella, Campylobacter jejuni, Campylobacter upsaliensis, Campylobacter coli, Clostridium difficile, Vibrio parahaemolyticus, Vibrio cholerae, Vibrio vulnificus, Yersinia enterocolitica, Enteropathogenic Escherichia coli (EPEC), Shiga toxin-producing Escherichia coli (STEC), Escherichia coli O157, Plesiomonas shigelloides, Helicobacter pylori, Staphylococcus aureus, Enterobacter cloacae, and Cronobacter sakazakii.

[0123] K13. Coupling of the first bacterial antibody magnetic beads and harmful bacteria to obtain a harmful bacteria coupling body

[0124] The mixed liquid processed by the filtration equipment enters the harmful bacteria magnetic bead tank 212 through the filtrate inlet pipe. Take the above-mentioned harmful bacteria antibody magnetic beads, mix the magnetic beads in a certain ratio (such as 1:1) to obtain the first antibody magnetic beads 211. Add the first antibody magnetic beads 211 into the harmful bacteria magnetic bead tank 212 from the first material port 2142. The first antibody magnetic beads 211 are mixed with the intestinal microorganism mixed liquid, and the blades connected to the first stirring motor 2141 rotate to mix evenly for 10 minutes, and then stand for incubation for 40 minutes. Let the first antibody magnetic beads 211 bind to harmful bacteria (such as Salmonella) to obtain harmful bacteria conjugates.

[0125] During actual operation, the first antibody magnetic beads 211 can also be referred to as harmful bacteria antibody magnetic beads. The method of obtaining the first antibody magnetic beads 211, and the method of coupling the first antibody magnetic beads 211 with harmful bacteria to form harmful bacteria conjugates are all conventional means in the art and will not be elaborated here.

[0126] K2. The negative pressure pipeline drives the harmful bacteria conjugates into the harmful bacteria adsorption bottle 213, and under the action of the first electromagnetic induction coil 2151, the harmful bacteria conjugates are reversely enriched, thereby removing harmful bacteria from the microorganism mixed liquid;

[0127] During actual operation, this step aims to remove extremely trace amounts of harmful bacteria that may exist in the donor, reduce the competition and potential risks of harmful bacteria, and ensure the safety of the bacterial liquid using biotechnology. Preferably, this step can remove multiple harmful bacteria simultaneously to further improve the safety of the bacterial liquid.

[0128] K3. Continue to let the microorganism mixed liquid obtained in step K2 enter the beneficial bacteria magnetic bead tank 222. The second antibody magnetic beads 221 enter the beneficial bacteria magnetic bead tank 222 from the second material port 2242. The second stirring motor 2241 rotates the second stirring blades 2244 connected to itself to mix evenly for 10 minutes, and then incubates at room temperature for 40 minutes, so as to couple the beneficial bacteria antibody magnetic beads with beneficial bacteria to obtain beneficial bacteria conjugates;

[0129] In this application, the beneficial bacteria magnetic beads that have been surface carboxyl-activated are coupled with the corresponding antibodies of the beneficial bacteria to obtain the second antibody magnetic beads 221. During actual operation, the second antibody magnetic beads 221 are prepared before being sent to the beneficial bacteria magnetic bead tank 222 to avoid affecting the beneficial bacteria conjugates obtained in this application. Taking Faecalibacterium prausnitzii as an example, this application further illustrates how to enrich beneficial bacteria in this application to make the technical solution of the present invention easier to understand.

[0130] K31. Carboxyl activation of the surface of beneficial bacteria magnetic beads

[0131] In the clean bench, take 2000mL Mag COOH sterile magnetic bead suspension (70113-5, Suzhou Beaver Biotechnology) (magnetic bead concentration of 10mg / mL) into a 5000mL sterile centrifuge bottle, remove the supernatant by magnetic separation, wash twice with 4000mL MEST solution (100mMMES, pH 5.0, 0.05% Tween 20), and then remove the supernatant; quickly add freshly prepared 2000mL EDC solution (10mg / mL, using the above MEST solution as a dispersant) and 2000mL NHS (10mg / mL, using the above MEST solution as a dispersant) solution to the centrifuge tube containing the magnetic beads, vortex mix to fully suspend the magnetic beads, and activate at room temperature for 30min. Transfer the magnetic bead suspension to a new sterile centrifuge bottle according to 200mL per tube. During this period, the magnetic beads are kept in suspension (they can be mixed by inversion using a vertical mixer); after the above steps, the carboxyl groups on the surface of the magnetic beads have been activated and can be covalently coupled with biological ligands with primary amino groups. (The activated state should not be stored for a long time, and it is recommended to couple immediately)

[0132] Covalent coupling of K32, magnetic beads and beneficial bacteria antibodies

[0133] Replace the monoclonal antibody buffer solution of Clostridium prausnitzii with 15mM MES buffer (pH=6.0), and dilute the antibody to 5mg / mL with MES buffer. Take 8mL of the antibody and mix thoroughly with 200mL (10μm in diameter) of the above-mentioned activated carboxyl magnetic beads. React at room temperature for 2h, and keep the magnetic beads suspended during coupling (vertical mixer can be used for inversion mixing). Magnetic separation, aspirate the supernatant and detect the remaining antibody content in the supernatant at the same time, calculate the amount and concentration of magnetic bead-coupled antibody, continue to wash the magnetic beads 2 to 3 times, and resuspend them with physiological saline to obtain monoclonal antibody magnetic beads of Clostridium prausnitzii. The monoclonal antibody magnetic beads of Clostridium prausnitzii can accurately enrich Clostridium prausnitzii.

[0134] In actual work, the staff can also replace Faecalibacterium prausnitzii with other beneficial bacteria, such as Bacteroides monomorphum and Roseburia, etc. At the same time, when performing the above step K32, the present application can simultaneously place multiple beneficial bacteria antibody magnetic beads (i.e., second antibody magnetic beads 221) into the beneficial bacteria magnetic bead tank 222 to simultaneously enrich multiple beneficial bacteria in the intestinal microbial mixture.

[0135] Further, in order to improve work efficiency, the beneficial bacteria of the present application can be one or more of Bacteroides monomorpha, Faecalibacterium, Coprococcus, Ruminococcus, Bifidobacterium, Lactobacillus, Reuteria, Akkermansia, Christensen, Weinberg non-permeable bacteria, Butyricum, Lachnospira, Spindle Linker, Butyrivibrio, Streptococcus thermophilus, Bacillus, Butyricoccus, Eubacterium, Roseborgia, and Bacillus.

[0136] K33. The second antibody magnetic beads 221 are combined with the beneficial bacteria to obtain a beneficial bacteria conjugate.

[0137] After the mixed solution is processed through step K32, it enters the beneficial bacteria magnetic bead tank 222. Take the above-mentioned beneficial bacteria antibody magnetic beads (the second bacterial antibody magnetic beads), mix the magnetic beads in a certain ratio (such as 1:1), add the second antibody magnetic beads 221 into the beneficial bacteria magnetic bead tank 222 from the second material port 2242. The second antibody magnetic beads 221 are mixed with the intestinal microorganism mixed solution, and the second stirring motor 2241 drives the second stirring blade 2244 connected to it to rotate and mix evenly for 10 min, and then stand and incubate for 40 min. Let the second antibody magnetic beads 221 bind to the beneficial bacteria to obtain a beneficial bacteria conjugate.

[0138] During actual operation, the second antibody magnetic beads 221 can also be referred to as harmful bacteria antibody magnetic beads. The method of obtaining the second antibody magnetic beads 221, as well as the method of coupling the second antibody magnetic beads 221 with harmful bacteria to form a harmful bacteria conjugate, are all conventional means in the art and will not be elaborated here.

[0139] K4. The negative pressure pipeline drives the beneficial bacteria conjugate into the beneficial bacteria adsorption bottle, and under the action of the second electromagnetic induction coil 2251, the beneficial bacteria conjugate is positively enriched, thereby adsorbing the beneficial bacteria in the mixed solution; at this time, the waste liquid can be collected by the waste liquid bottle;

[0140] The purpose of this step is to enrich the beneficial bacteria that may exist in the donor. Preferably, this step can enrich multiple beneficial bacteria simultaneously to further improve the collection efficiency of beneficial bacteria.

[0141] K5. Let the beneficial bacteria conjugate enter the dissociation bottle, add dissociation enzyme into the third material port 2342, mix evenly for 10 minutes, and incubate at room temperature for 30 minutes;

[0142] In this step, the beneficial bacteria conjugate enters the dissociation bottle, the beneficial bacteria conjugate is resuspended with physiological saline, and REAlease magnetic bead dissociation reagent is added into the third material port 2342 to dissociate the second antibody magnetic beads 221 and the beneficial bacteria, and then incubate at room temperature for 30 min to separate the beneficial bacteria and the antibody magnetic beads.

[0143] During actual operation, the method of separating the beneficial bacteria conjugate into the second antibody magnetic beads and the beneficial bacteria by using the dissociation enzyme belongs to the conventional means in the art and will not be elaborated here.

[0144] K6. The negative pressure pipeline pushes the mixed liquid in step K5 into the magnetic bead adsorption bottle 233, and under the action of the third electromagnetic induction coil 2351, adsorbs the magnetic beads with the second antibody 221 to remove the magnetic beads with the second antibody 221 in the mixed liquid of step K5, thereby obtaining pure beneficial bacteria;

[0145] K7. The beneficial bacteria enter the bacterial liquid collection bottle, thereby realizing the collection of beneficial bacteria.

[0146] During actual operation, people can also sequence the 16S rDNA gene sequence of the beneficial bacteria collected in step K7 to determine the antigen-capturing bacterial species.

[0147] When it is necessary to directly and selectively screen out the target beneficial bacteria from fecal samples to meet the needs of future precise transplantation of intestinal flora and precise treatment of specific intestinal-related diseases, the directional separation steps of the intestinal bacteria directional separation system based on the microbial antibody group technology of the present application include:

[0148] P1. Under the action of the first negative pressure component 160, the filtration and impurity removal device (100) performs filtration and impurity removal treatment on the fecal sample to obtain a microbial mixed liquid;

[0149] In this step, the method for obtaining the microbial mixed liquid is also as follows:

[0150] P1. The water source in the dilution bottle 150 first enters the sample tank 131 under the action of the first filtration peristaltic pump 161. At this time, the sample tank 131 contains a mixture of water source and fecal sample, which is recorded as the first mixture;

[0151] P12. The shredding motor 132 below the sample tank 131 performs centrifugal stirring on the first mixture to obtain a mixture after centrifugal crushing of the fecal sample, which is recorded as the second mixture;

[0152] P13. The second mixture, under the action of the second filtration peristaltic pump 162, passes through the primary filter 141, the second filtration peristaltic pump 162, and the secondary filter 142 in sequence, forms an intestinal microbial mixed liquid, and enters the bacterial liquid collection bottle 182;

[0153] Preferably, the above step P1 further includes a step of removing odors, including:

[0154] P14. Under the action of the first negative pressure pump 172, a negative pressure is formed in the bacterial liquid collection bottle 182, and the odor in the bacterial liquid collection bottle 182, under the action of the negative pressure, passes through the gas first filter 174, the first buffer bottle 1731, the second buffer bottle 1732, and the vacuum pump in sequence and enters the first purification bottle 171, and finally the waste gas is discharged from the first purification bottle 171.

[0155] P2. Under the action of the second negative pressure assembly 250, the harmful bacteria adsorption assembly 210 removes harmful bacteria from the microbial mixed solution;

[0156] In this step, the step of removing harmful bacteria from the microbial mixed solution includes:

[0157] P21. The microbial mixed solution enters the harmful bacteria magnetic bead tank 212, and the first antibody magnetic beads 211 enter the harmful bacteria magnetic bead tank 212 from the first material port 2142;

[0158] P22. The first stirring motor 2141 drives the first stirring blade 2144 connected to itself to rotate according to the first preset time (such as ten minutes) to mix the solution in the harmful bacteria magnetic bead tank 212;

[0159] P23. Incubate at room temperature for the second preset time (such as 40 minutes) to couple the first antibody magnetic beads with the harmful bacteria to obtain a solution containing harmful bacteria conjugates;

[0160] P24. The second negative pressure assembly 250 drives the solution containing harmful bacteria conjugates into the harmful bacteria adsorption bottle 213, and the harmful bacteria adsorbent 215 performs reverse enrichment on the harmful bacteria conjugates, thereby removing harmful bacteria from the microbial mixed solution.

[0161] P3. Under the action of the second negative pressure assembly 250, the beneficial bacteria enrichment assembly 220 performs positive enrichment on the beneficial bacteria in the microbial mixed solution to obtain a bacterial liquid mixed solution containing beneficial bacteria conjugates, and the beneficial bacteria conjugates are formed by coupling beneficial bacteria and second antibody magnetic beads;

[0162] In this step, the steps to obtain the bacterial liquid mixed solution containing beneficial bacteria conjugates are as follows:

[0163] P31. The microbial mixed solution enters the beneficial bacteria magnetic bead tank 222, and the second antibody magnetic beads 221 enter the beneficial bacteria magnetic bead tank 222 from the second material port 2242;

[0164] P32. The second stirring motor 2241 drives the second stirring blade 2244 connected to itself to rotate according to the first preset time (such as ten minutes) to mix the solution in the beneficial bacteria magnetic bead tank 222;

[0165] P33. Incubate at room temperature for the second preset time (such as 40 minutes) to couple the second antibody magnetic beads with the beneficial bacteria to obtain beneficial bacteria conjugates;

[0166] P34. The second negative pressure assembly 250 drives the beneficial bacteria conjugates into the beneficial bacteria enrichment bottle 223, and the beneficial bacteria enrichment member 225 performs positive enrichment on the beneficial bacteria conjugates, thereby obtaining a bacterial liquid mixed solution containing beneficial bacteria conjugates.

[0167] P4. Under the action of the second negative pressure assembly 250, the beneficial bacteria dissociation assembly 230 first dissociates the beneficial bacteria coupling body to obtain a beneficial bacteria solution containing the second antibody magnetic beads and beneficial bacteria; then the beneficial bacteria dissociation assembly 230 removes the second antibody magnetic beads in the beneficial bacteria solution to obtain a beneficial bacteria solution;

[0168] In this step, the steps of obtaining the beneficial bacteria solution from the bacterial liquid mixture include:

[0169] P41. Let the bacterial liquid mixture containing the beneficial bacteria coupling body enter the dissociation bottle, and add the dissociation enzyme into the third material port 2342;

[0170] P42. The third stirring motor 2341 drives the third stirring blade 2344 connected thereto to rotate according to the first preset time (such as ten minutes) to mix the solution in the dissociation bottle;

[0171] P43. Incubate at room temperature for the third preset time such as 30 minutes to dissociate the third antibody magnetic beads from the beneficial bacteria to obtain a mixture containing the third antibody magnetic beads and dissociated beneficial bacteria;

[0172] P44. The second negative pressure assembly 250 drives the above-mentioned mixture containing the third antibody magnetic beads and dissociated beneficial bacteria into the magnetic bead adsorption bottle 233, and the magnetic bead adsorbing member 235 adsorbs the second antibody magnetic beads 221 to obtain a beneficial bacteria solution.

[0173] P5. Under the action of the negative pressure driving mechanism, the beneficial bacteria collection bottle 240 collects the beneficial bacteria solution.

[0174] To make the technical solution of the present application easier to understand, the present application also separately lists the beneficial bacteria catalog and the harmful bacteria catalog, as follows:

[0175] Table 1: List of Beneficial Bacteria Catalog

[0176]

[0177]

[0178]

[0179]

[0180]

[0181] Table 2: List of Harmful Bacteria Catalog

[0182]

[0183]

[0184] In summary, compared with the prior art, the advantages of the present invention include:

[0185] 1. Avoid cross - contamination: This application can isolate intestinal microorganisms from fecal samples. Under the combined action of the first filter partition and the second filter partition, when it is necessary to replace the sampling consumables (including the shredding component, the filtering component, the sample collection rack, and the bacterial liquid collection bottle), this application only needs to open the first filter cover plate; when it is necessary to supplement or replace the diluent, this application only needs to open the second filter cover plate. In this way, the first pollution area, the first clean area, and the first backstage area cooperate with each other and do not interfere with each other, so that this application avoids cross - contamination and is more convenient to use.

[0186] 2. Improve the purity and safety of the bacterial liquid. This application can isolate intestinal microorganisms from fecal samples. The filtering and impurity - removing part can separate and collect intestinal microorganisms from feces to obtain a microbial mixture; the bacterial group enrichment part can remove harmful bacteria in the bacterial liquid, solving the problem of potential harmful bacteria remaining in the traditional physical screening technology. In this way, with the combination of the above - mentioned three parts of filtering and impurity - removing, harmful bacterial group adsorption, and beneficial bacterial group enrichment, this patent can ensure the purity of the bacterial liquid and provide a more safe and reliable bacterial source for subsequent bacterial group transplantation applications.

[0187] 3. Convenient and efficient use. The filtering and impurity - removing part and the enrichment and adsorption part of this application can be used separately or together; moreover, the filtering and impurity - removing part of this application has been further optimized in structure for the existing intestinal flora microorganisms, making it more convenient to use.

[0188] 4. Precise enrichment and standardized operation. This patent adopts a precise enrichment technology for beneficial bacteria. Through a screening mechanism of highly specific antibody - protein specific binding, the purpose of quickly and accurately separating and enriching the target beneficial bacteria is achieved. In addition, this patent has also realized a standardized operation process, can obtain bacterial liquid with stable quality and consistent performance, and greatly improves the repeatability of the bacterial liquid.

[0189] 5. This application combines a low - oxygen device to ensure the activity and stability of the bacterial group. Many beneficial bacteria can maintain the best activity and stability only under low - oxygen or anaerobic conditions. This patent device introduces a low - oxygen device, and by precisely controlling the oxygen content, it can create a relatively stable anaerobic environment during the bacterial liquid treatment process, thus significantly enhancing the activity of the bacterial group, extending the validity period of the bacterial liquid, and further ensuring the quality of the bacterial liquid.

[0190] Other parts not described in detail are all prior art. Although the above - mentioned embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, not all embodiments. People can also obtain other embodiments without creative efforts based on this embodiment, and these embodiments all belong to the protection scope of the present invention.

Claims

1. An intestinal bacteria directional isolation system based on microbial antibodyomics technology, characterized in that: It includes a negative pressure driving mechanism, a filtering and impurity removing device (100) for filtering and removing impurities from fecal samples, a beneficial bacteria enrichment component (220) for positively enriching beneficial bacteria in the microbial mixture, a beneficial bacteria dissociation component (230) for dissociating beneficial bacteria conjugates, and a beneficial bacteria collection bottle (240) for collecting beneficial bacteria. The negative pressure driving mechanism is used to enable the beneficial bacteria in the fecal sample to automatically enter the beneficial bacteria collection bottle (240) after passing through the filtering and impurity removing device (100), the beneficial bacteria enrichment component (220), and the beneficial bacteria dissociation component (230) in sequence.

2. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 1, wherein: The beneficial bacteria enrichment component (220) includes second antibody magnetic beads (221), a beneficial bacteria magnetic bead tank (222), and a beneficial bacteria enrichment bottle (223). A beneficial bacteria coupling member (224) for coupling beneficial bacteria and the second antibody magnetic beads (221) is provided on the harmful bacteria magnetic bead tank (212), and a beneficial bacteria enrichment member (225) for positively enriching beneficial bacteria conjugates is provided on the beneficial bacteria enrichment bottle (223).

3. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 2, characterized in that: The beneficial bacteria enrichment component (220) further includes a waste liquid collection bottle (226). A waste liquid guiding port is provided on the beneficial bacteria enrichment bottle (223), and the waste liquid collection bottle (226) is connected to the waste liquid guiding port to collect the waste liquid after positive enrichment treatment.

4. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 2, wherein: The beneficial bacteria coupling member (224) includes a second stirring motor (2241) and a second material port (2242). The second material port (2242) is used to introduce the second antibody magnetic beads (221) into the beneficial bacteria magnetic bead tank (222), and the output end of the second stirring motor (2241) is connected to a second stirring blade (2244) located inside the beneficial bacteria magnetic bead tank (222).

5. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 2, characterized in that: The beneficial bacteria enrichment member (225) includes a second electromagnetic induction coil (2251) and a second coil controller (2252). The second electromagnetic induction coil (2251) is sleeved on the beneficial bacteria enrichment bottle (223), and the second coil controller (2252) is electrically connected to the second electromagnetic induction coil (2251) to control the working state of the second electromagnetic induction coil (2251).

6. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 1, wherein: The beneficial bacteria dissociation component (230) includes a beneficial bacteria dissociation tank (232) and a magnetic bead adsorption bottle (233). A beneficial bacteria dissociation member (234) for dissociating beneficial bacteria conjugates is provided on the beneficial bacteria dissociation tank (232), and a magnetic bead adsorption member (235) for adsorbing the second antibody magnetic beads (221) is provided on the magnetic bead adsorption bottle (233).

7. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 6, characterized in that: The beneficial bacteria dissociation member (234) includes a third stirring motor (2341) and a third material port (2342). The third material port (2342) is used to introduce a dissociation enzyme into the beneficial bacteria dissociation tank (232), and the output end of the third stirring motor (2341) is connected to a third stirring blade (2344) located inside the beneficial bacteria dissociation tank (232).

8. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 6, characterized in that: The magnetic bead adsorbing component (235) includes a third electromagnetic induction coil (2351) and a third coil controller (2352). The third electromagnetic induction coil (2351) is sleeved on the magnetic bead adsorption bottle (233), and the third coil controller (2352) is electrically connected to the third electromagnetic induction coil (2351) to control the working state of the third electromagnetic induction coil (2351).

9. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 1, wherein: It further includes a harmful bacteria adsorption component (210) for removing harmful bacteria from the microbial mixture. The harmful bacteria adsorption component (210) is located between the filtration and impurity removal device and the beneficial bacteria enrichment component (220).

10. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 9, characterized in that: The harmful bacteria adsorption component (210) includes a first antibody magnetic bead (211), a harmful bacteria magnetic bead tank (212), and a harmful bacteria adsorption bottle (213). A harmful bacteria coupling component (214) for coupling harmful bacteria and the first antibody magnetic bead (211) together is provided on the harmful bacteria magnetic bead tank (212), and a harmful bacteria adsorbing component (215) for reversely enriching the harmful bacteria coupling body is provided on the harmful bacteria adsorption bottle (213).

11. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 10, wherein: The harmful bacteria coupling component (214) includes a first stirring motor (2141) and a first material port (2142). The first material port (2142) is used to introduce the first antibody magnetic bead (211) into the harmful bacteria magnetic bead tank (212), and the output end of the first stirring motor (2141) is a first stirring blade (2144) located inside the harmful bacteria magnetic bead tank (212).

12. The intestinal bacteria directed isolation system based on microbial antibodyomics technology according to claim 10, characterized in that: The harmful bacteria adsorbing component (215) includes a first electromagnetic induction coil (2151) and a first coil controller (2152). The first electromagnetic induction coil (2151) is sleeved on the harmful bacteria adsorption bottle (213), and the first coil controller (2152) is electrically connected to the first electromagnetic induction coil (2151) to control the working state of the first electromagnetic induction coil (2151).

13. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 9, characterized in that: It further includes an enrichment adsorption box (208). A first enrichment partition (261) and a second enrichment partition (262) are further provided inside the enrichment adsorption box (208). The first enrichment partition (261) divides the interior of the enrichment adsorption box (208) into a second operation area (263) and a second background area (264). The second enrichment partition (262) divides the second operation area (263) into a second clean area (265) and a second pollution area (266). The harmful bacteria adsorption component (210), the beneficial bacteria enrichment component (220), and the beneficial bacteria dissociation component (230) are all located in the second pollution area (266). The beneficial bacteria collection bottle (240) is located in the second clean area (265), and the second negative pressure component (250) is installed on the first enrichment partition (261).

14. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 8, wherein: It further includes a second purification bottle (271) and a second negative pressure pump (272). One end of the second negative pressure pump (272) is sequentially connected to the second purification bottle (271) and the harmful bacteria adsorption component (210), and the other end is sequentially connected to the second buffer (273), the second filter (274), and the beneficial bacteria collection bottle (240).

15. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 8, characterized in that: The second buffer (273) includes a third buffer bottle (2731) and a fourth buffer bottle (2372) arranged sequentially along the gas flow direction.

16. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 8, characterized in that: A second display controller (280) is further provided on the outer shell of the second clean area (265), and the harmful bacteria adsorption component (210), the beneficial bacteria enrichment component (220), and the beneficial bacteria dissociation component (230) are all electrically connected to the second display controller (280).

17. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 12, characterized in that: A second nitrogen cabinet (291) for placing nitrogen and a second consumable cabinet (292) for placing consumables are provided at the lower end of the enrichment and adsorption box (208), and the second nitrogen cabinet (291) is located between the filtration and impurity removal box (108) and the second consumable cabinet (292).

18. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 12, wherein: One or more of a second printer (201), a second fan (202), a second buzzer (203), a second spotlight (204), and a second lighting lamp (205) are further provided on the enrichment and adsorption box (208).

19. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to any one of claims 1 to 18, characterized in that: The filtration and impurity removal device (100) includes a dilution bottle (150) for providing water, a shredding component (130) for shredding the fecal sample, and a filtration component (140) for filtering the mixture after shredding. The negative pressure driving mechanism is used to make the water source in the dilution bottle (150) flow through the shredding component (130) and the filtration component (140) in sequence, thereby forming a microbial mixture.

20. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 19, characterized in that: The filtration and impurity removal device (100) further includes a filtration and impurity removal box (108), a first filtration partition (121), and a second filtration partition (122); the first filtration partition (121) divides the interior of the filtration and impurity removal box (108) into a first operation area (111) and a first backstage area (114), and the second filtration partition (122) divides the first operation area (111) into a first clean area (112) and a first pollution area (113); the shredding component (130) and the filtration component (140) are both arranged in the first pollution area (113), and the dilution bottle (150) is arranged in the first clean area (112).

21. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 20, wherein: The shredding component (130) includes a sample tank (131) and a shredding motor (132). A shredding shaft with shredding blades is arranged in the sample tank (131), and the output end of the shredding motor (132) is connected to the shredding shaft and drives the shredding shaft to rotate.

22. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 21, wherein: A first filter support plate (123) and a third filter support plate (125) for placing a sample tank (131) are installed in the first contamination area (113). The shredding motor (132) is installed below the first filter support plate (123). The third filter support plate (125) is installed on the first filter support plate (123). A shredding coupling (133) is arranged in the third filter support plate (125). The output end of the shredding motor (132) is connected to the shredding shaft through the shredding coupling (133).

23. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 20, characterized in that: The filter assembly (140) includes a primary filter (141) for performing primary filtration on the shredded mixture and a secondary filter (142) for performing secondary filtration on the shredded mixture. The dilution bottle (150), the sample tank (131), the primary filter (141), and the secondary filter (142) are sequentially connected through pipelines.

24. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 20, wherein: A second filter support plate (124) for placing the primary filter (141) and the secondary filter (142) is installed in the first contamination area (113). The first filter support plate (123) is located below the second filter support plate (124), and the first filter support plate (123) and the second filter support plate (124) are connected through a fourth filter support plate (126).

25. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 20, characterized in that: It further includes a first purification bottle (171) and a first negative pressure pump (172). One end of the negative pressure driving mechanism is sequentially communicated with the first purification bottle (171) and the shredding assembly (130), and the other end is sequentially communicated with a first buffer (173) and a first filter (174).

26. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 20, wherein: The first buffer (173) includes a first buffer bottle (1731) and a second buffer bottle (1732) arranged in sequence along the gas flow direction.

27. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 20, characterized in that: A sample collection rack (181) is further arranged in the first contamination area (113). A plurality of bacterial liquid collection bottles (182) are installed on the sample collection rack (181). The bacterial liquid collection bottles (182) are used for collecting the mixed liquid processed by the filter assembly (140).

28. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 20, characterized in that: A first display controller (191) is further arranged on the outer shell of the first clean area (112). The shredding assembly (130), the filter assembly (140), and the negative pressure assembly (160) are all electrically connected to the first display controller (191).

29. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 20, wherein: A first nitrogen cabinet (192) for placing nitrogen and a first consumable cabinet (193) for placing consumables are arranged at the lower end of the filter and impurity removal box (108). The first nitrogen cabinet (192) is located between the filter and impurity removal box (108) and the first consumable cabinet (193).

30. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 20, characterized in that: One or more of a first printer (101), a first fan (102), a first buzzer (103), a first spotlight (104), and a first lighting lamp (105) are further arranged on the filter and impurity removal box (108).

31. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 19, characterized in that: The negative pressure driving mechanism includes a first negative pressure component (160) and a second negative pressure component (250). The first negative pressure component (160) is used to sequentially transport the water source in the dilution bottle (150) to the shredding component (130) and the filtering component (140), and then form a microbial mixture; the second negative pressure component (250) is used to allow the microbial mixture to sequentially pass through the beneficial bacteria enrichment component (220) and the beneficial bacteria dissociation component (230), and then automatically enter the beneficial bacteria collection bottle (240).

32. The intestinal bacteria directed isolation system based on the microbial antibodyomics technology according to claim 31, characterized in that: The first negative pressure component (160) includes a first filtering peristaltic pump (161) installed between the dilution bottle (150) and the sample tank (131), and a second filtering peristaltic pump (162) installed between the primary filter (141) and the secondary filter (142).

33. The intestinal bacteria directional isolation system based on the microbial antibodyomics technology according to claim 31, characterized in that: The second negative pressure component (250) includes a first enrichment peristaltic pump (251) installed between the harmful bacteria adsorption component (210) and the beneficial bacteria enrichment component (220), a second enrichment peristaltic pump (252) installed between the beneficial bacteria enrichment component (220) and the beneficial bacteria dissociation component (230), and a third enrichment peristaltic pump (253) installed between the beneficial bacteria dissociation component (230) and the beneficial bacteria collection bottle (240).

34. A method for directional isolation of intestinal bacteria based on microbial antibodyomics technology, characterized in that, It includes the following steps: Under the action of the negative pressure driving mechanism, the filtration and impurity removal device (100) performs filtration and impurity removal treatment on the fecal sample to obtain a microbial mixture; Under the action of the negative pressure driving mechanism, the beneficial bacteria enrichment component (220) performs positive enrichment treatment on the beneficial bacteria in the microbial mixture to obtain a bacterial liquid mixture containing beneficial bacteria coupling bodies, and the beneficial bacteria coupling bodies are formed by coupling beneficial bacteria and second antibody magnetic beads; Under the action of the negative pressure driving mechanism, the beneficial bacteria dissociation component (230) first dissociates the beneficial bacteria coupling bodies to obtain a beneficial bacteria solution containing second antibody magnetic beads and beneficial bacteria; then the beneficial bacteria dissociation component (230) removes the second antibody magnetic beads in the beneficial bacteria solution to obtain a beneficial bacteria solution; Under the action of the negative pressure driving mechanism, the beneficial bacteria collection bottle (240) collects the beneficial bacteria solution.

35. The method for directional isolation of intestinal bacteria based on microbial antibodyomics technology according to claim 34, wherein The filtration and impurity removal device (100) performs filtration and impurity removal treatment on the fecal sample to obtain a microbial mixture, including: The water source in the dilution bottle (150) first enters the sample tank (131) under the action of the first filtering peristaltic pump (161). At this time, the sample tank (131) contains a mixture of water source and fecal sample, which is recorded as the first mixture; The shredding motor (132) below the sample tank (131) centrifugally stirs the first mixture to obtain a mixture after centrifugal crushing of the fecal sample, which is recorded as the second mixture; The second mixture, under the action of the second filtering peristaltic pump (162), sequentially passes through the primary filter (141), the second filtering peristaltic pump (162) and the secondary filter (142) for treatment, forms an intestinal microbial mixture, and enters the bacterial liquid collection bottle (182).

36. The method for directional isolation of intestinal bacteria based on microbial antibodyomics technology according to claim 35, wherein The beneficial bacteria enrichment component (220) performs a positive enrichment process on the beneficial bacteria in the microbial mixture to obtain a bacterial liquid mixture containing beneficial bacteria conjugates, including: The microbial mixture enters the beneficial bacteria magnetic bead tank (222), and the second antibody magnetic beads (221) enter the beneficial bacteria magnetic bead tank (222) from the second material port (2242); The second stirring motor (2241) drives the second stirring blade (2244) connected to itself to rotate according to a first preset time to mix the solution in the beneficial bacteria magnetic bead tank (222); Incubate at room temperature for a second preset time to couple the second antibody magnetic beads with the beneficial bacteria to obtain beneficial bacteria conjugates; The second negative pressure component (250) drives the beneficial bacteria conjugates into the beneficial bacteria enrichment bottle (223), and the beneficial bacteria enrichment member (225) performs positive enrichment on the beneficial bacteria conjugates to obtain a bacterial liquid mixture containing beneficial bacteria conjugates.

37. The method for directional isolation of intestinal bacteria based on microbial antibodyomics technology according to claim 36, wherein The steps of obtaining a beneficial bacteria solution from the bacterial liquid mixture include: Let the bacterial liquid mixture containing beneficial bacteria conjugates enter the dissociation bottle, and add dissociation enzyme through the third material port (2342); The third stirring motor (2341) drives the third stirring blade (2344) connected to itself to rotate according to a first preset time to mix the solution in the dissociation bottle; Incubate at room temperature for a third preset time to dissociate the third antibody magnetic beads from the beneficial bacteria to obtain a mixture containing the third antibody magnetic beads and dissociated beneficial bacteria; The second negative pressure component (250) drives the above-mentioned mixture containing the third antibody magnetic beads and dissociated beneficial bacteria into the magnetic bead adsorption bottle (233), and the magnetic bead adsorption member (235) adsorbs the second antibody magnetic beads (221) to obtain a beneficial bacteria solution.

38. The method for directional isolation of intestinal bacteria based on microbial antibodyomics technology according to claim 37, wherein After obtaining the microbial mixture, before the beneficial bacteria enrichment component (220) performs a positive enrichment process on the beneficial bacteria in the microbial mixture, it further includes the step of removing harmful bacteria in the microbial mixture through the harmful bacteria adsorption component (210), including: The microbial mixture enters the harmful bacteria magnetic bead tank (212), and the first antibody magnetic beads (211) enter the harmful bacteria magnetic bead tank (212) from the first material port (2142); The first stirring motor (2141) drives the first stirring blade (2144) connected to itself to rotate according to a first preset time to mix the solution in the harmful bacteria magnetic bead tank (212); Incubate at room temperature for a second preset time to couple the first antibody magnetic beads with the harmful bacteria to obtain a solution containing harmful bacteria conjugates; The second negative pressure component (250) drives the solution containing harmful bacteria conjugates into the harmful bacteria adsorption bottle (213), and the harmful bacteria adsorption member (215) performs reverse enrichment on the harmful bacteria conjugates to remove the harmful bacteria in the microbial mixture.

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