A sample adding card and a sample adding assembly

Abstract

The application provides a sample adding card and a sample adding assembly, which comprise a body, first, second and third sample adding holes arranged in the body, sample adding openings of the sample adding holes being located on a first surface of the body, first, second, third and fourth sample distribution channels arranged in the body, first ends of the first and second sample distribution channels being communicated with the corresponding first and second sample adding holes respectively, first ends of the third and fourth sample distribution channels being communicated with the corresponding third sample adding holes respectively, first mixing pools and second mixing pools being arranged in the body at intervals, each first mixing pool being communicated with a second end of the corresponding first sample distribution channel and a second end of the corresponding third sample distribution channel respectively, each second mixing pool being communicated with a second end of the corresponding second sample distribution channel and a second end of the corresponding fourth sample distribution channel respectively, and the first and second mixing pools being provided with first communication channels arranged at intervals on a first side of the body. The sample adding card and the sample adding assembly are simple to operate and can avoid cross contamination.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medical detection, and in particular to a sample adding card and a sample adding assembly. BACKGROUND

[0002] Blood transfusion is one of the common medical treatments in clinical practice. If the blood types are not matched during blood transfusion, it will cause serious damage to the health of the patient, and even endanger the life of the patient. Therefore, blood type identification before blood transfusion is very important. The gel microcolumn method is one of the common blood type identification methods. However, the existing gel microcolumn card has the following problems:

[0003] Various detection cards (such as blood type detection cards) often use soft film materials (such as aluminum foil and plastic film) to seal the fillers (such as various reagents and / or gel microspheres) therein. When used, the sealing film needs to be torn off or punctured, and then the sample can be added to the detection card. For example, the traditional blood type card needs to be punched using a puncher before use, which is easy to cause cross contamination. For a blood type card including multiple microcolumns (i.e., reaction cavities), multiple sample additions need to be performed for each blood type card, which is complicated and difficult to ensure accurate and equal sample addition. In addition, with the miniaturization of various reagent cards, the inner diameter of the sample addition port on the reagent card is only 1-2 mm or less, and manual sample addition is also difficult to perform.

[0004] In addition, the microcolumn gel card in the prior art is complicated to operate and has high requirements for operation standardization. Therefore, before using the microcolumn gel card, the operator usually needs to be trained, which to some extent limits the further promotion and application of the microcolumn gel method in the field of blood type detection.

[0005] Meanwhile, in the reverse type sample addition in the prior art, the detection reagent (for example, reverse type cell A and reverse type cell B) and the sample to be detected (for example, plasma) need to be operated in two steps, for example, the detection reagent is added first, and then the sample to be detected is added. Such step-by-step operation is not conducive to improving the detection efficiency. If multiple sample additions are required, the sample addition frequency is very high, and during manual sample addition, it is easy to cause misoperation. SUMMARY

[0006] The present application aims to provide a reverse type sample adding card to partially solve or alleviate the above-mentioned deficiencies in the prior art, and further simplify the operation process, which is conducive to the automation of sample addition and detection.

[0007] The first aspect of the present application provides a microfluidic sample adding card, comprising:

[0008] a body;

[0009] At least one first sample adding hole, at least one second sample adding hole and at least one third sample adding hole are arranged in the body in a spaced manner, and the sample adding ports of the first, second and third sample adding holes are located on the first surface of the body;

[0010] At least one first sample dividing channel, at least one second sample dividing channel, at least one third sample dividing channel and at least one fourth sample dividing channel are arranged in the body in a spaced manner, the first end of the first sample dividing channel is in communication with the corresponding first sample adding hole, the first end of the second sample dividing channel is in communication with the corresponding second sample adding hole, the first end of the third sample dividing channel is in communication with the corresponding third sample adding hole, and the first end of the fourth sample dividing channel is in communication with the corresponding third sample adding hole.

[0011] At least one first mixing pool and at least one second mixing pool are arranged in the body in a spaced manner, wherein each first mixing pool is in communication with the second end of the corresponding at least one first sample dividing channel and the second end of the corresponding at least one third sample dividing channel, and each second mixing pool is in communication with the second end of the corresponding at least one second sample dividing channel and the second end of the corresponding at least one fourth sample dividing channel.

[0012] The first and second mixing pools are each provided with at least one first communication channel, and the first communication channels are arranged in a spaced manner on the first side of the body.

[0013] In some embodiments, the second end of the first communication channel is provided with a sharp tip for puncture.

[0014] In some embodiments, at least one air hole is arranged on the first sample dividing channel; and / or, at least one air hole is arranged on the second sample dividing channel; and / or, at least one air hole is arranged on the third sample dividing channel; and / or, at least one air hole is arranged on the fourth sample dividing channel.

[0015] In some embodiments, the first sample dividing channel is in communication with the first mixing pool through a first through hole; and / or,

[0016] The second sample dividing channel is in communication with the second mixing pool through a first through hole.

[0017] In some embodiments, the third sample dividing channel is in communication with the first mixing pool through a second through hole; and / or,

[0018] The fourth sample dividing channel is in communication with the second mixing pool through a second through hole.

[0019] In some embodiments, the first communication channel is in communication with the first mixing pool through a third through-hole, and the first communication channel is in communication with the second mixing pool through a third through-hole.

[0020] In some embodiments, further comprising:

[0021] a first waste liquid cavity disposed inside the body, the first waste liquid cavity being in communication with the corresponding first sample adding hole; and / or,

[0022] a second waste liquid cavity disposed inside the body, the second waste liquid cavity being in communication with the corresponding second sample adding hole; and / or,

[0023] a third waste liquid cavity disposed inside the body, the third waste liquid cavity being in communication with the corresponding third sample adding hole.

[0024] In some embodiments, the first, second, third, and fourth sample distribution channels are disposed on the second surface of the body, and a hydrophilic layer is disposed on a first area of the second surface of the body, wherein the first area includes the area where the first, second, third, and fourth sample distribution channels are disposed.

[0025] In some embodiments, the first and second mixing pools are disposed on the first surface of the body, and a hydrophobic layer or a hydrophilic layer is disposed on a second area of the first surface of the body, wherein the second area includes the area where the first and second mixing pools are disposed.

[0026] In some embodiments, an inner surface area of the sample adding card is treated with paraffin oil, wherein the inner surface area includes the inner surface of the first, second, third, and fourth sample distribution channels, and / or the inner surface of the first communication channel.

[0027] In some embodiments, the sample adding card includes one first sample adding hole, one second sample adding hole, and three third sample adding holes, and the sample adding card includes three first mixing pools and three second mixing pools disposed in the body in a spaced manner.

[0028] In some embodiments, the orthographic projection of at least one first sample distribution channel and the orthographic projection of at least one first through-hole overlap, and / or the orthographic projection of at least one second sample distribution channel and the orthographic projection of at least one first through-hole overlap.

[0029] In some embodiments, the three third sample wells arranged inside the body are respectively a left third sample well, a middle third sample well, and a right third sample well, wherein the first sample well is located on the symmetry axis of the left third sample well and the middle third sample well, the second sample well is located on the symmetry axis of the middle third sample well and the right third sample well, and the horizontal direction of the first sample well and the horizontal direction of the second sample well are both above the three third sample wells.

[0030] In some embodiments, further comprising: two card connection openings formed by two side walls extending from two sides of the body, for inserting a detection card;

[0031] When the detection card is pushed into the card connection opening, the first communication channels of the sample card are respectively communicated with corresponding reaction cavities on the detection card.

[0032] In some embodiments, further comprising: a detection card integrated with the sample card, and reaction cavities arranged on the detection card for being communicated with corresponding first communication channels, wherein the reaction cavities are provided with air holes for solving pressure difference, and / or the first communication channels are provided with air holes for solving pressure difference.

[0033] The application further provides a microfluidic sample adding assembly, comprising: a sample card as described in any of the above embodiments, and a detection card, wherein the sample card is provided with a card connection opening for being inserted with the detection card, when the sample card is pushed into the card connection opening, the first communication channels of the sample card are respectively communicated with corresponding reaction cavities on the detection card, and there is a gap between the first communication channels and the reaction cavities for ventilation.

[0034] In some embodiments, the sample adding assembly comprises: a sample card as described in any of the above embodiments, and a detection card, wherein the sample card is provided with a card connection opening for being inserted with the detection card, and the first communication channels are provided with air holes for solving pressure difference, and / or the reaction cavities are provided with air holes for solving pressure difference,

[0035] When the detection card is pushed into the card connection opening, the second ends of the first communication channels of the sample card are respectively communicated with corresponding reaction cavities on the detection card, and the first communication channels and the reaction cavities are closely fitted.

[0036] The application provides a microfluidic sample adding assembly, comprising: a sample card as described in any of the above embodiments, and a detection card, wherein the sample card is provided with a card connection opening for being inserted with the detection card, and the side wall of the card connection opening is provided with a first card connection position and a second card connection position along the extension direction of the side wall.

[0037] The detection card is installed at the second card joint position in a manner that it can move relative to the card joint opening.

[0038] When the detection card is moved from the second card joint position to the first card joint position under the action of an external force, the first communication channels on the sample adding card are respectively in communication with the corresponding reaction cavities on the detection card, and there is a gap for ventilation between the first communication channels and the reaction cavities.

[0039] The application further provides another microfluidic sample adding assembly, which comprises a sample adding card as described in any of the above embodiments and a detection card, wherein the sample adding card is provided with a card joint opening that can be jointed with the detection card, and one side or both sides of the detection card are provided with a second card joint position and a first card joint position in a direction gradually away from the opening end of the reaction cavity.

[0040] The detection card is installed at the second card joint position in a manner that it can move relative to the card joint opening.

[0041] When the detection card is moved from the second card joint position to the first card joint position under the action of an external force, the first communication channels on the sample adding card are respectively in communication with the corresponding reaction cavities on the detection card, and there is a gap for ventilation between the first communication channels and the reaction cavities.

[0042] In some embodiments, the reagent pre-stored in the reaction cavity comprises a gel and a working solution, and the working solution comprises an antibody.

[0043] In some embodiments, the reagent comprises a gel and a working solution, and the working solution comprises an antibody.

[0044] The application further provides a detection method based on the sample adding assembly described in any of the above embodiments, the sample adding card in the sample adding assembly comprises a first sample adding hole, a second sample adding hole and three third sample adding holes, and the method comprises the following steps:

[0045] The first, second and third liquid samples are respectively added into the first, second and third sample adding holes of the sample adding card, and the first, second and third liquid samples enter into the corresponding first, second, third and fourth subsample channels respectively;

[0046] The sample adding assembly is subjected to a first horizontal centrifugation, and under the action of a first centrifugal force, the liquid samples in the first, second, third and fourth subsample channels enter into the corresponding first and second mixing pools respectively to obtain corresponding mixed liquids;

[0047] The sample adding assembly is subjected to a second horizontal centrifugation, under the action of a second centrifugal force, the mixed solution in the first and second mixing pools enters into the corresponding reaction cavities through the third through hole and the first communication channel;

[0048] The sample adding assembly is subjected to a third horizontal centrifugation, under the action of a third centrifugal force, the mixed solution entering into the reaction cavities is fully mixed with the reagent pre-stored in the reaction cavities and reacts;

[0049] The first, second and third centrifugal forces increase in size in sequence.

[0050] Advantages

[0051] The sample adding card can be applied to multiple-person reverse typing tests or multiple-person antibody screening tests, and can simultaneously realize sample adding of multiple persons on a single sample adding card, greatly reduces the sample adding times, simplifies the operation process, avoids the misoperation caused by too many sample additions, and effectively improves the sample adding and detection efficiency. Meanwhile, multiple-person detection is realized through a single sample adding card and a detection card, which reduces the detection cost to a certain extent.

[0052] Since the liquid samples in each sample channel can enter the corresponding first and second mixing pools at the same time (or the time difference of the liquid samples in each sample channel entering the first and second mixing pools is very small and can be ignored), the two liquids can be fully mixed, the insufficient mixing caused by different sample adding sequences is avoided, and the accuracy of the experimental results is improved. BRIEF DESCRIPTION OF DRAWINGS

[0053] In order to more clearly illustrate the technical solutions of the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. In all the drawings, similar elements or parts are generally identified by similar reference signs. In the drawings, each element or part is not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present application, and those skilled in the art can obtain other drawings according to these drawings without any creative labor.

[0054] Figure 1 The first perspective view of an exemplary embodiment of the present application;

[0055] Figure 2 The second perspective view of an exemplary embodiment of the present application;

[0056] Figure 3 The first structural schematic view of an exemplary embodiment of the present application;

[0057] Figure 4 The second structural schematic view of an exemplary embodiment of the present application;

[0058] Figure 5 Structure diagram of the first surface of an exemplary embodiment of the present application;

[0059] Figure 6 Structure diagram of the second surface of an exemplary embodiment of the present application;

[0060] Figure 7a Structure diagram of the detection card in an exemplary embodiment of the present application;

[0061] Figure 7b Structure diagram of the detection card in another exemplary embodiment of the present application;

[0062] Figure 8 Structure diagram of the third surface of an exemplary embodiment of the present application;

[0063] Figure 9 Structure diagram of the first communication channel and the opening end of the reaction cavity in an exemplary embodiment of the present application;

[0064] Figure 10 Liquid sample flow state diagram of the sample card in an exemplary embodiment of the present application after sample addition to the first sample addition hole during experiment operation;

[0065] Figure 11 Liquid sample flow state diagram of the sample card in an exemplary embodiment of the present application after sample addition to the first, second and third sample addition holes during experiment operation.

[0066] 1 is the main body, 21 is the first sample addition hole, 22 is the second sample addition hole, 23 is the third sample addition hole, 31 is the first sample distribution channel, 32 is the second sample distribution channel, 33 is the third sample distribution channel, 34 is the fourth sample distribution channel, 51 is the first through hole, 52 is the second through hole, 53 is the third through hole, 6 is the first communication channel, 61 is the matching part, 7 is the side wall, 71 is the limiting card protrusion, 8 is the air hole, 91 is the first waste liquid cavity, 92 is the second waste liquid cavity, 93 is the third waste liquid cavity, 10 is the limiting card slot, 11 is the reaction cavity, 11a is the reagent storage area, 11b is the opening end of the reaction cavity, 41 is the first mixing pool, 42 is the second mixing pool, 61a is the dashed triangle. DETAILED DESCRIPTION

[0067] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0068] In the present document, unless explicitly specified and limited, the dotted lines in the drawings represent invisible edge lines and invisible contour lines, and the solid lines in the drawings represent visible edge lines and visible contour lines.

[0069] In the present document, unless explicitly specified and limited, "through" means that two structures are connected so that liquid can flow unidirectionally or bidirectionally between the two structures; "through hole" means a channel for connecting two structures, and the length of the channel is usually set to be short, and the cross section of the channel can be circular, oval or square, and the cross section of the channel is preferably circular for the convenience of processing. For example, the "first through hole 51" in the present document can be regarded as a channel for connecting the third aliquot channel 33 and the second mixing pool 42, and the length of the channel is very small.

[0070] In the present document, unless explicitly specified and limited, "three-stage centrifugation" means a centrifugation method including three steps of first horizontal centrifugation, second horizontal centrifugation and third horizontal centrifugation, wherein the centrifugal speed / force of the first, second and third horizontal centrifugation is different.

[0071] In the present document, unless explicitly specified and limited, "tapered structure" means a structure that is large at the top and small at the bottom, and the cross section of which is trapezoidal or similar to trapezoidal.

[0072] In the present document, the suffix such as "module", "part" or "unit" used to indicate an element is only for the convenience of description of the present application, and has no specific meaning by itself. Therefore, "module", "part" or "unit" can be used mixedly.

[0073] In the present document, the terms "upper", "lower", "inner", "outer", "front", "back", "one end", "the other end", "left", "middle", "right" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application. In addition, the terms "first", "second" are only for the purpose of description, and cannot be understood as indicating or implying relative importance.

[0074] In the present document, "equal" or "same" or "simultaneous" does not mean that the absolute value of the difference between the two or more items specified is zero, but means that the difference between the items specified is very small, which can be ignored in practice. For example, in the present document, "the amount of liquid sample that the third aliquot channel and the fourth aliquot channel can hold is equal" means that the amount of liquid sample that the third aliquot channel and the fourth aliquot channel can hold is exactly equal, or the difference between the two is so small that it has little effect on the result and can be ignored in practical application.

[0075] In this document, unless otherwise explicitly specified and limited, the terms "mount", "provided with", "connected" and the like should be interpreted broadly, for example, "connected" can be fixedly connected, or detachably connected, or integrally connected; can be mechanically connected, can be directly connected, or indirectly connected through an intermediate medium, can be internal connection of two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0076] In this document, the side surface on which the opening (or sample adding port) of the sample adding hole (for example, the first, second and third sample adding holes) is located is referred to as the first surface (or upper surface), and the other side is referred to as the second surface (or lower surface).

[0077] In this document, the "first side" of the body refers to the side of the sample adding card close to the opening end of the reaction cavity of the detection card when the sample adding card is matched with the detection card, that is, the side close to the second end of the sample dividing channel.

[0078] In this document, when referring to a structure located "inside" the sample adding card, it means that it is surrounded by the body material or other materials of the sample adding card and is not directly in contact with the external space. For example, "at least one first sample dividing channel arranged in the interior of the body" can be understood as the first sample dividing channel being surrounded by the body material of the sample adding card and not directly in contact with the external space, or it can be understood that the first sample dividing channel is arranged on the first side or the second side of the body, and it is surrounded by the body material and other materials (for example, a hydrophilic layer or a hydrophobic layer) of the sample adding card and not directly in contact with the external space.

[0079] In this document, "horizontal centrifugation" refers to the state that when the detection card is centrifuged, the detection card is placed horizontally or nearly horizontally, that is, the horizontal plane on which the first side or the second side of the detection card is located is at an angle of 90 degrees or close to 90 degrees with the centrifugal shaft of the centrifugal device. "Horizontal centrifuge" refers to the state that when the centrifugal device is working, the detection card placed in the centrifugal device is in a horizontal or nearly horizontal state.

[0080] In this article, "close fit" means that when two structures are matched, the surface distance between the inner / outer surface of one structure and the outer / inner surface of the other structure is very small, or the inner / outer surface of one structure and the outer / inner surface of the other structure are in contact with each other, so that there is no gap or the gap is very small between the adjacent surfaces of the two structures, so that gas or liquid cannot enter the detection card or the reaction chamber through the gap between the two structures. For example, when "the second end of the first connecting channel is in close fit with the open end of the reaction chamber", it can be understood that the surface distance between the outer surface of the second end of the first connecting channel and the inner surface of the open end of the reaction chamber is very small or the two surfaces are in contact with each other, so that air or debris in the external space (dust or water droplets in the air) cannot enter the first connecting channel or the reaction chamber, and the liquid inside the sample loading device (i.e., the sample loading card) or the detection card cannot splash out to the external space.

[0081] Herein, "orthographic projection" refers to the projection of a certain structure (eg, the first sample separation channel) perpendicular to the plane of the body (eg, the plane where the first surface or the second surface is located).

[0082] In this article, “sample separation channel” is a general term for the first, second, third and fourth sample separation channels, “mixing pool” is a general term for the first and second mixing pools, and “through hole” is a general term for the first, second and third through holes.

[0083] Herein, the "hydrophilic layer" refers to a film or barrier having hydrophilic properties, for example, a hydrophilic membrane, and the "hydrophobic layer" refers to a film or barrier having hydrophobic properties.

[0084] Example 1

[0085] See also Figure 1 The present invention provides a microfluidic sampling card, comprising:

[0086] Ontology 1;

[0087] At least one first sample loading well 21, at least one second sample loading well 22, and at least one third sample loading well 23 are spaced apart inside the body 1, and the sample loading ports of the first, second, and third sample loading wells are located on the first surface (i.e., the upper surface) of the body 1;

[0088] At least one first sample splitting channel 31, at least one second sample splitting channel 32, at least one third sample splitting channel 33, and at least one fourth sample splitting channel 34 are spaced apart within the body 1. The first end of the first sample splitting channel 31 is connected to the corresponding first sample loading well 21, the first end of the second sample splitting channel 32 is connected to the corresponding second sample loading well 22, the first end of the third sample splitting channel 33 is connected to the corresponding third sample loading well 23, and the first end of the fourth sample splitting channel 34 is connected to the corresponding third sample loading well 23.

[0089] At least one first mixing pool 41 and at least one second mixing pool 42 are arranged in the body, wherein each first mixing pool 41 is in communication with the second end of the corresponding at least one first sub-sample channel 31 and the second end of the corresponding at least one third sub-sample channel 33, respectively; and each second mixing pool 42 is in communication with the second end of the corresponding at least one second sub-sample channel 32 and the second end of the corresponding at least one fourth sub-sample channel 34, respectively.

[0090] The first and second mixing pools are each provided with at least one first communication channel 6, and the first communication channels 6 are arranged in the first side of the body.

[0091] Preferably, in some embodiments, the first, second, third and fourth sub-sample channels are capillary sub-sample channels, and when liquid samples (detection reagents or samples to be detected) are added to the first, second and third sample addition holes, the liquid samples fill the corresponding sub-sample channels (i.e., the first, second, third and fourth sub-sample channels) under capillary action.

[0092] The sample addition card needs to be used in cooperation with a detection card (i.e., a micro-column gel detection card), wherein the detection card includes a plurality of reaction cavities 11 arranged at intervals on the detection card (it can be understood that the number of reaction cavities is the same as the number of first communication channels of the sample addition card), and the reaction cavities are provided with reaction cavity opening ends 11b for sample addition. In addition, reagents are pre-stored in the reaction cavities of the detection card, and the reagents are sealed by arranging a sealing film on the reaction cavity opening ends or arranging paraffin oil on the upper ends of the reaction cavities, so as to prevent the reagents from drying.

[0093] In order to realize the cooperation between the sample addition card and the detection card, further, in some embodiments, referring to Figures 2-6 The sample addition card further includes two card joint openings formed by two side walls 7 extending from the two sides of the body 1 and used for plugging with the detection card; when the detection card is pushed into the card joint openings, the second ends of the first communication channels of the sample addition card are in communication with the corresponding reaction cavities of the detection card; and when the sample addition card and the detection card are subjected to centrifugation, under the action of centrifugal force, the liquid samples in the first, second and third sample addition holes sequentially pass through the sub-sample channels (i.e., the first, second, third and fourth sub-sample channels) and enter the corresponding first and second mixing pools, and then enter the corresponding reaction cavities through the first communication channels connected with the first and second mixing pools.

[0094] When the detection card is pushed into the card joint opening, if the second end of the first communication channel closely fits the open end of the reaction chamber, when the liquid sample enters into each sample distribution channel or the first communication channel and reaches a certain length, the liquid sample in the channel will extrude the air in the sample distribution channel, the first communication channel and the reaction chamber, resulting in an increase in air pressure in the sample distribution channel, the first communication channel and the reaction chamber, and a pressure difference between the inside and the outside, so that the liquid sample cannot enter into the reaction chamber even under the combined action of capillary action and centrifugal force.

[0095] Therefore, when the sample adding card cooperates with the detection card, there is a gap between the second end of the first communication channel and the open end of the reaction chamber, so that the air in the external space can enter into the first communication channel and the reaction chamber through the gap, so that the air pressure at the second end opening of the first communication channel and the open end of the reaction chamber is equal to or close to the atmospheric pressure of the external space, so that the liquid sample entering into the first communication channel can smoothly enter into the corresponding reaction chamber under the action of centrifugal force.

[0096] Specifically, in some embodiments, referring to Figure 1 , the second end of the first communication channel 6 is spaced apart from the first side of the body 1 and extends to the outside of the body, when the sample adding card cooperates with the detection card, the second end of the first communication channel 6 extends into the corresponding open end of the reaction chamber of the detection card and communicates with the corresponding reaction chamber.

[0097] Referring to Figure 3 , in some embodiments, since part of the detection card is provided with a sealing film at the open end, in order to make the first communication channel communicate with the reaction chamber, the second end of the first communication channel is provided with a cooperating part 61 (preferably, the cooperating part is a sharp end for piercing the sealing film) for piercing the sealing film, when the sample adding card cooperates with the detection card, the cooperating part (i.e. the sharp end) pierces the sealing film at the open end of the reaction chamber, so that the first communication channel communicates with the corresponding reaction chamber, respectively.

[0098] Of course, in other embodiments, referring to Figure 7b , the open end of the reaction chamber 11 on the detection card extends outward in a direction away from the bottom end of the reaction chamber by a certain length, accordingly, the second end of the first communication channel on the sample adding card can not extend to the outside of the body, when the sample adding card cooperates with the detection card, the open end of the reaction chamber extends into the inside of the first communication channel, so that the first communication channel communicates with the reaction chamber.

[0099] In some embodiments, a plurality of sample distribution channels are provided on the body, therefore, there can be one or more sample distribution channels with a length that is too long, and a gas hole 8 for at least one drainage is provided on each of these sample distribution channels with a length that is too long.

[0100] Specifically, in some embodiments, one or more of the first, second, third, and fourth sample distribution channels are provided with an air hole 8. For example, see Figure 3 An air hole 8 is provided on the longest sample distribution channel (i.e., one of the second sample distribution channels 32) for draining. Since part of the sample distribution channel is longer, the inner diameter of the corresponding sample distribution channel is set to be smaller, and thus the flow of gas in the longer sample distribution channel can be blocked, causing the gas in the sample distribution channel to be unable to smoothly drain when the liquid enters a certain length of the sample distribution channel, and thus being pressed by the liquid sample, resulting in a pressure difference between the internal gas pressure of the sample distribution channel and the external atmosphere. The air hole can solve the problem of pressure difference, thereby playing a role in draining the liquid sample.

[0101] Further, in some embodiments, see Figure 2 The first and second sample distribution channels are respectively connected to the first and second mixing pools through the first through hole 51.

[0102] Further, in some embodiments, see Figure 2 The third and fourth sample distribution channels are respectively connected to the first and second mixing pools through the second through hole 52.

[0103] Further, in some embodiments, the first communication channel 6 is connected to the first mixing pool 41 through the third through hole, and is connected to the second mixing pool 42 through the third through hole 53.

[0104] In some embodiments, one or more of the first, second, and third through holes are channel structures with a certain length, which play a role in connecting the mixing pools and the sample distribution channels or the first communication channel. Specifically, the first and second through holes connected to the sample distribution channels can be capillary channels, which can be considered as an extension of the sample distribution channels. Of course, the first and second through holes can also be non-capillary channels, which can play a role in stopping the flow of liquid sample when the liquid sample fills the sample distribution channels (e.g., the first, second, third, and fourth sample distribution channels).

[0105] Further, in order to accommodate excess liquid sample added to the first sample addition hole 21, see Figure 3 A first waste liquid chamber 91 can be provided near the first sample addition hole 21 and inside the body.

[0106] Further, in order to prevent the sample (i.e., liquid sample) from entering the first waste liquid chamber 91 when it is not excessive, and to avoid cross-infection caused by the sample in the first waste liquid chamber 91 returning to the first sample addition hole, a baffle can be provided near the sample addition hole in the first waste liquid chamber.

[0107] In some embodiments, the baffle is a raised sheet in the first waste liquid chamber 91, and the sample can only overflow the upper end of the baffle to enter the first waste liquid chamber when the sample is too much.

[0108] In addition, in order to facilitate the liquid sample to enter the first waste liquid chamber, the first waste liquid chamber can have an opening on the upper surface of the body 1. When the body 1 is made of a hydrophilic material, the baffle can be treated to have hydrophobic properties, for example, a hydrophobic layer is added, to further prevent the liquid sample in the first waste liquid chamber from returning to the first sample addition hole.

[0109] Further, in some embodiments, in order to accommodate the excess liquid sample added to the second sample addition hole, referring to Figure 3 the sample addition card further comprises a second waste liquid chamber 92 arranged inside the body, which is in communication with the corresponding second sample addition hole 22.

[0110] Further, in some embodiments, in order to accommodate the excess liquid sample added to the third sample addition hole, referring to Figure 4 the sample addition card further comprises a third waste liquid chamber 93 arranged inside the body, which is in communication with the corresponding third sample addition hole 23.

[0111] The arrangement of the second and third waste liquid chambers can refer to the first waste liquid chamber.

[0112] In some embodiments, the first, second, third, and fourth sample splitting channels are arranged on the second surface (i.e., the lower surface) of the body, and the first region of the second surface of the body is provided with a hydrophilic layer, wherein the first region includes the region provided with the first, second, third, and fourth sample splitting channels.

[0113] Specifically, referring to Figure 4 , Figure 4 The structure shown can be considered as a cross-sectional view of the lower surface (each sample splitting channel) removed to show the internal structure of the sample addition card of the present application. In addition, for the convenience of processing (especially each sample splitting channel) and / or hydrophilic treatment, the structure shown can also be processed first, and then a lower surface layer (e.g., a hydrophilic layer, specifically, a hydrophilic film or a baffle with hydrophilic properties) can be covered by pasting or bonding, etc. to achieve the same effect that the first, second, third, and fourth sample splitting channels are located inside the body 1. Therefore, the sample addition card, whether it is integrally injection molded, prepared by layered processing, or the main structure processed by layered processing but does not include the lower surface layer, should be covered within the protection scope of the present application.

[0114] In the present embodiment, the first, second, third, and fourth sample splitting channels are covered by pasting a hydrophilic film layer or a baffle with hydrophilic properties to facilitate the flow of aqueous liquids such as blood therein.

[0115] Further, to avoid the liquid sample in the first and second mixing pools splashing out during centrifugation and thus contaminating the test card or the test environment, in some embodiments, the first and second mixing pools are arranged on the first surface of the body (the part where the first and second mixing pools are arranged is referred to as the second region), and correspondingly, a hydrophobic layer (for example, a hydrophobic film or a baffle with hydrophobic properties) or a hydrophilic layer is arranged on the second region of the first surface of the body.

[0116] Further, to reduce the residue of the liquid sample inside the sample loading card (for example, the respective sample distribution channels or the mixing pools), in some embodiments, the inner surface region of the sample loading card is treated with paraffin oil, wherein the inner surface region includes: the inner surface of the first, second, third and fourth sample distribution channels, and / or the inner surface of the first communication channel.

[0117] Specifically, in some embodiments, the paraffin oil treatment step includes: adding paraffin oil into the first, second and third sample loading holes respectively, the paraffin oil enters the respective sample distribution channels under capillary action (or the combined action of capillary action and centrifugal force), then passes through the sample distribution channels and enters the respective mixing pools, and finally flows out from the second end of the first communication channel. An empty test card (i.e., a test card without stored reagents) can be used to cooperate with the sample loading card to receive the excess paraffin oil.

[0118] In an exemplary embodiment of the present application, referring to Figure 1 The sample loading card includes: a first sample loading hole 21, a second sample loading hole 22 and three third sample loading holes 23, and correspondingly, the sample loading card includes: three first mixing pools 41 and three second mixing pools 42 arranged in the body in a spaced manner.

[0119] Specifically, in this exemplary embodiment, the first sample loading hole 21 is connected to the corresponding first mixing pool 41 through three first sample distribution channels 31 respectively, the second sample loading hole 22 is connected to the corresponding second mixing pool 42 through three second sample distribution channels 32 respectively, and the three third sample loading holes 23 are connected to the first mixing pools 41 through three corresponding third sample distribution channels 33 respectively, and at the same time, the three third sample loading holes 23 are connected to the second mixing pools 42 through three corresponding fourth sample distribution channels 34 respectively.

[0120] When each sample distribution channel is arranged on the first surface of the body, if the first and second through holes are also arranged on the first surface, at least one through hole and at least one sample distribution channel will intersect and interfere with each other. Therefore, to avoid the interference between the first through hole or the second through hole and the sample distribution channel, at least one of the first through holes or at least one of the second through holes is arranged on the second surface of the body, and at this time, the orthographic projection of the through hole arranged on the second surface and the orthographic projection of the sample distribution channel have an overlapping part.

[0121] For example, in this exemplary embodiment, see Figure 1 There is an overlapping area between the orthographic projection of at least one second sample separation channel (ie, the projection perpendicular to the plane of the main body) and the orthographic projection of at least one first through hole.

[0122] For another example, in some other exemplary embodiments, the orthographic projection of at least one first sample dividing channel and the orthographic projection of at least one first through hole have an overlapping area. Furthermore, in this exemplary embodiment, Figure 1 As shown, Figure 1 The positive direction of the X axis is left, the positive direction of the Y axis is upward, and the three third loading wells spaced apart inside the body are: a left third loading well, a middle third loading well, and a right third loading well. The first loading well is located on the symmetric axis between the left third loading well and the middle third loading well, the second loading well is located on the symmetric axis between the middle third loading well and the right third loading well, and the horizontal direction where the first loading well is located and the horizontal direction where the second loading well is located are both above the three third loading wells.

[0123] In this embodiment, the spatial structure of the first, second and third sample loading holes cooperates with each other to achieve sample loading for multiple people in a limited space, for example, three-person sample loading for a reverse typing test or two-person sample loading for an antibody screening test.

[0124] Specifically, when the sampling card is used for anti-typing test sampling, anti-typing cells A are added to the first sampling well, anti-typing cells B are added to the second sampling well (or anti-typing cells B are added to the first sampling well and anti-typing cells A are added to the second sampling well), and three samples to be tested (from three subjects) are added to the three third sampling wells respectively. After adding the sample, the sample card with the test card is placed in the centrifugal card slot of the centrifuge for centrifugation. Under the combined action of capillary action and centrifugal force, the liquid samples in each sample addition hole pass through the corresponding sample separation channel and through hole in turn and enter the corresponding mixing pool. Among them, the anti-fixed cell A is respectively mixed with the three samples to be tested in the first mixing pool, and the anti-fixed cell B is respectively mixed with the three samples to be tested in the second mixing pool; further, the mixed liquid obtained by the intersection and mixing of the first and second mixing pools enters the corresponding reaction chamber through the third through hole and the first connecting channel under the action of centrifugal force, and reacts with the reagent in the reaction chamber. After the centrifugation is completed, the reaction results in the reaction chamber are observed, and the test data of the three samples to be tested are recorded.

[0125] When the sample card is applied to the antibody screening test, the corresponding test samples (derived from two subjects) are added to the first and second sample wells respectively, and the first anti-screening cells, the second anti-screening cells, and the third anti-screening cells are added to the three third sample wells respectively (it can be understood that there is no corresponding relationship between the first, second, and third anti-screening cells and the three third sample wells). After adding the sample, the sample card with the detection card is placed in the centrifugal card slot of the centrifuge for centrifugation. Under the combined action of capillary action and centrifugal force, the liquid samples in the first and third sample wells pass through the corresponding sample separation channels and through holes in turn into the corresponding first mixing pool for intersection and mixing, and the liquid samples in the second and third sample wells pass through the corresponding sample separation channels and through holes in turn into the corresponding second mixing pool for intersection and mixing; further, the mixed liquid obtained after the intersection and mixing in the first and second mixing pools enters the corresponding reaction chamber through the third through hole and the first connecting channel under the action of centrifugal force, and reacts with the reagent in the reaction chamber. After the centrifugation is completed, the reaction results in the reaction chamber are observed, and the test data of the two samples to be tested are recorded.

[0126] The usage process is described here in a sequential order. Those skilled in the art will understand that it is not necessary to operate in this order. For example, the sample loading card can be placed in the centrifuge, and then the test card can be plugged in and the sample can be loaded, or the sample can be loaded and the test card can be plugged in.

[0127] In this embodiment, the sampling card can realize the sampling of multiple people, and the operation is simple, which is conducive to the automation of the operation process. For example, in the prior art, four samplings are required to perform a reverse typing test on a single person's sample (two samplings of the sample to be tested and two samplings of the detection reagent). Accordingly, 12 samplings are required to complete the reverse typing test of three people. If manual sampling is used, the large number of sampling times may increase the probability of misoperation and reduce the detection efficiency. The sampling card in this exemplary embodiment only needs 5 samplings to complete the reverse typing test of three people, which reduces the number of samplings, simplifies the operation process, reduces the possibility of misoperation, and greatly improves the detection efficiency.

[0128] Furthermore, in some embodiments, by designing and adjusting the inner diameter and length of the sample separation channel, uniform separation of the sample to be tested and the detection reagent can be achieved, thereby improving the accuracy of the test results.

[0129] Preferably, the sample loading card is tested by three-stage centrifugation method, see Figure 8 , Figure 8 Shows the flow direction of the liquid sample after adding the sample to the second sample well ( Figure 8 The direction indicated by the arrow is the flow direction of the liquid sample). It can be understood that in order to more clearly illustrate the characteristics of the liquid sample flowing inside the sample card,Figure 8 The drawing is not drawn in the actual proportion of the product, but only briefly shows the necessary structure of the product.

[0130] Specifically, when liquid sample (e.g. anti-cell A or B) is added to the second sample well, the liquid sample enters the second sample channel under capillary action. In some embodiments, since the inner diameter of the first through hole 51 is larger than the inner diameter of the second sample channel 32, the liquid will not continue to enter the first through hole 51. Alternatively, in some embodiments, the first through hole 51 adopts a capillary structure (i.e. the first through hole 51 is equivalent to an extension of the second sample channel), and the liquid sample enters the first through hole 51, but because the inner diameter of the first mixing pool 41 is larger than the inner diameter of the first through hole 51, the liquid sample will not continue to enter the first mixing pool 41 through the first through hole 51. In addition, since the sample card is placed horizontally or nearly horizontally when in use, at this time, the liquid sample in the second sample channel 32 needs to overcome its own gravity to enter the first through hole 51 and then enter the second mixing pool 42 through the first through hole 51, which further restricts the flow of the liquid sample, that is, the first through hole 51 has a flow-stopping effect; similarly, the liquid samples (e.g. plasma or detection reagent) of other sample wells enter the corresponding sample channels under capillary action, and cannot enter the first or second mixing pool without external force.

[0131] After the sample addition of each sample well is completed, the sample card and the matched detection card are subjected to centrifugation, and preferably a three-stage centrifugation method is adopted. In the first horizontal centrifugation process, the liquid samples (detection reagent or sample to be tested) in the first, second, third and fourth sample channels enter the corresponding first and second mixing pools through the first and second through holes under the action of the first centrifugal force, and are mixed to obtain the corresponding mixed liquid; in the second horizontal centrifugation process, the mixed liquid in the first and second mixing pools enters the corresponding reaction chamber through the third through hole under the action of the second centrifugal force; in the third horizontal centrifugation process, the mixed liquid in the reaction chamber and the reagent in the reaction chamber are mixed and reacted under the action of the third centrifugal force.

[0132] It can be understood that the above-mentioned three horizontal centrifugation processes are continuous, and there is no interval or a very short interval between adjacent centrifugation processes.

[0133] In addition, in the present embodiment, the first through-hole 51 has a flow-stopping effect on the liquid sample in the second sample channel 32, which is equivalent to a first resistance to the flow of the liquid sample. Meanwhile, when the liquid sample enters the third through-hole 53, the liquid sample is stopped in the third through-hole 53 because the inner diameter of the third through-hole 53 is smaller than that of the first communication channel 6, which is equivalent to a second resistance to the flow of the liquid sample, and the second resistance is set to be greater than the first resistance. Therefore, the first centrifugal force is set to be greater than the first resistance and smaller than the second resistance, so that the liquid sample in the second sample channel 32 can only enter the second mixing pool 42 under the action of the first centrifugal force and cannot enter the first communication channel 6.

[0134] Similarly, the first centrifugal force is set to meet the requirement that the liquid sample in the first, third and fourth sample channels enters the first mixing pool or the second mixing pool, and the liquid sample in the first and second mixing pools cannot continue to enter the first communication channel.

[0135] Further, in some embodiments, when the volume of the first and second mixing pools is greater than the volume of the mixed liquid actually to be contained, for example, the volume of the first mixing pool is greater than about 20 μL, and the volume of the mixed liquid is about 10 μL, the mixed liquid can only occupy about half of the volume of the mixing pool, and when the sample card is placed horizontally, the liquid level of the mixed liquid in the first and second mixing pools can be lower than the height of the connection between the third through-hole and the first mixing pool, and the mixed liquid needs to be forced into the third through-hole by an external force, which further resists the flow of the liquid sample.

[0136] Of course, in some embodiments, a hydrophobic layer with hydrophobic properties can also be covered on the third through-hole, which also resists the flow of the liquid sample.

[0137] Preferably, in some embodiments, the centrifugal speed / centrifugal force set in the first, second and third horizontal centrifugation processes gradually increases.

[0138] Further, in some embodiments, the first centrifugal force is about 9-16 g, the centrifugation time is about 10 s-1 min, the second centrifugal force is about 55 g, the centrifugation time is about 1 min 50 s-1 min, and the third centrifugal force is about 200 g, and the centrifugation time is about 3 min.

[0139] In a specific experimental operation process, see Figure 10 ( Figure 10 the physical diagram of the sample card), Figure 10The sample card in the embodiment is horizontally placed, and liquid samples are added into the first sample wells 21. The liquid samples fill the first sample channels under capillary action (at this time, a very small amount of liquid sample may enter the connection between the through hole and the first sample channel), and do not enter the first mixing pool, as shown in FIG. 2. Figure 11 Figure 11 The sample card in the embodiment is horizontally placed, and liquid samples are added into the first sample wells 21. The liquid samples fill the first sample channels under capillary action (at this time, a very small amount of liquid sample may enter the connection between the through hole and the first sample channel), and do not enter the first mixing pool, as shown in FIG. 2. Figure 11 The sample card in the embodiment is horizontally placed, and liquid samples are added into the first sample wells 21. The liquid samples fill the first sample channels under capillary action (at this time, a very small amount of liquid sample may enter the connection between the through hole and the first sample channel), and do not enter the first mixing pool, as shown in FIG. 2.

[0140] Further, in some embodiments, two or more air holes can be arranged on the longer sample channel for liquid drainage.

[0141] Embodiment Two

[0142] Based on the microfluidic sample card in the above-described embodiment one, the present application further provides a microfluidic sample card, which comprises the components in the above-described embodiments, wherein the inner diameter of the first sample channel is between about 0.2 mm and 0.8 mm, the inner diameter of the second sample channel is between about 0.1 mm and 0.5 mm, and the inner diameter of the third and fourth sample channels is between about 0.2 mm and 1.5 mm.

[0143] It can be understood that, in order to ensure that the sample channels can achieve quantitative sample distribution, the volume of the liquid sample contained in each first sample channel is equal or similar, the volume of the liquid sample contained in each second sample channel is equal or similar, and the volume of the liquid sample that can be contained in the third and fourth sample channels is equal.

[0144] Further, in order to enable multiple sample channels to be arranged on the same sample card, the length and width of each sample channel are different. For example, the length of some sample channels is relatively long, and the inner diameter (i.e., the width) of the sample channel is relatively small, and the capillary action of the sample channel is relatively strong. The length of some sample channels is relatively short, and the inner diameter (i.e., the width) of the sample channel is relatively large, and the capillary action of the sample channel is relatively weak.

[0145] ​To avoid cross-talk between the sample channels (i.e. liquid exchange between adjacent sample channels), the spacing between adjacent sample channels is greater than about 0.3 mm.

[0146] Further, to avoid cross-talk between the first and second mixing pools (i.e. mixed liquid in the first mixing pool entering the second mixing pool, or mixed liquid in the second mixing pool entering the first mixing pool), the spacing between adjacent first and second mixing pools is greater than about 0.5 mm.

[0147] In some embodiments, the volume (amount of liquid sample that can be contained) of the first and second mixing pools is greater than about 10 μL.

[0148] Of course, in other embodiments, the volume (amount of liquid sample that can be contained) of the first and second mixing pools is greater than about 20 μL.

[0149] Preferably, to avoid interference between different first communication channels, in some embodiments, the spacing between the second ends of adjacent first communication channels is greater than about 4 mm.

[0150] Embodiment Three

[0151] Based on the microfluidic sample loading card in the above embodiments, the present application further provides a microfluidic sample loading assembly, comprising: the sample loading card in any of the above embodiments, and a detection card (see the above embodiments for the specific structure), wherein the sample loading card is provided with a card joint opening that can be plugged with the detection card, when the sample loading card is pushed into the card joint opening, the first communication channels of the sample loading card are respectively connected with the corresponding reaction cavities on the detection card.

[0152] For example, in some embodiments, the card joint opening of the sample loading card is provided with a limiting card protrusion 71 (as shown in Figure 4 and the detection card is provided with a limiting card slot 10 (as shown in Figure 7a and Figure 7b When the detection card is pushed into the card joint opening, and the limiting card protrusion on the sample loading card cooperates with the limiting card slot on the detection card, the second ends of the first communication channels on the sample loading card are respectively connected with the open ends of the corresponding reaction cavities on the detection card. Of course, the limiting card slot can also be provided at the card joint opening, and the limiting card protrusion corresponding thereto can be provided on the detection card.

[0153] Preferably, in some embodiments, the sample loading assembly further comprises: reagents pre-stored in the reaction cavities.

[0154] Further, in some embodiments, the reagents pre-stored in the reaction cavities comprise: a gel and a working solution, wherein the working solution comprises: an antibody. For example, when the sample loading assembly is applied to ABO blood group positive typing, anti-A blood group typing reagents and anti-B blood group typing reagents are respectively pre-stored in the reaction cavities.

[0155] Further, in order to prevent the reagent from drying, in some embodiments, the detection assembly (i.e. the sample adding assembly) further comprises paraffin oil arranged in the reaction cavity for sealing the reagent. It can be understood that, in order to seal the reagent, the paraffin oil is arranged above the reagent, so that the paraffin oil and the inner wall of the reaction cavity form a closed structure for preserving the reagent.

[0156] Specifically, in some embodiments, in the production process of the sample adding assembly, firstly, the corresponding reagent is added into the reaction cavity, and then the melted paraffin oil is added into the opening end of the reaction cavity. It can be understood that, when the detection assembly is used, the paraffin oil is in a solid state, and at this time, the detection card is heated to melt the paraffin oil.

[0157] Of course, in other embodiments, the reagent is sealed by arranging a film on the opening end of the reaction cavity.

[0158] Preferably, the sample adding assembly is subjected to horizontal centrifugation, i.e. the sample adding assembly is subjected to centrifugation by using a horizontal centrifuge.

[0159] Embodiment four

[0160] Based on the microfluidic sample adding card (i.e. the sample adding device) in the above embodiments, the present application further provides a microfluidic sample adding assembly, comprising the sample adding card in any one of the above embodiments, and a detection card (for specific structure, refer to the above embodiments), wherein the sample adding card is provided with a card joint opening which can be inserted with the detection card, and the side wall of the card joint opening is provided with a first card joint position and a second card joint position which are spaced apart along the extension direction of the side wall;

[0161] Wherein, the detection card is installed at the second card joint position in a movable manner relative to the card joint opening.

[0162] When the detection card is moved from the second card joint position to the first card joint position under the action of an external force, the first communication channel on the sample adding card is in communication with the corresponding reaction cavity on the detection card, respectively.

[0163] In some embodiments, the first card joint position is provided with a limiting card protrusion or a limiting card slot.

[0164] In some embodiments, the second card joint position is provided with a limiting card protrusion or a limiting card slot.

[0165] In an exemplary embodiment, the detection card is installed at the second clamping position of the clamping opening of the sample adding card when the assembly is shipped (or in the initial state). For example, by setting a limiting clamping protrusion on the detection card in fixed connection with the second clamping position (specifically, a breakable line is arranged at the connection). In this state, the second end of the first communication channel has a certain distance e from the opening end of the reaction chamber, and the distance is slightly less than or equal to the interval distance between the first clamping position and the second clamping position, that is, the first communication channel and the reaction chamber are not in a communication state. When the detection card is pushed towards the first clamping position, and when the detection card moves from the second clamping position to the first clamping position (for example, the limiting clamping protrusion is buckled with the limiting clamping groove), the second end of the first communication channel is communicated with the reaction chamber.

[0166] In the embodiment, the sample adding card and the detection card can be saved in the form of an assembly, and since the sample adding card and the detection card are fixedly connected through the second clamping position, damage of the product caused by collision during storage and transportation can be avoided.

[0167] Embodiment five

[0168] Based on the sample adding card described in the above embodiments, the application further provides a microfluidic sample adding assembly, comprising: the sample adding card described in any one of the above embodiments, and a detection card (see the above embodiments for specific structures), and different from the above embodiments, wherein the sample adding card is provided with a clamping opening which can be inserted with the detection card, and the detection card is provided with a second clamping position and a first clamping position on one side or both sides thereof in a direction gradually away from the opening end of the reaction chamber;

[0169] Wherein, the detection card is installed at the second clamping position in a manner that can move relative to the clamping opening;

[0170] When the detection card moves from the second clamping position to the first clamping position under the action of an external force, the first communication channel of the sample adding card is respectively communicated with the corresponding reaction chamber on the detection card.

[0171] When the detection card moves from the second clamping position to the first clamping position under the action of an external force, the second end of the first communication channel of the sample adding card is respectively communicated with the corresponding reaction chamber.

[0172] Specifically, in some embodiments, the first and second clamping positions are respectively provided with a limiting clamping protrusion or a limiting clamping groove.

[0173] For example, in some embodiments, one side or both sides of the detection card is respectively provided with a first limiting card slot and a second limiting card slot at the first and second card joint positions; correspondingly, the card joint opening comprises a guide rail (specifically, two side walls extending from the body, or a guide rail provided on the two side walls along the length direction thereof) for providing a sliding path for the detection card, and the guide rail is provided with a limiting card convex that can cooperate with the first and second limiting card slots;

[0174] When the limiting card convex cooperates with the second limiting card slot on the detection card under the action of an external force, the detection card is card-jointed with the card joint opening at the second card joint position, and at this time, the first communication channel of the sample adding card is not communicated with the reaction cavity of the detection card, that is, the opening end of the reaction cavity is actually still a certain distance from the second end of the first communication channel;

[0175] When the limiting card convex is separated from the second limiting card slot under the action of an external force, the detection card can move along the guide rail towards the second end of the first communication channel; and when the first limiting card slot on the detection card moves to the position of the limiting card convex on the guide rail and cooperates with the limiting card convex, the detection card is card-jointed with the card joint opening at the first card joint position, and at this time, the second end of the first communication channel 6 is communicated with the corresponding reaction cavity.

[0176] Embodiment six

[0177] Based on the above embodiments, the application further provides a sample adding assembly, comprising the above sample adding card and a detection card, wherein the detection card is described as follows Figure 7b , comprising a plurality of reaction cavities, and the reaction cavity comprises a reagent storage area 11a and a reaction cavity opening end 11b. In order to realize the cooperation between the sample adding device and the detection card, preferably, in some embodiments, the second end of the first communication channel extends to the outside of the body, and the second end of the first communication channel is provided with a cooperating part that cooperates with the reaction cavity opening end 11b of the detection card. When the cooperating part cooperates with the reaction cavity opening end 11b, the second flow communication channel is communicated with the corresponding reaction cavity.

[0178] In some embodiments, the inside of the reaction cavity opening end of the detection card is a tapered structure that is large at the top and small at the bottom (i.e., a trapezoidal or approximately trapezoidal structure), and correspondingly, the cooperating part of the second end of the first communication channel 6 is also a tapered structure that is large at the top and small at the bottom, and the outer diameter of the cooperating part of the first communication channel is slightly smaller than the inner diameter of the reaction cavity opening end, so that the cooperating part can extend into the inside of the reaction cavity opening end.

[0179] It can be understood that the structure of the cooperating part can only cooperate with the inside of the opening end of the reaction cavity, for example, in some embodiments, when the reaction cavity opening end is a tapered structure that is large at the top and small at the bottom, the cooperating part is also correspondingly provided with a tapered structure that is large at the top and small at the bottom, and the outer diameter of the cooperating part is slightly smaller than the inner diameter of the reaction cavity opening end.

[0180] For example, in some embodiments, when the opening end of the reaction cavity is in a cylindrical structure, the fitting portion is also in a cylindrical structure, and the outer diameter of the fitting portion is slightly smaller than the inner diameter of the opening end of the reaction cavity.

[0181] In some embodiments, paraffin oil is also provided at the edge of the fitting portion to prevent liquid from hanging at the edge of the fitting portion and flowing into the reaction cavity.

[0182] Preferably, in some embodiments, in order to prevent the liquid sample in the reagent storage area from flowing back into the sample loading card, the inner diameter of the reagent storage area is about 1.2mm-1.4mm, so that when the detection card is placed horizontally or nearly horizontally, the reagent or liquid sample in the reagent storage area will hardly flow without external force. Of course, the inner diameter of the reagent storage area can also be set to other sizes as long as the liquid sample in the reagent storage area hardly flows when the detection card is placed horizontally or nearly horizontally.

[0183] Further, in some embodiments, in order to prevent the reagent in the reaction cavity from flowing back into the sample loading device when the detection card is pushed, the liquid surface of the reagent in the detection card is usually at a certain distance from the opening end of the reaction cavity, so that even if the reagent flows, it will not flow back into the sample loading device. For example, in a specific embodiment, the length of the reagent storage area is 14-15mm, and after a certain amount of reagent is added, the reagent is located in the area close to the lower end of the reagent storage area (i.e. the side away from the opening end of the reaction cavity), and the liquid surface of the reagent is about 7-8mm away from the opening end of the reaction cavity.

[0184] Preferably, in a specific embodiment, the volume of the storage area in the reaction cavity is greater than about 20μL.

[0185] Embodiment Seven

[0186] Based on the above embodiments, the present application also provides a sample loading and detection integrated detection card / sample loading card, which is different from the above embodiments in that the detection card / sample loading card further comprises a detection area matched with the sample loading card, and a plurality of reaction cavities are provided on the detection area, and the reaction cavities are respectively connected with the second ends of the corresponding first communication channels, wherein a gas hole for solving the internal and external pressure difference is provided on the reaction cavities (close to the opening end).

[0187] The air hole is used to solve the pressure difference when the sample is separated. In the embodiment, the air hole plays a role in solving the pressure difference between the first communication channel and the reaction cavity. In actual application, each separation channel in the sample adding card, the first and second mixing pools, the first communication channel and the reaction cavity all adopt a closed structure (for example, covered and sealed by a hydrophilic layer). For example, the first separation channel 31, the first mixing pool connected with the separation channel, and the corresponding first communication channel and reaction cavity form a closed space. When the liquid sample added into the first sample adding hole enters the corresponding separation channel or the corresponding mixing pool to a certain amount, the air in the closed space is extruded by the liquid sample, so that the air pressure in the closed space is greater than the atmospheric pressure at the opening of each sample adding hole, that is, a pressure difference is formed between the closed space and the atmospheric pressure, so that the liquid sample cannot continue to flow. Therefore, the air hole is opened to make the air pressure in the closed space equal to or close to the atmospheric pressure, that is, equal to or close to the air pressure at the opening of the sample adding hole, so that the liquid sample to be tested can flow smoothly into the reaction cavity under the action of external force.

[0188] Of course, in other embodiments, an air hole can be provided on the first communication channel to solve the pressure difference problem in the reaction cavity and the first communication channel.

[0189] Specifically, the reaction cavity and the corresponding first communication channel are integrally formed, that is, the reaction cavity can extend a certain length in the direction away from the through hole through the first communication channel.

[0190] In the embodiment, the sample adding card adopts an integrated structure design of sample adding and detection. On the one hand, the steps of matching the sample adding card with the detection card are reduced, the operation process of blood type detection is further simplified, the detection efficiency is improved, and the automation of the detection process is more favorable. On the other hand, compared with the split type detection card in the prior art, the integrated sample adding card (that is, the detection card) has a simpler structure, thereby simplifying the production process and reducing the production cost to a certain extent. In addition, compared with the two relatively independent parts in the split type detection card, the integrated sample adding card is more convenient to store and transport.

[0191] Meanwhile, in the embodiment, by adopting the closed structure of the sample distribution channel, the first and second flow-through channels and the closed structure of the reaction cavity, a closed area is provided for sample adding and reaction of blood type detection, which reduces or avoids the contact between the liquid or reagent inside the detection card / sample adding card and the outside world, that is, avoids the interference of the outside pollutants (such as dust, water droplets, etc.) into the detection card to cause interference to the experimental results; meanwhile, since each sample adding channel and the reaction cavity are provided in a closed manner, the spatter of the liquid inside the detection card under the action of external force is avoided, thereby causing environmental pollution and cross contamination, and the accuracy of the detection result is improved; in addition, the differential pressure problem of the closed structure is solved by setting the corresponding air hole, so that the air pressure inside the closed structure is equal to or close to the atmospheric pressure, and the liquid sample inside the closed structure can flow down smoothly under the action of external force.

[0192] Embodiment Eight

[0193] Based on the above embodiment, the application further provides another sample adding assembly, which comprises a sample adding card and a detection card matched with the sample adding card, and is different from the above embodiment in that the first communication channel of the sample adding card is provided with an air hole for solving the internal and external pressure difference, so that when the detection card is completely pushed into the sample adding opening, the second end of the first communication channel of the sample adding card is closely attached to the opening end of the reaction cavity of the detection card.

[0194] Of course, in other embodiments, the air hole can be provided on the reaction cavity to solve the pressure difference problem in the reaction cavity.

[0195] Embodiment Nine Based on the above embodiment, the application further provides a sample adding assembly comprising the components or structures of the sample adding assembly in the above embodiment, and further, referring to Figure 9 (understandably, in order to more clearly illustrate the technical solutions and effects of the application, Figure 9 the actual proportions of the products are not drawn according to the actual proportions, and only the main structures meeting the detection application in the sample adding assembly are shown), when the matching part 61 is matched with the opening end 11b of the reaction cavity, the lower end surface (the end with a smaller opening in the conical structure) of the matching part 61 is not directly in contact with the lower end surface of the opening end of the reaction cavity, but a certain space is left, so that even if a small part of the reagent in the reagent storage area flows into the opening end of the reaction cavity under the action of external force, it can only stay in the area shown by the dashed triangle 61a and cannot enter the first communication channel 6.

[0196] Embodiment Ten

[0197] Based on the sample adding assembly of the above embodiment, the application further provides a detection method, wherein the sample adding card in the selected sample adding assembly comprises a first sample adding hole, a second sample adding hole and three third sample adding holes, and correspondingly, the method comprises the following steps:

[0198] adding the first, second and third liquid samples into the first, second and third sample wells of the sample card respectively, and the first, second and third liquid samples enter the corresponding first, second, third and fourth sub-sample channels respectively;

[0199] carrying out a first horizontal centrifugation on the sample adding assembly, under the action of a first centrifugal force, the liquid samples in the first, second, third and fourth sub-sample channels enter the corresponding first and second mixing pools respectively, and corresponding mixed liquids are obtained;

[0200] carrying out a second horizontal centrifugation on the sample adding assembly, under the action of a second centrifugal force, the mixed liquids in the first and second mixing pools enter the corresponding reaction cavities through the third through hole and the first communication channel;

[0201] carrying out a third horizontal centrifugation on the sample adding assembly, under the action of a third centrifugal force, the mixed liquids in the reaction cavities fully mix with the reagents pre-stored in the reaction cavities and react;

[0202] wherein the first, second and third centrifugal forces increase in turn.

[0203] In some embodiments, the first liquid sample is anti-cell A, the second liquid sample is anti-cell B (or the first liquid sample is anti-cell B, and the second liquid sample is anti-cell A), and the third liquid sample is plasma of different subjects, for example, plasma of subjects A, B and C respectively, and the plasma of A, B and C is added into the three third sample wells respectively.

[0204] It should be noted that in this document, the terms "comprising", "including", or any other variant thereof are intended to cover non-exclusive inclusions, so that processes, methods, articles or devices including a series of elements not only include those elements, but also include other elements not explicitly listed, or include elements inherent to such processes, methods, articles or devices. Without more limitations, the element defined by the statement "including a" does not exclude the presence of another identical element in the process, method, article or device including the element.

[0205] The embodiments of the present application are described above in conjunction with the drawings, but the present application is not limited to the above specific embodiments, and the above specific embodiments are only illustrative, not restrictive, and those skilled in the art can make many forms under the inspiration of the present application without departing from the purpose of the present application and the scope protected by the claims. These are all within the protection of the present application.

Claims

1. A microfluidic sample loading card, characterized in that, The application relates to a sample adding card, which comprises the following parts: a body; at least one first sample adding hole, at least one second sample adding hole and at least one third sample adding hole which are arranged in the body in a spaced manner, and the sample adding mouths of the first, second and third sample adding holes are located on the first surface of the body; at least one first sample distribution channel, at least one second sample distribution channel, at least one third sample distribution channel and at least one fourth sample distribution channel which are arranged in the body in a spaced manner, the first end of the first sample distribution channel is communicated with the corresponding first sample adding hole, the first end of the second sample distribution channel is communicated with the corresponding second sample adding hole, the first end of the third sample distribution channel is communicated with the corresponding third sample adding hole, and the first end of the fourth sample distribution channel is communicated with the corresponding third sample adding hole; at least one first mixing pool and at least one second mixing pool which are arranged in the body in a spaced manner, wherein each first mixing pool is communicated with the second end of the corresponding at least one first sample distribution channel and the second end of the corresponding at least one third sample distribution channel respectively, and each second mixing pool is communicated with the second end of the corresponding at least one second sample distribution channel and the second end of the corresponding at least one fourth sample distribution channel respectively; the first sample distribution channel is communicated with the first mixing pool through a first through hole, the third sample distribution channel is communicated with the first mixing pool through a second through hole, the fourth sample distribution channel is communicated with the second mixing pool through a second through hole, the second sample distribution channel is communicated with the second mixing pool through a first through hole, and the inner diameter of the first through hole (51) is larger than that of the second sample distribution channel (32); the inner diameter of the first mixing pool is larger than that of the first through hole; each of the first and second mixing pools is provided with at least one first communication channel which is arranged on the first side of the body in a spaced manner, and the second end of the first communication channel is provided with a sharp end for puncture; the first communication channel is communicated with the first mixing pool through a third through hole, the first communication channel is communicated with the second mixing pool through a third through hole, and the inner diameter of the third through hole is smaller than that of the first communication channel.

2. The sample application card of claim 1, wherein, at least one air hole is arranged on the first sample distribution channel, at least one air hole is arranged on the second sample distribution channel, at least one air hole is arranged on the third sample distribution channel, and / or at least one air hole is arranged on the fourth sample distribution channel.

3. The sample application card of claim 1, wherein, the sample adding card further comprises a first waste liquid cavity which is arranged in the body and communicated with the corresponding first sample adding hole; and / or the sample adding card further comprises a second waste liquid cavity which is arranged in the body and communicated with the corresponding second sample adding hole; and / or the sample adding card further comprises a third waste liquid cavity which is arranged in the body and communicated with the corresponding third sample adding hole.

4. The sample application card of claim 1, wherein, The first, second, third and fourth sample channels are arranged on the second surface of the body, and a hydrophilic layer is arranged on a first area of the second surface of the body, wherein the first area includes an area where the first, second, third and fourth sample channels are arranged.

5. The sample application card of claim 1, wherein, The first and second mixing pools are arranged on the first surface of the body, and a hydrophobic layer or a hydrophilic layer is arranged on a second area of the first surface of the body, wherein the second area includes an area where the first and second mixing pools are arranged.

6. The sample application card of claim 1, wherein, An inner surface area of the sample card is treated with paraffin oil, wherein the inner surface area includes inner surfaces of the first, second, third and fourth sample channels and / or an inner surface of the first communication channel.

7. The sample application card of claim 1, wherein, The sample card includes one first sample well, one second sample well and three third sample wells, and the sample card includes three first mixing pools and three second mixing pools arranged at intervals in the body.

8. The sample application card of claim 3, wherein, The first sample channel and the first through hole have an overlapping area in the orthographic projection.

9. The sample application card of claim 3, wherein, The second sample channel and the first through hole have an overlapping area in the orthographic projection.

10. The sample application card of claim 3, wherein, The three third sample wells arranged at intervals in the body are a left third sample well, a middle third sample well and a right third sample well, wherein the first sample well is located on a symmetry axis of the left third sample well and the middle third sample well, the second sample well is located on a symmetry axis of the middle third sample well and the right third sample well, and a horizontal direction of the first sample well and a horizontal direction of the second sample well are both above the three third sample wells.

11. The sample card of any one of claims 1-9, wherein, Further comprising: Two side walls extending from two sides of the body to form a card insertion opening for inserting a detection card; When the detection card is pushed into the card insertion opening, the first communication channel of the sample card is in communication with a corresponding reaction chamber of the detection card.

12. The sample application card of claim 1, wherein, Further comprising: A detection card integrally formed with the sample card, the detection card being provided with reaction chambers for being in communication with the first communication channels, and / or the first communication channels being provided with air holes for solving pressure difference.

13. The sample card of any one of claims 1-9, wherein, The first, second, third and fourth sample channels are capillary sample channels.

14. A microfluidic dosing assembly, characterized in that Further comprising: The sample card of any one of claims 1-11 and a detection card, wherein the sample card is provided with a card insertion opening for being inserted into the detection card, when the sample card is pushed into the card insertion opening, the first communication channel of the sample card is in communication with a corresponding reaction chamber of the detection card, and a gap for ventilation is formed between the first communication channel and the reaction chamber; Alternatively, the sample card of any one of claims 1-11 and a detection card, wherein the sample card is provided with a card insertion opening for being inserted into the detection card, the first communication channel is provided with air holes for solving pressure difference, and / or the reaction chamber is provided with air holes for solving pressure difference. When the detection card is pushed into the card joint opening, the second ends of the first communication channels on the sample card are respectively communicated with the corresponding reaction cavities on the detection card, and the first communication channels and the reaction cavities are closely attached.

15. The sample application assembly of claim 14, wherein, Also comprising: A reagent pre-stored in the reaction cavity, wherein the reagent comprises: a gel and a working solution, wherein the working solution comprises: an antibody.

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