CN211206531U - Coated magnetic microsphere biochemical detection system with microporous membrane for intercepting and gathering - Google Patents

Abstract

The coated magnetic microsphere biochemical detection system intercepted and gathered by the microporous membrane comprises a microporous filter cup, a coated magnetic microsphere reaction reagent and a magnetic device; the capture reagent fixed on the surface of the coated magnetic microsphere in the coated magnetic microsphere reaction reagent specifically identifies the substance to be detected and is in direct or indirect ligand relationship with the marking reagent. The magnetic device transfers the coated magnetic microsphere composite to the microporous filter bowl. The pore diameter of the microporous filter membrane is basically uniform and is smaller than the diameter of the coated magnetic microsphere and larger than the diameter of the labeled reagent, and the structural solution of the microporous filter cup can only flow out through the microporous filter membrane. The microporous filter membrane precisely intercepts the coated magnetic microsphere compound for detection.

Description

Coated magnetic microsphere biochemical detection system with microporous membrane for intercepting and gathering

Technical Field

The utility model relates to a liquid phase immunity or ligand detection technology in the field of biomedical diagnosis, in particular to a biochemical detection system and a detection method thereof, wherein the coated magnetic microspheres are gathered and dispersed in solution, and the microporous membrane with uniform pore diameter intercepts and gathers the coated magnetic microspheres simultaneously.

Background

The immunoassay technology utilizes high-specificity recognition and binding reaction between an antigen and an antibody to realize detection of biomolecules, has the advantages of high sensitivity, strong specificity, wide application range, simple required equipment, wider linear range and the like, becomes one of the most competitive and challenging analysis and test technologies at present, and is widely applied to the fields of life science, clinical medicine, environment, food, medicine and the like.

The widely used enzyme label plate coats protein, so that the stability of the protein is enhanced, the detected signal molecules are combined with the protein coated on the enzyme label plate, impurities in a solution can be conveniently removed, but the multi-step operations of embedding, elution, separation and the like are required, the analysis process is complicated, the analysis time is long, and the requirements of rapid detection and diagnosis cannot be met.

The protein is coated by soluble polymer particles to enhance the stability of the protein, and the patent document CN201110306613.4 discloses that the HCV-cAg in human serum is quantitatively detected by using a polystyrene microsphere coated HCV-cAg primary antibody, an enzyme-labeled antibody HCV-cAg secondary antibody and a 96-hole filter plate with a permeable filter membrane at the bottom, compared with a commercially available E L ISA kit, the sensitivity is high (4.3 pg/ml vs 50 pg/ml), and the linear range (13 pg-50 ng/ml) is wide (more than three orders of magnitude vs is less than two orders of magnitude).

Patent document CN201310228708.8 discloses a quantitative detection device based on fiber membrane trapping and separation and a detection method thereof: the latex microspheres for fixing the capture reagent are placed in a reaction cup, static mixing incubation reaction is carried out on the latex microspheres and the labeled reagent, the reaction mixture is transferred into a detection cup provided with a deep-hole filter plate, then a reaction solution and a washing solution are drained from a filter membrane of the filter plate through the action of a water absorption cup, small molecules such as the labeled reagent and the like or substances with small particle sizes flow out of the deep-hole filter plate along with the solution, the coated microspheres are partially intercepted when passing through the bottom filter membrane, and then the surface of the membrane is detected through optics to obtain an experiment of related detection signals. The method has the significance that the immunoreaction reagent is statically incubated in a homogeneous environment, so that the requirement on the affinity of the immune antibody antigen can be reduced; meanwhile, after incubation reaction, the coated microspheres carrying the reaction compound in the solution are gathered in the filter membrane, so that a good reaction substance concentration effect is achieved. However, the relationship between the particle size of the coated microspheres and the pore size of the filter membrane in the patent document is not clearly defined, and the microspheres will penetrate into the filter membrane to a certain depth, which will cause part of the microspheres to penetrate through the filter membrane, thereby causing inaccurate detection results. At the same time, differences in the spectral signals reflected by the microspheres at different depths of the filter will also result.

Impurities in the sample may affect the filtration operation, such as clogging of the pores of the filter membrane or adhesion and resistance generation. To solve this problem, a pretreatment step is usually added, and a certain degree of purification is performed during the pretreatment.

The magnetic components can be controlled by the magnetic frame and fixed on the set surface to complete the operations of separation, impurity removal, transfer and the like. Patent documents CN201520303370.2 and CN201620401672.8 each disclose a magnetic rack for separating, purifying and extracting nucleic acids, respectively.

SUMMERY OF THE UTILITY MODEL

The utility model discloses hold back the millipore filtration membrane of coating microballon and gather the separation preliminary treatment that the coating microballon increases magnetic force mediation among the fluorescence labeling detection scheme to the millipore filtration membrane filters the step and optimizes. The utility model aims at providing a microporous membrane holds back biochemical detecting system of peridium magnetic microsphere that gathers.

A coated magnetic microsphere biochemical detection system with microporous membrane for trapping and collecting includes an incubation container for specific binding reaction. The binding reaction of the specific and highly sensitive capture reagent with the substance to be detected is the detection basis of the immunoassay. At least one group of capture reagent and labeling reagent which are directly or indirectly in ligand relationship with each other are arranged in the incubation container, and the ratio of the binding state and the free state of the labeling reagent is determined by the amount of the highly specific complex of the capture reagent and the substance to be detected. When a proper amount of capture reagent and labeling reagent exist and the concentration of the substance to be detected is in a detection range, a simple and definite quantitative relation exists between the amount of the substance to be detected and the amount of the labeling reagent in the compound, namely, the quantitative relation between the amount of the substance to be detected and the detection signal intensity can be obtained through concentration curve or simplified theoretical calculation, and the amount of the substance to be detected is obtained according to the quantitative relation and the detection signal intensity.

The product of the utility model can accurately remove the free labeled reagent and accurately determine the amount of the labeled reagent in the compound.

The incubation container is a container with an upper opening. The capture reagent is fixed on the surface of a coated magnetic microsphere with uniform diameter.

The coated magnetic microsphere biochemical detecting system with microporous membrane for trapping and collecting also includes magnetic unit for fixing magnetic particle under strong magnetic force, microporous filtering cup for separating the specific reaction compound and equipment for detecting the fluorescence or color signal of the labeled reagent.

The microporous filter cup is of an upper open structure, the volume of the microporous filter cup is not more than 900 microliters, the center of the bottom of the microporous filter cup is provided with a first through hole with the area of less than 12 square millimeters, a microporous filter membrane with basically uniform aperture and the aperture range of 0.02-1.2um is attached to the outer surface of the wall of the first through hole, and the microporous filter membrane covers the first through hole; the inner surface of the bottom of the microporous filter cup which takes the center of the first through hole as an axis and is the lowest point is set into an arc surface, a spherical surface or a conical surface with a certain inclination angle.

The pore diameter of the microporous filter membrane of the microporous filter cup is uniform, and the pore diameter of the microporous filter membrane is smaller than the diameter of the coated magnetic microspheres. The capture reagent-coated magnetic microsphere compound containing the marking reagent is trapped and gathered on the surface of the microporous filter membrane. The solution in the microporous filter cup is coated with magnetic microspheres to enrich the microporous filter membrane from the whole volume to a plane and from the section of the larger microporous filter cup to the section of the smaller first through hole.

Impurities in the reaction solution are removed from the reaction complex solution in the incubation container to the coated magnetic microspheres in the microporous filter cup. The volume of the incubation container is not limited by the detection device, and a relatively large amount of sample or a sample having a high viscosity and requiring a large amount of solution treatment can be processed, but is limited by the effective acting distance of the magnetic force of the magnetic device (the processing time is about 10s, the effective distance is millimeter, and the volume is about 500 microliters).

Both permanent magnets and electromagnets may provide magnetic force. The utility model provides two kinds of magnetic means schemes that contain magnet: magnetic force frame and bar magnet add stick cover. And both protocols allow batch processing of samples. The magnet adopted by the magnetic frame is in a ring shape, a round shape or a strip shape, the two surfaces of the slice are respectively provided with N/S poles, and the two ends of the magnetic bar are respectively provided with the N/S poles. The magnetic frame fixes the particles with magnetism on the surface of the wall of the incubation container, then removes the solution containing impurities, and disperses the particles with magnetism in the new solution in the incubation container; the combination of the magnetic rod and the rod sleeve enables the particles with magnetism to be fixed on the surface of the rod sleeve and then dispersed in the solution in the incubation container or the micropore filter cup.

The solution of the utility model can only flow out through the microporous filter membrane. In order to fix the microporous filter membrane, a microporous sleeve cup is arranged outside the microporous filter cup, and the microporous sleeve cup provides a second through hole and clamps the microporous filter membrane together with the first through hole of the microporous filter cup. The outer wall of the microporous filter cup and the inner wall of the microporous sleeve cup are provided with a first groove or a first convex ring buckle to connect the microporous filter cup and the microporous sleeve cup, so that the microporous filter membrane is tightly clamped by the microporous filter cup and the microporous sleeve cup. And a second groove or a second convex ring is arranged below the outer wall of the microporous filter cup and below the inner wall of the microporous sleeve cup to be in sealing connection with the microporous filter cup and the microporous sleeve cup, so that the solution cannot flow between the microporous filter cup and the microporous sleeve cup, only can pass through the microporous filter membrane, and flows out of the microporous filter cup through the first through hole and the second through hole.

The utility model discloses it reaches the collection sleeve pipe to set for solution to flow out from the second through-hole. The collecting sleeve comprises a sleeve on the upper part and a collecting pipe arranged on the lower part, the inner part of the sleeve is fixedly connected with the microporous filter bowl or the microporous sleeve bowl, the sleeve is thick, the collecting pipe is thin, gas in the collecting pipe escapes from a gap between the sleeve and the microporous filter bowl or the microporous sleeve bowl, and the sleeve is connected with the collecting pipe in a sealing way.

Except that the solution passes through the microporous filter membrane under the action of gravity, the utility model also provides two schemes of centrifugal force and vacuum pressure.

The coated magnetic microsphere biochemical detection system intercepted and gathered by the microporous membrane also comprises a centrifuge, wherein an arc-shaped groove is arranged on a centrifugal rotating head of the centrifuge and is a collecting sleeve mounting groove; the collecting sleeve can slide relative to the collecting sleeve mounting groove. The sliding of the collecting sleeve ensures that the stress direction of the solution in the microporous filter cup points to the microporous filter membrane, the solution in the microporous filter cup can quickly and uniformly flow out through the microporous filter membrane, and the coated magnetic microspheres (containing compounds) in the solution are intercepted.

The centrifugal rotor arc collection sleeve mounting groove of the centrifugal machine has the following application conditions: the shapes and weights of the centrifugal tubes are consistent. The centrifuging tube is vertical when the centrifugation turn round is static, can use rotational position sensor location centrifuging tube, and the cooperation manipulator snatchs and carries out subsequent full-automatic (detection) operation.

The coated magnetic microsphere biochemical detection system intercepted and gathered by the microporous membrane also comprises a vacuum kit, namely, the vacuum kit is communicated with the air exhaust function or vacuum low pressure of a second through hole of the microporous sleeve cup or the outer gap of the microporous filter membrane of the microporous filter cup and is communicated with the air inlet function or constant atmospheric pressure of the microporous filter cup.

The vacuum kit comprises an air suction and inlet cylinder arranged above the opening of the microporous filter cup and a sealing sleeve which is sleeved outside the microporous filter cup and is communicated with the collection sleeve and an air outlet pipeline in the air suction and inlet cylinder; the upper part of the central position in the air extracting and inlet cylinder is provided with an air extracting column, the lower part is provided with an air inlet column, and the air inlet column is provided with one or more air inlet pipelines and one or more air outlet pipelines, and the air inlet column is butted and sealed with the opening of the microporous filter cup or the microporous sleeve cup; one end of the air inlet pipeline is communicated with the air inlet column, and the other end of the air inlet pipeline is communicated with the atmosphere; the air inlet column maintains the atmospheric pressure; the air pumping column is connected with a vacuum air path of the vacuum pumping equipment, one end of the air outlet pipeline is communicated with the air pumping column, and the other end of the air outlet pipeline is communicated with a gap between the sealing sleeve and the microporous filter cup or the microporous sleeve cup. The air suction cylinder is in sealing fit with the sealing sleeve, and the lower part of the sealing sleeve is in sealing fit connection with the collecting sleeve, so that the vacuum sleeve and the collecting sleeve form a closed space. The upper edge of the wall of the microporous filter cup or the microporous sleeve cup is hermetically connected with an air inlet column of the air suction and inlet column body; the air inlet channel passes through the air inlet pipeline and the air inlet column of the air suction and air inlet column from the atmosphere to the microporous filter cup, the air pressure of the microporous filter cup is larger than the air pressure of the collecting sleeve and the sealing sleeve, and the liquid passes through the microporous filter membrane from the first through hole to reach the second through hole and the collecting pipe. The air pumping channel is arranged outside the microporous filter cup or the microporous sleeve cup and is connected to the air outlet pipeline/the air pumping column in the air pumping inlet column from the second through hole to the collecting sleeve and the sealing sleeve and then to the vacuum pump. The liquid enters the collection tube after exiting the second through hole. The pumping channel is kept at a low pressure.

Under the action of air pressure, the liquid in the microporous filter cup can quickly pass through the microporous filter membrane. The atmospheric pressure is kept in front of the microporous filter membrane, and the air exhaust channel behind the microporous filter membrane is free of liquid, so that the phenomenon of aerial fog formed by the surface breakage of liquid drops is avoided, and the pollution is reduced. In contrast, centrifugal forces act uniformly on the surface and interior of the droplets, theoretically avoiding turbulence is only necessary to avoid aerosol.

Furthermore, the outer wall of the air suction and inlet cylinder is provided with one or more fourth convex rings or fourth grooves, the inner wall of the sealing sleeve is provided with one or more fourth grooves or fourth convex rings, and the fourth convex rings are in sealing fit connection with the fourth grooves. The sealing sleeve simultaneously seals the pipeline leading the air suction and inlet cylinder to the surface, so that the processing of the air suction and inlet cylinder can be simplified.

The pore diameter of the microporous filter membrane is larger than the diameter of the labeled reagent, and the free labeled reagent can pass through the microporous filter membrane; the aperture of the microporous filter membrane is smaller than the diameter of the coated magnetic microspheres, and the coated magnetic microspheres and the compound thereof are trapped and gathered on the surface of the microporous filter membrane. The aperture of the coated magnetic microspheres and the compound accumulation layer trapped and collected on the surface of the microporous filter membrane is larger than the diameter of the marking reagent, and the free marking reagent can trap the coated magnetic microspheres and the compound accumulation layer collected and collected on the surface of the microporous filter membrane. The microscopic size relationship between the coated magnetic microspheres and the labeling reagent in the microporous filter cup is as follows: when the free coated magnetic microspheres and the coated magnetic microspheres which form a reaction compound with the labeled reagent are stacked on the surface of the microporous membrane, the free labeled reagent can be carried by the solution from the pores among the stacked coated magnetic microspheres to pass through the micropores of the microporous membrane again and flow out, and no blocking residue is formed. It can be determined by experiment whether the microscopic dimensions of the labeled reagent and the coated magnetic microsphere complex are consistent.

The model is that a capture reagent small ball is fixed on a large coated magnetic microsphere ball, and a marking reagent small ball is freely put in and out. The capture reagent is not considered, the simple geometric relationship is that four same large spheres are close to each other, four centers of the four spheres form four vertexes of a regular tetrahedron, and the maximum diameter of the sphere which can freely go in and out among the four spheres is solved; three same circles are close to each other, the centers of the three circles form three vertexes of a regular triangle, and the largest circle which can be drawn between the three circles is obtained. The ratio of the diameter of the small labeling reagent ball to the diameter of the large coated magnetic microsphere is less than

(approximately equal to 0.15). Assuming that the diameters of the capture reagent and the labeling reagent are the same, the above ratio should be less than about 0.09. In practice, the test results shall be used as the standard.

The utility model provides a microporous membrane holds back the biochemical detecting system's of the coating magnetic microsphere that gathers detection method. To detect a label signal related to the amount of the substance to be detected,

the selected labeling reagent comprises a fluorescent substance with a spectral signal or a color particle directly visible to the naked eye, and the color particle is a nanoparticle with a color signal; the fluorescent substance is selected from one or more of fluorescent molecules and microspheres thereof, quantum dot particles and microspheres thereof, up-conversion luminescent particles and microspheres thereof, time-resolved fluorescent molecules and microspheres thereof, and fluorescent protein.

The selected detection device optically detects at least one wavelength signal from the spectral signal of the labeled reagent gathered on the surface of the microporous filter membrane of the first through hole at the bottom of the microporous filter cup from the upper part of the microporous filter cup.

The detection method comprises the following steps:

putting a solution to be tested into the incubation container provided with the coated magnetic microspheres and the labeled reagent;

enriching, gathering and fixing the coated magnetic microspheres in the solution in the incubation container by using a magnetic device; discarding the solution containing impurities in the incubation container; transferring the coated magnetic microspheres into a microporous filter cup;

after the solution in the microporous filter cup flows out from the first through hole attached to the microporous filter membrane, the microspheres gathered on the surface of the microporous filter membrane are subjected to spectral signal detection.

The utility model discloses a microporous membrane holds back the beneficial effect of the biochemical detecting system of packet magnetic microsphere who gathers is:

1. low background and high sensitivity. And removing impurities from the step of transferring from the incubation container, completely and strictly obtaining the labeled reagent in the compound by the filter membrane interception, wherein the labeled reagent has high signal intensity and is matched with a detection method.

2. The detection is convenient, and the operation is easy and fast. The liquid phase reaction is easy, and the operation steps are easy. Vacuum and centrifugation protocols accelerate filtration.

3. The pretreatment is convenient. An incubation container is arranged, primary purification is carried out under magnetic force mediation, the treatment is convenient, and impurities blocking the microporous filter membrane in the subsequent steps can be removed along with the waste liquid.

4. Uniform operation and automation are easily realized.

5. Avoiding pollution.

Drawings

FIG. 1, coated magnetic microsphere biochemical detection system with microporous membrane trapped and aggregated

FIG. 2, appearance and internal Structure of microporous Filter bowl

FIG. 3, microporous filter bowl and microporous sleeve bowl clamping microporous filter membrane

FIG. 4, the projection outside the microporous sleeve cup (together with the collecting sleeve or the sealing sleeve to form the gas channel)

FIG. 5 three examples of magnetic force frames

FIG. 6 examples of magnetic rods and rod sleeves

FIG. 7, microporous sleeve and collection sleeve

FIG. 8, microporous sleeve, microporous filter bowl and collection sleeve

FIG. 9, centrifugal rotor

FIG. 10 shows the structure of the vacuum kit (air inlet cylinder and sealing sleeve), microporous sleeve cup, microporous filter cup, and collecting sleeve (left: air inlet section and right: air inlet section)

FIG. 11 Assembly of vacuum kit (air suction cylinder + sealing sleeve), microporous sleeve cup and microporous filter cup, Collection sleeve

FIG. 12, the structure diagrams of the vacuum set (air suction inlet cylinder + sealing sleeve), microporous sleeve, microporous filter bowl, and collecting sleeve (left: air suction section and right: air inlet section) of FIG. 10 without drawing microporous sleeve bowl and microporous filter bowl

The black six stars indicate that the vacuum sleeve and the collection sleeve are hermetically connected at two places

FIG. 13 is a schematic drawing of the vacuum kit (air suction inlet cylinder + sealing sleeve), microporous sleeve cup and microporous filter cup of FIG. 10 without drawing the wall of the right collection sleeve, and the collection sleeve (left: air suction cross-section and right: air inlet cross-section). Hollow arrows indicate gas passages, black six stars indicate microporous sleeve cups and microporous filter cups and air suction and inlet cylinders

FIG. 14 shows an example of the inlet and exhaust passages, two inlet pipes and six outlet pipes

FIG. 15, bleed air and intake air passages (with internal structure of all relevant parts as background)

FIG. 16 shows the result of the coated magnetic microsphere biochemical detection system for intercepting and aggregating microporous membrane for detecting the content of benzopyrene in edible oil

100. A microporous filter bowl; 110. a micropore retainer cup; 101. a microporous filtration membrane; 102. a first through hole; 112. a second through hole; 103. the inner bottom surface of the microporous filter cup; 104. 114, a first groove convex ring which is connected with the microporous filter cup and the microporous sleeve cup in a buckling way; 105. 115 and a second groove convex ring which is hermetically connected with the microporous filter cup and the microporous sleeve cup; 117. the outer wall of the micropore retainer cup is raised;

200. a magnetic device; 210. a magnetic frame; 220. the magnetic rod and the rod sleeve; 211. a magnetic assembly; 221. a magnetic bar; 222. a rod sleeve; 213. 223, container holder;

300. incubating the container;

400. collecting the cannula; 410. a collection pipe; 420. a sleeve; 427. a sleeve-micropore sleeve cup gap;

500. a vacuum bushing; 510. air extraction and inlet column; 520 sealing the sleeve; 511. pumping a gas column; 512. an air inlet column; 513. an air outlet pipe; 514. an air intake duct;

600. and (5) centrifuging and rotating the head.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, embodiments accompanied with figures are described in detail below, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person having ordinary skill in the art without creative efforts shall belong to the protection scope of the present invention.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be implemented in other ways different from the specific details set forth herein, and one skilled in the art may similarly generalize the present invention without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.

In the description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "upper, lower, inner and outer" and the like are based on the directions or positional relationships shown in the drawings, and are only for convenience of description and simplification of description, but do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and thus, should not be construed as limiting the present invention.

One of the purposes of the embodiment design of the utility model is to realize large flux and automatic detection.

Example 1

As shown in FIG. 1, a coated magnetic microsphere biochemical detection system with microporous membrane trapping and accumulating comprises a microporous filter bowl 100, an incubation container (test tube) and a magnetic device. The left side of fig. 1 is provided with two test tube racks, the upper left test tube rack is provided with a magnet and is a magnetic rack (a magnetic device), the lower left test tube rack is a common test tube rack, and one test tube is internally provided with a rod sleeve and a magnetic rod (another magnetic device). The right side is an enlarged view of the microporous filter bowl structure.

The incubation container (test tube) is provided with coated magnetic microspheres with uniform diameters, a capture reagent which is fixed on the surfaces of the coated magnetic microspheres and has the functions of specifically recognizing and combining substances to be detected, and a labeling reagent which is in direct or indirect ligand relationship with a capture reagent-substance to be detected compound. The quantity of the coated magnetic microspheres, the capture reagent and the marking reagent of the coated magnetic microsphere biochemical detection system (product) intercepted and gathered by the microporous membrane is fixed, and the quantity relationship between the quantity of the substance to be detected and the quantity of the compound of the substance to be detected and the capture reagent and the quantity relationship between the quantity of the substance to be detected and the quantity of the compound of the marking reagent and the coated magnetic microspheres are determined in a certain interval of the concentration of the substance to be detected and have simple and accurate formulas. In the incubation process, the optimal state of the coated magnetic microspheres is that the coated magnetic microspheres are uniformly dispersed in the solution, the magnetic force action is not needed, and the incubation container test tubes are inserted into a common test tube rack.

After incubation, to remove impurities from the solution and transfer the coated magnetic microsphere-labeled reagent complex to a microwell,

scheme 1: the incubation container test tube is inserted into the magnetic frame shown at the upper left of the figure 1, the magnet is tightly attached to the wall of the test tube, and the coated magnetic microspheres in the solution are gathered and fixed at the position, close to the magnet, of the inner wall of the test tube under the action of the magnetic force. The solution in the tube may be removed (aspirated, poured off, or otherwise). The test tube is moved out of the magnetic frame and added with a proper amount of solution without impurities, the coated magnetic microspheres attracted and fixed by the magnet on the wall of the test tube are dispersed in the newly added solution, the process can be repeated to remove the impurities and the free labeled reagent, the coated magnetic microsphere-labeled reagent compound formed by incubation is kept stable in a short time, and the coated magnetic microsphere solution is transferred into the microporous filter cup.

Scheme 2: as shown in the left lower part of fig. 1, the magnetic rod is inserted into the bottom of the rod sleeve, and is inserted into the incubation container test tube together with the rod sleeve, under the action of magnetic force, the coated magnetic microspheres in the solution are gathered and fixed on the outer wall of the rod sleeve of the magnetic rod, the magnetic rod/rod sleeve is taken out, the solution in the incubation container test tube is poured out, a proper amount of solution without impurities is added into the incubation container or the microporous filter cup, the magnetic rod/rod sleeve is inserted into the newly replaced solution, the magnetic rod is pulled out from the rod sleeve, the coated magnetic microspheres fixed on the rod sleeve are dispersed in the solution, the above process can be repeated to remove impurities, and finally the coated magnetic microspheres are dispersed in the solution in the microporous filter cup.

Alternative schemes are as follows: as shown in the left lower part of fig. 1, the magnetic rod is inserted into the bottom of the rod sleeve, and is inserted into the incubation container test tube together with the rod sleeve, under the action of magnetic force, the coated magnetic microspheres in the solution are gathered and fixed on the outer wall of the rod sleeve of the magnetic rod, the magnetic rod/rod sleeve is taken out, the solution in the incubation container test tube is poured out, a proper amount of solution without impurities is added into the incubation container or the microporous filter cup, the magnetic rod/rod sleeve is inserted into the newly replaced solution, the magnetic rod is pulled out from the rod sleeve, the coated magnetic microspheres fixed on the rod sleeve are dispersed in the solution, the above process can be repeated to remove impurities, finally, the coated magnetic microspheres are dispersed in the solution in the incubation container, and the solution containing the coated magnetic microspheres is transferred into the microporous filter cup.

The center of the bottom surface 103 of the microporous filter cup is provided with a first through hole 102, and the microporous filter membrane is attached to the outer wall of the first through hole. The bottom surface of the microporous filter cup is arranged into an arc surface, a spherical surface or a conical surface with a certain inclination angle by taking the center of the first through hole as an axis and taking the lowest point as the lowest point; the junction of the bottom surface and the side wall of the microporous filter cup and the first through hole is a fillet, so that the solution is not retained. The diameter of the coated magnetic microspheres is uniform, the aperture of the microporous filter membrane is uniform, the diameter of the coated magnetic microspheres is larger than that of the microporous filter membrane, and the diameter of the labeled reagent is smaller than that of the microporous filter membrane. The coated magnetic microsphere solution flows out through the microporous filter membrane in the microporous filter cup. The free labeling reagent flows out along with the solution, the labeling reagent combined with the coated magnetic microspheres is trapped in the first through hole microporous filter membrane, and the amount of the substance to be detected is calculated according to the detection result.

The following table is summarized:

generally, the two ends of the bar magnet are N or S poles, and the upper and lower surfaces of the magnet piece are N or S poles, limited by the magnet. The magnet pieces are stronger than the magnetic field of the magnetic bar. The magnetic frame can adopt a magnet piece with a stronger magnetic field, so that the magnetic frame is suitable for treating solution with larger volume, and the scheme of matching the magnetic frame and the magnetic rod is as follows: the magnetic particles are fixed in a smaller range, and after being removed, the magnetic rod is used for gathering and fixing the magnetic particles in a relay way.

Example 2

Three magnetic frames are shown in fig. 5. The upper and middle magnetic assemblies 211 of fig. 5 can move, the upper magnetic assembly 211 of fig. 5 rotates around the fixed shaft at the upper part of the magnetic frame, and is positioned at the bottom of the container placed on the container support 213 at the position A, the magnetic force is strong enough to adsorb the particles coated with magnetism and fix the particles coated with magnetism on the inner surface of the bottom wall of the container, the magnetic assembly 211 is far away from the position A at the position B, the magnetic force is weakened, and the particles coated with magnetism are dispersed; in FIG. 5, the magnetic assembly can be far away from, farthest to the B position, or close to, nearest to A, when the incubation container is placed on the support 213, the magnetic force of the magnetic assembly at the A position is so strong that the coated magnetic particles can be fixed on the side wall of the incubation container, and when the magnetic assembly is at the B position, the coated magnetic particles are dispersed; moving incubation container (cuvette) 300 in fig. 5, when cuvette 300 is in position a, coated magnetic particles are collected on the sidewall of the cuvette, and when the cuvette is even removed in position B, coated magnetic particles are dispersed. The magnetic frame is designed in a way that the requirements of cost, processing difficulty, incubation reaction volume and automatic operation design are comprehensively considered.

Example 3

As shown in FIG. 6, the magnetic rod and the rod cover together form a surface independent of the incubation container, which should be free of substances interfering with biochemical detection, sometimes with a magnetic force so strong that the particles coated with magnetic particles are aggregated and immobilized, and sometimes with no magnetic force at all, and the particles coated with magnetic particles are dispersed. The specific scheme is that the magnetic rod is inserted into the rod sleeve to the bottom or the magnetic rod is pulled out of the rod sleeve. Obviously, an electromagnet rod with an inert material surface that does not affect biochemical reactions and biochemical detection would achieve the same result if the electromagnet could better eliminate residual magnetism.

Example 4: the microfiltration bowl 100 and the microporous sleeve 110 hold the microporous filter membrane 101.

As shown in fig. 2, the microporous filter membrane 101 is attached to the outer surface of the bottom of the first through hole 102 of the microporous filter cup 100; as shown in FIG. 3, the microfiltration membrane 101 is held by the microfiltration cup 100 and the microporous sleeve 110. The labeled reagent bound to the coated magnetic microspheres is entrapped within the first through-hole microporous filter membrane as described in example 1. Placing the microporous filter membrane together with the microporous filter cup (as shown in figure 2) or together with the microporous filter cup and the microporous sleeve cup (as shown in figure 3) on a position to be detected of a detection instrument to detect the coated magnetic microsphere-signal molecule compound intercepted on the surface of the microporous filter membrane. And calculating the amount of the substance to be detected according to the detection result.

The microporous filter membrane can be attached to the outer surface of the first through hole by glue or the like (as shown in fig. 2), but it is difficult to obtain a good effect because the attachment area is small. It is necessary to protect its position under the microfiltration membrane. Or fixing the microporous filter membrane by using glue and the microporous sleeve cup at the same time. When the glue is used, the microporous filter membrane is required to be firmly adhered, the common microporous filter membrane is discarded after use, and if the microporous filter cup is cleaned and reused, the discarded microporous filter membrane is required to be easily and completely removed, so that the microporous sleeve cup which is only used without the glue is better in the condition.

As shown in fig. 3, the microporous filter bowl and the microporous sleeve bowl are sealingly connected by a second lower groove or second raised ring (105 and 115, 105 being shown as a protrusion in fig. 2 and 3, and 105 also being a groove). The purpose of sealing connection is to prevent the solution from entering the narrow space between the microporous filter cup and the microporous sleeve cup from the side surface of the microporous filter membrane, and if the solution with the labeled reagent stays at the periphery of the microporous filter membrane, the solution is not easy to be washed and removed, thereby influencing the experimental result. The sealing of the second groove or the second convex ring of the microporous filter cup and the microporous sleeve cup and the clamping of the first through hole and the second through hole on the microporous filter membrane form a minimum space between the microporous filter cup and the microporous sleeve cup, and the minimum space can be considered to accord with the setting of sealing in the process that the solution passes through the microporous filter membrane: the product PV of pressure and volume is constant. The smaller the volume of the sealed space is, the same volume change rate corresponding to the same pressure change is, the smaller the volume change is, that is, the absolute value of the space with the small volume which gives out the liquid due to the external pressure is small, while the larger the volume change rate corresponding to the same volume change is, the larger the pressure change is, that is, the greater the difficulty and resistance of the liquid with the same volume to permeate. Therefore, a second groove or a second collar of the seal is designed below. But for a longer time the pressure of this very small space is the same as the outside, i.e. the space is not absolutely sealed.

The microporous filter cup and the microporous sleeve cup are connected by a first groove or a first convex ring (104 and 114, which are matched with each other and can be concave or convex) on the upper part in a buckling way, and the purpose of the buckling connection is to ensure that the force for clamping the microporous filter membrane by the first through hole and the second through hole is enough. The strength that ensures the first through-hole of millipore filtration laminating is mainly two: the stress and the friction force determined by the surface properties of the microporous filter membrane, the microporous filter cup and the microporous sleeve cup are very slightly elastically deformed when the first through hole is abutted against the second through hole. The snap connection needs to be positioned accurately and does not need to be sealed.

Example 5

To avoid contamination or the need for collection of the solution that has passed through the microporous membrane, a collection cannula consisting of a cannula 420 and a collection tube 410 is designed (FIG. 7). The collecting sleeve is connected with the micropore sleeve cup. The microporous sleeve outer wall and bottom (outer) surface are designed with protrusions 117 as shown in figure 4, four protrusions extending through the outer wall to the bottom surface, and gas passages 427 as shown in figures 7 and 8 cooperating with the smooth inner surface of the collection sleeve.

In order to make solution cross millipore filtration more fast, the utility model provides a two schemes, centrifugal force and evacuation to for avoiding producing the aerial fog of solution in the scheme of evacuation, the design is to receiving the pipe downwards after millipore filtration, and the gas circuit of evacuation upwards connects evacuating device outside the micropore retainer cup.

Example 6

The design of a centrifuge rotor as shown in fig. 9 is very different from a conventional centrifuge rotor (the conventional centrifuge rotor is designed to tolerate imbalance caused by different centrifuge tubes).

The utility model provides a centrifuging tube is identical, and characteristics are: 1. when the centrifuge is static, the centrifuge tube is vertical and fixed in position, so that the centrifuge tube can be conveniently positioned and clamped; 2. When the centrifugal machine rotates, the central axis of the centrifugal tube can be infinitely close to the direction of centrifugal force and gravity resultant force, so that the centrifugal tube solution with holes at the bottom can be thrown away completely.

Example 7

As shown in fig. 11, the assembly of the vacuum assembly (the suction inlet column 510+ the sealing sleeve 520), the microporous bowl 110, the microporous filter bowl 100 and the collection sleeve 400, with the sealing sleeve 520 removed, as shown in the lower left and lower right of fig. 11, is, in order from top to bottom: the micro-porous filtering cup comprises a suction air inlet column 510, a micro-porous filtering cup 100, a micro-porous sleeve 110 and a collecting sleeve 400, wherein a sealing sleeve 520 is arranged outside the micro-porous filtering cup 100 and the micro-porous sleeve 110, and the upper end and the lower end of the sealing sleeve are respectively connected with the suction air inlet column 510 and the sealing sleeve 520. As shown in the black hexagram of figure 12, the sealing sleeve and the suction inlet cylinder are snap-fit connected by a fourth groove or a fourth raised ring, where the connection is sealed. The sealing sleeve is connected with the collecting sleeve in a sealing way. The vacuum kit and the collection cannula constitute an enclosed space.

The assembled vacuum kit, microporous filter bowl, microporous sleeve bowl and collecting sleeve are shown in figure 10, the left side shows an air exhaust passage, and the right side shows an air intake passage. The difference between fig. 15 and fig. 10 is that the suction passage and the intake passage are indicated by thick lines. Fig. 12 and 13 show the sealing connection with a hexagon star. FIG. 12 does not show the microporous filter bowl, the microporous sleeve bowl, the suction passage and the intake passage are indicated by thick lines and the air passage direction is indicated by hollow arrows, FIG. 13 indicates the sealing connection with the microporous filter bowl, the microporous sleeve bowl, the suction passage and the intake passage and the air passage direction are indicated by hollow arrows, and the right inner wall of the collection sleeve on the left side of FIG. 13 is not shown so that there is a space to draw a hollow arrow indicating the suction passage.

As shown in FIG. 12, the extraction column 511 and the intake column 512 are centered within the extraction intake column 510. The microporous filter bowl 100 is in communication with an air inlet column 512, the air inlet column 512 being on the lower half of the extraction air inlet column 510 and the extraction column 511 being above. An outlet pipe 513 and an inlet pipe 514 which are communicated with the exhaust column 511 and the inlet column 512 are arranged at the periphery. From the microporous filter membrane 101, the path of air extraction is microporous filter membrane 101, collection tube 400, gap 427 between microporous sleeve cup and collection tube, gap between microporous sleeve cup and sealing sleeve, air outlet pipeline 513, air extraction column 511, and vacuum extractor. The path of the intake air is the intake air duct 514, the intake air column 512, the microporous filter bowl 100. The sealing sleeve 520 is in sealing connection with the extraction air inlet column 510, the sealing sleeve 520 and the sleeve 420 of the collecting sleeve (shown as a solid six-pointed star).

As shown in fig. 13, the microporous sleeve cup, the microporous filter cup and the air suction/intake column are hermetically connected. The sealed connection of the microporous filter bowl to the microporous sleeve bowl discussed in example 4 ensures that the solution does not become trapped between the microporous filter bowl and the microporous sleeve bowl by side passage. The sealed connection of the microporous filter cup and the air suction and inlet cylinder ensures that the air inlet passage is from the air inlet cylinder to the microporous filter cup, and the air outlet passage is from the outside of the microporous sleeve cup (in the sealing sleeve) to the air outlet pipeline. The microporous filter cup and the microporous sleeve cup are as high as each other or one of the microporous filter cup and the microporous sleeve cup is higher than each other. One solution for sealing here: the third convex ring is inserted into an opening above the microporous filter cup or the microporous sleeve cup and is in sealing connection with the microporous filter cup or the microporous sleeve cup, so that the gas in the gas inlet column only flows out of the first through hole of the microporous filter cup and/or the gas in the gas inlet column only communicates with the second through hole of the microporous sleeve cup. Another solution for sealing here: the gasket is arranged between the air inlet column and the microporous filter cup or the microporous sleeve cup, the lower part of the microporous sleeve cup props against the collecting sleeve, and the collecting sleeve is connected with the sealing sleeve and the air suction and inlet cylinder in a sealing way, namely the air inlet column is tightly pressed with the microporous filter cup or the microporous sleeve cup to form sealing connection.

The utility model provides a vacuum external member scheme, the route of admitting air keeps unanimous with external atmospheric pressure, and pressure is less than atmospheric pressure behind the millipore filtration membrane, and the pressure of collecting pipe is only slightly less than atmospheric pressure, and the power that provides on the one hand is less, and on the other hand can not destroy the interface of liquid drop, can not form aerial fog. The liquid reaches the collecting pipe under the action of gravity, and the gas passes through a gap 427 between the micropore sleeve cup and the collecting pipe and a gap between the micropore sleeve cup and the sealing sleeve pipe, passes through an air outlet pipeline 513 and an air exhaust column 511, and is connected with a vacuum pumping device.

Example 8

An example of an extraction inlet cylinder that facilitates machining is shown in FIG. 14. The extraction column 511 and the intake column 512 are centered within the extraction intake column 510. The air pumping column and the air inlet column are respectively communicated with a plurality of horizontal channels to reach the side surface of the air pumping and inlet column, and the horizontal channels connected with the air pumping column or the air inlet column are connected with the vertical channels to reach the opposite surface. The opening of the horizontal channel is closed off by a sealing sleeve 520 in the assembly according to fig. 7.

Example 9A coated magnetic microsphere biochemical detection system with microporous membrane trapped and accumulated detects benzopyrene BaP in vegetable oil.

1. Sample pretreatment: vegetable oil (dilution factor 20). Accurately sucking a 500 mu l plant oil sample into a 15ml centrifuge tube, and adding 9.5ml 70% ethanol water; oscillating and mixing evenly, centrifuging at room temperature of 5000rpm for 5 min; taking 50 mu l of the supernatant solution for analysis.

2. Specific reaction: the supernatant was added to an incubation container (containing microspheres that had been coated with BSA-Baq-coupled antigen and fluorescent microspheres coupled with Baq murine monoclonal antibody) and mixed well and incubated for 15 min.

3. Washing and removing impurities: collecting and coating the magnetic microspheres by using a magnetic rod and a rod sleeve, removing the supernatant solution, adding a proper amount of aqueous solution, oscillating and uniformly mixing, collecting and coating the magnetic microspheres by using the magnetic rod and the rod sleeve, and removing the aqueous solution.

4. Transferring and removing impurities: the magnetic rod and the rod sleeve are used for gathering and coating the magnetic microspheres and transferring the magnetic microspheres into the microporous filter cup, the magnetic rod is pulled out, and the coated magnetic microspheres are dissolved in the aqueous solution in the microporous filter cup.

5. Centrifuging or vacuum filtering: the microporous membrane intercepts, gathers and coats the magnetic microspheres.

6. And taking out the microporous filter cup, and putting the microporous filter cup into a fluorescence detection system to read a fluorescence value.

The mean OD values of the standards were plotted against the concentration index (y = log10< mean OD >, < x = log10< concentration (ppb) >, points with concentration of 0 not counted), as in fig. 16, resulting in a straight line, slope-0.2567 error 0.00215, and intercept 3.08478 error 0.00213. From the mean OD values of the samples, sample concentrations (ppb) of 0.585 (sample 1) and 0.0602 (sample 2) were calculated based on the fitted straight lines.

In step 3, when the magnetic rod adsorbs the magnetic microspheres in the solution within twenty seconds or ten seconds, the substance in the solution is limited to be about several millimeters from the rod sleeve, the volume of an incubation tube used in actual operation is 500u L, and the solution is about 50-100u L. in step 3 and step 4, the added aqueous solution does not contain target molecules or substances capable of influencing the combination of the target molecules and a capture reagent.

Claims (11)

1. A coated magnetic microsphere biochemical detection system for intercepting and gathering by a microporous membrane is characterized by comprising a microporous filter cup, a coated magnetic microsphere reaction reagent and a magnetic device;

the upper part of the microporous filter cup is of an open structure, the volume of the microporous filter cup is not more than 900 microliters, a first through hole with the area of less than 12 square millimeters is arranged at the center of the bottom of the microporous filter cup, a microporous filter membrane with basically uniform pore diameter and the pore diameter range of 0.02-1.2um is attached to the outer surface of the bottom wall of the first through hole, and the microporous filter membrane covers the first through hole; the inner surface of the bottom of the microporous filter cup which takes the center of the first through hole as an axis and is the lowest point is set into an arc surface, a spherical surface or a conical surface with a certain inclination angle;

the coated magnetic microsphere reaction reagent comprises at least one coated magnetic microsphere with uniform diameter and larger than the aperture of the microporous filter membrane, a capture reagent fixed on the surface of the coated magnetic microsphere and a marking reagent with diameter smaller than the aperture of the microporous filter membrane; substances having a direct or indirect ligand relationship with each other in the capturing reagent and the labeling reagent;

the magnetic device is used for providing enough strong magnetic force to enable the particles with magnetism, namely the coated magnetic microspheres and the compound thereof to be gathered and fixed at gathering positions when needed; when the magnetic force is weakened or disappears, the fine particles having magnetism are dispersed; when the fine particles having magnetism are aggregated and fixed at the aggregation positions, the portions of the solution not affected by the magnetic force can be easily removed.

2. The coated magnetic microsphere biochemical detection system of claim 1, wherein the magnetic means is a magnetic frame; the magnetic frame is provided with a position for installing the incubation container and a component for providing magnetic force, the magnetic component is a magnet, and the magnetic force of the N pole or the S pole of the magnet is strongest; the magnetic assembly is outside the position of the incubation container, and the relative position of the magnetic assembly and the incubation container can be changed; when the N pole or the S pole of the magnet is close enough to the bottom wall or the side wall of the incubation container, the magnetic particles and the reaction compound thereof in the solution in the incubation container are gathered and fixed at the position of the incubation container close to the N pole or the S pole of the magnet, when the magnet is far away from the incubation container, the magnetic action on the solution disappears, and the magnetic particles and the reaction compound thereof are dispersed in the solution; when the magnetic particles and their reactive complexes aggregate and are immobilized, the solution in the incubation container can be removed and the aggregated magnetic particles and their reactive complexes left behind.

3. The coated magnetic microsphere biochemical detection system intercepted and accumulated by the microporous membrane as claimed in claim 1, wherein the magnetic device comprises a magnetic rod and a rod sleeve, the magnetic rod is a component for providing magnetic force, and N or S poles with the strongest magnetic force can be two ends of the magnetic rod or two sides of a rod body; the outer wall of the rod sleeve does not react with the solution; when the magnetic rod is inserted into the bottom of the rod sleeve, the surface of the rod sleeve tightly attached to the N pole or the S pole of the magnetic rod has strong enough magnetic force, and the rod sleeve is inserted into the solution in the incubation container, so that the magnetic particles and the reaction compound thereof are gathered and fixed on the surface of the rod sleeve close to the N pole or the S pole of the magnetic rod; when the magnetic rod is pulled out and is far away from the bottom of the rod sleeve, the magnetic particles and the reaction compound thereof are dispersed and suspended in the solution; when the magnetic particles and their reaction complexes are aggregated and immobilized, the magnetic particles and their reaction complexes immobilized on the surface of the sleeve may be transferred or the solution in the incubation container may be removed.

4. The coated magnetic microsphere biochemical detection system intercepted and accumulated by the microporous membrane as claimed in claim 1, wherein the magnetic device comprises a magnetic frame, a magnetic rod and a rod sleeve, the magnetic frame is provided with a position for installing the incubation container and a component for providing magnetic force, the magnetic component is a magnet, and the magnetic force is strongest at the N pole or the S pole of the magnet; the magnetic assembly is outside the position of the incubation container, and the relative position of the magnetic assembly and the incubation container can be changed; when the N pole or the S pole of the magnet is close enough to the bottom wall or the side wall of the incubation container, the magnetic particles and the reaction compound thereof in the solution in the incubation container are gathered and fixed at the position of the incubation container close to the N pole or the S pole of the magnet, when the magnet is far away from the incubation container, the magnetic action on the solution disappears, and the magnetic particles and the reaction compound thereof are dispersed in the solution; the magnetic rod in the combination of the magnetic rod and the rod sleeve is a component for providing magnetic force, and the N pole or the S pole with the strongest magnetic force can be two ends of the magnetic rod or two sides of the rod body; the outer wall of the rod sleeve does not react with the solution; when the magnetic rod is inserted into the bottom of the rod sleeve, the surface of the rod sleeve tightly attached to the N pole or the S pole of the magnetic rod has strong enough magnetic force, and the rod sleeve is inserted into the solution in the incubation container, so that the magnetic particles and the reaction compound thereof are gathered and fixed on the surface of the rod sleeve close to the N pole or the S pole of the magnetic rod; when the magnetic rod is pulled out and is far away from the bottom of the rod sleeve, the magnetic particles and the reaction compound thereof are dispersed and suspended in the solution; the magnetic force frame and the magnetic rod-rod sleeve are matched to ensure that all the magnetic particles in the solution are gathered and fixed in a short time so as to separate the magnetic particles from the solution.

5. The coated magnetic microsphere biochemical detection system trapped and accumulated by the microporous membrane as claimed in claim 1, wherein a microporous sleeve cup is arranged outside the microporous filter cup, the microporous sleeve cup is of an upper open structure, and a second through hole is arranged at the center of the bottom of the microporous sleeve cup; when the microporous filter cup is inserted into the upper position and the lower position of the microporous sleeve cup to be fixed, and the inner surface of the second through hole of the microporous sleeve cup and the outer surface of the first through hole of the microporous filter cup are clamped and pressed on the microporous filter membrane.

6. The coated magnetic microsphere biochemical detection system trapped and accumulated by the microporous membrane as claimed in claim 5, wherein a first groove or a first convex ring is arranged on the outer wall of the microporous filter cup, a first convex ring or a first groove is arranged on the inner wall of the microporous filter cup, the microporous filter cup and the microporous filter cup are connected with the first convex ring in a buckling manner through the first groove, and the microporous filter membrane is clamped and compressed by the inner surface of the second through hole of the microporous filter cup and the outer surface of the first through hole of the microporous filter cup.

7. The coated magnetic microsphere biochemical detection system trapped and accumulated by the microporous membrane as claimed in claim 5, further comprising a collection sleeve, wherein the collection sleeve comprises a sleeve arranged at the upper part and a collection pipe arranged at the lower part to form a step structure, the diameter of the collection pipe is smaller than that of the sleeve, and the inner part of the sleeve is fixedly connected with the microporous filter cup or the microporous sleeve cup.

8. The coated magnetic microsphere biochemical detection system trapped and accumulated by the microporous membrane as claimed in claim 7, further comprising a centrifuge, wherein the collection sleeve is connected with a centrifugal rotor of the centrifuge in a matching manner, and a plurality of collection sleeve mounting grooves are arranged on the centrifugal rotor of the centrifuge; the collecting sleeve is matched with the collecting sleeve mounting groove, the collecting sleeve mounting groove is an arc-shaped groove, and the collecting sleeve can slide relative to the collecting sleeve mounting groove.

9. The coated magnetic microsphere biochemical detection system of claim 7,

the biochemical detection system is characterized in that a sleeve of the collecting sleeve is hermetically connected with the collecting pipe, and the biochemical detection system also comprises a vacuum suite, wherein the vacuum suite comprises an air suction and inlet cylinder and a sealing sleeve sleeved outside the air suction and inlet cylinder; the air suction cylinder is in sealing fit with the sealing sleeve, the lower part of the sealing sleeve is in sealing fit connection with the collecting sleeve, and the sleeve of the collecting sleeve is in sealing connection with the collecting pipe, so that a sealed space is formed by the vacuum sleeve and the collecting sleeve;

if the microporous sleeve cup is arranged, the microporous filter cup and the microporous sleeve cup are hermetically connected, so that the fluid can only reach the collecting sleeve from the microporous filter cup through the first through hole, the microporous filter membrane and the second through hole; gaps for circulating gas are arranged between the outer wall of the microporous filter cup or the microporous sleeve cup and the inner walls of the collecting sleeve and the sealing sleeve;

one or more air inlet pipelines and air inlet columns and one or more air outlet pipelines and air outlet columns are arranged in the air extracting and air inlet cylinder body, and the air inlet columns are butted and sealed with the openings of the microporous filter cups or the microporous sleeve cups; one end of the air inlet pipeline is communicated with the air inlet column, and the other end of the air inlet pipeline is communicated with the atmosphere; the air pumping column is connected with a vacuum air path of the vacuum pumping equipment, one end of the air outlet pipeline is communicated with the air pumping column, and the other end of the air outlet pipeline is communicated with a gap between the sealing sleeve and the microporous filter cup or the microporous sleeve cup.

10. The biochemical detection system of the coated magnetic microspheres trapped and accumulated by the microporous membrane according to claim 9, wherein one or more fourth convex rings or fourth grooves are arranged on the outer wall of the air suction and inlet cylinder, one or more fourth grooves or fourth convex rings are arranged on the inner wall of the sealing sleeve, and the fourth convex rings and the fourth grooves are in sealing fit with the air suction and inlet cylinder and the sealing sleeve.

11. The coated magnetic microsphere biochemical detection system of claim 1, wherein the coated magnetic microspheres in the microporous filter cup have the following microscopic dimensional relationship with the labeled reagent: when the free coated magnetic microspheres and the coated magnetic microspheres forming a reaction compound with the labeled reagent are stacked on the surface of the microporous membrane, the free labeled reagent can be carried by the solution from the pores among the stacked coated magnetic microspheres to pass through the micropores of the microporous membrane again and flow out, and no blocking residue is formed.

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