Biological reaction chip, biological reaction equipment and biological detection method

Abstract

The invention discloses a biological reaction chip, biological reaction equipment 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, and the material cavity is provided with a ventilation structure; the material chamber comprises a sample containing area, a diluent sample adding area, a sample diluting and mixing area, a pre-reaction area and a reaction detection area, a detection sample entering the sample containing area from the first sample adding port and a diluent entering the diluent sample adding area from the second sample adding port can enter the sample diluting and mixing area to be mixed, and then the mixed liquid enters the pre-reaction area; reacting with a first reaction substance in the pre-reaction area to form pre-reaction liquid, and enabling the reaction liquid to enter the reaction detection area to react with a second reaction substance to form a test sample. According to the biological reaction chip provided by the invention, the labor intensity of workers during detection is effectively reduced.

Description

Technical Field

[0001] The present invention relates to the field of materials and biotechnology, and in particular to a bioreaction chip, a bioreaction device and a biodetection method. Background Art

[0002] When conducting biological two-step reaction detection, for example, there are many biochemical indicators involved in renal function testing. Traditional cystatin C detection mostly uses immunological methods. The detection process has very high requirements on the mixing effect of reagents, and the reaction is divided into two steps. At the same time, the two-step reactions are required to occur independently of each other and to be completed on independent structures.

[0003] In the specific implementation, the staff is required to transfer the pre-reaction liquid after the first step reaction is completed to the structure required for the second step reaction, and then carry out the second step reaction. The operation is cumbersome and the labor intensity of the staff is high.

[0004] Therefore, how to reduce the labor intensity of the staff when performing the test is a technical problem that needs to be solved urgently by those skilled in the art. Summary of the invention

[0005] The object of the present invention is to provide a bioreactor chip, which can reduce the labor intensity of the staff when performing the detection. Another object of the present invention is to provide a bioreactor device including the above-mentioned bioreactor chip. Another object of the present invention is to provide a biodetection method.

[0006] To achieve the above object, the present invention provides a bioreactor chip, comprising:

[0007] A chip body, wherein a material chamber is provided on the chip body, and the material chamber has a ventilation structure, the material chamber comprises a sample accommodating area, a diluent adding area, a sample dilution mixing area, a pre-reaction area and a reaction detection area, the sample accommodating area and the diluent adding area can both be connected to the sample dilution mixing area, the sample dilution mixing area is connected to the reaction detection area through the pre-reaction area, the pre-reaction area and the reaction detection area are respectively preset with a first reaction substance and a second reaction substance, the sample accommodating area is provided with a first adding port, and the diluent adding area is provided with a second adding port;

[0008] A sealing layer, wherein the sealing layer is capable of sealing the material chamber;

[0009] The test sample entering the sample containing area from the first sample adding port and the diluent entering the diluent adding area from the second sample adding port can both enter the sample dilution mixing area and mix, and the mixed liquid enters the pre-reaction area to react with the first reaction substance 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 substance to form a test sample.

[0010] Optionally, in the above-mentioned bioreactor chip, the bioreactor chip is a centrifugal chip, the sample containing area includes a sample collection chamber, a sample quantitative separation chamber, a sample waste liquid chamber and a sample siphon tube for transporting the liquid in the sample quantitative separation chamber to the sample dilution and mixing area, the sample collection chamber is used to receive the test sample, the sample quantitative separation chamber includes a first cavity, a first connecting tube, a second cavity, a second connecting tube and a third cavity connected in sequence from the proximal end to the distal end, the cross-sectional area along the direction perpendicular to the rotation radius, the cross-sectional area of ​​the first connecting tube and the cross-sectional area of ​​the second connecting tube 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, the sample siphon tube connects the second connecting tube and the sample dilution and mixing area, the sample waste liquid chamber is connected to the third cavity through a waste liquid outlet tube, and the distance from the closest point of the sample siphon tube to the rotation center to the rotation center is smaller than the distance from the closest point of the waste liquid outlet tube to the rotation center;

[0011] The ventilation structure includes a first ventilation tube connected to the sample waste liquid chamber and a second ventilation tube connected to the sample collection chamber.

[0012] Optionally, in the above bioreactor chip, the inner cavity of the second cavity is a spindle-shaped cavity that is thick in the middle and thin at both ends along the fluid flow direction.

[0013] Optionally, in the above bioreactor chip, the sample collection cavity is a hydrophilic cavity.

[0014] Optionally, in the above-mentioned bioreactor chip, the diluent loading area includes a diluent loading chamber, a diluent loading port for loading samples into the diluent loading chamber, and a diluent transfer tube connecting the diluent loading chamber and the sample dilution mixing area, the diluent loading port is located at the proximal end of the diluent loading chamber, and the diluent transfer tube is connected to the distal end of the diluent loading chamber.

[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 mixing chamber outlet and is used to transport the mixed solution to the pre-reaction area, the mixing chamber siphon is a hydrophilic tube body, and the first connection port connecting the diluent adding area with the mixing chamber and the second connection port connecting the sample containing area with the mixing chamber are located on the same side of the mixing chamber outlet;

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

[0017] Optionally, in the above bioreaction chip:

[0018] The pre-reaction zone includes a diluted sample temporary storage chamber, a pre-reaction chamber, a third connecting pipe, a first siphon tube and a second siphon tube, the first reaction substance is preset in the pre-reaction chamber, the third connecting pipe connects the proximal end liquid outlet of the diluted sample temporary storage chamber and the liquid inlet of the pre-reaction chamber, the first siphon tube is connected to the liquid outlet of the pre-reaction chamber, and the second siphon tube is connected to the distal end liquid outlet of the diluted sample temporary storage chamber; the ventilation structure includes a fourth ventilation pipe connected to the pre-reaction chamber;

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

[0020] Optionally, in the above-mentioned bioreactor 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 cavity structure, the pre-reaction detection chamber and the first detection chamber are both connected to the first distribution tube, the liquid inlet of the first distribution tube is connected to the liquid outlet of the first siphon tube; the ventilation structure includes a ventilation structure connected to an end of the first distribution tube away from the liquid inlet of the first distribution tube, and the second reaction substance is located in the first detection chamber.

[0021] Optionally, in the above-mentioned bioreactor chip, along the flow direction of the fluid in the first distribution tube, the connecting end of the first detection chamber and the first distribution tube is located downstream of the connecting end of the pre-reaction detection chamber and the first distribution tube, the first detection chamber 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 tube, and along the direction perpendicular to the rotation center radius, 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, and 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 liquid inlet of the second distribution tube is connected to the liquid outlet of the second siphon tube, the third reaction substance is located in the second detection chamber, along the fluid flow direction in the second distribution tube, the connecting port of the waste liquid chamber and the second distribution tube is located downstream of the connecting port of the second detection chamber and the second distribution tube, and the ventilation structure includes a sixth ventilation tube connected to an end of the second distribution tube away from the liquid inlet section of the second distribution tube.

[0023] Optionally, in the above-mentioned bioreactor chip, a positioning structure is provided on the chip body, and the positioning structure includes a positioning tooth arranged at the distal end of the chip body and a positioning hole arranged on the chip body.

[0024] A bioreaction device comprises a bioreaction chip and a driving device for driving the bioreaction chip to centrifugally rotate. The bioreaction chip is any of the bioreaction chips described above.

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

[0026] A biological detection method, according to any of the above-mentioned biological reaction chips, comprises the following steps:

[0027] Send the test sample into the sample holding area, and the diluent into the diluent adding area;

[0028] The bioassay chip is centrifugally rotated, and the particles in the test sample in the sample containing area are separated and enter the sample dilution and mixing area, and the diluent enters the sample dilution and mixing area at the same time;

[0029] The biological detection chip reciprocates, and the diluent is mixed with the detection sample to form a mixed liquid;

[0030] The bioassay chip is centrifugally rotated, and the mixed liquid enters the pre-reaction area;

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

[0032] The bioassay chip is centrifugally rotated, and the pre-reaction liquid enters the reaction detection area and reacts with the second reaction substance to form a test sample.

[0033] Optionally, in the above biological detection method, when the biological detection chip swings back and forth, the biological detection chip swings at a non-uniform speed.

[0034] In the above technical scheme, the bioreactor chip provided by the present invention includes a chip body and a sealing layer, a material chamber is provided on the chip body, and the material chamber has a ventilation structure, the material chamber includes a sample accommodating area, a diluent adding area, a sample dilution mixing area, a pre-reaction area and a reaction detection area, the accommodating area and the diluent adding area can be connected with the sample dilution mixing area, the sample dilution mixing area is connected with the reaction detection area through the pre-reaction area, the pre-reaction area and the reaction detection area are respectively preset with a first reaction substance and a second reaction substance, the sample accommodating area is provided with a first adding port, and the diluent adding area is provided with a second adding port. The sealing layer can seal the material chamber; the test sample entering the sample accommodating area from the first adding port and the diluent entering the diluent adding area from the second adding port can both enter the sample dilution mixing area for mixing, the mixed liquid enters the pre-reaction area, reacts with the first reaction substance 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 substance to form a test sample.

[0035] It can be seen from the above description that in the bioreactor chip provided by the present application, the material chamber includes a sample accommodating area, a diluent adding area, a sample dilution mixing area, a pre-reaction area and a reaction detection area, and different fluids can be mixed during the liquid flow process. The pre-reaction area and the reaction detection area are respectively preset with a first reaction substance and a second reaction substance, and the one-step reaction to form the pre-reaction liquid and the two-step reaction to form the test sample are both implemented in the chip body, without the need for staff to transfer the reaction liquid, which effectively reduces the labor intensity of the staff in conducting test detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0037] Figure 1 A schematic diagram showing the sample receiving area, the first reaction detection area and the second reaction detection area in the bioreactor chip provided by an embodiment of the present invention;

[0038] Figure 2 A schematic diagram of the marking of the pre-reaction area in the bioreactor chip provided in an embodiment of the present invention;

[0039] Figure 3 A schematic diagram showing the marking of the diluent adding area and the sample dilution mixing area in the bioreactor chip provided in an embodiment of the present invention;

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

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

[0042] Figure 6 A schematic diagram of the structure of a sample collection chamber provided by an embodiment of the present invention;

[0043] Figure 7 A schematic diagram of the structure of the back side of the bioreactor chip provided by an embodiment of the present invention;

[0044] Figure 8 A schematic diagram of the fluid structure of adding sample liquid and diluent into the bioreactor chip provided by an embodiment of the present invention;

[0045] Fig. 9 A schematic diagram of the flow of the diluent entering the sample dilution and mixing area and the sample liquid entering the sample quantitative separation chamber in the bioreactor chip provided by an embodiment of the present invention;

[0046] Fig.10 A schematic diagram of the flow of sample liquid entering the sample dilution and mixing area in the bioreactor chip provided by an embodiment of the present invention;

[0047] Fig.11 A schematic diagram of the fluid flow of the mixed liquid entering the pre-reaction zone in the bioreactor chip provided by an embodiment of the present invention;

[0048] Fig.12 This is a schematic diagram of the flow of mixed liquid entering the first reaction detection area and the second reaction detection area in the bioreactor chip provided by an embodiment of the present invention.

[0049] in Figure 1-12 middle:

[0050] The sample containing area 10, the sample collecting chamber 11, the sample quantitative separation chamber 12, the first chamber 121, the first connecting tube 122, the second chamber 123, the second connecting tube 124, the third chamber 125, the sample waste liquid chamber 13, the sample siphon 14, the first ventilation tube 15, the second ventilation tube 16, and the waste liquid outlet tube 17;

[0051] diluent sample adding area 20, diluent sample adding chamber 21, diluent sample adding port 22, diluent transfer tube 23;

[0052] Sample dilution and mixing area 30, mixing chamber 31, third ventilation pipe 32, exhaust hole 33, mixing chamber siphon 34;

[0053] Pre-reaction area 40, diluted sample temporary storage chamber 41, pre-reaction chamber 42, third connecting pipe 43, fourth ventilation pipe 44, second siphon 45, first siphon 46;

[0054] A first reaction detection area 50, a first distribution tube 51, a pre-reaction detection chamber 52, a first detection chamber 53, a blank control chamber 54, and a ventilation structure 55;

[0055] A second reaction detection area 60, a second distribution pipe 61, a second detection chamber 62, and a waste liquid chamber 63;

[0056] Positioning structure 70 , positioning hole 71 , positioning tooth 72 . DETAILED DESCRIPTION

[0057] The core of the present invention is to provide a bioreactor chip, which reduces the labor intensity of the staff when performing the detection. Another core of the present invention is to provide a bioreactor device including the above-mentioned bioreactor chip. Another core of the present invention is to provide a biodetection method.

[0058] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and implementation modes.

[0059] Please refer to Figures 1 to 12 .

[0060] In a specific embodiment, the bioreactor chip provided by the specific embodiment of the present 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), PS (polystyrene) by injection molding or machining.

[0061] Specifically, the sealing layer can be made of PC (polycarbonate), PMMA (polymethyl methacrylate), PS (polystyrene), PP (polypropylene), PET (polyethylene terephthalate) and other materials bonded by single-sided or double-sided adhesive. Figure 1 The sealing layer can also be directly welded to the chip body, for example, by ultrasonic welding, laser welding, hot pressing welding, etc.

[0062] The chip body is provided with a material chamber, the sealing layer can seal the material chamber, the material chamber has a ventilation structure, the material chamber includes a sample accommodating area 10, a diluent adding area 20, a sample dilution mixing area 30, a pre-reaction area 40 and a reaction detection area, the sample accommodating area 10 and the diluent adding area 20 can be connected with the sample dilution mixing area 30, that is, the material in the sample accommodating area 10 and the diluent adding area 20 can flow to the sample dilution mixing area 30. The sample dilution mixing area 30 is connected with the reaction detection area through the pre-reaction area 40, and the pre-reaction area 40 and the reaction detection area are respectively preset with a first reaction substance and a second reaction substance, and the first reaction substance and the second reaction substance are determined according to the reaction substances required. Specifically, the sample dilution mixing area 30 can be used for serum dilution and mixing.

[0063] The test sample entering the sample containing area 10 from the first sample adding port and the diluent entering the diluent adding area 20 from the second sample adding 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 reaction substance 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 reaction substance to form a test sample.

[0064] The sample receiving area 10 is provided with a first sample loading port. Specifically, the first sample loading port is arranged at the side end of the chip body, specifically at the proximal end of the chip body, and the diluent loading area 20 is provided with a second sample loading port. Specifically, an independent liquid receiving chamber can be provided at the position where the second reaction substance is located before entering the reaction detection area. Different liquid receiving chambers are arranged along the liquid flow direction, and the liquids are filled one by one, that is, before entering the reaction detection area, a part of the liquid flows preferentially to the solution receiving chamber, and the other part flows to the position where the second reaction substance is located. Specifically, the reaction detection area is provided with a pre-reaction detection chamber after the primary reaction and a test sample detection chamber after the secondary reaction. The reactant that has only reacted with the first reaction substance is detected by the pre-reaction detection chamber, and the reactant that has reacted with the first reaction substance and the second reaction substance is detected by the test sample detection chamber.

[0065] The bioreactor chip is a centrifugal chip, and the sample containing area 10 includes a sample collection chamber 11, a sample quantitative separation chamber 12, a sample waste liquid chamber 13, and a sample siphon 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, and 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 in sequence from the proximal end to the distal end. In terms of the cross-sectional area along the direction perpendicular to the rotation radius, the cross-sectional area of ​​the first connecting tube 122 and the cross-sectional area of ​​the second connecting tube 124 are both smaller than the cross-sectional area of ​​the first cavity 121, the cross-sectional area of ​​the second cavity 123, and the cross-sectional area of ​​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 thick in the middle and thin 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 tube 124 and the first connecting tube 122 connect the two opposite corner positions of the second cavity 123.

[0066] like Figure 6 As shown, the sample siphon 14 connects the second connecting tube 124 and the sample dilution and mixing area 30, and the distance from the closest point a of the sample siphon 14 to the rotation center is smaller 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 14 is a V-shaped tube structure protruding toward the proximal end, and preferably, the angle position of the inner cavity of the tube body is smoothly transitioned. The sample waste liquid cavity 13 is connected to the third cavity 125 through the waste liquid outlet tube 17.

[0067] The ventilation structure includes a first ventilation tube 15 connected to the sample waste liquid chamber 13 and a second ventilation tube 16 connected to the sample collection chamber 11. Specifically, the outlet ends of the first ventilation tube 15 and the second ventilation tube 16 can both be opened on the side wall 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 with blood / serum / plasma and other samples through its first sample loading port located on the side of the proximal end of the bioreactor chip to produce capillary action, thereby completing the collection and storage of the sample.

[0069] like Figure 6As shown, the sample quantitative separation chamber 12 has a "double dumbbell" structure with three cavities connected in sequence, that is, it has two contracted "thin waist" structures (first connecting tube 122 and second connecting tube 124), which realize particle separation and quantification of the sample under the action of centrifugal force. The thin waist structure (second connecting tube 124) at the lower end 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 cavity 125 and the waste liquid outlet tube 17; the first ventilation tube 15 is used for ventilation during liquid transfer.

[0070] Two ends of the waste liquid outlet tube 17 are connected to the proximal end of the third cavity 125 and the proximal end of the sample waste liquid cavity 13 respectively.

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

[0072] The diluent injection cavity 21 is used to contain a fixed volume of diluent required for biochemical detection on the chip; the diluent injection port 22 is specifically matched with the size of the pipette tip for diluent injection from Figure 7 The chip body shown is loaded with samples on the back side.

[0073] The sample dilution and mixing area 30 includes a mixing chamber 31 and a mixing chamber siphon 34 for conveying the mixed solution to the pre-reaction area 40, and the liquid outlet of the mixing chamber 31 is connected to the mixing chamber siphon 34. The mixing chamber siphon 34 is a hydrophilic tube body, and the first connection port between the diluent adding area 20 and the mixing chamber 31 and the second connection port between the sample containing area 10 and the mixing chamber 31 are located on the same side of the liquid outlet of the mixing chamber 31; the ventilation structure includes a third ventilation tube 32 connected to the mixing chamber 31, and specifically, the exhaust hole 33 at the end of the third ventilation tube 32 is opened on the front of the chip body.

[0074] During specific operation, the mixing chamber 31 is used to mix the transferred test sample and the diluent; the exhaust hole 33 ensures that the mixing chamber 31 is connected to the outside atmosphere.

[0075] The pre-reaction zone 40 includes a diluted sample temporary 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 is preset with a first reaction substance. The third connecting tube 43 connects the proximal end liquid outlet of the diluted sample temporary storage chamber 41 and the liquid inlet of the pre-reaction chamber 42. The first siphon tube 46 connects the liquid outlet of the pre-reaction chamber 42. The second siphon tube 45 connects the distal end liquid outlet of the diluted sample temporary storage chamber 41. The distance from the proximal end liquid outlet to the rotation center is smaller than the distance from the distal end liquid outlet to the rotation center. 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 pipe 44 connected to the pre-reaction chamber 42. Specifically, the pre-reaction zone 40 may be a pre-reaction zone 40 for performing a cystatin C reaction.

[0077] The reaction detection area includes a first reaction detection area 50 and a second reaction detection area 60. The first reaction detection area 50 and the second reaction detection area 60 are independently arranged. The first siphon 46 is connected to the liquid inlet of the first reaction detection area 50, and the second siphon 45 is connected to the liquid inlet of the second reaction detection area 60. The second reaction substance is located in the first reaction detection area 50, and the second reaction detection area 60 is provided with a third reaction substance. The first reaction detection area 50 can be a detection area for cystatin C reaction, and the second reaction detection area 60 can be a detection area for other renal function indicators.

[0078] Specifically, the first reaction substance is a freeze-dried reagent for the first step pre-reaction in the cystatin C biochemical test, and the sample diluted by the diluent enters the freeze-dried reagent for the first step reaction of the cystatin C biochemical test to re-dissolve and realize the pre-reaction. The fourth ventilation pipe 44 is connected to the pre-reaction chamber 42 to realize ventilation with the outside; the second siphon 45 connects the pre-reaction chamber 42 and the downstream first reaction detection area 50; the first siphon 46 connects the diluted sample temporary storage chamber 41 and the downstream second reaction detection area 60.

[0079] The first reaction detection area 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 pre-reaction detection chamber 52 and the first detection chamber 53 are both connected to the first distribution tube 51. Of course, the blank control chamber 54 can also be connected to the first distribution tube 51. Specifically, the pre-reaction detection chamber 52, the first detection chamber 53 and the blank control chamber 54 can all be hole structures. Along the flow direction of the fluid in the first distribution tube 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 tube 51, the connection end of the first detection chamber 53 and the first distribution tube 51 can be located downstream of the connection end of the pre-reaction detection chamber 52 and the first distribution tube 51. The fluid entering the first distribution tube 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 tube 51 away from the liquid inlet of the first distribution tube 51. The liquid inlet of the first distribution tube is connected to the liquid outlet of the first siphon 46, and the second reaction substance is located in the first detection chamber 53. The bioreaction chip provided in the present application can detect cystatin C by two-step method. The final detection result is achieved with the pre-reaction detection chamber 52 and the first detection chamber 53 on the same centrifugal radius. The two-step reaction of cystatin C has been fully mixed to ensure stable reaction and reliable detection. The blank control chamber 54 is a cavity structure, that is, the blank control chamber 54 is not provided with any 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. In the direction perpendicular to the rotation center radius, the cross-sectional area of ​​the expansion chamber is larger than the cross-sectional area of ​​the reaction chamber. That is, the first detection chamber 53 is in a shape with a large circumferential dimension at the proximal end and a small circumferential dimension at the distal end, and is wide at the upper (proximal end) and narrow at the lower (distal end). 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 the reagents, thereby ensuring a fully stable reaction.

[0082] The blank control chamber 54 is an empty hole, and the pre-reaction detection chamber 52 is used to accommodate the pre-reaction product in the pre-reaction chamber 42 to detect the pre-reaction result; specifically, the first detection chamber 53 is fixed with the freeze-dried reagent of the second step cystatin C biochemical reaction as the second reaction substance, which can be re-dissolved by the liquid from the pre-reaction chamber 42 to complete the second step biochemical reaction of cystatin C.

[0083] The second reaction detection area 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. Specifically, the second detection chamber 62 and the waste liquid chamber 63 can be a hole structure with an opening on the upper surface. The liquid inlet of the second distribution pipe 61 is connected to the liquid outlet of the second siphon tube 45, and the third reaction substance is located in the second detection chamber 62. Along the flow direction of the fluid in the second distribution pipe 61, the connection between the waste liquid chamber 63 and the second distribution pipe 61 is located downstream of the connection 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 an end of the second distribution pipe 61 away from the liquid inlet section of the second distribution pipe 61.

[0084] Specifically, multiple second detection chambers 62 can be provided, each of which is preset with different reaction substances to perform multiple tests. Preferably, the fluid entering the second distribution pipe 61 first enters each second detection chamber 62 in sequence, and finally enters the waste liquid chamber 63.

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

[0086] In order to avoid mutual interference of structures, 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 ventilation tube 44 can be located at the side end of the chip body, and there is no need to set a separate ventilation hole.

[0087] The second reaction detection area 60 can be other renal function index detection areas. The second detection cavity 62 stores the freeze-dried substrate required for the biochemical detection of renal function detection indexes as the third reaction substance. The detection indexes include urea nitrogen, blood creatinine, uric acid and the like.

[0088] The chip body is provided with a positioning structure 70, which includes a positioning tooth 72 provided at the distal end of the chip body and a positioning hole 71 provided on the chip body. The positioning hole 71 may be one or at least two. Specifically, a corresponding mounting seat is provided with a limiting column matched with the positioning hole 71. Specifically, the positioning tooth 72 is provided in a serrated shape along the periphery of the chip body. Specifically, the mounting seat is provided with a limiting tooth engaged with the positioning tooth 72, or an optical sensor is provided for judging the position of the detection hole through the positioning tooth 72 when the chip rotates.

[0089] The bioreaction chip can realize the biochemical detection of various renal function-related indicators including cystatin C. All substrates of the biochemical reaction are frozen and stored in the chip in advance. Among them, the biochemical reaction of cystatin C is a two-step method, while other indicators are all one-step methods. Through the specific chip structure and centrifugal workflow, the one-step method and the two-step method can be automatically and controllably carried out simultaneously.

[0090] Specifically, the biochemical reaction chip is arranged on a rotating platform, and one rotating platform is preferably provided with a plurality of circumferentially arranged biochemical reaction chips.

[0091] The present application provides a bioreactor device, including a bioreactor chip and a driving device for driving the bioreactor chip to rotate centrifugally, wherein the bioreactor chip is any of the above-mentioned bioreactor chips. The above-mentioned specific structure of the bioreactor chip is described, and the present application includes the above-mentioned bioreactor chip and also has the above-mentioned technical effects.

[0092] In a specific embodiment, the bioreactor device further comprises a control device connected to the driving device, and the control device can control the driving device to drive the bioreactor chip to swing and mix the substances in the chip body.

[0093] The present application provides a biological detection method, according to the above-mentioned biological reaction chip, comprising the following steps:

[0094] The test sample is sent to the sample containing area 10, and the diluent is sent to the diluent adding area 20. Specifically, the side of the sample collection cavity 11 contacts the sample to absorb the test sample, and a fixed volume of diluent is added through the diluent adding port 22 on the back of the chip body, and then the bioreactor chip is placed on the centrifugal platform of the matching detection instrument.

[0095] The biological detection chip is centrifuged, and the particles in the detection sample in the sample containing area 10 are separated, and then enter the sample dilution mixing area 30, and the diluent enters the sample dilution mixing area 30. Specifically, the biological detection chip is centrifuged at high speed for a period of time, and the detection sample enters the sample quantitative separation chamber 12 from the sample collection chamber 11 under the action of centrifugal force to achieve quantification, and the excess sample enters the sample waste liquid chamber 13. The sample realizes particle separation in the sample under the action of centrifugal force, such as serum and blood cell separation; the diluent sample addition chamber 21 enters the mixing chamber through the diluent transfer tube 23; the centrifugation is stopped or the centrifugation speed is reduced, and the separated serum or plasma sample fills the sample siphon 14 under capillary action, and the mixing chamber siphon 34 will not be filled with liquid because it has not been hydrophilically modified.

[0096] During specific operation, the biological detection chip rotates centrifugally, and the sample enters the sample dilution and mixing area 30 through the siphon tube 14 under the action of siphon and contacts the diluent therein. The biological detection chip swings back and forth, and the diluent is mixed with the detection sample to form a mixed liquid. Among them, when the biological detection chip swings back and forth, the biological detection chip can swing at a uniform speed. In order to improve the mixing efficiency, it is preferred that when the biological detection chip swings back and forth, the biological detection chip swings at a non-uniform speed. Specifically, the swinging of the biological detection chip is: reducing the rotation speed of the biological detection chip, and using a high acceleration and low speed forward and reverse mode (such as a rotation speed of ±300rpm, an acceleration of 10000rpm / s) to swing the biological detection chip, so as to achieve sufficient mixing of the detection sample and the diluent. In addition, the high acceleration back and forth swinging allows the liquid to enter and fill the mixing chamber siphon tube 34 while mixing.

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

[0098] Acceleration range: 100rpm / s~30000rpm / s. Specifically, when the purpose is to mix the liquid in the container, symmetrical rotation speed and acceleration are usually used (for example, ±1000rpm, acceleration 20000rpm / s); and when it is necessary to fill the corresponding siphon pipe by high-acceleration liquid impact, asymmetric acceleration will be used, and the speed may also be asymmetric (for example, rotation speed ±200rpm, clockwise acceleration 500rpm / s, counterclockwise acceleration 20000rpm / s, at this time the liquid gradually flows into the siphon pipe). Specifically, when the motor changes speed, there must be a clockwise or counterclockwise force on the liquid. This force is used to make the liquid have the effect of rushing to the left or right, that is, the impact force on the liquid is generated by the speed change of the motor, and then the liquid impacts the siphon pipe on the biological detection chip until the siphon pipe is finally filled.

[0099] The biological detection chip rotates centrifugally, and the mixed liquid enters the pre-reaction area 40. The biological detection chip swings back and forth, and the mixed liquid mixes with the first reaction substance to form a pre-reaction liquid.

[0100] Specifically, the biological detection chip is centrifugally rotated at a high speed, and the mixed sample in the mixing chamber enters the diluted sample temporary storage chamber 41 through the mixing chamber siphon tube 34 under the action of siphon. After the diluted sample temporary storage chamber 41 is filled, the excess liquid enters the pre-reaction chamber 42 through the third connecting tube 43; thereafter, the rotation speed of the biological reaction chip is reduced, and the chip can also be swung 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 freeze-dried reagent therein 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 swing of the biological detection chip.

[0101] The biological detection chip is centrifugally rotated, and the pre-reaction liquid enters the reaction detection area and reacts with the second reaction substance to form a test sample. The biological detection chip is centrifugally rotated at high speed, and the liquid in the dilution sample temporary storage chamber 41 and the pre-reaction chamber 42 enters the first distribution tube 51 and the second distribution tube 61 through the second siphon 45 and the first siphon 46 respectively under the action of centrifugal force, and fills the pre-reaction detection chamber 52 and partially fills the first detection chamber 53, and fills the second detection chamber 62 and partially fills the waste liquid chamber 63 respectively. Afterwards, the rotation speed of the biological detection chip is reduced, and the biological detection chip is swung in a high acceleration and low speed forward and reverse mode so that the liquid in the first detection chamber 53 as the cystatin C detection hole and the second detection chamber 62 as the renal function index reaction detection hole is fully mixed with the freeze-dried substrate, and the absorbance in the pre-cystatin C pre-reaction detection hole, the cystatin C detection hole and the renal function index reaction detection hole is detected respectively, and the concentration of the index to be detected can be calculated according to the standard curve.

[0102] Specifically, the centrifugal speed and the swinging speed of the biological detection chip are determined according to actual needs, and this application does not make specific limitations.

[0103] The biological detection chip in the above embodiment can realize two-step and one-step biochemical detection at the same time. The substrate required for the biochemical reaction is pre-stored in the corresponding cavity and reaction hole of the chip in a freeze-dried form. The user can adjust the freeze-dried substrate placed in the chip to realize the combined detection of other indicators as needed. The pre-storage of the substrate for the biochemical reaction greatly simplifies the operation of the biological detection chip user.

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

[0105] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one 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 present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A bioreactor chip, characterized in that: include: A chip body, wherein a material chamber is provided on the chip body, and the material chamber has a ventilation structure, the material chamber comprises a sample accommodating area (10), a diluent adding area (20), a sample dilution mixing area (30), a pre-reaction area (40) and a reaction detection area, the sample accommodating area (10) and the diluent adding area (20) can both be 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 reaction substance and a second reaction substance, the sample accommodating area (10) is provided with a first sample adding port, and the diluent adding area (20) is provided with a second sample adding port; A sealing layer, wherein the sealing layer is capable of sealing the material chamber; The test sample entering the sample containing area (10) from the first sample adding port and the diluent entering the diluent adding area (20) from the second sample adding port can both enter the sample dilution mixing area (30) for mixing. The mixed liquid then enters the pre-reaction area (40) and reacts with the first reaction substance in the pre-reaction area (40) to form a pre-reaction liquid. The pre-reaction liquid then enters the reaction detection area and reacts with the second reaction substance to form a test sample.

2. The bioreactor chip according to claim 1, characterized in that: The bioreactor chip is a centrifugal chip. The sample containing area (10) comprises a sample collection chamber (11), a sample quantitative separation chamber (12), a sample waste liquid chamber (13), and a sample siphon (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 a test sample. The sample quantitative separation chamber (12) comprises a first cavity (121), a first connecting tube (122), a second cavity (123), a second connecting tube (124), and a third cavity (125) which are sequentially connected from the proximal end to the distal end. The cross-sectional area along the direction perpendicular to the rotation radius 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 and mixing area (30); the sample waste liquid cavity (13) is connected to the third cavity (125) via a waste liquid outlet tube (17); the distance from the closest point of the sample siphon tube (14) to the rotation center 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 comprises a first ventilation tube (15) connected to the sample waste liquid chamber (13) and a second ventilation tube (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 along the fluid flow direction, which is thick in the middle and thin at both ends.

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 sample loading area (20) comprises a diluent sample loading chamber (21), a diluent sample loading port (22) for loading samples into the diluent sample loading chamber (21), and a diluent transfer tube (23) connecting the diluent sample loading chamber (21) and the sample dilution mixing area (30), wherein the diluent sample loading port (22) is located at the proximal end of the diluent sample loading chamber (21), and the diluent transfer tube (23) is connected to the distal end of the diluent sample loading chamber (21).

6. The bioreactor chip according to claim 1, characterized in that: The sample dilution and mixing area (30) comprises a mixing chamber (31) and a mixing chamber siphon (34); the mixing chamber siphon (34) is connected to the liquid outlet of the mixing chamber (31) and is used to transport the mixed liquid to the pre-reaction area (40); the mixing chamber siphon (34) is a hydrophilic tube; a first connection port connecting the diluent adding area (20) and the mixing chamber (31) and a second connection port connecting the sample containing area (10) and the mixing chamber (31) are located on the same side of the liquid outlet of the mixing chamber (31); The ventilation structure comprises a third ventilation pipe (32) connected to the mixing chamber (31).

7. The bioreactor chip according to claim 1, characterized in that: The pre-reaction zone (40) comprises a diluted sample temporary storage chamber (41), a pre-reaction chamber (42), a third connecting pipe (43), a first siphon pipe (46) and a second siphon pipe (45); the first reaction substance is preset in the pre-reaction chamber (42); the third connecting pipe (43) is connected to the proximal end liquid outlet of the diluted sample temporary storage chamber (41) and the liquid inlet of the pre-reaction chamber (42); the first siphon pipe (46) is connected to the liquid outlet of the pre-reaction chamber (42); and the second siphon pipe (45) is connected to the distal end liquid outlet of the diluted sample temporary storage chamber (41); the ventilation structure comprises a fourth ventilation pipe (44) connected to the pre-reaction chamber (42); The reaction detection zone comprises a first reaction detection zone (50) and a second reaction detection zone (60) independent of the first reaction detection zone (50); the first siphon tube (46) is connected to the liquid inlet of the first reaction detection zone (50); the second siphon tube (45) is connected to the liquid inlet of the second reaction detection zone (60); the second reaction substance is located in the first reaction detection zone (50); and a third reaction substance is preset in the second reaction detection zone (60).

8. The bioreactor chip according to claim 7, characterized in that: The first reaction detection area (50) comprises 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 hollow structure; the pre-reaction detection chamber (52) and the first detection chamber (53) are both connected to the first distribution tube (51); the liquid inlet of the first distribution tube (51) is connected to the liquid outlet of the first siphon tube (46); the ventilation structure comprises a ventilation structure (55) connected to an end of the first distribution tube (51) away from the liquid inlet of the first distribution tube (51); and the second reaction substance is located in the first detection chamber (53).

9. The bioreactor chip according to claim 8, characterized in that: Along the flow direction of the fluid in the first distribution tube (51), the connection end between the first detection chamber (53) and the first distribution tube (51) is located downstream of the connection end between the pre-reaction detection chamber (52) and the first distribution tube (51), and the first detection chamber (53) includes an expansion chamber and a reaction chamber connected to the distal end of the expansion chamber, and the proximal end of the expansion chamber is connected to the first distribution tube (51). Along the direction perpendicular to the rotation center radius, 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.

10. The bioreactor chip according to claim 7, characterized in that: The second reaction detection area (60) comprises 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 liquid inlet of the second distribution pipe (61) is connected to the liquid outlet of the second siphon tube (45); the third reaction substance is located in the second detection chamber (62); along the fluid flow direction in the second distribution pipe (61), the connection port of the waste liquid chamber (63) and the second distribution pipe (61) is located downstream of the connection port of the second detection chamber (62) and the second distribution pipe (61); the ventilation structure comprises a sixth ventilation pipe connected to an end of the second distribution pipe (61) away from the liquid inlet section of the second distribution pipe (61).

11. The bioreactor chip according to claim 1, characterized in that: A positioning structure (70) is provided on the chip body, and the positioning structure (70) comprises a positioning tooth (72) arranged at the distal end of the chip body and a positioning hole (71) arranged on the chip body.

12. A bioreactor device, comprising a bioreactor chip and a driving device for driving the bioreactor chip to rotate centrifugally, characterized in that: The bioreactor chip is the bioreactor chip according to any one of claims 1 to 11.

13. The bioreactor according to claim 12, characterized in that: The invention also comprises a control device connected to the driving device, wherein the control device can control the driving device to drive the bioreactor chip to swing and mix the substances in the chip body.

14. A biological detection method, characterized in that: The bioreactor chip according to any one of claims 1 to 11, comprising the following steps: The test sample is sent into the sample containing area (10), and the diluent is sent into the diluent adding area (20); The biological detection chip is centrifugally rotated, and particles in the detection sample in the sample containing area (10) are separated and enter the sample dilution mixing area (30), and at the same time, the diluent enters the sample dilution mixing area (30); The biological detection chip reciprocates, and the diluent is mixed with the detection sample to form a mixed liquid; The bioassay chip is centrifugally rotated, and the mixed liquid enters the pre-reaction zone (40); The bio-detection chip swings back and forth, and the mixed liquid mixes with the first reaction substance to form a pre-reaction liquid; The bioassay chip is centrifugally rotated, and the pre-reaction liquid enters the reaction detection area and reacts with the second reaction substance to form a test sample.

15. The biological detection method according to claim 14, characterized in that: When the biological detection chip swings back and forth, the biological detection chip swings at a non-uniform speed.

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