Bioraction chip, bioraction device and biological detection method

Abstract

The application discloses a biological reaction chip, a biological reaction device and a biological detection method. The biological reaction chip comprises a chip body and a sealing layer. The chip body is provided with a material cavity. The material cavity is provided with a ventilation structure. The material cavity comprises a sample containing area, a diluent sample adding area, a sample dilution mixing area, a pre-reaction area and a reaction detection area. The detection sample from the first sample adding port and the diluent from the second sample adding port can be mixed in the sample dilution mixing area. The mixed liquid can be reacted with the first reaction material in the pre-reaction area to form a pre-reaction liquid. The reaction liquid can be reacted with the second reaction material in the reaction detection area to form a test sample. The biological reaction chip provided by the application effectively reduces the labor intensity of the staff during the detection.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of materials and biotechnology, and particularly relates to a biological reaction chip, a biological reaction device and a biological detection method. BACKGROUND

[0002] In the process of biological two-step reaction detection, for example, the renal function detection involves various biochemical indexes. The traditional detection of cystatin C adopts an immune method, which has a high requirement for the mixing effect of reagents in the detection process, and the reaction is divided into two steps, and the two-step reaction is required to be independent of each other and to be completed on independent structures.

[0003] In the specific implementation, the staff needs to transfer the pre-reaction liquid after the completion of the first-step reaction to the structure required for the second-step reaction, and then perform the second-step reaction, which is complicated and labor-intensive.

[0004] Therefore, how to reduce the labor intensity of the staff in the detection process is a technical problem to be solved by the person skilled in the art. SUMMARY

[0005] The present application aims to provide a biological reaction chip to reduce the labor intensity of the staff in the detection process. Another object of the present application is to provide a biological reaction device comprising the above biological reaction chip. Still another object of the present application is to provide a biological detection method.

[0006] To achieve the above object, the present application provides a biological reaction chip, comprising:

[0007] A chip body is provided with a material chamber, and the material chamber has a ventilation structure. The material chamber includes a sample containing area, a diluent sample adding area, a sample dilution mixing area, a pre-reaction area and a reaction detection area. The sample containing area and the diluent sample adding area can communicate with the sample dilution mixing area. The sample dilution mixing area communicates with the pre-reaction area and the reaction detection area. The pre-reaction area and the reaction detection area are respectively provided with first and second reaction substances. The sample containing area is provided with a first sample adding port, and the diluent sample adding area is provided with a second sample adding port.

[0008] A sealing layer is capable of sealing the material chamber.

[0009] The detection sample from the first sample inlet and the diluent from the second diluent inlet can enter the sample dilution mixing area to mix, and the mixed liquid enters the pre-reaction area to react with the first reaction material in the pre-reaction area to form a pre-reaction liquid, and the pre-reaction liquid enters the reaction detection area to react with the second reaction material to form a test sample.

[0010] Optionally, in the above-mentioned biological reaction chip, the biological reaction chip is a centrifugal chip, the sample containing area comprises a sample collection cavity, a sample quantitative separation cavity, a sample waste liquid cavity, and a sample siphon pipe for conveying liquid in the sample quantitative separation cavity to the sample dilution mixing area, the sample collection cavity is used for receiving a detection sample, the sample quantitative separation cavity comprises a first cavity, a first connecting pipe, a second cavity, a second connecting pipe and a third cavity connected in sequence from a proximal end to a distal end, the cross-sectional area of the first connecting pipe and the cross-sectional area of the second connecting pipe are both smaller than the cross-sectional area of the first cavity, the cross-sectional area of the second cavity and the cross-sectional area of the third cavity along a direction perpendicular to the radius of rotation, the sample siphon pipe is connected to the second connecting pipe and the sample dilution mixing area, the sample waste liquid cavity is connected to the third cavity through a waste liquid outlet pipe, and the distance from the closest part of the sample siphon pipe to the rotation center to the rotation center is smaller than the distance from the closest part of the waste liquid outlet pipe to the rotation center to the rotation center.

[0011] The ventilation structure comprises a first ventilation pipe connected to the sample waste liquid cavity and a second ventilation pipe connected to the sample collection cavity.

[0012] Optionally, in the above-mentioned biological reaction chip, the inner cavity of the second cavity is a spindle-shaped cavity with a middle thick part and two end thin parts along the fluid flow direction.

[0013] Optionally, in the above-mentioned biological reaction chip, the sample collection cavity is a hydrophilic cavity.

[0014] Optionally, in the above-mentioned biological reaction chip, the diluent sample inlet area comprises a diluent sample cavity, a diluent sample inlet for adding diluent to the diluent sample cavity, and a diluent transfer pipe connected to the diluent sample cavity and the sample dilution mixing area, the diluent sample inlet is located at the proximal end of the diluent sample cavity, and the diluent transfer pipe is connected to the distal end of the diluent sample cavity.

[0015] Optionally, in the above-mentioned bioreactor chip, the sample dilution and mixing area includes a mixing chamber and a mixing chamber siphon. The mixing chamber siphon is connected to the outlet of the mixing chamber and is used to deliver the mixture to the pre-reaction area. The mixing chamber siphon is a hydrophilic tube. The first connection port connecting the dilution sample addition area to the mixing chamber and the second connection port connecting the sample holding area to the mixing chamber are located on the same side of the outlet of the mixing chamber.

[0016] The ventilation structure includes a third ventilation tube connected to the mixing chamber.

[0017] Optionally, in the above-mentioned bioreactor chip:

[0018] The pre-reaction zone includes a sample dilution storage chamber, a pre-reaction chamber, a third connecting tube, a first siphon tube, and a second siphon tube. The first reactant is pre-filled in the pre-reaction chamber. The third connecting tube connects the proximal outlet of the sample dilution storage chamber to the inlet of the pre-reaction chamber. The first siphon tube connects to the outlet of the pre-reaction chamber, and the second siphon tube connects to the distal outlet of the sample dilution storage chamber. The ventilation structure includes a fourth ventilation tube connected to the pre-reaction chamber.

[0019] The reaction detection area includes a first reaction detection area and a second reaction detection area independent of the first reaction detection area. The first siphon is connected to the liquid inlet of the first reaction detection area, the second siphon is connected to the liquid inlet of the second reaction detection area, the second reactant is located in the first reaction detection area, and a third reactant is preset in the second reaction detection area.

[0020] Optionally, in the above-mentioned bioreaction chip, the first reaction detection area includes a first distribution tube, a pre-reaction detection chamber, a first detection chamber, and a blank control chamber. The blank control chamber is a hollow structure. Both the pre-reaction detection chamber and the first detection chamber are connected to the first distribution tube. The inlet of the first distribution tube is connected to the outlet of the first siphon tube. The ventilation structure includes a ventilation structure connected to the end of the first distribution tube away from the inlet of the first distribution tube. The second reactant is located in the first detection chamber.

[0021] Optionally, in the above-mentioned bioreactor chip, along the fluid flow direction in the first distribution tube, the connection end of the first detection chamber and the first distribution tube is located downstream of the connection end of the pre-reaction detection chamber and the first distribution tube. The first detection chamber includes an expansion chamber and a reaction chamber communicating with the distal end of the expansion chamber. The proximal end of the expansion chamber is connected to the first distribution tube. Along the direction perpendicular to the radius of the rotation center, the cross-sectional area of ​​the expansion chamber is larger than the cross-sectional area of ​​the reaction chamber, so that the liquid fills the reaction chamber, while the expansion chamber is in an unfilled state.

[0022] Optionally, in the above-mentioned bioreactor chip, the second reaction detection area includes a second distribution tube, a second detection chamber, and a waste liquid chamber. The second detection chamber and the waste liquid chamber are both connected to the second distribution tube. The inlet of the second distribution tube is connected to the outlet of the second siphon tube. The third reactant is located in the second detection chamber along the fluid flow direction in the second distribution tube. The connection between the waste liquid chamber and the second distribution tube is located downstream of the connection between the second detection chamber and the second distribution tube. The ventilation structure includes a sixth ventilation tube connected to the end of the second distribution tube away from the inlet section of the second distribution tube.

[0023] Optionally, in the above-mentioned bioreactor chip, the chip body is provided with a positioning structure, the positioning structure including positioning teeth disposed at the distal end of the chip body and positioning holes disposed on the chip body.

[0024] A bioreactor device includes a bioreactor chip and a drive device for centrifugally rotating the bioreactor chip, wherein the bioreactor chip is any of the bioreactor chips described above.

[0025] Optionally, the above-mentioned bioreactor also includes a control device connected to the drive device, which can control the drive device to drive the bioreactor chip to swing and mix the substances in the chip body.

[0026] A biological detection method, based on the bioreactor chip described in any one of the preceding claims, includes the following steps:

[0027] The test sample is sent into the sample holding area, and the diluent is sent into the diluent addition area;

[0028] The biodetection chip is centrifuged and rotated. Particles in the test sample in the sample holding area are separated and enter the sample dilution and mixing area. At the same time, the diluent enters the sample dilution and mixing area.

[0029] The bio-detection chip oscillates back and forth, and the diluent mixes with the detection sample to form a mixed liquid.

[0030] The bio-detection chip rotates centrifugally, and the mixed liquid enters the pre-reaction zone;

[0031] The bio-detection chip oscillates back and forth, and the mixed liquid mixes with the first reactant to form a pre-reaction liquid;

[0032] The bio-detection chip rotates centrifugally, and the pre-reaction liquid enters the reaction detection zone, where it reacts with the second reactant to form a test sample.

[0033] Optionally, in the above-described biological detection method, when the biological detection chip reciprocates, the biological detection chip oscillates at a non-uniform speed.

[0034] In the above technical solution, the bioreactor chip provided by the present invention includes a chip body and a sealing layer. The chip body has a material chamber with a ventilation structure. The material chamber includes a sample holding area, a diluent loading area, a sample dilution mixing area, a pre-reaction area, and a reaction detection area. The holding area and the diluent loading area are both connected to the sample dilution mixing area. The sample dilution mixing area is connected to the pre-reaction area and the reaction detection area. The pre-reaction area and the reaction detection area are respectively pre-loaded with a first reactant and a second reactant. The sample holding area has a first loading port, and the diluent loading area has a second loading port. The sealing layer can seal the material chamber. The test sample entering the sample holding area from the first loading port and the diluent entering the diluent loading area from the second loading port can both enter the sample dilution mixing area for mixing. The mixed liquid enters the pre-reaction area and reacts with the first reactant in the pre-reaction area to form a pre-reaction liquid. The pre-reaction liquid enters the reaction detection area and reacts with the second reactant to form a test sample.

[0035] As described above, the bioreactor chip provided in this application includes a sample containment area, a diluent addition area, a sample dilution and mixing area, a pre-reaction area, and a reaction detection area. During liquid flow, different fluids can be mixed. The pre-reaction area and the reaction detection area are pre-set with a first reactant and a second reactant, respectively. Both the one-step reaction to form the pre-reaction liquid and the two-step reaction to form the test sample are performed within the chip itself, eliminating the need for personnel to transfer the reaction liquid and effectively reducing the workload of personnel conducting experiments. Attached Figure Description

[0036] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0037] Figure 1 This is a schematic diagram illustrating the sample containment area, the first reaction detection area, and the second reaction detection area in the bio-reaction chip provided in an embodiment of the present invention.

[0038] Figure 2 This is a schematic diagram illustrating the pre-reaction zone in the bioreactor chip provided in an embodiment of the present invention;

[0039] Figure 3 This is a schematic diagram illustrating the diluent addition area and the sample dilution mixing area in the bioreactor chip provided in this embodiment of the invention;

[0040] Figure 4This is a schematic diagram illustrating the positioning structure in the bioreactor chip provided in an embodiment of the present invention;

[0041] Figure 5 This is a schematic diagram illustrating the components of the bioreactor chip provided in an embodiment of the present invention;

[0042] Figure 6 This is a schematic diagram of the sample collection cavity provided in an embodiment of the present invention;

[0043] Figure 7 This is a schematic diagram of the back structure of the bioreactor chip provided in an embodiment of the present invention;

[0044] Figure 8 This is a schematic diagram of the fluid structure of the bioreactor chip provided in an embodiment of the present invention, in which sample solution and diluent are added;

[0045] Figure 9 This is a fluid flow diagram showing the diluent entering the sample dilution and mixing zone and the sample solution entering the sample quantitative separation chamber in the bioreactor chip provided in this embodiment of the invention.

[0046] Figure 10 This is a schematic diagram of the fluid flow from the sample solution into the sample dilution and mixing zone in the bioreactor chip provided in this embodiment of the invention;

[0047] Figure 11 This is a schematic diagram of the fluid flow in the pre-reaction zone of the bioreactor chip provided in an embodiment of the present invention;

[0048] Figure 12 This is a schematic diagram of the fluid flow in the bioreactor chip provided in this embodiment of the invention, showing the mixed liquid entering the first and second reaction detection zones.

[0049] in Figures 1-12 middle:

[0050] Sample containment area 10, sample collection chamber 11, sample quantitative separation chamber 12, first chamber 121, first connecting tube 122, second chamber 123, second connecting tube 124, third chamber 125, sample waste liquid chamber 13, sample siphon tube 14, first vent tube 15, second vent tube 16, waste liquid outlet tube 17;

[0051] Diluent addition area 20, diluent addition chamber 21, diluent addition port 22, diluent transfer tube 23;

[0052] Sample dilution and mixing zone 30, mixing chamber 31, third vent tube 32, exhaust port 33, mixing chamber siphon tube 34;

[0053] Pre-reaction zone 40, dilution sample storage chamber 41, pre-reaction chamber 42, third connecting tube 43, fourth vent tube 44, second siphon tube 45, first siphon tube 46;

[0054] First reaction detection zone 50, first distribution tube 51, pre-reaction detection chamber 52, first detection chamber 53, blank control chamber 54, ventilation structure 55;

[0055] Second reaction detection zone 60, second distribution pipe 61, second detection chamber 62, waste liquid chamber 63;

[0056] Positioning structure 70, positioning hole 71, positioning tooth 72. Detailed Implementation

[0057] The core of this invention is to provide a bioreactor chip that reduces the workload of workers during the detection process. Another core aspect of this invention is to provide a bioreactor device including the aforementioned bioreactor chip. Yet another core aspect of this invention is to provide a biological detection method.

[0058] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0059] Please refer to Figures 1 to 12 .

[0060] In one specific embodiment, the bioreactor chip provided by this invention includes a chip body and a sealing layer, such as... Figure 1 As shown, the sealing layer seals the front side of the chip body. Specifically, the chip body can be made of high molecular polymers such as PC (polycarbonate), PMMA (polymethyl methacrylate), and PS (polystyrene) by injection molding or machining.

[0061] Specifically, the sealing layer can be made of materials such as PC (polycarbonate), PMMA (polymethyl methacrylate), PS (polystyrene), PP (polypropylene), and PET (polyethylene terephthalate) bonded with single-sided or double-sided adhesive, and can be sealed by adhesive bonding. Figure 1 The diagram shows the structural surface of the chip body. The sealing layer can also be directly welded to the chip body, for example, through ultrasonic welding, laser welding, or thermocompression welding.

[0062] The chip body has a material chamber, which is sealed by a sealing layer. The material chamber has a ventilated structure and includes a sample holding area 10, a diluent loading area 20, a sample dilution mixing area 30, a pre-reaction area 40, and a reaction detection area. The sample holding area 10 and the diluent loading area 20 are both connected to the sample dilution mixing area 30, meaning that substances in the sample holding area 10 and the diluent loading area 20 can flow to the sample dilution mixing area 30. The sample dilution mixing area 30 is connected to the reaction detection area through the pre-reaction area 40. The pre-reaction area 40 and the reaction detection area are respectively pre-set with a first reactant and a second reactant, the latter depending on the required reactant. Specifically, the sample dilution mixing area 30 can be used for serum dilution and mixing.

[0063] The test sample entering the sample holding area 10 from the first sample dispensing port and the diluent entering the diluent dispensing area 20 from the second sample dispensing port can both enter the sample dilution mixing area 30 for mixing. After mixing, the mixed liquid enters the pre-reaction area 40 and reacts with the first reactant in the pre-reaction area 40 to form a pre-reaction liquid. The reaction liquid enters the reaction detection area and reacts with the second reactant to form the test sample.

[0064] The sample container area 10 is provided with a first sample dispensing port, specifically located on the side end of the chip body, near the center of the chip body. The diluent dispensing area 20 is provided with a second sample dispensing port. Specifically, before entering the reaction detection area, the location of the second reactant can be provided with an independent liquid container. These different liquid containers are arranged along the liquid flow direction, and the liquid is filled sequentially. That is, before entering the reaction detection area, a portion of the liquid preferentially flows to the solution container, and the other portion flows to the location of the second reactant. Specifically, the reaction detection area is provided with a pre-reaction detection chamber after the first reaction and a test sample detection chamber after the second reaction. The pre-reaction detection chamber detects reactants that have only reacted with the first reactant, and the test sample detection chamber detects reactants that have reacted with both the first and second reactants.

[0065] The bioreactor chip is a centrifugal chip. The sample holding area 10 includes a sample collection chamber 11, a sample quantitative separation chamber 12, a sample waste liquid chamber 13, and a sample siphon tube 14 that transports the liquid in the sample quantitative separation chamber 12 to the sample dilution and mixing area 30. The sample collection chamber 11 is used to receive the test sample. The sample quantitative separation chamber 12 includes a first cavity 121, a first connecting tube 122, a second cavity 123, a second connecting tube 124, and a third cavity 125 connected sequentially from the proximal end to the distal end. Along the cross-sectional area perpendicular to the radius of rotation, the cross-sectional areas of the first connecting tube 122 and the second connecting tube 124 are both smaller than the cross-sectional areas of the first cavity 121, the second cavity 123, and the third cavity 125. Specifically, the cross-sectional area of ​​the second connecting tube 124 is smaller than the cross-sectional area of ​​the first connecting tube 122. Preferably, the inner cavity of the second cavity 123 is a spindle-shaped cavity that is thicker in the middle and thinner at both ends along the fluid flow direction, as shown in the figure. The inner cavity of the second cavity 123 can be arranged in a parallelogram shape, and the second connecting pipe 124 and the first connecting pipe 122 connect the two opposite corner positions of the second cavity 123.

[0066] like Figure 6 As shown, the sample siphon tube 14 connects the second connecting tube 124 and the sample dilution and mixing zone 30. The distance from the closest point a of the sample siphon tube 14 to the rotation center is less than the distance from the closest point b of the waste liquid outlet tube 17 to the rotation center. Specifically, the middle part of the sample siphon tube 14 has a V-shaped tube structure that protrudes towards the proximal end. Preferably, the bend in the inner cavity of the tube is smoothly transitioned. The sample waste liquid chamber 13 is connected to the third chamber 125 through the waste liquid outlet tube 17.

[0067] The ventilation structure includes a first ventilation pipe 15 connected to the sample waste liquid chamber 13 and a second ventilation pipe 16 connected to the sample collection chamber 11. Specifically, the outlet end of the first ventilation pipe 15 and the outlet end of the second ventilation pipe 16 can both be located on the sidewall of the chip.

[0068] Preferably, the sample collection chamber 11 is a hydrophilic chamber, that is, the sample collection chamber 11 has hydrophilic properties and can directly contact samples such as blood / serum / plasma through the first sample application port located on the side of the proximal end of the bioreactor chip to generate capillary action, thereby completing the collection and containment of the sample.

[0069] like Figure 6As shown, the sample quantitative separation chamber 12 has a "double dumbbell" structure with three sequentially connected chambers, namely, two constricted "waist" structures (first connecting tube 122 and second connecting tube 124). Under the action of centrifugal force, it realizes the separation and quantification of sample particles. The lower end of the waist structure (second connecting tube 124) is connected to the sample siphon tube 14, and the distal end of the second connecting tube 124 is connected to the sample waste liquid chamber 13 through the third chamber 125 and the waste liquid outlet tube 17. The first vent tube 15 is used for ventilation during liquid transfer.

[0070] The two ends of the waste liquid outlet tube 17 are connected to the proximal end of the third chamber 125 and the proximal end of the sample waste liquid chamber 13, respectively.

[0071] The diluent loading area 20 includes a diluent loading chamber 21, a diluent loading port 22 for loading samples into the diluent loading chamber 21, and a diluent transfer tube 23 connecting the diluent loading chamber 21 and the sample dilution mixing area 30. The diluent loading port 22 is located at the proximal end of the diluent loading chamber 21, and the diluent transfer tube 23 is connected to the distal end of the diluent loading chamber 21. Specifically, the diluent loading port 22 can be located on the back of the chip body.

[0072] The diluent dispensing chamber 21 is used to hold a fixed volume of diluent required for chip biochemical assays; the diluent dispensing port 22 is specifically matched to the size of the pipette tip for dispensing diluent from the pipette tip. Figure 7 The chip body shown is sampled on the back side.

[0073] The sample dilution mixing zone 30 includes a mixing chamber 31 and a mixing chamber siphon 34 for delivering the mixture to the pre-reaction zone 40. The outlet of the mixing chamber 31 is connected to the mixing chamber siphon 34. The mixing chamber siphon 34 is a hydrophilic tube. The first connection port between the dilution sample addition zone 20 and the mixing chamber 31 and the second connection port between the sample holding zone 10 and the mixing chamber 31 are located on the same side as the outlet of the mixing chamber 31. The ventilation structure includes a third ventilation pipe 32 connected to the mixing chamber 31. Specifically, the exhaust port 33 at the end of the third ventilation pipe 32 is located on the front side of the chip body.

[0074] In actual operation, the mixing chamber 31 is used to mix the transferred test sample with the diluent; the exhaust port 33 ensures that the mixing chamber 31 is connected to the outside atmosphere.

[0075] The pre-reaction zone 40 includes a sample dilution storage chamber 41, a pre-reaction chamber 42, a third connecting tube 43, a first siphon tube 46, and a second siphon tube 45. The pre-reaction chamber 42 contains a first reactant. The third connecting tube 43 connects the proximal outlet of the sample dilution storage chamber 41 to the inlet of the pre-reaction chamber 42. The first siphon tube 46 connects to the outlet of the pre-reaction chamber 42, and the second siphon tube 45 connects to the distal outlet of the sample dilution storage chamber 41. The distance from the proximal outlet to the center of rotation is less than the distance from the distal outlet to the center of rotation. Figure 5 As shown, the distance from the closest point c of the third connecting pipe 43 to the rotation center is greater than the distance from the closest point d of the second siphon pipe 45 to the rotation center.

[0076] The ventilation structure includes a fourth ventilation tube 44 connected to the pre-reaction chamber 42. Specifically, the pre-reaction zone 40 can be a pre-reaction zone for performing the cystatin C reaction.

[0077] The reaction detection area includes a first reaction detection area 50 and a second reaction detection area 60, which are independently set up. A first siphon tube 46 is connected to the inlet of the first reaction detection area 50, and a second siphon tube 45 is connected to the inlet of the second reaction detection area 60. The second reactant is located in the first reaction detection area 50, and the second reaction detection area 60 contains a third reactant. The first reaction detection area 50 can be used for the detection of the cystatin C reaction, and the second reaction detection area 60 can be used for the detection of other renal function indicators.

[0078] Specifically, the first reactant is the lyophilized reagent used in the first step of the cystatin C biochemical assay. The diluted sample, when placed in the diluent, re-dissolves the lyophilized reagent used in the first step of the cystatin C biochemical assay, thus achieving the pre-reaction. The fourth vent tube 44 connects to the pre-reaction chamber 42, enabling ventilation to the outside environment; the second siphon tube 45 connects the pre-reaction chamber 42 to the downstream first reaction detection area 50; and the first siphon tube 46 connects the diluted sample storage chamber 41 to the downstream second reaction detection area 60.

[0079] The first reaction detection zone 50 includes a first distribution pipe 51, a pre-reaction detection chamber 52, a first detection chamber 53, and a blank control chamber 54. The pre-reaction detection chamber 52 and the first detection chamber 53 are both connected to the first distribution pipe 51. Of course, the blank control chamber 54 can also be connected to the first distribution pipe 51. Specifically, the pre-reaction detection chamber 52, the first detection chamber 53, and the blank control chamber 54 can all be porous structures. Along the flow direction of the fluid in the first distribution pipe 51, the blank control chamber 54 is located downstream of the first detection chamber 53.

[0080] Along the flow direction of the fluid in the first distribution pipe 51, the connection end between the first detection chamber 53 and the first distribution pipe 51 can be located downstream of the connection end between the pre-reaction detection chamber 52 and the first distribution pipe 51. The fluid entering the first distribution pipe 51 first enters the pre-reaction detection chamber 52 and then enters the first detection chamber 53. The ventilation structure includes a ventilation structure 55 connected to the end of the first distribution pipe 51 away from the liquid inlet of the first distribution pipe 51. The liquid inlet of the first distribution pipe is connected to the liquid outlet of the first siphon tube 46. The second reactant is located in the first detection chamber 53. The bioreaction chip provided in this application can detect cystatin C in a two-step method. The final detection result is achieved using the pre-reaction detection chamber 52 and the first detection chamber 53 at the same centrifugation radius. Both steps of the cystatin C reaction are thoroughly mixed to ensure stable reaction and reliable detection. The blank control chamber 54 is an empty cavity structure, that is, the blank control chamber 54 has no structure and is only used for comparison with other chambers.

[0081] The first detection chamber 53 includes an expansion chamber and a reaction chamber connected to the distal end of the expansion chamber. The proximal end of the expansion chamber is connected to the first distribution pipe 51. Along the direction perpendicular to the radius of the rotation center, the cross-sectional area of ​​the expansion chamber is larger than that of the reaction chamber. That is, the first detection chamber 53 has a shape that is wider at the top (proximal end) and narrower at the bottom (distal end) than the proximal end (circumferential dimension). After the liquid from the pre-reaction chamber 42 fills the pre-reaction detection chamber 52, it can only fill 1 / 2 to 1 / 3 of the volume of the first detection chamber 53, leaving part of the cavity to facilitate the mixing of reagents, thereby ensuring a fully stable reaction.

[0082] The blank control chamber 54 is an empty cavity. The pre-reaction detection chamber 52 is used to contain the pre-reaction products from the pre-reaction chamber 42 and to detect the pre-reaction results. Specifically, the first detection chamber 53 contains a lyophilized reagent for the second step of the cystatin C biochemical reaction as a second reaction substance, which can be reconstituted by the liquid from the pre-reaction chamber 42 to complete the second step of the cystatin C biochemical reaction.

[0083] The second reaction detection zone 60 includes a second distribution pipe 61, a second detection chamber 62, and a waste liquid chamber 63. Both the second detection chamber 62 and the waste liquid chamber 63 are connected to the second distribution pipe 61. Specifically, the second detection chamber 62 and the waste liquid chamber 63 can be perforated structures with open upper surfaces. The inlet of the second distribution pipe 61 is connected to the outlet of the second siphon pipe 45. The third reactant is located within the second detection chamber 62. Along the fluid flow direction within the second distribution pipe 61, the connection point between the waste liquid chamber 63 and the second distribution pipe 61 is located downstream of the connection point between the second detection chamber 62 and the second distribution pipe 61. That is, the fluid entering the second distribution pipe 61 first enters the second detection chamber 62 and then enters the waste liquid chamber 63. The ventilation structure includes a ventilation structure 55 connected to one end of the second distribution pipe 61 away from the inlet section of the second distribution pipe 61.

[0084] Specifically, multiple second detection chambers 62 can be provided, each pre-set with different reactive substances to perform multiple tests. Preferably, the fluid entering the second distribution pipe 61 first enters each of the second detection chambers 62 sequentially, and finally enters the waste liquid chamber 63.

[0085] In one specific embodiment, the first distribution pipe 51 and the second distribution pipe 61 are arc-shaped pipe structures with the same radius of rotation but not concentric.

[0086] To avoid structural interference, preferably, the first siphon tube 46 and the second siphon tube 45 are located at opposite ends of the chip body surface. In this case, the opening of the fourth vent tube 44 can be located at the side end of the chip body, without the need to set a separate vent hole.

[0087] The second reaction detection zone 60 can be used to detect other renal function indicators. The second detection chamber 62 stores lyophilized substrates required for biochemical detection of renal function indicators as the third reaction substances. The detection indicators include blood urea nitrogen, serum creatinine, uric acid, etc.

[0088] The chip body is provided with a positioning structure 70, which includes positioning teeth 72 disposed at the distal end of the chip body and positioning holes 71 disposed on the chip body. There can be one or at least two positioning holes 71. Specifically, a limiting post that mates with the positioning hole 71 is provided on the corresponding mounting base. Specifically, the positioning teeth 72 are arranged in a serrated shape along the periphery of the chip body. Specifically, the mounting base is provided with limiting teeth that engage with the positioning teeth 72, or an optical sensor that determines the position of the detection hole by means of the positioning teeth 72 when the chip rotates.

[0089] This bioreactor chip enables the biochemical detection of various renal function-related indicators, including cystatin C. All substrates for the biochemical reactions are pre-stored in the chip via cryoablation. The cystatin C biochemical reaction is a two-step process, while the other indicators are detected using a one-step process. Through a specific chip structure and centrifugation workflow, both one-step and two-step reactions are simultaneously and automatically controlled.

[0090] Specifically, the biochemical reaction chip is set on a rotating platform, and preferably, multiple circumferentially arranged biochemical reaction chips are set on a rotating platform.

[0091] This application provides a bioreactor device, including a bioreactor chip and a drive device for centrifugally rotating the bioreactor chip, wherein the bioreactor chip is any of the aforementioned bioreactor chips. The specific structure of the bioreactor chip has been described above; this application includes the aforementioned bioreactor chip and also possesses the aforementioned technical effects.

[0092] In one specific embodiment, the bioreactor further includes a control device connected to the drive device, which can control the drive device to swing the bioreactor chip to mix the substances inside the chip body.

[0093] This application provides a biological detection method based on the aforementioned biological reaction chip, comprising the following steps:

[0094] The test sample is delivered into the sample holding area 10, and the diluent is delivered into the diluent loading area 20. Specifically, the sample collection chamber 11 contacts the sample and draws in the test sample. A fixed volume of diluent is added through the diluent loading port 22 on the back of the chip body. Then, the bioreactor chip is placed on the centrifuge platform of the matching detection instrument.

[0095] The biodetector chip is centrifuged, separating particles in the sample within the sample holding area 10. The sample then enters the sample dilution and mixing area 30, where diluent is also introduced. Specifically, the biodetector chip is centrifuged at high speed for a period of time. Under centrifugal force, the sample enters the sample quantitative separation chamber 12 from the sample collection chamber 11 for quantification. Excess sample enters the sample waste chamber 13. The sample undergoes particle separation under centrifugal force, such as serum blood cell separation. The diluent is added to the sample chamber 21 and then transferred to the mixing chamber via the diluent transfer tube 23. When centrifugation is stopped or the centrifugation speed is reduced, the separated serum or plasma sample fills the sample siphon tube 14 under capillary action. The mixing chamber siphon tube 34, being unmodified for hydrophilicity, is not filled with liquid.

[0096] In operation, the biodetector chip rotates centrifugally, and the sample, under the influence of siphon, enters the sample dilution and mixing zone 30 through the siphon tube 14 and comes into contact with the diluent therein. The biodetector chip oscillates back and forth, and the diluent and the test sample mix to form a mixed liquid. While the biodetector chip can oscillate at a uniform speed, to improve mixing efficiency, it is preferable that the biodetector chip oscillates at a non-uniform speed. Specifically, the oscillation of the biodetector chip is achieved by reducing the chip's rotational speed and using a high-acceleration, low-speed forward and reverse rotation mode (e.g., a rotational speed of ±300 rpm and an acceleration of 10000 rpm / s) to allow the chip to oscillate, thereby achieving thorough mixing of the test sample and the diluent. Furthermore, the high-acceleration back-and-forth oscillation allows the liquid to enter and fill the mixing chamber siphon tube 34 simultaneously with the mixing.

[0097] Specifically, in the high acceleration low speed forward and reverse rotation mode: speed range: ±2000rpm (i.e., alternating between counterclockwise and clockwise 2000rpm, with 2000rpm being the highest speed within the usable range. In actual operation, there may be speed settings of -100rpm / +400rpm, 0rpm / +300rpm).

[0098] Acceleration range: 100 rpm / s to 30000 rpm / s. Specifically, when the purpose is to mix liquids in a container, symmetrical rotational speed and acceleration are usually used (e.g., ±1000 rpm, acceleration 20000 rpm / s). However, when it is necessary to fill the corresponding siphon tube by impacting the liquid with high acceleration, asymmetrical acceleration is used, and the speed may also be asymmetrical (e.g., rotational speed ±200 rpm, clockwise acceleration 500 rpm / s, counterclockwise acceleration 20000 rpm / s, in which case the liquid gradually flows into the siphon tube). Specifically, when the motor speed changes, there is inevitably a force that causes the liquid to rotate clockwise or counterclockwise. This force is used to make the liquid have the effect of rushing to the left or right, that is, the impact force generated by the motor speed change on the liquid, and then the liquid impacts the siphon tube on the biodetection chip until the siphon tube is finally filled.

[0099] The biodetector chip rotates centrifugally, and the mixed liquid enters the pre-reaction zone 40. The biodetector chip oscillates back and forth, and the mixed liquid mixes with the first reactant to form a pre-reaction liquid.

[0100] Specifically, the bio-detection chip rotates at high speed, and the sample mixed in the mixing chamber enters the dilution sample storage chamber 41 through the mixing chamber siphon tube 34 under the siphon effect. After the dilution sample storage chamber 41 is filled, the excess liquid enters the pre-reaction chamber 42 through the third connecting tube 43. Then, the rotation speed of the bio-detection chip is reduced, and the chip can also be oscillated in a high acceleration and low speed forward and reverse mode, so that the liquid in the pre-reaction chamber 42 can fully dissolve the lyophilized reagent and realize the pre-reaction of cystatin C. At the same time, the liquid will fill the second siphon tube 45 and the first siphon tube 46 respectively under the action of the bio-detection chip oscillation.

[0101] The biodetector chip is centrifuged, and the pre-reaction liquid enters the reaction detection zone, reacting with the second reactant to form a test sample. The biodetector chip is centrifuged at high speed, and the liquid in the sample dilution chamber 41 and the pre-reaction chamber 42, under centrifugal force, flows through the second siphon tube 45 and the first siphon tube 46 into the first distribution tube 51 and the second distribution tube 61, respectively, sequentially filling the pre-reaction detection chamber 52, partially filling the first detection chamber 53, and completely filling the second detection chamber 62 and partially filling the waste liquid chamber 63. Subsequently, the rotation speed of the biodetector chip is reduced, and a high-acceleration, low-speed forward and reverse rotation mode is used to allow the biodetector chip to oscillate, ensuring that the liquid in the first detection chamber 53 (used as the cystatin C detection well) and the second detection chamber 62 (used as the renal function indicator reaction detection well) thoroughly mixes the lyophilized substrate. The absorbance in the pre-cystatin C pre-reaction detection well, the cystatin C detection well, and the renal function indicator reaction detection well are measured, and the concentration of the target indicator can be calculated based on the standard curve.

[0102] Specifically, the centrifugation speed and oscillation speed of the biodetection chip are determined according to actual needs, and this application does not impose specific limitations.

[0103] The biodetection chip in the above embodiments can simultaneously perform two-step and one-step biochemical detection. The substrates required for the biochemical reaction are pre-stored in the corresponding cavities and reaction wells of the chip in lyophilized form. Users can adjust the lyophilized substrates placed in the chip to achieve combined detection of other indicators as needed. The pre-storage of substrates for biochemical reactions greatly simplifies the operation for users of the biodetection chip.

[0104] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0105] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A bioreactor chip, characterized in that, include: The chip body has a material chamber with a ventilation structure. The material chamber includes a sample holding area (10), a diluent loading area (20), a sample dilution mixing area (30), a pre-reaction area (40), and a reaction detection area. The sample holding area (10) and the diluent loading area (20) are both connected to the sample dilution mixing area (30). The sample dilution mixing area (30) is connected to the reaction detection area through the pre-reaction area (40). The pre-reaction area (40) and the reaction detection area are respectively pre-set with a first reactant and a second reactant. The sample holding area (10) has a first loading port, and the diluent loading area (20) has a second loading port. A sealing layer capable of sealing the material chamber; The test sample entering the sample holding area (10) from the first sample dispensing port and the diluent entering the diluent dispensing area (20) from the second sample dispensing port can both enter the sample dilution mixing area (30) for mixing. The mixed liquid enters the pre-reaction area (40) and reacts with the first reactant in the pre-reaction area (40) to form a pre-reaction liquid. The pre-reaction liquid enters the reaction detection area and reacts with the second reactant to form a test sample. The pre-reaction zone (40) includes a sample dilution storage chamber (41), a pre-reaction chamber (42), a third connecting tube (43), a first siphon tube (46), and a second siphon tube (45). The first reactant is pre-set in the pre-reaction chamber (42). The third connecting tube (43) connects the proximal outlet of the sample dilution storage chamber (41) to the inlet of the pre-reaction chamber (42). The first siphon tube (46) connects to the outlet of the pre-reaction chamber (42). The second siphon tube (45) connects to the distal outlet of the sample dilution storage chamber (41). The ventilation structure includes a fourth ventilation tube (44) connected to the pre-reaction chamber (42). The reaction detection area includes a first reaction detection area (50) and a second reaction detection area (60) independent of the first reaction detection area (50). The first siphon (46) is connected to the inlet of the first reaction detection area (50), the second siphon (45) is connected to the inlet of the second reaction detection area (60), the second reactant is located in the first reaction detection area (50), and a third reactant is preset in the second reaction detection area (60).

2. The bioreactor chip according to claim 1, characterized in that, The bioreactor chip is a centrifugal chip. The sample holding area (10) includes a sample collection chamber (11), a sample quantitative separation chamber (12), a sample waste liquid chamber (13), and a sample siphon tube (14) for transporting the liquid in the sample quantitative separation chamber (12) to the sample dilution and mixing area (30). The sample collection chamber (11) is used to receive the test sample. The sample quantitative separation chamber (12) includes a first cavity (121), a first connecting tube (122), a second cavity (123), a second connecting tube (124), and a third cavity (125) connected sequentially from the proximal end to the distal end. The cross-sectional area along the direction perpendicular to the radius of rotation is... The cross-sectional areas of the first connecting tube (122) and the second connecting tube (124) are both smaller than the cross-sectional areas of the first cavity (121), the second cavity (123), and the third cavity (125). The sample siphon tube (14) connects the second connecting tube (124) and the sample dilution mixing zone (30). The sample waste liquid cavity (13) is connected to the third cavity (125) through the waste liquid outlet tube (17). The distance from the closest point of the sample siphon tube (14) to the rotation center is smaller than the distance from the closest point of the waste liquid outlet tube (17) to the rotation center. The ventilation structure includes a first ventilation pipe (15) connected to the sample waste liquid chamber (13) and a second ventilation pipe (16) connected to the sample collection chamber (11).

3. The bioreactor chip according to claim 2, characterized in that, The inner cavity of the second cavity (123) is a spindle-shaped cavity that is thick in the middle and thin at both ends along the fluid flow direction.

4. The bioreactor chip according to claim 2, characterized in that, The sample collection cavity (11) is a hydrophilic cavity.

5. The bioreactor chip according to claim 1, characterized in that, The diluent loading area (20) includes a diluent loading chamber (21), a diluent loading port (22) for loading the diluent loading chamber (21), and a diluent transfer tube (23) connecting the diluent loading chamber (21) and the sample dilution mixing area (30). The diluent loading port (22) is located at the proximal end of the diluent loading chamber (21), and the diluent transfer tube (23) is connected to the distal end of the diluent loading chamber (21).

6. The bioreactor chip according to claim 1, characterized in that, The sample dilution mixing zone (30) includes a mixing chamber (31) and a mixing chamber siphon (34). The mixing chamber siphon (34) is connected to the outlet of the mixing chamber (31) and is used to deliver the mixture to the pre-reaction zone (40). The mixing chamber siphon (34) is a hydrophilic tube. The first connection port of the dilution sample addition zone (20) connected to the mixing chamber (31) and the second connection port of the sample holding zone (10) connected to the mixing chamber (31) are located on the same side of the outlet of the mixing chamber (31). The ventilation structure includes a third ventilation tube (32) connected to the mixing chamber (31).

7. The bioreactor chip according to claim 1, characterized in that, The first reaction detection zone (50) includes a first distribution tube (51), a pre-reaction detection chamber (52), a first detection chamber (53), and a blank control chamber (54). The blank control chamber (54) is a cavity structure. The pre-reaction detection chamber (52) and the first detection chamber (53) are both connected to the first distribution tube (51). The inlet of the first distribution tube (51) is connected to the outlet of the first siphon tube (46). The ventilation structure includes a ventilation structure (55) connected to the end of the first distribution tube (51) away from the inlet of the first distribution tube (51). The second reactant is located in the first detection chamber (53).

8. The bioreactor chip according to claim 7, characterized in that, Along the fluid flow direction in the first distribution pipe (51), the connection end of the first detection chamber (53) and the first distribution pipe (51) is located downstream of the connection end of the pre-reaction detection chamber (52) and the first distribution pipe (51). The first detection chamber (53) includes an expansion chamber and a reaction chamber connected to the distal end of the expansion chamber. The proximal end of the expansion chamber is connected to the first distribution pipe (51). Along the direction perpendicular to the radius of the rotation center, the cross-sectional area of ​​the expansion chamber is larger than the cross-sectional area of ​​the reaction chamber, so that the liquid fills the reaction chamber, while the expansion chamber is in an unfilled state.

9. The bioreactor chip according to claim 1, characterized in that, The second reaction detection zone (60) includes a second distribution pipe (61), a second detection chamber (62), and a waste liquid chamber (63). The second detection chamber (62) and the waste liquid chamber (63) are both connected to the second distribution pipe (61). The inlet of the second distribution pipe (61) is connected to the outlet of the second siphon pipe (45). The third reactant is located in the second detection chamber (62) along the fluid flow direction in the second distribution pipe (61). The connection port between the waste liquid chamber (63) and the second distribution pipe (61) is located downstream of the connection port between the second detection chamber (62) and the second distribution pipe (61). The ventilation structure includes a sixth ventilation pipe connected to one end of the second distribution pipe (61) away from the inlet section of the second distribution pipe (61).

10. The bioreactor chip according to claim 1, characterized in that, The chip body is provided with a positioning structure (70), which includes a positioning tooth (72) disposed at the distal end of the chip body and a positioning hole (71) disposed on the chip body.

11. A bioreactor device, comprising a bioreactor chip and a drive device for centrifugally rotating the bioreactor chip, characterized in that, The bioreactor chip is the bioreactor chip according to any one of claims 1-10.

12. The bioreactor according to claim 11, characterized in that, It also includes a control device connected to the drive device, which can control the drive device to drive the bioreactor chip to swing and mix the substances in the chip body.

13. A biological detection method, characterized in that, The bioreactor chip according to any one of claims 1-10 comprises the following steps: The test sample is sent into the sample holding area (10), and the diluent is sent into the diluent addition area (20). The biodetection chip is centrifuged and rotated. After the particles in the detection sample in the sample holding area (10) are separated, they enter the sample dilution and mixing area (30), and at the same time, the diluent enters the sample dilution and mixing area (30). The bio-detection chip oscillates back and forth, and the diluent mixes with the detection sample to form a mixed liquid. The bio-detection chip is centrifugally rotated, and the mixed liquid enters the pre-reaction zone (40). The bio-detection chip oscillates back and forth, and the mixed liquid mixes with the first reactant to form a pre-reaction liquid; The bio-detection chip rotates centrifugally, and the pre-reaction liquid enters the reaction detection zone, where it reacts with the second reactant to form a test sample.

14. The biological detection method according to claim 13, characterized in that, When the biodetector chip oscillates back and forth, the oscillation is non-uniform.

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