A multi-channel synchronous detection reagent card

Abstract

The application discloses a kind of multi-channel synchronous detection reagent cards, comprising: outer shell;Sample port is used to fill sample;Shunt module is configured as several microcolumn arrays, for according to size size shunt target component in sample;Barrier tape is arranged at the entrance side of shunt module, for blocking non-target component in sample;Detection channel is configured as several, detection channel is built-in individual detection reagent strip;Drainage channel is configured as several, each drainage channel is independently connected shunt module and detection channel respectively;Drainage channel includes: long-distance drainage channel and short-distance drainage channel.The application utilizes microcolumn array in shunt module to separate target component according to size, and guides component flow by designing drainage channel of different lengths, and different sizes of component can reach detection area in different paths simultaneously, solve the problem that detection is not synchronized due to chromatography speed difference in traditional technology, and detection time is shortened to 1 / 2-2 / 3 of traditional method.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of immunodiagnosis, in particular to a multi-channel synchronous detection reagent card. BACKGROUND

[0002] As an innovative analysis method developed at the end of the 20th century, the immunochromatographic detection technology is an organic integration of immunology principles and chromatography technology, which is developed on the basis of immunofiltration technology and forms an immunological detection system with the characteristics of rapid response and simple operation.

[0003] In the current design of multi-target synchronous detection cards, although different types of targets such as antibodies, bacteria, viruses, and toxins are recognized through a multi-channel architecture, and each channel is configured with an independent target recognition reagent strip, there are inherent technical bottlenecks. When mixed samples pass through multi-porous media including a water absorption pad, a sample pad, a labeled pad, and an analysis film, biological particles of different sizes show significant differences in chromatographic rates. Specifically, bacteria with larger diameters have the slowest migration speed due to the largest pore resistance, while toxins with smaller molecular weights migrate quickly due to the smallest flow resistance. This size-dependent migration characteristic makes it difficult to achieve multi-target synchronous detection.

[0004] In addition, non-target components (such as red blood cells, microbial fragments, protein polymers, and contaminants) widely exist in complex matrix samples, which significantly increase the viscosity of the sample processing fluid. The pore blocking effect of macromolecular substances on the chromatographic medium further exacerbates the instability of fluid dynamics. These factors together cause the analysis time of the existing detection system to be generally extended to 10-15 minutes, severely restricting the improvement of detection efficiency.

[0005] Therefore, it is necessary to provide a multi-channel synchronous detection reagent card to solve the above technical problems. SUMMARY

[0006] The present application overcomes the shortcomings of the prior art and provides a multi-channel synchronous detection reagent card.

[0007] To achieve the above purpose, the technical solution adopted by the present application is as follows: a multi-channel synchronous detection reagent card, comprising:

[0008] an outer shell,

[0009] a sample addition port for adding a sample;

[0010] a shunt module configured as an array of microcolumns for separating target components in the sample according to size;

[0011] a barrier band arranged on the inlet side of the shunt module for blocking non-target components in the sample;

[0012] Detection channels are configured as several, and a separate detection reagent strip is built in each detection channel;

[0013] Drainage channels are configured as several, and each of the drainage channels is independently connected to the shunt module and the detection channel respectively; the drainage channels include long-distance drainage channels and short-distance drainage channels;

[0014] The shunt module includes a large molecule flow channel and a small molecule flow channel; the large molecule flow channel is connected to the short-distance drainage channel, and the small molecule flow channel is connected to the long-distance drainage channel.

[0015] In a preferred embodiment of the application, the outer shell includes a snap-connected upper shell and a lower shell.

[0016] The detection channels are parallel to each other and equidistantly distributed, and are all rectangular grooves arranged on the surface of the lower shell.

[0017] In a preferred embodiment of the application, the long-distance drainage channels are distributed on both sides, the short-distance drainage channels are distributed at the middle position, and the length of the long-distance drainage channels connected between the shunt module and the detection channel is greater than that of the short-distance drainage channels.

[0018] In a preferred embodiment of the application, the long-distance drainage channel includes a circular arc segment extending towards the large molecule flow channel, and a connecting segment corresponding to the detection channel.

[0019] The drainage channel is provided with a drainage pad.

[0020] In a preferred embodiment of the application, one side of the barrier strip corresponds to the sample addition port, and the other side is connected to the inlet of the shunt module.

[0021] The barrier strip is configured as a 3D microporous polyurethane foam with a pore size of 20-80 μm.

[0022] In a preferred embodiment of the application, the barrier strip includes:

[0023] An outer layer, the pore size of the outer layer is 70-80 μm, and the minimum thickness is 0.6 mm;

[0024] A middle layer, the pore size of the middle layer is 40-50 μm, and the minimum thickness is 0.5 mm;

[0025] An inner layer, the pore size of the inner layer is 20-30 μm, and the minimum thickness is 0.4 mm.

[0026] In a preferred embodiment of the present application, the thickness of the barrier strip is 1.6-2.0 mm, and the distance between the barrier strip and the sample inlet of the flow splitting module is controlled to be 1.0-1.5 mm; the sample inlet side of the barrier strip adopts a tapered transition structure with a taper angle of 15-20°.

[0027] In a preferred embodiment of the present application, the flow splitting module further comprises:

[0028] an inlet area arranged at the upper part of the flow splitting module, the size of the inlet area being consistent with that of the barrier strip;

[0029] a sorting area, the central part of which is a microcolumn array, the small molecule flow channels being symmetrically arranged on both sides, and the large molecule flow channels being arranged at the end of the straight path;

[0030] the cross section of the microcolumn is an equilateral triangle, and the microcolumn array is arranged at a certain inclination angle in the horizontal direction, the inclination angle θ being 5.7°, and the minimum gap between adjacent microcolumns being 8-10 μm.

[0031] In a preferred embodiment of the present application, the width of the microcolumn array is 1-1.5 times the length of the inlet area, and the aspect ratio of the microcolumn array is 1.5-2:1.

[0032] In a preferred embodiment of the present application, the sorting area further comprises a primary sorting area in the middle part and secondary sorting areas on both sides; the minimum gap between adjacent microcolumns in the primary sorting area is 8-10 μm, and the minimum gap between adjacent microcolumns in the secondary sorting area is 2-5 μm.

[0033] The present application solves the defects in the background art and has the following beneficial effects:

[0034] (1) The present application proposes a multi-channel synchronous detection reagent card, which separates target components according to size by using the microcolumn array in the flow splitting module, and guides the components to flow by designing different lengths of drainage channels - large molecules are guided to short paths, and small molecules are guided to long paths. In this way, components of different sizes (such as bacteria, viruses and toxins) can reach the detection area in different paths at the same time. This method ensures that components of various sizes can reach the detection area synchronously, solves the problem of asynchronous detection caused by the difference in chromatography speed in traditional technology, and shortens the detection time to 1 / 2-2 / 3 of that of traditional methods.

[0035] (2) The barrier strip of the present application is made of 3D micro-porous polyurethane foam, and the designed gradient pore size is optimized by a hierarchical filtration mechanism and parameters, which realizes efficient interception of non-target components and lossless passage of target objects, purifies the sample and reduces the viscosity, and indirectly improves the flowability of the sample and the speed of immunochromatography.

[0036] (3) The present application realizes the directional separation of large and small cells in the sample by the setting of the shunt module, the setting of the micro column array, and the control of the minimum gap between adjacent micro columns, so that the part entering the long distance drainage channel has the largest proportion of small cells, and the part entering the short distance drainage channel has the largest proportion of large cells. Compared with the prior art, the proportion of the presence of corresponding size cells in the long and short distance drainage channels is opposite, that is, the proportion of small cells in the short distance drainage channel is large, and the proportion of large cells in the long distance drainage channel is large, which leads to the fact that the designated detection reagent strip cannot accurately measure the target component.

[0037] (4) The present application significantly optimizes the target distribution by the size exclusion and flow field disturbance of the micro column array, solves the channel crosstalk problem caused by natural random distribution. Moreover, the aspect ratio of the micro column array is within 1.5-2:1, which is the best balance point of separation and time cost, combined with the pre-blocking belt, the detection time can be greatly reduced. BRIEF DESCRIPTION OF DRAWINGS

[0038] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments described in the present application, and other drawings can be obtained by those skilled in the art without creative labor;

[0039] Figure 1 is the overall top view of the shell body of the preferred embodiment of the present application;

[0040] Figure 2 is the internal structure diagram of the lower shell of the preferred embodiment of the present application;

[0041] Figure 3 is the spatial distribution schematic diagram of the sorting area, large molecule flow channel and small molecule flow channel of embodiment 1 of the present application;

[0042] Figure 4 is the micro column array schematic diagram of the preferred embodiment of the present application.

[0043] In the figure: 1, upper shell; 2, lower shell; 3, sample inlet; 4, blocking belt; 5, detection channel; 6, shunt module; 61, sorting area; 62, large molecule flow channel; 63, small molecule flow channel; 64, baffle; 7, drainage channel; 8, long distance drainage channel; 9, short distance drainage channel; 10, micro column array. DETAILED DESCRIPTION

[0044] Clearly, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present application.

[0045] In the following description, many specific details are set forth in order to provide a thorough understanding of the present application. However, the present application can be practiced without the specific details, other than those described herein, and therefore the scope of the present application is not limited to the details of the following description.

[0046] In the description of the present application, it should be understood that the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the scope of protection of the present application.

[0047] In the description of the present application, it should be noted that unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connection" should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or integrally connected; it can be mechanically connected, or it can be electrically connected; it can be directly connected, or it can be indirectly connected through an intermediate medium; it can be the communication between the two elements inside. For those of ordinary skill in the art, the specific meaning of the above terms in the present application can be understood through specific circumstances.

[0048] Embodiment 1

[0049] As shown in Figure 1 The present application provides a multi-channel synchronous detection reagent card, which is a multi-channel synchronous detection reagent card based on biological probe immunochromatography, comprising:

[0050] An outer shell, which comprises a snap-on connected upper shell 1 and lower shell 2.

[0051] A sample port 3 for adding sample.

[0052] A shunt module 6 configured as a plurality of micro-column arrays 10 for shunting target components in the sample according to size.

[0053] A barrier tape 4 arranged at the inlet side of the shunt module 6 for blocking non-target components in the sample.

[0054] The detection channels 5 are 3-20 in number, and each detection channel 5 is internally provided with a separate detection reagent strip; the detection channels 5 are parallel to each other and equidistantly distributed, and are all rectangular groove structures arranged on the surface of the lower shell 2; when the outer shell is in the connected state, the surface of the upper shell 1 is in close contact with the surface of the lower shell 2, effectively blocking the sample solution from flowing out of the gaps between different detection channels 5 between the upper shell 1 and the lower shell 2, which can effectively prevent the sample solution from flowing between different detection channels 5 and disturbing the flow of the sample solution after backflow.

[0055] The drainage channels 7 are 3-20 in number, and each drainage channel 7 independently communicates the shunt module 6 and the detection channel 5; the drainage channels 7 include long-distance drainage channels 8 and short-distance drainage channels 9; the long-distance drainage channels 8 are distributed on both sides, the short-distance drainage channels 9 are distributed at the middle position, and the length of the long-distance drainage channels 8 connecting the shunt module 6 and the detection channel 5 is greater than that of the short-distance drainage channels 9.

[0056] The shunt module 6 further includes a large molecule flow channel 62 and a small molecule flow channel 63; the large molecule flow channel 62 communicates with the short-distance drainage channel 9, and the small molecule flow channel 63 communicates with the long-distance drainage channel 8.

[0057] It is worth noting that the detection reagent strips arranged in the detection channels 5 are as follows: the detection reagent strips for detecting small-size toxin targets are arranged in the detection channels 5 close to the edges, and these detection channels 5 correspond to the long-distance drainage channels 8; the detection reagent strips for detecting larger-size bacterial targets are arranged in the detection channels 5 at the central position, and these detection channels 5 correspond to the short-distance drainage channels 9.

[0058] The upper shell 1 and the lower shell 2 cooperate with each other, and both the upper shell 1 and the lower shell 2 are made of hard plastic; the upper shell 1 and the lower shell 2 can be spliced together to form a reagent card for detection work. Specifically, the upper surface of the lower shell 2 is provided with a plurality of sleeve pipes, which are uniformly distributed on both sides of the detection channels 5 and at the top end position in the length direction; the lower surface of the upper shell 1 is provided with a plurality of clamping columns corresponding to the sleeve pipes, and the sleeve pipes and the clamping columns match with each other; the upper shell 1 and the lower shell 2 are spliced together by nesting the clamping columns into the sleeve pipes.

[0059] It also includes an observation port, which is arranged on the outer surface of the upper shell 1 and used for observing the chromatographic state and the final detection result of the detection reagent strips in the detection channels 5; the sample inlet 3 is also arranged on the outer surface of the upper shell 1, and both the sample inlet 3 and the observation port are arranged on the central axis of the upper shell 1 in the length direction.

[0060] Further, the drainage pads are arranged in the drainage channels 7. The long-distance drainage channel 8 includes a circular arc segment extending towards the macromolecular flow channel 62 of the shunt module 6, and a connecting segment towards the corresponding detection channel 5.

[0061] The barrier strip 4 is opposite to the sample port 3, and the other side is connected with the inlet of the shunt module 6. The barrier strip 4 is consistent with the sample port 3 in shape and size or slightly larger. The barrier strip 4 is used to block non-target components in the sample, including blood cells, large-sized microorganisms, cell fragments, impurities, and pollutants. These non-target components have a large molecular volume, which affects the viscosity and flowability of the sample, thereby indirectly affecting the moving speed of the target components. As a result, the sample needs to be detected for a long time, mostly 10-15 minutes, before the target components in the sample can penetrate / move to the designated detection area, and the efficiency is low.

[0062] The barrier strip 4 is specifically a 3D microporous polyurethane foam. The micropores on the 3D microporous polyurethane foam have a pore size of 20-80 μm, which is used to intercept large-sized non-target components such as cell fragments / fibrin clots with a size greater than 20 μm, while allowing bacteria with a size of 0.2-10 μm and viruses with a size of 20-400 nm to pass freely.

[0063] It is worth noting that the thickness of the barrier strip 4 is 1.6-2.0 mm, and the barrier strip 4 is tightly attached to the inlet plane of the shunt module 6, with a distance of 1.0-1.5 mm from the sample port 3. The side of the sample port 3 of the barrier strip 4 adopts a tapered transition structure with a taper angle of 15-20°, which guides the sample to diffuse uniformly.

[0064] The main body of the barrier strip 4 is made of 3D microporous polyurethane foam. Due to its unique microporous structure, it can effectively intercept blood cells, large-sized microorganisms, cell fragments, and various impurities and pollutants in the sample. The sample is purified and the viscosity is reduced, which indirectly improves the flowability of the sample and the speed of the immunochromatography.

[0065] Further, the 3D microporous polyurethane foam is provided with a gradient pore size, and the pore size decreases layer by layer from the outside to the inside, and the non-target components are filtered out layer by layer. The 3D microporous polyurethane foam specifically includes:

[0066] The outer layer has a pore size of 70-80 μm and a minimum thickness of 0.6 mm, which is used to intercept large-particle impurities (such as cell fragments, fibrin clots, etc.)

[0067] The middle layer has a pore size of 40-50 μm and a minimum thickness of 0.5 mm, which is used to further intercept medium-sized particles (such as part of the microorganisms or cell fragment residues)

[0068] The inner layer has a pore size of 20-30 μm and a minimum thickness of 0.4 mm, which is used to intercept the remaining non-target components with a size greater than 20 μm.

[0069] The outer layer of the barrier band intercepts larger particles, reducing the load of the subsequent layers; the inner layer precisely filters to avoid the interception of target components. The design of the gradient pore size in the barrier band balances interception and flow rate, and the thickness and corresponding pore size of each gradient are set to achieve the optimal ratio of filtration and passing time, thereby reducing the total detection time to the maximum extent. Compared with the prior art, even if the time of the sample processing liquid passing through the barrier band is increased, the overall detection time is reduced.

[0070] Further, the 3D microporous polyurethane foam is subjected to plasma cleaning treatment for 3 min, and the plasma cleaning process is controlled as follows: oxygen flow rate 50 sccm, power 100 W. Then, the cleaned polyurethane foam is immersed in a 0.1% polydopamine PDA solution and oscillated for 2 hours to form an adhesive interface layer. It should be noted that the surface contact angle of the polyurethane foam after plasma cleaning treatment is reduced to below 30°, which ensures the interception effect while minimizing the hindering ability of the sample processing liquid.

[0071] The barrier band of the present application is designed and verified as follows.

[0072] Prepare multiple identical sample simulators for control experiments. The sample simulators contain fluorescently labeled bacteria (size 1-3 pm) and virus-like particles (size 100-200 nm), as well as fluorescently labeled microspheres (simulating non-target components such as cell fragments, fibrin clots, etc.) with sizes of 100 pm, 60 pm, 50 pm, 30 pm, 20 pm, and 10 pm.

[0073] Ordinary, a single-channel reagent card for detecting labeled bacteria.

[0074] Experiment group 1: a barrier band is arranged at sample inlet 3 of the single-channel reagent card, and the barrier band comprises:

[0075] The outer layer has a pore size of 80 pm and a thickness of 0.6 mm;

[0076] The middle layer has a pore size of 40 pm and a thickness of 0.5 mm;

[0077] The inner layer has a pore size of 20 pm and a thickness of 0.5 mm.

[0078] Experiment group 2: a barrier band is arranged at sample inlet 3 of the single-channel reagent card, and the barrier band comprises:

[0079] The outer layer has a pore size of 80 pm and a thickness of 0.6 mm;

[0080] The middle layer has a pore size of 30 pm and a thickness of 0.5 mm; (the pore size of the middle layer is reduced)

[0081] The inner layer has a pore size of 20 pm and a thickness of 0.5 mm.

[0082] Experimental group 3: A barrier tape is arranged at the sample inlet 3 of the single-channel reagent card, which includes:

[0083] The outer layer has a pore size of 80 pm and a thickness of 0.6 mm;

[0084] The middle layer has a pore size of 40 pm and a thickness of 1.1 mm; (The thickness of the middle layer is increased)

[0085] The inner layer has a pore size of 20 pm and a thickness of 0.5 mm.

[0086] Experimental group 4: A barrier tape is arranged at the sample inlet 3 of the single-channel reagent card, which includes:

[0087] The outer layer has a pore size of 80 pm and a thickness of 0.6 mm;

[0088] The middle layer has a pore size of 40 pm and a thickness of 0.8 mm;

[0089] The inner layer has a pore size of 20 pm and a thickness of 0.6 mm.

[0090] Experimental group 5: A barrier tape is arranged at the sample inlet 3 of the single-channel reagent card, which includes:

[0091] The outer layer has a pore size of 80 pm and a thickness of 0.6 mm;

[0092] The middle layer has a pore size of 40 pm and a thickness of 0.5 mm;

[0093] The inner layer has a pore size of 10 pm and a thickness of 0.5 mm. (The pore size of the inner layer is reduced)

[0094] Control group 1: A barrier tape is arranged at the sample inlet 3 of the single-channel reagent card, which is a single pore size of 20 pm and has a thickness of 1.6 mm.

[0095] Control group 2: A common single-channel reagent card is directly used.

[0096] The sample simulation agent was randomly divided into 7 groups, 5 samples in each group. A quantitative pipette was used to control the sample volume of each sample to be 500 m L, and the sample simulation agent was injected into the reagent card through the sample inlet 3 of the experimental groups 1-5 and the control groups 1-2 above, and the timer was started at the same time. The time from sample addition to complete color development of the detection line was recorded, and the average value and standard deviation of the detection time of each group were calculated. The experimental data is shown in Table 1.

[0097] Table 1. Control experiment summary

[0098] Group Average detection time (min) Standard deviation Experiment group 1 8.5 ±0.4 Experiment group 2 9.2 ±0.6 Experiment group 3 10.1 ±0.8 Experiment group 4 8.9 ±0.5 Experiment group 5 12.3 ±1.2 Control group 1 13.8 ±1.5 Control group 2 14.1 ±1.8

[0099] From the above experimental results, it can be seen that the gradient pore size design of the experimental group 1 exhibits the best performance, and the chromatography completion time is 8.5±0.4 min, which is shortened by 43% compared with the conventional reagent card of the control group 2. The results show that the gradient pore size structure significantly reduces the sample simulation agent viscosity and improves the fluidity through the step-by-step interception mechanism (the outer layer 80 μm retains 100-60 μm microspheres, the middle layer 40 μm filters 50-30 μm particles, and the inner layer 20 μm removes 20 μm impurities), thereby accelerating the chromatography process of the target (bacteria / virus).

[0100] The average detection time of the experimental group 2 is increased to 9.2 minutes (8.2% longer than the experimental group 1), which indicates that although the reduction of the middle layer pore size to 30 μm can enhance the interception efficiency of 30 μm microspheres, the fluid resistance will be increased due to the decrease of porosity, and the balance between filtration precision and flow rate is needed.

[0101] The detection time of the experimental group 3 is further extended to 10.1 minutes (18.8% longer than the experimental group 1) due to the increase of the middle layer thickness to 1.1 mm, which indicates that the too thick middle layer will increase the sample migration path and offset the efficiency advantage of the gradient design.

[0102] The average detection time of the experimental group 4 is 8.9 minutes (not significantly different from the experimental group 1), which indicates that the appropriate increase of the inner layer thickness (from 0.5 mm to 0.6 mm) can improve the filtration stability by extending the impurity adsorption path, and the influence on the flow rate is controllable.

[0103] It is worth noting that the experimental group 5 also has a 15% bacterial interception rate, and the detection time is increased to 12.3 minutes (44.7% longer than the experimental group 1). This result confirms that the too small inner layer pore size will cause the target (1-3 μm bacteria) to be misintercepted due to the space hindering effect, resulting in the risk of false negative.

[0104] The average detection time of the control group 1 is 13.8 minutes, which is significantly worse than the gradient design. The homogeneous structure causes the large particles (such as 100 μm microspheres) to quickly accumulate at the inlet to form a filter cake layer, which hinders the subsequent sample to pass through, highlighting the anti-clogging advantage of the gradient design.

[0105] In summary, the gradient pore size barrier band realizes the efficient interception of non-target components and the lossless passage of the target through the hierarchical filtration mechanism and parameter optimization, and the detection time is compressed to 56% of the traditional design. The parameter combination of the experimental group 1 (80 / 40 / 20 μm pore size + 0.6 / 0.5 / 0.5 mm thickness) can be used as a standardized design benchmark for multi-channel reagent cards.

[0106] As Figure 3As shown, the flow splitting module 6 includes: an inlet area, a sorting area 61, a large molecule flow channel 62 and a small molecule flow channel 63; wherein the inlet area is arranged at the upper part of the flow splitting module 6, the sorting area 61 is centrally provided with a micro-pillar array 10, and the two sides are symmetrically provided with small molecule flow channels 63 connected to long-distance drainage channels 8, and the end of the straight path is a large molecule flow channel 62 connected to a short-distance drainage channel 9.

[0107] It is worth noting that the size of the inlet area is consistent with that of the barrier band 4, the width of the micro-pillar array 10 is 1-1.5 times the length of the inlet area, and the aspect ratio of the micro-pillar array 10 is 1.5-2:1.

[0108] The length of the micro-pillar array 10 determines the residence time of the particles in the sorting area 61, the longer the length, the higher the probability of collision between the particles and the micro-pillars, and the sorting accuracy is improved. The width is related to the uniformity of the flow field, and too small width will lead to uneven flow rate and uncontrollable particle path deviation; too large width will reduce the sorting efficiency and further affect the overall detection time.

[0109] As shown in FIG. 2, Figure 4 The micro-pillar cross-section is an equilateral triangle with a side length of 30 μm, but it can also be circular, "I" shaped or other shapes. The micro-pillar array 10 is arranged at a certain inclination in the horizontal direction, the inclination angle θ = 5.7°, and the minimum gap between adjacent micro-pillars is 8-10 μm, Figure 3 The middle curve represents the fluid direction. Further, according to the maximum particle size of the detection target, 1-5 μm can be set to perform targeted flow splitting.

[0110] It is worth noting that the channel height in the micro-pillar array 10 is 20 μm, which realizes 3D flow focusing and prevents molecules and cells from escaping vertically.

[0111] In the micro-pillar array 10, particles larger than the critical size will migrate along the inclined arrangement direction, because large cell particles cannot pass through the micro-pillar gap, collide with the micro-pillars in the flow, and are pushed by the fluid shear force to move along the inclined direction of the micro-pillar arrangement in the main flow direction, and enter the large molecule flow channel 62-short distance drainage channel 9; and particles smaller than the critical size can bypass the micro-pillar gap, and are subjected to periodic flow field disturbance to produce lateral displacement, and after passing through N rows of micro-pillars, the total deviation reaches the set value, and significantly deviates from the main flow direction, and enters the two sides. Small molecule channel-long distance drainage channel 8.

[0112] Further, a baffle plate 64 is arranged in the sorting area 61 to divide the positions of the large molecule flow channel 62 and the small molecule flow channel 63; the long-distance drainage channel 8 extends to the small molecule flow channel 63, and based on the drainage pad arranged therein, particles in the small molecule flow channel 63 flow into the long-distance drainage channel 8, and the proportion of small-size particles is high; correspondingly, the short-distance drainage channel 9 extends to the large molecule flow channel 62, and based on the drainage pad arranged therein, particles in the large molecule flow channel 62 flow into the short-distance drainage channel 9, and the proportion of large-size particles is high.

[0113] It should be noted that the baffle plate 64 is arranged at the end of the path of the sorting area 61, between the short-distance drainage channel 9 and the long-distance drainage channel 8, to avoid confusion of particles flowing to the end of the sorting area 61.

[0114] By arranging the shunt module 6, the minimum gap between adjacent microcolumns in the microcolumn array 10 is controlled, the large cells and small cells in the sample are directionally shunted, and thus the proportion of small cells in the part entering the long-distance drainage channel 8 is the largest, and the proportion of large cells in the part entering the short-distance drainage channel 9 is the largest. Compared with the prior art, the proportion of large cells and small cells in the long-distance drainage channel 8 and the short-distance drainage channel 9 is opposite, that is, the proportion of small cells in the short-distance drainage channel 9 is large, and the proportion of large cells in the long-distance drainage channel 8 is large, which causes the designated detection reagent strip to be unable to accurately measure the target component.

[0115] Embodiment 2

[0116] Based on the above embodiment 1, a detection reagent strip for detecting a virus target between two sizes is arranged in the detection channel 5 corresponding to the drainage channel 7 between the long-distance drainage channel 8 and the short-distance drainage channel 9, that is, a medium-distance drainage channel, which is not shown in the figure.

[0117] The corresponding shunt module 6 includes: an inlet area, a sorting area 61, a large molecule flow channel 62, a medium molecule flow channel, and a small molecule flow channel 63; wherein the inlet area is arranged at the upper part of the shunt module 6; the sorting area 61 has a microcolumn array 10 in the center, which is divided into a first-level sorting area 61 in the middle and second-level sorting areas 61 on both sides. The microcolumn array 10 is arranged symmetrically on both sides to connect the long-distance drainage channel 8 through the small molecule flow channel 63, the end of the straight path of the first-level sorting area 61 is connected to the short-distance drainage channel 9 through the large molecule flow channel, and the end of the straight path of the second-level sorting area 61 is connected to the medium-distance drainage channel through the medium molecule flow channel.

[0118] It should be noted that the size of the inlet area is consistent with that of the blocking belt 4, the width of the microcolumn array 10 is 1-1.5 times the length of the inlet area, and the length-width ratio of the microcolumn array 10 is 1.5-2:1.

[0119] Specifically, the micro-pillar cross-section is an equilateral triangle with a side length of 30 pm, but it can also be circular, "I" shaped, or other shapes. The micro-pillar array 10 is arranged at a certain angle in the horizontal direction, with an inclination angle θ = 5.7°, and is divided into a middle primary sorting area 61 and two secondary sorting areas 61 on both sides; wherein the minimum gap between adjacent micro-pillars in the primary sorting area 61 is 8-10 pm; the minimum gap between adjacent micro-pillars in the secondary sorting area 61 is 2-5 pm.

[0120] It is worth noting that the channel height in the micro-pillar array 10 is 20 pm, achieving 3D flow focusing to prevent molecules and cells from escaping vertically.

[0121] The micro-pillar array 10 is arranged, in the primary sorting area 61, particles larger than the critical size will migrate along the inclined arrangement direction, because large cell particles cannot pass through the micro-pillar gap, collide with the micro-pillar in the flow, and are pushed by the fluid shear force to move along the inclined direction of the micro-pillar arrangement in the main flow direction, entering the large molecule flow channel 62 - short distance flow channel 9; and particles smaller than the critical size can bypass the micro-pillar gap, and are laterally displaced by periodic flow field disturbance, entering the secondary sorting area 61. In the secondary sorting area 61, particles larger than the critical size will migrate along the inclined arrangement direction, entering the medium molecule flow channel - medium distance flow channel. Particles smaller than the critical size of the secondary sorting area 61, after passing through N rows of micro-pillars, the total offset reaches the setting, significantly offsetting the main flow direction, entering the small molecule channels on both sides - long distance flow channel 8.

[0122] Further, the baffle 64 is arranged in the sorting area 61 to divide the positions of the large molecule flow channel 62, the small molecule flow channel 63, and the medium molecule flow channel; the long distance flow channel 8 extends to the small molecule flow channel 63, based on the built-in flow guide pad, so that the particles in the small molecule flow channel 63 flow into the long distance flow channel 8, with a high proportion of small size particles; correspondingly, the short distance flow channel 9 extends to the large molecule flow channel 62, based on the built-in flow guide pad, so that the particles in the large molecule flow channel 62 flow into the short distance flow channel 9, with a high proportion of large size particles; the medium distance flow channel extends to the medium molecule flow channel, based on the built-in flow guide pad, so that the particles in the medium molecule flow channel flow into the medium distance flow channel, with a high proportion of medium size particles.

[0123] It should be noted that the baffle 64 is arranged at the end of the path in the sorting area 61, between the short distance flow channel 9 and the medium distance flow channel, and between the medium distance flow channel and the long distance flow channel 8, to avoid confusion of particles flowing to the end of the sorting area 61.

[0124] Through the setting of the shunt module 6, especially the first-stage sorting area 61 and the second-stage sorting area 61, the gradient control of the minimum gap between the micro-pillars, the directional shunting of large, medium and small cells in the sample is realized, specifically the directional shunting of large-size bacteria, medium-size viruses and small-size toxins, so that the corresponding target accounts for a large proportion in the designated drainage channel 7. Compared with the prior art, the proportion of the presence of the corresponding cells in the drainage channel 7 is avoided, which leads to the situation that the designated detection reagent strip cannot accurately measure the target component.

[0125] The design of the shunt module 6 is compared below.

[0126] An 8-channel reagent card is prepared, and 4 long-distance drainage channels are arranged on both sides of the short-distance drainage channel. A sample simulation agent containing fluorescent markers is prepared: large molecule target: 5 μm fluorescent microspheres (simulate bacteria), small molecule target: 200 nm fluorescent virus-like particles.

[0127] Experimental group 6: 8-channel reagent card with shunt module 6, the micro-pillar array 10 in the shunt module 6 is set to a micro-pillar gap of 8-10 μm, an inclination angle θ = 5.7°, and a channel height of 20 μm. Among them, the micro-pillar array 10 is 6 mm wide and 12 mm long.

[0128] Experimental group 7: based on experimental group 6, wherein the micro-pillar array 10 is 6 mm wide and 9 mm long.

[0129] Control group 3: 8-channel reagent card without shunt module 6.

[0130] Control group 4: based on experimental group 6, wherein the micro-pillar array 10 is 6 mm wide and 18 mm long.

[0131] 500 μL of sample simulation agent is added to the inlet area of experimental groups 6, 7 and control groups 3, 4, respectively, the shunting efficiency is evaluated by detecting the fluorescence intensity proportion of the target target in the designated detection channel 5, and the detection time (the average time from sample addition to all detection line coloration) is recorded, which is shown in Table 2.

[0132] Table 2. Shunt control experiment table

[0133]

[0134] From the above experimental data, it can be seen that the shunt module 6 in the experimental group 6 significantly optimizes the target distribution through the size exclusion and flow field disturbance of the microcolumn array 10 (aspect ratio 2:1), and solves the channel crosstalk problem caused by natural random distribution; the shunt efficiency of the microcolumn array 10 in the experimental group 7 with an aspect ratio of 1.5:1 (microcolumn length 9 mm) has a lower increase, and because the shunt time is less, the overall detection time is reduced. In the control group 4, the large molecule sorting efficiency is abnormally high (85.1%), but the small molecule efficiency is only 73.4%, indicating that the ultra-long microcolumn (18 mm) enhances the large molecule sorting, but causes partial misjudgment due to small molecule diffusion, and because the sorting time is longer, the detection time is basically the same as that of the control group 3. Therefore, the aspect ratio of the microcolumn array 10 in the shunt module 6 is within the range of 1.5-2:1, which is the best balance point of shunt and time cost, and combined with the pre-preceding barrier, the detection time required can be greatly reduced.

[0135] The above is based on the ideal embodiment of the application, and through the above description, relevant personnel can make various changes and modifications without deviating from the technical idea of the application. The technical scope of the application is not limited to the content of the specification, and the technical scope must be determined according to the scope of the claims.

Claims

1. A multi-channel synchronous detection reagent card, characterized in that, include: outer shell, Sample dispensing port, used for adding samples; The splitting module is configured as an array of micropillars to split the target components in the sample according to their size. A barrier strip, located on the inlet side of the diversion module, is used to block non-target components in the sample; The detection channel is configured to have several channels, and each detection channel has a built-in individual detection reagent strip. The diversion channel is configured as several, and each diversion channel is independently connected to the diversion module and the detection channel; The drainage channels include: long-distance drainage channels and short-distance drainage channels; The diversion module includes a macromolecular flow channel and a small molecule flow channel; the macromolecular flow channel is connected to the short-distance drainage channel, and the small molecule flow channel is connected to the long-distance drainage channel. The cross-section of the micropillar is an equilateral triangle, and the array of micropillars is arranged at an inclination angle θ=5.7° in the horizontal direction, with a minimum gap of 8-10μm between adjacent micropillars; The barrier strip is configured as a 3D microporous polyurethane foam with a pore size between 20-80 μm, comprising: The outer layer has a pore size of 70-80μm and a minimum thickness of 0.6mm; The intermediate layer has a pore size of 40-50 μm and a minimum thickness of 0.5 mm; and The inner layer has a pore size of 20-30 μm and a minimum thickness of 0.4 mm.

2. The multi-channel synchronous detection reagent card according to claim 1, characterized in that: The outer casing includes: an upper casing and a lower casing connected by snap-fit; The detection channels are parallel to each other and equidistant from each other, and each is a rectangular groove provided on the surface of the lower housing.

3. The multi-channel synchronous detection reagent card according to claim 1, characterized in that: The long-distance drainage channels are distributed on both sides, and the short-distance drainage channels are distributed in the middle. The length of the long-distance drainage channels connecting the diversion module and the detection channel is greater than that of the short-distance drainage channels.

4. The multi-channel synchronous detection reagent card according to claim 1, characterized in that: The long-distance drainage channel includes: an arc segment extending toward the direction of the macromolecular flow channel, and a connecting segment toward the corresponding detection channel; each drainage channel is provided with a drainage pad.

5. A multi-channel synchronous detection reagent card according to claim 1, characterized in that: One side of the barrier band corresponds to the sample inlet, and the other side connects to the inlet of the diversion module.

6. The multi-channel synchronous detection reagent card according to claim 1, characterized in that: The barrier band is 1.6-2.0 mm thick, closely attached to the inlet plane of the diversion module, and the distance between it and the sample inlet is controlled at 1.0-1.5 mm; the sample inlet side of the barrier band adopts a conical transition structure with a cone angle of 15-20°.

7. A multi-channel synchronous detection reagent card according to claim 1, characterized in that, The traffic splitting module further includes: An inlet area is located at the top of the diversion module, and the size of the inlet area is the same as that of the barrier strip; The sorting area has a central array of micropillars, with small molecule channels symmetrically arranged on both sides, while large molecule channels are located at the end of the straight path.

8. A multi-channel synchronous detection reagent card according to claim 7, characterized in that, The width of the micropillar array is 1-1.5 times the length of the entrance region, and the aspect ratio of the micropillar array is 1.5-2:

1.

9. A multi-channel synchronous detection reagent card according to claim 7, characterized in that, The sorting area further includes: a primary sorting area in the middle and secondary sorting areas on both sides; the minimum gap between adjacent microcolumns in the primary sorting area is 8-10 μm, and the minimum gap between adjacent microcolumns in the secondary sorting area is 2-5 μm.

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