CN114324958B

CN114324958B - Method for detecting based on sample adding assembly

Info

Publication number: CN114324958B
Application number: CN202210261160.6A
Authority: CN
Inventors: 王丽; 王伟权; 黄志刚; 王秀柱; 郝进学; 岳晓宇
Original assignee: Tianjin Dexiang Biotechnology Co ltd
Current assignee: Tianjin Texiang Biotechnology Co Ltd
Priority date: 2022-03-17
Filing date: 2022-03-17
Publication date: 2022-05-24
Anticipated expiration: 2042-03-17
Also published as: CN114324958A; CN114924090A

Abstract

The invention provides a sample adding card and a sample adding assembly, comprising: a body; the sample adding holes are arranged in the body, and the sample adding ports of the sample adding holes are positioned on the first surface of the body; the first ends of the first and second sample dividing channels are respectively communicated with the corresponding first and second sample adding holes, and the first ends of the third and fourth sample dividing channels are respectively communicated with the corresponding third sample adding holes; the first mixing pool and the second mixing pool are arranged in the body at intervals, wherein each first mixing pool is respectively communicated with the second end of the corresponding first sample dividing channel and the second end of the corresponding third sample dividing channel; each second mixing pool is respectively communicated with the second end of the corresponding second sampling channel and the second end of the corresponding fourth sampling channel; first, two mix the pond and all be provided with first communication channel, and this first communication channel interval sets up the first side at the body. The sample adding card and the sample adding assembly are simple to operate, and cross contamination is avoided.

Description

Method for detecting based on sample adding assembly

Technical Field

The invention relates to the field of medical detection, in particular to a sample adding card and a sample adding assembly.

Background

Blood transfusion is one of the clinical commonly used medical means, and if the blood types are not matched during blood transfusion, the health of a patient is seriously damaged, even the life of the patient is threatened, so the blood grouping before blood transfusion is very important, and the gel microcolumn method is one of the commonly used blood grouping means. However, the existing gel micro-column card has the following problems:

various test cards, such as blood type test cards, often have a flexible film material (e.g., aluminum foil, plastic film) that is sealed to the contents (e.g., various reagents and/or gel beads). When the detection card is used, the sealing film needs to be torn off or punctured, and then the sample can be added to the detection card. For example, a traditional blood type card needs to be punched by a puncher before being used, which is easy to cause cross contamination; to the blood group card including a plurality of microcolumns (be reaction chamber), every blood group card need carry out the application of sample many times, complex operation also is difficult to guarantee accurate equivalent application of sample. In addition, with the miniaturization of various reagent cards, when the inner diameter of a sample addition port on a reagent card is only 1-2mm or less, manual sample addition is difficult.

In addition, the operation of the prior art microcolumn gel card is complex, and the requirement on the operation normative is high, so that the operator is generally required to be trained before using the microcolumn gel card, which limits the further popularization and application of the microcolumn gel method in the field of blood type detection to a certain extent.

Meanwhile, in the reverse typing sample application in the prior art, the detection reagent (e.g., the reverse cell a and the reverse cell B) and the sample to be detected (e.g., the 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, so that the step-by-step operation is not beneficial to the improvement of the detection efficiency. If the sample adding is carried out by a plurality of samples, the sample adding times are very frequent, and the misoperation is easily caused when the samples are added manually.

Disclosure of Invention

The present invention provides a reverse-shaped sample adding card, which partially solves or alleviates the above-mentioned deficiencies in the prior art, further simplifies the operation process, and facilitates the automation of sample adding and detection.

In a first aspect, the present invention provides a microfluidic sample addition card comprising:

a body;

the sample adding ports of the first, second and third sample adding holes are positioned on the first surface of the body;

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;

the at least one first mixing pool and the at least one second mixing pool are arranged in the body at intervals, 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; each second mixing pool is respectively communicated with the second end of at least one corresponding second sub-sampling channel and the second end of at least one corresponding fourth sub-sampling channel;

the first mixing pool and the second mixing pool are both provided with at least one first communication channel, and the first communication channels are arranged on the first side of the body at intervals.

In some embodiments, the second end of the first communication channel is provided with a pointed tip for puncturing.

In some embodiments, the first sample channel is provided with at least one air hole; and/or at least one air hole is arranged on the second sampling channel; and/or 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.

In some embodiments, the first sample distribution channel is in communication with the first mixing well through a first through-hole; and/or the presence of a gas in the gas,

the second sample separation channel is communicated with the second mixing pool through a first through hole.

In some embodiments, the third aliquot passage is in communication with the first mixing well via a second through-hole, and/or,

and the fourth sample distribution channel is communicated with the second mixing pool through a second through hole.

In some embodiments, the first communicating channel communicates with the first mixing tank through a third through hole, and the first communicating channel communicates with the second mixing tank through a third through hole.

In some embodiments, further comprising:

the first waste liquid cavity is arranged in the body and communicated with the corresponding first sample adding hole; and/or the presence of a gas in the gas,

the second waste liquid cavity is arranged in the body and communicated with the corresponding second sample adding hole; and/or the presence of a gas in the gas,

the third waste liquid cavity is arranged in the body and communicated with the corresponding third sample adding hole.

In some embodiments, the first, second, third and fourth sample distribution channels are disposed on the second surface of the body, and accordingly, a hydrophilic layer is disposed on a first region of the second surface of the body, wherein the first region comprises: the area is provided with a first sample distribution channel, a second sample distribution channel, a third sample distribution channel and a fourth sample distribution channel.

In some embodiments, the first and second mixing tanks are disposed on the first surface of the body, and correspondingly, a hydrophobic layer or a hydrophilic layer is disposed on a second region of the first surface of the body, wherein the second region includes: a region provided with a first mixing tank and a second mixing tank.

In some embodiments, an inner surface region of the sample application card is treated with paraffin oil, wherein the inner surface region comprises: the inner surface of the first, second, third, fourth sample distribution channel, and/or the inner surface of the first communication channel.

In some embodiments, the sample application card comprises: one said first well, one said second well and three said third wells, respectively, said sample card comprising: the three first mixing tanks and the three second mixing tanks are arranged inside the body at intervals.

In some embodiments, there is an overlap region between the orthographic projection of at least one of the first sampling channels and the orthographic projection of at least one of the first through holes, and/or,

an overlapping area exists between the orthographic projection of at least one second sampling channel and the orthographic projection of at least one first through hole.

In some embodiments, the three third sampling holes spaced apart inside the body are respectively: a left side third application of sample hole, well third application of sample hole, right third application of sample hole, wherein, first application of sample hole is located a left side third application of sample hole with on the symmetry axis of well third application of sample hole, second application of sample hole is located well third application of sample hole with on the symmetry axis of right side third application of sample hole, just the horizontal direction at first application of sample hole place with the horizontal direction at second application of sample hole place all is located three the top of third application of sample hole.

In some embodiments, further comprising: the clamping opening is formed by side walls extending from two sides of the body and is used for being inserted with the detection card;

when the detection card is pushed into the clamping opening, the first communication channel of the sample adding card is respectively communicated with the corresponding reaction cavity on the detection card.

In some embodiments, further comprising: the detection card is integrally formed with the sample adding card, reaction cavities which are respectively communicated with the corresponding first communication channels are arranged on the detection card, air holes used for solving the pressure difference are arranged on the reaction cavities, and/or air holes used for solving the pressure difference are arranged on the first communication channels.

The invention also provides a microfluidic sample adding assembly, comprising: the sample adding card and the detection card in the above embodiments, wherein the sample adding card is provided with a clamping opening capable of being inserted into the detection card, when the sample adding card is pushed into the clamping opening, the first communicating channels of the sample adding card are respectively communicated with the corresponding reaction chambers on the detection card, and a gap for ventilation exists between the first communicating channels and the reaction chambers.

In some embodiments, the sample adding assembly comprises the sample adding card in any one of the above embodiments, and a detection card, the sample adding card is provided with a clamping opening capable of being in clamping connection with the detection card, wherein the first communication channel is provided with an air hole for solving the pressure difference, and/or the reaction cavity is provided with an air hole for solving the pressure difference,

when the detection card is pushed into the clamping opening, the second end of the first communicating channel on the sample adding card is respectively communicated with the corresponding reaction cavity on the detection card, and the first communicating channel is closely attached to the reaction cavity.

The invention provides a microfluidic sample adding assembly, which comprises: the sample adding card and the detection card in any one of the above embodiments, wherein the sample adding card is provided with a clamping opening capable of being inserted into the detection card, and a side wall of the clamping opening is provided with a first clamping position and a second clamping position at intervals along the extending direction of the side wall;

the detection card is movably arranged at the second clamping position relative to the clamping opening;

when under the effect of external acting force, make the detection card follow the second joint position removes to when first joint position, add on the appearance card first intercommunication passageway respectively with corresponding reaction chamber is linked together on the detection card, just first intercommunication passageway with there is the gap that is used for ventilating between the reaction chamber.

The invention also provides another microfluidic sample addition assembly comprising: the sample adding card and the detection card in the embodiment, wherein the sample adding card is provided with a clamping opening capable of being inserted into the detection card, and one side or two sides of the detection card are provided with a second clamping position and a first clamping position at intervals along a direction gradually far away from the opening end of the reaction cavity;

when under the effect of external acting force, make the detection card follow the second joint position removes to during the first joint position, add the appearance card first intercommunication passageway respectively with corresponding reaction chamber is linked together on the detection card, just first intercommunication passageway with there is the gap that is used for ventilating between the reaction chamber.

In some embodiments, further comprising: reagents pre-stored in the reaction chamber.

In some embodiments, the reagent comprises: a gel and a working fluid, wherein the working fluid comprises: an antibody.

The present invention also provides a method for performing detection based on the sample adding assembly described in any of the above embodiments, where a sample adding card in the sample adding assembly includes a first sample adding hole, a second sample adding hole, and three third sample adding holes, and accordingly, the method includes the steps of:

respectively adding a first liquid sample, a second liquid sample and a third liquid sample into a first sample adding hole, a second sample adding hole and a third sample adding hole of the sample adding card, and respectively enabling the first liquid sample, the second liquid sample and the third liquid sample to enter a corresponding first sample adding channel, a corresponding second sample adding channel, a corresponding third sample adding channel and a corresponding fourth sample adding channel;

carrying out first horizontal centrifugation on the sample adding assembly, and respectively enabling liquid samples in the first, second, third and fourth sample dividing channels to enter the corresponding first and second mixing tanks under the action of first centrifugal force to obtain corresponding mixed liquid;

carrying out second horizontal centrifugation on the sample adding assembly, wherein under the action of second centrifugal force, the mixed liquid in the first mixing pool and the second mixing pool enters the corresponding reaction cavities through the third through holes and the first communication channels;

carrying out third horizontal centrifugation on the sample adding assembly, and under the action of third centrifugal force, fully mixing the mixed solution entering the reaction cavity with a reagent prestored in the reaction cavity and reacting;

wherein the magnitudes of the first, second and third centrifugal forces increase in sequence.

Advantageous effects

The sample adding card can be applied to a multi-person reverse sizing test or a multi-person antibody screening test, can realize sample adding of multiple persons on a single sample adding card simultaneously, greatly reduces the sample adding times, simplifies the operation process, avoids misoperation possibly caused by multiple samples adding, thereby effectively improving the sample adding and detecting efficiency, and simultaneously realizes multi-person detection by matching the single sample adding card with the detecting card, and reduces the detecting cost to a certain extent.

And because the liquid sample in each sample separation channel can enter the corresponding first mixing pool and the second mixing pool at the same time (or the time difference of the liquid sample in each sample separation channel entering the first mixing pool and the second mixing pool is very small and can be ignored), the two liquids can be fully mixed, the situation that the two liquids are not fully mixed due to different sampling sequences is avoided, and the accuracy of the experimental result is improved.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. Throughout the drawings, like elements or portions are generally identified by like reference numerals. In the drawings, elements or portions are not necessarily drawn to scale. It is obvious that the drawings in the following description are some embodiments of the invention, and that for a person skilled in the art, other drawings can be derived from them without inventive exercise.

FIG. 1 is a first perspective view of an exemplary embodiment of the present invention;

FIG. 2 is a second perspective view of an exemplary embodiment of the present invention;

FIG. 3 is a schematic diagram of a first configuration of an exemplary embodiment of the present invention;

FIG. 4 is a schematic diagram of a second surface in accordance with an exemplary embodiment of the present invention;

FIG. 5 is a schematic view of a first surface of an exemplary embodiment of the present invention;

FIG. 6 is a second structural schematic of an exemplary embodiment of the present invention;

FIG. 7a is a diagram illustrating a structure of a test card according to an exemplary embodiment of the present invention;

FIG. 7b is a schematic diagram of a test card according to another exemplary embodiment of the present invention;

FIG. 8 is a third structural diagram in accordance with an exemplary embodiment of the present invention;

FIG. 9 is a schematic view of a configuration of a first communicating channel in cooperation with an open end of a reaction chamber in an exemplary embodiment of the invention;

FIG. 10 is a schematic view illustrating a flow state of a liquid sample after the sample application card applies the sample to the first sample application hole during an experimental operation according to an exemplary embodiment of the present invention;

fig. 11 is a schematic view illustrating a flow state of a liquid sample after the sample application card applies samples to the first, second, and third sample application holes during an experimental operation process according to an exemplary embodiment of the invention.

The device comprises a body 1, a first sample adding hole 21, a second sample adding hole 22, a third sample adding hole 23, a first sample dividing channel 31, a second sample dividing channel 32, a third sample dividing channel 33, a fourth sample dividing channel 34, a first through hole 51, a second through hole 52, a third through hole 53, a first communicating channel 6, a matching part 61, a side wall 7, a limiting clamping protrusion 71, an air hole 8, a first waste liquid cavity 91, a second waste liquid cavity 92, a third waste liquid cavity 93, a limiting clamping groove 10, a reaction cavity 11, a reagent storage area 11a, a reaction cavity opening end 11b, a first mixing pool 41, a second mixing pool 42 and a triangle-shaped dotted line 61 a.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Herein, unless otherwise explicitly specified and limited, the dotted lines in the drawings represent invisible edge lines and invisible outline lines, and the solid lines in the drawings represent visible edge lines and visible outline lines.

Herein, unless otherwise expressly specified and limited, "pass-through" means to communicate two structures such that a one-way or two-way flow of liquid between the two structures can be achieved; the "through hole" means a passage for connecting two structures and the length of the passage is usually set short, wherein the cross section of the passage can be selected from circular, oval and square, and the cross section of the passage is preferably circular for the sake of convenience of processing. For example, the "first through hole 51" herein can be regarded as a channel for connecting the third fractionating passage 33 and the second mixing well 42, and the channel length is small.

Herein, unless otherwise specifically specified and limited, "three-stage centrifugation" refers to a centrifugation method comprising three steps of first horizontal centrifugation, second horizontal centrifugation and third horizontal centrifugation, wherein the centrifugation speeds/centrifugal forces of the first, second and third horizontal centrifugation are different.

Herein, unless otherwise expressly specified and limited, "tapered structure" refers to a structure that is large at the top and small at the bottom, and has a trapezoidal or trapezoid-like cross-section.

Herein, suffixes such as "module", "part", or "unit" used to denote elements are used only for facilitating the description of the present invention, and have no specific meaning in itself. Thus, "module", "component" or "unit" may be used mixedly.

Herein, the terms "upper", "lower", "inner", "outer", "front", "rear", "one end", "the other end", "left", "center", "right", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

"equal" or "the same" or "simultaneously" herein does not mean that the absolute value of the difference between two or more items specified is zero, but that the difference between the items specified is very small, and in practice the difference is negligible. For example, the phrase "the third sample distribution channel and the fourth sample distribution channel can contain equal amounts of liquid sample" means that the third sample distribution channel and the fourth sample distribution channel can contain completely equal amounts of liquid sample, or the difference between the third sample distribution channel and the fourth sample distribution channel has little effect on the result and can be ignored in practical applications.

As used herein, unless otherwise expressly specified or limited, the terms "mounted," "disposed," "connected," and the like are to be construed broadly, such that the terms "connected," or "connected," as used herein, may be fixedly connected, detachably connected, or integrally connected; they may be mechanically coupled, directly coupled, indirectly coupled through intervening media, or may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

Herein, a surface of the sample application card on which the opening (or sample application port) of the sample application well (e.g., first, second, and third sample application wells) is located is referred to as a first surface (or upper surface), and the other surface is referred to as a second surface (or lower surface).

As used herein, the "first side" of the body refers to the side of the sample application card that is adjacent to the open end of the reaction chamber of the test card, i.e., the side that is adjacent to the second end of the sample distribution channel, when the sample application card is mated with the test card.

Herein, when referring to a structure that is "internal" to the sample application card, it is meant that the structure is surrounded by the bulk material of the sample application card or other material and is not in direct contact with the external space. For example, "at least one first sample channel spaced apart inside the body" may be understood as a first sample channel that is surrounded by the bulk material of the sample application card and not in direct contact with the external space, or may be understood as a first sample channel that is provided on a first side or a second side of the body and is surrounded by the bulk material of the sample application card and other materials (e.g., a hydrophilic layer or a hydrophobic layer) and not in direct contact with the external space.

Herein, the term "horizontal centrifugation" means that when the detection card is centrifuged, the detection card is horizontally placed or nearly horizontally placed, that is, the horizontal plane of the first side surface or the second side surface of the detection card is at an angle equal to or nearly 90 degrees with the centrifugal axis of the centrifugal device. "horizontal centrifuge" means a state in which a test card placed in the centrifuge is horizontal or nearly horizontal when the centrifuge is operated.

Herein, "closely fit" means that when two structures are mated, the inner/outer surface of one structure is closely spaced from the outer/inner surface of the other structure, or the inner/outer surface of one structure is in contact with the outer/inner surface of the other structure, so that there is no gap or a small gap between the adjacent surfaces of the two structures, so that gas or liquid cannot enter the inside of the detection card or the reaction chamber through the gap between the two structures, for example, when "the second end of the first communicating channel is closely fitted to the open end of the reaction chamber", it can be understood that the surface spacing between the outer surface of the second end of the first communicating channel and the inner surface of the open end of the reaction chamber is small, or the two surfaces are in contact with each other, so that air or foreign matter (dust or water droplets in the air) in the outer space cannot enter the first communicating channel or the inside of the reaction chamber, and the liquid in the sample adding device (i.e. sample adding card) or the detection card can not be splashed to the external space.

Herein, "orthographic projection" refers to a projection of a structure (e.g., a first sample channel) in a direction perpendicular to the plane of the body (e.g., the plane of the first surface or the second surface).

Herein, the "sample distribution channel" is a general term for the first, second, third and fourth sample distribution channels, the "mixing well" is a general term for the first and second mixing wells, and the "through hole" is a general term for the first, second and third through holes.

Herein, "hydrophilic layer" refers to a film or a baffle having hydrophilic properties, for example, a hydrophilic membrane, and "hydrophobic layer" refers to a film or a baffle having hydrophobic properties.

Example one

Referring to fig. 1, the present invention provides a microfluidic sample application card comprising:

a body 1;

at least one first sample adding hole 21, at least one second sample adding hole 22 and at least one third sample adding hole 23 which are arranged in the body 1 at intervals, and the sample adding ports of the first, second and third sample adding holes are positioned on the first surface (namely the upper surface) of the body 1;

at least one first sample distribution channel 31, at least one second sample distribution channel 32, at least one third sample distribution channel 33 and at least one fourth sample distribution channel 34 which are arranged in the body 1 at intervals, wherein the first end of the first sample distribution channel 31 is communicated with the corresponding first sample adding hole 21, the first end of the second sample distribution channel 32 is communicated with the corresponding second sample adding hole 22, the first end of the third sample distribution channel 33 is communicated with the corresponding third sample adding hole 23, and the first end of the fourth sample distribution channel 34 is communicated with the corresponding third sample adding hole 23;

at least one first mixing pool 41 and at least one second mixing pool 42 which are arranged in the body at intervals, wherein each first mixing pool 41 is respectively communicated with the second end of the corresponding at least one first sample distribution channel 31 and the second end of the corresponding at least one third sample distribution channel 33; each second mixing cell 42 is respectively communicated with the second end of the corresponding at least one second sub-sampling channel 32 and the second end of the corresponding at least one fourth sub-sampling channel 34;

first, two mix the pond and all be provided with at least one

first communication channel

6, and 6 intervals of first communication channel set up the first side at the body.

Preferably, in some embodiments, the first, second, third and fourth sample distribution channels are capillary sample distribution channels, and when a liquid sample (detection reagent or sample to be detected) is added to the first, second and third sample addition holes, the liquid sample fills the corresponding sample distribution channels (i.e., the first, second, third and fourth sample distribution channels) under capillary action.

This application of sample card needs to cooperate with the detection card (also micro-column gel detects the card) and uses, wherein, include in the detection card that the interval sets up a plurality of reaction chambeies 11 on detecting the card (can understand, reaction chamber number is the same with the first communication channel number of application of sample card), is equipped with the reaction chamber open end 11b that is used for the application of sample on the reaction chamber. And the reaction cavity of the detection card is pre-stored with reagent, a sealing film is arranged on the opening end of the reaction cavity, or paraffin oil is arranged at the upper end of the reaction cavity to seal the reagent, thereby preventing the reagent from drying.

To achieve the interaction between the sample application card and the detection card, further, in some embodiments, referring to fig. 2 to 6, the sample application card further includes: two clamping openings formed by side walls 7 extending from two sides of the body 1 and used for being inserted with a detection card;

when the detection card is pushed into the card-connecting opening, the second end of the first communicating channel of the sample adding card is communicated with the corresponding reaction cavity on the detection card; and when the sample adding card and the detection card are centrifuged, under the action of centrifugal force, liquid samples in the first sample adding hole, the second sample adding hole and the third sample adding hole sequentially pass through the sample dividing channels (the first sample dividing channel, the second sample dividing channel, the third sample dividing channel and the fourth sample dividing channel) to enter the corresponding first mixing pool and the corresponding second mixing pool, and then enter the corresponding reaction cavities through the first communication channels connected with the first mixing pool and the second mixing pool.

When the detection card is pushed in the joint opening, if the second end of first intercommunication passageway is laminated closely with the open end of reaction chamber, enter into each branch appearance passageway or first intercommunication passageway when liquid sample, and when getting into certain length, liquid sample in the passageway will extrude the air of branch appearance passageway, first intercommunication passageway and reaction chamber, lead to dividing the appearance passageway, first intercommunication passageway and the increase of reaction intracavity atmospheric pressure, and there is pressure differential between the outside air, thereby make liquid sample even under capillary action and centrifugal force's combined action, also can't enter into the reaction chamber.

Therefore, when the sample adding card is matched with the detection card, the second end of the first communication channel is not closely attached to the opening end of the reaction cavity, but a certain gap exists, so that air in an external space can enter the first communication channel and the reaction cavity through the gap, the opening of the second end of the first communication channel is formed, the air pressure at the opening end of the reaction cavity is equal to or close to the atmospheric pressure of the external space, and a liquid sample entering the first communication channel can smoothly enter the corresponding reaction cavity under the action of centrifugal force.

Specifically, in some embodiments, referring to fig. 1, the second end of the first communicating channel 6 is disposed at the first side of the body 1 at an interval and extends to the outside of the body, and when the sample adding card is matched with the detection card, the second end of the first communicating channel 6 extends into the open end of the corresponding reaction cavity on the detection card and is communicated with the corresponding reaction cavity.

Referring to fig. 3, in some embodiments, since part of the test card is provided with the sealing membrane at the open end, in order to make the first communicating channel communicate with the reaction chamber, the second end of the first communicating channel is provided with a fitting portion 61 for piercing the sealing membrane (preferably, the fitting portion is a pointed tip for piercing the sealing membrane), and when the sample adding card is fitted with the test card, the fitting portion (i.e., the pointed tip) pierces the sealing membrane at the open end of the reaction chamber, so that the first communicating channel communicates with the corresponding reaction chamber, respectively.

Of course, in other embodiments, referring to fig. 7b, the open end of the reaction cavity 11 on the detection card extends outward for a certain length in a direction away from the bottom end of the reaction cavity, accordingly, the second end of the first communicating channel on the sample adding card may not extend to the outside of the body, and when the sample adding card is matched with the detection card, the open end of the reaction cavity extends into the inside of the first communicating channel, so that the first communicating channel is communicated with the reaction cavity.

In some embodiments, the body is provided with a plurality of sample distribution channels, so that there may be one or more sample distribution channels of too long a length, and the sample distribution channels of the longer length are provided with air holes 8 for at least one flow.

Specifically, in some embodiments, one or more of the first, second, third, and fourth sampling channels are provided with air holes 8. For example, referring to fig. 3, the air hole 8 for drainage is provided on one of the sampling channels having the longest length (i.e., one of the second sampling channels 32). Because partial sample separation channel is longer, the internal diameter of corresponding sample separation channel sets for littleer, therefore gaseous flow in longer sample separation channel can be obstructed, leads to when liquid gets into this sample separation channel in certain length, thereby the extrusion that receives liquid sample is not discharged smoothly to the gas in the sample separation channel, leads to consequently leading the inside atmospheric pressure of sample separation channel and outside atmosphere to form the pressure differential, and can solve the pressure differential problem through setting up the gas pocket to play the drainage effect to liquid sample.

Further, in some embodiments, referring to fig. 2, the first and second half-sampling channels are respectively communicated with the first and second mixing pools through the first through hole 51.

Further, in some embodiments, referring to fig. 2, the third and fourth sample distribution channels are respectively communicated with the first and second mixing pools through the second through hole 52.

Further, in some embodiments, the first communicating path 6 communicates with the first mixing tank 41 through a third through hole, and communicates with the second mixing tank 42 through a third through hole 53.

In some embodiments, one or more of the first, second, and third through holes are channel structures having a certain length, and function to connect each mixing pool with each sample distribution channel or the first communicating channel, and specifically, the first and second through holes connected with the sample distribution channel may be capillary channels, which may be regarded as extensions of the sample distribution channel; of course, the first and second through holes may also be non-capillary channels, which may play a role of stopping flow, and when the liquid sample fills the sample distribution channels (e.g., the first, second, third, and fourth sample distribution channels), the liquid sample may be prevented from entering the through holes continuously by capillary action.

Further, in order to accommodate the excessive liquid sample added to the first well 21, referring to fig. 3, a first waste liquid chamber 91 may be provided near the first well 21 and inside the body.

Further, in order to prevent the sample (i.e., the liquid sample) from entering the first waste liquid chamber 91 when it is not in excess and to avoid cross-contamination caused by the sample in the first waste liquid chamber 91 returning to the first well, a baffle plate may be provided in the first waste liquid chamber near the well.

In some embodiments, the baffle acts as a raised flap in first waste chamber 91, and only when the sample is loaded is excess and spills over its upper end can it enter the first waste chamber.

In addition, in order to facilitate the entry of the liquid sample into the first waste liquid chamber, the first waste liquid chamber may have an opening on the upper surface of the body 1. When the body 1 is made of a hydrophilic material, it is contemplated that the baffle may be treated to have a hydrophobic property, for example, a hydrophobic layer may be added to further prevent the liquid sample entering the first waste liquid chamber from returning to the first well.

Further, in some embodiments, to accommodate the excess liquid sample added to the second well, referring to fig. 3, the sample addition card further comprises: and a second waste liquid chamber 92 provided inside the body, the second waste liquid chamber 92 being communicated with the corresponding second sample addition hole 22.

Further, in some embodiments, to accommodate the excess liquid sample added to the third well, referring to fig. 4, the sample addition card further comprises: and a third waste liquid cavity 93 arranged in the body and communicated with the corresponding third sample adding hole 23.

Wherein, the arrangement mode of the second three-waste liquid cavity is referred to the first waste liquid cavity.

In some embodiments, the first, second, third and fourth sampling channels are disposed on the second surface (i.e. the lower surface) of the body, and accordingly, a hydrophilic layer is disposed on a first region of the second surface of the body, wherein the first region includes: the area is provided with a first sample distribution channel, a second sample distribution channel, a third sample distribution channel and a fourth sample distribution channel.

Specifically, referring to fig. 4, the structure shown in fig. 4 can be considered as a sectional view with a portion of the lower surface (each of the sample application channels) removed to show the internal structure of the sample application card of the present invention. In addition, for the convenience of processing (especially, each sample distribution channel) and/or hydrophilic treatment, the structures shown in the drawings can be processed first, and then the structures such as the first, second, third, and fourth sample distribution channels can be covered by a lower surface layer (for example, a hydrophilic layer, specifically, a hydrophilic film or a baffle with hydrophilic property) through pasting or bonding, etc., so as to achieve the same effect that the structures such as the first, second, third, and fourth sample distribution channels are located inside the body 1. Therefore, the sample adding card, whether manufactured by integral injection molding or layered processing, or manufactured by layered processing without the main structure of the lower surface layer, should be covered in the protection scope of the present application.

In this embodiment, the structures such as the first, second, third and fourth sampling channels are covered by adhering a hydrophilic film layer or a baffle with hydrophilic property, so that the aqueous liquid such as blood can flow conveniently.

Further, in order to prevent the liquid sample in the first and second mixing wells from splashing during centrifugation and thus contaminating the test card or the test environment, in some embodiments, the first and second mixing wells are disposed on a first surface of the body (a portion where the first and second mixing wells are disposed is referred to as a second region), and accordingly, a hydrophobic layer (e.g., a hydrophobic film or a baffle having hydrophobic properties) or a hydrophilic layer is disposed on the second region of the first surface of the body.

Further, to reduce the retention of liquid sample inside the sample application card (e.g., each sample application channel or mixing well), in some embodiments, an inner surface area of the sample application card is treated with paraffin oil, wherein the inner surface area comprises: the inner surface of the first, second, third, fourth sample distribution channel, and/or the inner surface of the first communication channel.

Specifically, in some embodiments, the paraffin oil treatment step comprises: paraffin oil is added into the first sample adding hole, the second sample adding hole and the third sample adding hole respectively, the paraffin oil enters into each corresponding sample dividing channel under the capillary action (or the combined action of the capillary action and the centrifugal force), then enters into each mixing pool through the sample dividing channels, finally flows out from the second end of the first communicating channel, and can adopt an empty detection card (namely a detection card which does not store reagents) to be matched with the sample adding card to receive redundant paraffin oil.

In an exemplary embodiment of the present invention, referring to fig. 1, a sample application card comprises: a first well 21, a second well 22 and three third wells 23, and accordingly, the sample card comprises: three first mixing pools 41 and three second mixing pools 42 are arranged at intervals inside the body.

Specifically, in this exemplary embodiment, the first well 21 communicates with the corresponding first mixing well 41 through three first branch channels 31, respectively, the second well 22 communicates with the corresponding second mixing well 42 through three second branch channels 32, respectively, the third well 23 communicates with the first mixing well 41 through three third branch channels 33, respectively, and, at the same time, the third well 23 communicates with the second mixing well 42 through three fourth branch channels 34, respectively.

When each sample separation 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 separation channel will intersect and interfere with each other, therefore, in order to avoid the interference between the first through hole or the second through hole and the sample separation 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 separation channel have an overlapping part.

For example, in this exemplary embodiment, referring to fig. 1, there is an overlapping area of the orthographic projection (i.e., the projection perpendicular to the body plane direction) of the at least one second sampling channel and the orthographic projection of the at least one first through hole.

For another example, in further exemplary embodiments, there is an overlap region between the orthographic projection of the at least one first sampling channel and the orthographic projection of the at least one first through hole,

further, in this exemplary embodiment, as shown in fig. 1, taking the positive direction of the X axis in fig. 1 as the left direction and the positive direction of the Y axis as the up direction, the three third sampling holes disposed at intervals inside the body are: the third sample adding hole in the left side, the third sample adding hole in the middle, the third sample adding hole in the right side, wherein, first sample adding hole is located on the symmetry axis of the third sample adding hole in the left side and the third sample adding hole in the middle, the second sample adding hole is located on the symmetry axis of the third sample adding hole in the middle and the third sample adding hole in the right side, and the horizontal direction at first sample adding hole place and the horizontal direction at second sample adding hole place all are located the top of three third sample adding holes.

In this embodiment, the first, second, and third wells are mutually matched in spatial structure, so that multiple-person sample adding for detection is realized in a limited space, for example, three-person sample adding for a reverse typing test or two-person sample adding for an antibody screening test.

Specifically, when the sample adding card is applied to sample adding of a reverse typing test, the retrofixed cell A is added into the first sample adding hole, the retrofixed cell B is added into the second sample adding hole (or the retrofixed cell B is added into the first sample adding hole, the retrofixed cell A is added into the second sample adding hole), and three samples to be tested (from three subjects) are added into the three third sample adding holes at the same time. After sample adding, placing the sample adding card with the detection card into a centrifugal card slot of a centrifuge for centrifugation, under the combined action of capillary action and centrifugal force, enabling liquid samples in each sample adding hole to sequentially pass through corresponding sample dividing channels and through holes and enter corresponding mixing tanks, wherein the anti-fixed cells A are respectively mixed with three samples to be detected in a first mixing tank in an intersecting manner, and the anti-fixed cells B are respectively mixed with the three samples to be detected in a second mixing tank in an intersecting manner; further, mixed liquor obtained by intersection and mixing in the first mixing pool and the second mixing pool enters the corresponding reaction cavity through the third through hole and the first communicating channel under the action of centrifugal force, reacts with a reagent in the reaction cavity, and after centrifugation is finished, reaction results in the reaction cavity are observed and test data of three samples to be tested are recorded.

When the sample adding card is applied to an antibody screening test, corresponding samples to be tested (derived from two subjects) are respectively added into a first sample adding hole and a second sample adding hole, and a first anti-sieve cell, a second anti-sieve cell and a third anti-sieve cell are respectively added into a third sample adding hole (it can be understood that no corresponding relation exists between the first anti-sieve cell, the second anti-sieve cell and the third sample adding hole). After sample adding, placing the sample adding card with the detection card into a centrifugal card slot of a centrifuge for centrifugation, under the combined action of capillary action and centrifugal force, enabling liquid samples in the first and third sample adding holes to sequentially pass through corresponding sample dividing channels and through holes to enter corresponding first mixing pools for mixing, and enabling liquid samples in the second and third sample adding holes to sequentially pass through corresponding sample dividing channels and through holes to enter corresponding second mixing pools for mixing; further, mixed liquor obtained after intersection and mixing in the first mixing pool and the second mixing pool enters the corresponding reaction cavity through the third through hole and the first communicating channel under the action of centrifugal force, reacts with a reagent in the reaction cavity, and after centrifugation is finished, a reaction result in the reaction cavity is observed and test data of two samples to be tested are recorded.

The use process is described in sequence, and it is understood by those skilled in the art that the operation need not be performed in this sequence, for example, the sample adding card may be placed in a centrifuge, and then the detection card may be inserted and added in sequence, or the sample adding card and the detection card may be inserted in sequence.

In this embodiment, this application of sample card can realize the application of sample of many people, and easy operation, be favorable to realizing the automation of operation flow, for example, in prior art, carry out anti-design test to the sample of single person, need carry out 4 applications of sample (the application of sample of the sample that awaits measuring twice, and the application of sample of twice detect reagent), correspondingly, accomplish the blood type of three people and prevent design test and need carry out 12 applications of sample, if adopt artifical manual application of sample, application of sample number of times is various and probably increases the probability of maloperation, reduce detection efficiency, and application of sample card in this exemplary embodiment accomplishes three people's anti-design test only need carry out 5 applications of sample can, application of sample number of times has been reduced, operation flow has both been simplified, the condition of maloperation has been reduced simultaneously, very big improvement detection efficiency.

Further, in some embodiments, by designing and adjusting the inner diameter and the length of the sample separation channel, uniform sample separation of the sample to be detected and the detection reagent can be realized, and the accuracy of the test result is improved.

Preferably, the sample loading card performs sample loading detection by three-stage centrifugation, referring to fig. 8, fig. 8 shows a flow direction of the liquid sample after the sample is loaded to the second sample loading hole (the direction indicated by an arrow in fig. 8 is the flow direction of the liquid sample), and it can be understood that, in order to more clearly illustrate the flowing characteristic of the liquid sample inside the sample loading card, fig. 8 is not drawn according to the actual proportion of the product, but only the necessary structure of the product is briefly shown.

Specifically, when a liquid sample (e.g., a reciprocal cell a or B) is added to the second well, the liquid sample enters the second sampling channel 32 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 sampling channel 32, the liquid does not continue into the first through hole 51. Alternatively, in some embodiments, the first through hole 51 is in a capillary structure (i.e., the first through hole 51 corresponds to an extension of the second sampling 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 that of the first through hole 51, the liquid sample does not continue to enter the first mixing pool 41 through the first through hole 51. In addition, because the sample adding card is horizontally placed or placed in a state close to the horizontal state when in use, at this time, the liquid sample in the second sample distribution channel 32 can enter the first through hole 51 only by overcoming the self gravity effect, and then enters the second mixing pool 42 through the first through hole 51, so that the flow of the liquid sample is further limited, that is, the first through hole 51 has a flow stopping effect; similarly, the liquid sample (e.g., plasma or detection reagent) in each of the other wells enters the corresponding sample separation channel under capillary action and cannot enter the first mixing well or the second mixing well without external force.

After the sample is added into each sample adding hole, the sample adding card and the matched detection card start to be centrifuged, and preferably a three-stage centrifugation method is adopted, in the first horizontal centrifugation process, liquid samples (detection reagents or samples to be detected) in the first, second, third and fourth sample distribution channels simultaneously enter the corresponding first and second mixing tanks through the first and second through holes under the action of first centrifugal force, and are fully mixed to obtain corresponding mixed liquid; in the secondary horizontal centrifugation process, the mixed liquid in the first mixing pool and the second mixing pool enters the corresponding reaction cavity through the third through hole under the action of a second centrifugal force; in the third horizontal centrifugation process, the mixed liquid entering the reaction cavity and the reagent in the reaction cavity are fully mixed and react under the action of a third centrifugal force.

It is understood that the above three horizontal centrifugation processes are continuously performed, and there is no interval or a short interval between the adjacent centrifugation processes.

In the present embodiment, since the first through-hole 51 has a flow stopping function for the liquid sample in the second fractionating passage 32, it also corresponds to a resistance (i.e., a first resistance) against the flow of the liquid sample. Meanwhile, when the liquid sample enters the third through-hole 53, since the inner diameter of the third through-hole 53 is smaller than the inner diameter of the first communication channel 6, the flow of the liquid sample is prevented in the third through-hole 53, that is, the third through-hole 53 also functions as a flow stopper, which is equivalent to a resistance (i.e., a second resistance) generated to the flow of the liquid sample, and the second resistance is set to be larger than the first resistance. Therefore, the set first centrifugal force needs to be larger than the first resistance and smaller than the second resistance, so that the liquid sample in the second sampling channel 32 can only overcome the first resistance and can not overcome the second resistance to enter the first communicating channel 6 under the action of the first centrifugal force.

Likewise, the setting of the first centrifugal force needs to be satisfied such that the liquid sample in the first, third and fourth sample distribution channels enters the first mixing pool or the second mixing pool, while the liquid sample in the first and second mixing pools cannot continue to enter the first communication channel.

Further, in some embodiments, when the volume of the first and second mixing wells is larger than the volume of the mixed solution to be actually contained, for example, the volume of the first mixing well is larger than about 20 μ L, and the volume of the mixed solution is about 10 μ L, the mixed solution can only occupy about half of the volume of the mixing well, and when the sample adding card is horizontally placed, the liquid level of the mixed solution in the first and second mixing wells may be lower than the height of the connection portion of the third through hole and the first mixing well, and a certain external force is required to enter the third through hole, which further provides a certain resistance to the flow of the liquid sample.

Of course, in some embodiments, it is also possible to cover the third through-going hole with a hydrophobic layer having hydrophobic properties, which will also provide some resistance to the flow of the liquid sample.

Preferably, in some embodiments, the centrifugal speed/force set during the first, second, and third horizontal centrifugation is gradually increased.

Further, in some embodiments, the first centrifugal force is about 9-16g, the centrifugation time is about 10s-1min, the second centrifugal force is about 55g, the centrifugation time is about 1min50s-1min, the third centrifugal force is about 200g, and the centrifugation time is about 3 min.

In a specific experimental operation process, referring to fig. 10 (fig. 10 is a schematic diagram of an application card), the application card in fig. 10 is horizontally placed, after a liquid sample is applied to a first application hole 21, the liquid sample fills a first application channel under capillary action (at this time, a very small portion of the liquid sample may enter a connection portion between a through hole and the first application channel), and does not enter a first mixing pool, referring to fig. 11 (fig. 11 is a schematic diagram of an application card), the application card in fig. 11 is horizontally placed, and liquid samples are applied to a second application hole 22 and a third application hole 23 respectively, wherein the liquid samples in the second application hole and the third application hole respectively enter corresponding second application channel and third application channel under capillary action, but because the length of one of the second application channels is long, although the second application channel has strong capillary action, the flow rate of the liquid sample is still small, even the added liquid sample may not fill the whole second sample distribution channel under the capillary action, but in the first centrifugation process, the liquid sample in the second sample distribution channel can rapidly enter the corresponding second mixing pool under the combined action of the first centrifugal force and the stronger capillary action, so as to achieve the effect that the liquid sample which is introduced into the second mixing pool and the third mixing pool enters the first mixing pool at the same time.

Further, in some embodiments, two or more air holes may be provided in the longer sample distribution channel for fluid drainage.

Example two

Based on the microfluidic sample application card in the first embodiment, the invention further provides a microfluidic sample application card, which comprises the components in the first embodiment, wherein the inner diameter width of the first sample distribution channel is between about 0.2 mm and about 0.8mm, the inner diameter width of the second sample distribution channel is between about 0.1 mm and about 0.5mm, and the inner diameter width of the third sample distribution channel and the inner diameter width of the fourth sample distribution channel are between about 0.2 mm and about 1.5 mm.

It is understood that, in order to ensure that each sample distribution channel can realize quantitative sample distribution, the volumes of the liquid samples contained in the first sample distribution channels are equal or similar, the volumes of the liquid samples contained in the second sample distribution channels are equal or similar, and the volumes of the liquid samples contained in the third sample distribution channels and the fourth sample distribution channels are equal.

Further, in order to enable a plurality of sample channels to be arranged on the same sample application card, there is a difference in the arrangement of the length and the width of each sample channel, for example, the length of some sample channels is longer, and accordingly, the inner diameter (i.e., the width) of the sample channel is smaller, the capillary action of the sample passing through the sample channel is stronger, the length of part of the sample channel is shorter, the inner diameter (i.e., the width) of the sample channel is relatively larger, and accordingly, the capillary action is relatively smaller.

To avoid fluid channeling between sample presentation channels (i.e., fluid exchange between adjacent sample presentation channels), the spacing between adjacent sample presentation channels is greater than about 0.3 mm.

Further, in order to prevent liquid channeling between the first and second mixing pools (i.e., the mixed liquid in the first mixing pool enters the second mixing pool, or the mixed liquid in the second mixing pool enters the first mixing pool), the interval between the adjacent first and second mixing pools is greater than about 0.5 mm.

In some embodiments, the volume of the first and second mixing wells (the amount of liquid sample that can be contained) is greater than about 10 μ L.

Of course, in other embodiments, the volume of the first and second mixing wells (which can hold a volume of liquid sample) needs to be greater than about 20 μ L.

Preferably, to avoid interference between different first communication channels, in some embodiments, the second ends of adjacent first communication channels are spaced more than about 4mm apart.

EXAMPLE III

Based on the microfluidic sample adding card in the above embodiment, the present invention further provides a microfluidic sample adding assembly, including: the sample adding card and the detection card in any of the above embodiments (see the above embodiments for specific structures), wherein the sample adding card is provided with a clamping opening capable of being inserted into the detection card, and when the sample adding card is pushed into the clamping opening, the first communication channels of the sample adding card are respectively communicated with the corresponding reaction cavities on the detection card.

For example, in some embodiments, a position-limiting protrusion 71 (shown in fig. 4) is disposed at the clamping opening of the sample-adding card, a position-limiting slot 10 (shown in fig. 7a and 7 b) is disposed on the detection card, and when the detection card is pushed into the clamping opening and the position-limiting protrusion on the sample-adding card is matched with the position-limiting slot on the detection card, the second ends of the first communicating channels on the sample-adding card are respectively communicated with the open ends of the corresponding reaction chambers on the detection card. Of course, a limit slot may be provided at the clamping opening, and a limit protrusion corresponding to the limit slot may be provided on the detection card.

Preferably, in some embodiments, the sample application assembly further comprises: reagents pre-stored in the reaction chamber.

Further, in some embodiments, the reagents pre-existing in the reaction chamber include: a gel and a working fluid, wherein the working fluid comprises: an antibody. For example, when the sample adding assembly needs to be applied to ABO blood group positive typing, an anti-A blood group typing reagent and an anti-B blood group typing reagent are pre-stored in the reaction cavity respectively.

Further, to prevent the reagent from drying out, in some embodiments, the detection assembly (i.e., the sample application assembly) further comprises: the paraffin oil is arranged in the reaction cavity and used for sealing the reagent, and can be understood as being arranged above the reagent for achieving the purpose of sealing the reagent, so that the paraffin oil and the inner wall of the reaction cavity are enclosed to form a closed structure for storing the reagent.

Specifically, in some embodiments, in the production process of the sample adding assembly, after the corresponding reagent is added into the reaction cavity, the melted paraffin oil is added into the open end of the reaction cavity, and 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, so that the paraffin oil is melted.

Of course, in other embodiments, the reagents are sealed by placing a thin film over the open end of the reaction chamber.

Preferably, the sample application component is centrifuged horizontally, i.e. the sample application component is centrifuged by means of a horizontal centrifuge.

Example four

Based on the microfluidic sample application card (i.e., the sample application device) in the above embodiment, the present invention further provides a microfluidic sample application assembly, including any one of the sample application cards in the above embodiments, and a detection card (see the above embodiment for a specific structure), where the sample application card is provided with a clamping opening capable of being inserted into the detection card, and a sidewall of the clamping opening is provided with a first clamping position and a second clamping position at intervals along a sidewall extending direction;

the detection card is arranged at the second clamping position in a mode of moving relative to the clamping opening;

when the detection card moves from the second clamping position to the first clamping position under the action of external acting force, the first communication channels on the sample adding card are respectively communicated with the corresponding reaction cavities on the detection card.

In some embodiments, the first clamping position is provided with a limiting clamping protrusion or a limiting clamping groove.

In some embodiments, a limiting clamping protrusion or a limiting clamping groove is arranged on the second clamping position.

In an exemplary embodiment, the test card is mounted on the sample card at the second clamping position of the clamping opening when the assembly is shipped (or in the initial state). For example, the detection card is fixedly connected with the second clamping position through a limit card protrusion arranged on the detection card (specifically, the connection part is provided with a breaking line which is easy to break). Under the state, a certain distance e is arranged between the second end of the first communicating channel and the open end of the reaction cavity, and the distance is slightly smaller than or equal to the spacing distance between the first clamping position and the second clamping position, namely the first communicating channel and the reaction cavity are not in a communicating state. And when the detection card is pushed to the first clamping position, and the detection card is moved from the second clamping position to the first clamping position (for example, the limiting clamping protrusion is in clamping fit with the limiting clamping groove), the second end of the first communication channel is communicated with the reaction cavity.

In this embodiment, add appearance card and detection card can be preserved with the subassembly form, and because add appearance card and detection card second joint position department fixed connection, thereby can avoid this product to produce the collision and take place to damage in the transportation or transportation.

EXAMPLE five

Based on the sample adding card described in the above embodiment, the present invention also provides a microfluidic sample adding assembly, including: the sample adding card and the detection card (see the above embodiment for specific structure) according to any of the above embodiments, different from the above embodiment, wherein the sample adding card is provided with a clamping opening capable of being inserted into the detection card, and one side or two sides of the detection card are provided with a second clamping position and a first clamping position at intervals along a direction gradually far away from the opening end of the reaction cavity;

when the detection card moves from the second clamping position to the first clamping position under the action of external acting force, the first communication channel of the sample adding card is respectively communicated with the corresponding reaction cavity on the detection card.

When the detection card moves from the second clamping position to the first clamping position under the action of external acting force, the second ends of the first communication channels of the sample adding card are respectively communicated with the corresponding reaction cavities.

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

For example, in some embodiments, a first limiting clamping groove and a second limiting clamping groove are respectively arranged at one side or two sides of the detection card corresponding to the first clamping position and the second clamping position; accordingly, the snap opening comprises: a guide rail (specifically, two side walls extending from the body, or guide rails arranged on the two side walls along the length direction thereof) for providing a sliding path for the detection card, and a limit card protrusion capable of being matched with the first limit card slot and the second limit card slot is arranged on the guide rail;

when the limiting card bulge is matched with a second limiting card slot on the detection card under the action of external acting force, the detection card and the card-connecting opening are connected at the second connecting position in a clamping manner, and at the moment, the first communicating channel of the sample-adding card is not communicated with the reaction cavity of the detection card, namely, the opening end of the reaction cavity is still at a certain distance from the second end of the first communicating channel;

when the limiting clamp is separated from the second limiting clamp groove under the action of an external acting force, the detecting clamp can move towards the direction close to the second end of the first communicating channel along the guide rail; and when the first limit clamping groove on the detection card moves to the limit clamping convex position on the guide rail and is matched with the limit clamping convex position, the detection card is clamped at the first clamping position with the clamping opening, and at the moment, the second end of the first communication channel 6 is communicated with the corresponding reaction cavity.

EXAMPLE six

Based on the above embodiments, the present invention further provides a sample adding assembly, which includes the sample adding card and a detection card, wherein the detection card, referring to fig. 7b, includes a plurality of reaction chambers, and the reaction chambers include a reagent storage region 11a and a reaction chamber open end 11b, in order to realize the mutual cooperation between the sample adding device and the detection card, preferably, in some embodiments, the second end of the first communicating channel extends to the outside of the body, and the second end of the first communicating channel is provided with a matching portion matching with the reaction chamber open end 11b of the detection card; when the fitting portion is fitted to the open end 11b of the reaction chamber, the second flow passage communicates with the corresponding reaction chamber.

In some embodiments, the interior of the open end of the reaction chamber of the detection card is a tapered structure with a large top and a small bottom (i.e. the cross section is a trapezoidal or approximately trapezoidal structure), and correspondingly, the mating portion of the second end of the first communication channel 6 is also a tapered structure with a large top and a small bottom, and the outer diameter of the mating portion of the first communication channel is slightly smaller than the inner diameter of the open end of the reaction chamber, so that the mating portion can extend into the interior of the open end of the reaction chamber.

It will be understood that the structure of the fitting portion is configured to be capable of fitting with the inside of the open end of the reaction chamber, for example, in some embodiments, when the open end of the reaction chamber is a tapered structure with a large top and a small bottom, the fitting portion is suitably configured to be a tapered structure with a large top and a small bottom, and the outer diameter of the fitting portion is slightly smaller than the inner diameter of the open end of the reaction chamber.

For another example, in some embodiments, when the open end of the reaction chamber is a cylindrical structure, the fitting portion is configured as a cylindrical structure adapted thereto, and the outer diameter of the fitting portion is slightly smaller than the inner diameter of the open end of the reaction chamber.

In some embodiments, paraffin oil is further provided at the edge of the fitting portion to prevent liquid from being suspended at the edge of the fitting portion without flowing into the reaction chamber.

Preferably, in some embodiments, in order to prevent the liquid sample entering the reagent storage region in the reaction chamber from flowing back into the sample application card, the reagent storage region has an inner diameter with a width of about 1.2mm to 1.4mm, so that when the detection card is horizontally placed or nearly horizontally placed, the reagent or liquid sample in the reagent storage region hardly flows when the detection card is not subjected to an external force. Of course, the inner diameter of the reagent storage region may be set to other dimensions so long as it is satisfied that the liquid sample in the reagent storage region hardly flows when the card is horizontally placed or is placed close to horizontally.

Further, in some embodiments, in order to prevent the reagent in the reaction chamber from flowing back to the sample application device due to external force when the detection card is pushed, the liquid level of the reagent in the detection card is usually at a distance from the open end of the reaction chamber, so that the reagent does not flow back to the sample application device even though the reagent flows. For example, in one embodiment, the reagent storage region is 14-15mm in length, and when a specified amount of reagent is added, the reagent is located in a region near the lower end of the reagent storage region (i.e., the side away from the open end of the reaction chamber), and the reagent level is located at a distance of about 7-8mm from the open end of the reaction chamber.

Preferably, in a specific embodiment, the volume of the storage region in the reaction chamber is greater than about 20 μ L.

EXAMPLE seven

Based on the above embodiment, the present invention further provides a sample-adding and detecting integrated detection card/sample-adding card, which is different from the above embodiment in that the detection card/sample-adding card further comprises: with the matched with detection zone in adding the appearance card, be provided with a plurality of reaction chambers on the detection zone, and this reaction chamber is linked together with the second end of corresponding first intercommunication passageway respectively, wherein, be provided with the gas pocket that is used for solving inside and outside pressure differential on the reaction chamber (be close to open end department).

Wherein, this gas pocket is used for solving the sample and divides the pressure differential when the appearance, and in this embodiment, the gas pocket has played and has solved first communicating channel to and the effect of the interior external pressure differential of reaction chamber, because in practical application, each branch appearance passageway in the sample card, first, two mixed ponds, first communicating channel and reaction chamber all adopt enclosed construction (for example, cover through hydrophilic layer and seal), for example, first branch appearance passageway 31 and this first mixed pond that divides the appearance passageway to be linked together to and corresponding first communicating channel and reaction chamber form a confined space jointly. When the liquid sample added into the first sample adding hole enters the corresponding sample dividing channel or the corresponding mixing pool for 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, namely, the pressure difference is formed between the closed space and the atmospheric pressure, and the liquid sample cannot continue to flow.

Of course, in other embodiments, an air hole may be provided on the first communicating channel to solve the problem of the pressure difference between the reaction chamber and the first communicating channel.

Specifically, the reaction chamber and the corresponding first communication channel are integrally formed, that is, the reaction chamber may be formed by extending a certain length along a direction away from the through hole through the first communication channel.

In this embodiment, this application of sample card has adopted application of sample and the design that detects the integral type structure, on the one hand, to add the application of sample card and detect card matched with step has been reduced, the operation flow that the blood type detected has further been simplified, the detection efficiency is improved, be favorable to detecting the automation of flow more, on the other hand, the application of sample card (also detect the card) of integral type is compared in split type detection card among the prior art, the structure is simpler, thereby the production technology has been simplified to a certain extent, the production cost is reduced, in addition, compare in two relatively independent parts among the split type detection card, the integral type adds the application of sample card and is more convenient for deposit and transport.

Meanwhile, in the embodiment, by adopting the sample distribution channel with the closed structure, the first and second flow channels with the closed structure and the reaction cavity with the closed structure, a closed area is provided for sample addition and reaction of blood type detection, so that contact between liquid or reagent inside the detection card/sample addition card and the outside is reduced or avoided to a certain extent, that is, interference on an experimental result caused by external pollutants (such as dust, water drops and the like) entering the detection card is avoided; meanwhile, as each sample adding channel and each reaction cavity are sealed, the liquid in the detection card is prevented from splashing under the action of external acting force, so that the environmental pollution and cross pollution are caused, and the accuracy of the detection result is improved; in addition, the invention solves the problem of pressure difference of the closed structure by arranging the corresponding air holes, ensures that the air pressure in the closed structure is equal to or close to the atmospheric pressure, and ensures that the liquid sample in the closed structure can smoothly flow down under the action of external acting force.

Example eight

Based on the above embodiment, the present invention further provides another sample adding assembly, including: the difference with the above embodiment is that the air hole for solving the internal and external pressure difference is arranged on the first communicating channel of the sample adding card, so that when the detecting card is completely pushed into the sample adding opening, the second end of the first communicating channel of the sample adding card is closely attached to the opening end of the reaction cavity on the detecting card.

Of course, in other embodiments, air holes may be provided in the reaction chamber to solve the problem of pressure difference in the reaction chamber.

Example nine

Based on the above embodiments, the present invention further provides a sample adding assembly, which includes each component or structure of the sample adding assembly in the above embodiments, and further, referring to fig. 9 (it can be understood that, in order to more clearly illustrate the technical solution and effect of the present invention, fig. 9 is not drawn according to the actual scale of the product, and only shows the main structure of the sample adding assembly that meets the detection application), when the fitting portion 61 is fitted to the reaction chamber open end 11b, the lower end surface (the end having a smaller opening in the tapered structure) of the fitting portion 61 is not in direct contact with the lower end surface of the reaction chamber open end, but a certain space is left, so that even if a small part of the reagent in the reagent storage region flows into the opening end of the reaction cavity under the action of an external acting force, the reagent can only stay in the region shown by the dotted triangle 61a and can not enter the first communication channel 6.

Example ten

Based on the sample adding assembly of the above embodiment, the present invention further provides a detection method, wherein the sample adding card in the selected sample adding assembly includes a first sample adding hole, a second sample adding hole, and three third sample adding holes, and accordingly, the method includes the steps of:

respectively adding a first liquid sample, a second liquid sample and a third liquid sample into a first sample adding hole, a second sample adding hole and a third sample adding hole of the sample adding card, wherein the first liquid sample, the second liquid sample and the third liquid sample respectively enter a corresponding first sample adding channel, a corresponding second sample adding channel, a corresponding third sample adding channel and a corresponding fourth sample adding channel;

carrying out first horizontal centrifugation on the sample adding component, and respectively enabling liquid samples in the first, second, third and fourth sample dividing channels to enter the corresponding first and second mixing pools under the action of first centrifugal force to obtain corresponding mixed liquor;

performing second horizontal centrifugation on the sample adding component, wherein under the action of second centrifugal force, the mixed liquid in the first mixing pool and the second mixing pool enters the corresponding reaction cavity through the third through hole and the first communication channel;

In some embodiments, the first fluid sample is retrograded cell a, the second fluid sample is retrograded cell B (or the first fluid sample is retrograded cell B, the second fluid sample is retrograded cell a), the third fluid sample is plasma from a different subject, e.g., plasma from subjects three and three, respectively, and plasma from ethylene propylene is added to three, respectively, third addition wells.

It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.

While the present invention has been described with reference to the embodiments shown in the drawings, the present invention is not limited to the embodiments, which are illustrative and not restrictive, and it will be apparent to those skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A method for detecting based on a sample adding component is characterized in that the sample adding component comprises: add sample card and detection card, add the sample card and include: a body;

the at least one first mixing pool and the at least one second mixing pool are arranged in the body at intervals, 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; each second mixing pool is respectively communicated with the second end of at least one corresponding second sample distribution channel and the second end of at least one corresponding fourth sample distribution channel;

the first mixing tank and the second mixing tank are both provided with at least one first communication channel, and the first communication channels are arranged on the first side of the body at intervals;

the first communication channels of the sample adding card are respectively communicated with corresponding reaction cavities on the detection card; accordingly, the method comprises the steps of:

carrying out third horizontal centrifugation on the sample adding component, and under the action of third centrifugal force, fully mixing the mixed solution entering the reaction cavity with a reagent prestored in the reaction cavity and carrying out reaction;

2. The method according to claim 1, wherein at least one of the first sampling channels is provided with at least one air hole; and/or at least one air hole is arranged on at least one second sampling channel; and/or at least one third sample distribution channel is provided with at least one air hole, and/or at least one fourth sample distribution channel is provided with at least one air hole.

3. The method of claim 1, wherein the first sample channel is in communication with the first mixing well through a first through-hole; and/or the presence of a gas in the gas,

the second sample separation channel is communicated with the second mixing tank through a first through hole; and/or the presence of a gas in the gas,

the third sample distribution channel is communicated with the first mixing pool through a second through hole, and/or,

the fourth sample distribution channel is communicated with the second mixing pool through a second through hole; and/or the presence of a gas in the gas,

the first communicating channel is communicated with the first mixing tank through a third through hole, and the first communicating channel is communicated with the second mixing tank through a third through hole; and/or the presence of a gas in the gas,

the sample adding card further comprises: the first waste liquid cavity is arranged in the body and communicated with the corresponding first sample adding hole; and/or the presence of a gas in the gas,

the sample adding card further comprises: the second waste liquid cavity is arranged in the body and communicated with the corresponding second sample adding hole; and/or the presence of a gas in the atmosphere,

the sample adding card further comprises: the third waste liquid cavity is arranged in the body and communicated with the corresponding third sample adding hole.

4. The method of claim 1, wherein first, second, third and fourth spline channels are provided on a second surface of the body, and correspondingly, a hydrophilic layer is provided on a first region of the second surface of the body, wherein the first region comprises: a region provided with a first, a second, a third and a fourth sampling channels; and/or the presence of a gas in the gas,

the first mixing pool and the second mixing pool are arranged on the first surface of the body, correspondingly, a hydrophobic layer or a hydrophilic layer is arranged on the second area of the first surface of the body, wherein the second area comprises: a region provided with a first mixing tank and a second mixing tank; and/or the presence of a gas in the gas,

an inner surface region of the sample application card is treated with paraffin oil, wherein the inner surface region comprises: the inner surfaces of the first, second, third and fourth sample distribution channels and/or the inner surface of the first communication channel; and/or the presence of a gas in the gas,

the sample adding card comprises: one said first well, one said second well and three said third wells, and accordingly, said sample card comprises: the three first mixing tanks and the three second mixing tanks are arranged inside the body at intervals.

5. The method according to claim 3, wherein there is an overlap area between the orthographic projection of at least one of the first sampling channels and the orthographic projection of at least one of the first through holes, and/or,

an overlapping area exists between the orthographic projection of at least one second sampling channel and the orthographic projection of at least one first through hole; and/or the presence of a gas in the gas,

the interval sets up the body is inside three the third application of sample hole is respectively: the third sample adding hole in a left side, well third sample adding hole, right third sample adding hole, wherein, first sample adding hole is located third sample adding hole in a left side with on the symmetry axis of well third sample adding hole, the second sample adding hole is located well third sample adding hole with on the symmetry axis of third sample adding hole in a right side, just the horizontal direction at first sample adding hole place with the horizontal direction at second sample adding hole place all is located three the top of third sample adding hole.

6. The method of any of claims 1-5, wherein the sample card further comprises: the clamping opening is formed by side walls extending from two sides of the body and is used for being inserted with the detection card;

7. The method according to claim 1, wherein the sample adding card and the detection card are integrally formed, and the detection card is provided with reaction chambers respectively communicated with the corresponding first communication channels, wherein the reaction chambers are provided with air holes for solving the pressure difference, and/or the first communication channels are provided with air holes for solving the pressure difference.

8. The method according to claim 1, wherein the sample adding card is provided with a clamping opening capable of being inserted into and removed from the detection card, when the sample adding card is pushed into the clamping opening, the first communication channels of the sample adding card are respectively communicated with corresponding reaction cavities on the detection card, and a gap for ventilation is formed between the first communication channels and the reaction cavities;

or, add be provided with on the appearance card can with the joint opening of grafting is opened to the detection card, wherein, be provided with the gas pocket that is used for solving pressure differential on the first intercommunication passageway, and/or, be provided with the gas pocket that is used for solving pressure differential on the reaction chamber, work as the detection card is pushed during the joint opening, the second end of the first intercommunication passageway on the appearance card respectively with corresponding reaction chamber is linked together on the detection card, just first intercommunication passageway with laminate closely between the reaction chamber.

9. The method of claim 8, wherein the sample application assembly further comprises: reagents pre-stored in the reaction chamber, wherein the reagents comprise: a gel and a working fluid, wherein the working fluid comprises: an antibody.

10. The method of claim 1, wherein the second end of the first communication channel is provided with a pointed tip for puncturing.