Microfluidic fully-automated platelet test method and platelet analysis and stirring system

Abstract

The present application relates to a microfluidic fully-automated platelet test method and a platelet analysis and stirring system. The microfluidic fully-automated platelet test method integrates both the platelet-rich plasma test sampling process and the platelet-poor plasma test sampling process on the same platelet analysis and stirring apparatus, thus the whole apparatus is small in size, the additional arrangement of a centrifuge apparatus for preparing platelet-poor plasma is not needed, and the additional arrangement of a magnetic stirring apparatus for stirring is also not needed, thereby simplifying test steps, reducing costs and improving test efficiency.

Description

Microfluidic full-automatic platelet detection method and platelet analysis stirring system TECHNICAL FIELD

[0001] The present application relates to the technical field of IVD, in particular to a microfluidic full-automatic platelet detection method and a platelet analysis stirring system. BACKGROUND

[0002] LTA (light transmission aggregometry) is the most classic platelet function detection method, which has low detection cost, good correlation with clinical events, and is relatively popular in clinical application. The basic principle is that under specific continuous stirring conditions, an inducer is added to platelet-rich plasma (PRP) to cause the platelets in the PRP to aggregate, the turbidity of the PRP decreases, and a photoelectric tube converts the turbidity change into an electrical signal and records the aggregation curve on a recorder, thereby calculating the degree of platelet aggregation.

[0003] The traditional LTA detection method is to add pre-prepared platelet-poor plasma (PPP) to a cuvette, then perform measurement. After the measurement is completed, the platelet-poor plasma (PPP) in the cuvette is sucked out, and platelet-rich plasma (PRP) and an inducer are added to the cuvette. The detection data is obtained by stirring with a magnetic stirring device.

[0004] However, platelet-poor plasma (PPP) needs to be prepared by an additional centrifuge, which cannot complete platelet-rich plasma (PRP) detection, platelet-rich plasma (PRP) detection, and stirring of platelet-rich plasma (PRP) and an inducer in the same device, resulting in a complex detection procedure, high cost, and low detection efficiency of the traditional LTA detection method.

[0005] SUMMARY

[0006] Therefore, it is necessary to provide a microfluidic full-automatic platelet detection method and a platelet analysis stirring system to solve the problems of complex steps, high cost, and low detection efficiency of using the traditional LTA detection method to detect platelet function.

[0007] In one aspect, the present application provides a microfluidic full-automatic platelet detection method, which comprises:

[0008] Obtaining platelet-rich plasma and introducing the platelet-rich plasma into a disc;

[0009] Centrifuging the disc and transferring the platelet-rich plasma to a detection area of the disc;

[0010] introducing the inducing reagent into the disc;

[0011] centrifuging the disc to transfer the inducing reagent to the detection area of the disc;

[0012] centrifuging the disc to mix and react the platelet-rich plasma and the inducing reagent, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area;

[0013] centrifuging the detection area of the disc to obtain the platelet-poor plasma;

[0014] centrifuging the disc, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area.

[0015] In another aspect, the application further provides a microfluidic automatic platelet detection method, which comprises:

[0016] obtaining a whole blood sample, and introducing the whole blood sample into a sample adding area of a disc;

[0017] centrifuging the disc to separate the whole blood sample into platelet-rich plasma and blood cell precipitate;

[0018] centrifuging the disc to transfer the platelet-rich plasma to a detection area of the disc;

[0019] introducing an inducing reagent into the disc;

[0020] centrifuging the disc to transfer the inducing reagent to the detection area of the disc;

[0021] centrifuging the disc to mix and react the platelet-rich plasma and the inducing reagent, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area;

[0022] centrifuging the detection area of the disc to obtain the platelet-poor plasma;

[0023] centrifuging the disc, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area.

[0024] In another aspect, the application further provides a microfluidic automatic platelet detection method, which comprises:

[0025] obtaining a whole blood sample, and introducing the whole blood sample into a sample adding area of a disc;

[0026] centrifuging the disc to separate the whole blood sample into platelet-rich plasma and blood cell precipitate, and moving the platelet-rich plasma to a plasma area and the blood cell precipitate to a precipitate area;

[0027] centrifuging the disc to transfer the platelet-rich plasma from the plasma area to a quantification area of the disc;

[0028] centrifuging the disc, the platelet-rich plasma in the quantitative area is transferred to the detection area of the disc;

[0029] introducing an inducing agent into the disc;

[0030] centrifuging the disc, the inducing agent is transferred to the detection area of the disc;

[0031] centrifuging and oscillating the disc to mix and react the platelet-rich plasma and the inducing agent, and in the process of centrifuging and oscillating, the optical assembly is controlled to collect the light transmission data of the detection area;

[0032] centrifuging the detection area of the disc to obtain platelet-poor plasma;

[0033] centrifuging the disc, and in the process of centrifuging, the optical assembly is controlled to collect the light transmission data of the detection area.

[0034] In another aspect, the application also provides a platelet analysis and stirring system, comprising:

[0035] a disc, a first through hole is formed in the center of the disc;

[0036] a bearing device, comprising a tray, a rotating disc and a support component, the rotating disc is embedded in the groove on the top surface of the tray, and the tray is provided with at least one light transmission hole; a second through hole is formed in the center of the rotating disc; the rotating disc is further provided with a plurality of positioning holes, and the disc is fixedly installed on the rotating disc through the positioning holes; when installed, the first through hole is aligned with the second through hole;

[0037] a centrifugal device, comprising a motor and a motor shaft, the motor shaft passes through the first through hole and the second through hole at the same time, so that the motor shaft drives the rotating disc and the disc to rotate cooperatively when the motor shaft rotates;

[0038] a light source, arranged inside the tray, and the light source is located below the light transmission hole;

[0039] an optical assembly, arranged above the rotating disc, and the optical assembly is arranged opposite to the light transmission hole;

[0040] a first control device, electrically connected with the motor;

[0041] a second control device, electrically connected with the optical assembly.

[0042] The application relates to a microfluidic full-automatic platelet detection method and a platelet analysis and stirring system, wherein the microfluidic full-automatic platelet detection method carries out the detection sampling process of platelet-rich plasma and the detection sampling process of platelet-poor plasma on the same platelet analysis and stirring device, the whole device has a small volume, does not need to additionally set a centrifuge device for preparing platelet-poor plasma, and does not need to additionally set a magnetic stirring device for stirring, the detection steps are simplified, the cost is reduced, and the detection efficiency is improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG1 is a schematic flow chart of a microfluidic fully automatic platelet detection method provided in one embodiment of the present application.

[0044] Figure 2 is a schematic flow chart of the microfluidic fully automatic platelet detection method provided in Example 1 of the present application.

[0045] Figure 3 is a schematic flow chart of the microfluidic fully automatic platelet detection method provided in Example 2 of the present application.

[0046] Figure 4 is a schematic flow chart of the microfluidic fully automatic platelet detection method provided in Example 3 of the present application.

[0047] FIG5 is a schematic diagram of the structure of a single detection unit in disk A. FIG.

[0048] FIG6 is a schematic diagram of the structure of a single detection unit in disk B. FIG.

[0049] FIG. 7 is a schematic structural diagram of a single detection unit in the disk C. FIG.

[0050] FIG8 is a schematic structural diagram of a single detection unit in the disk A1.

[0051] FIG9 is a schematic structural diagram of the disk A1.

[0052] FIG10 is a schematic diagram of the structure of a single detection unit in the disk A2.

[0053] FIG11 is a schematic structural diagram of a single detection unit in the disk B1.

[0054] FIG12 is a schematic structural diagram of a single detection unit in the disk C1.

[0055] FIG13 is a schematic structural diagram of a platelet analysis and homogenization system provided in one embodiment of the present application (the motor shaft is not assembled).

[0056] FIG14 is a schematic structural diagram of a platelet analysis and homogenization system according to an embodiment of the present application (the motor shaft is assembled).

[0057] 10 - disc; 11 - positioning groove; 12 - first through hole; 100 - detection unit; 110 - sample adding area; 120 - reagent adding area; 130 - detection area; 131 - blocking area; 132 - partition; 133 - detection hole; 140 - plasma area; 150 - precipitation area; 160 - quantification area; 170 - channel; 180 - waste pool; 20 - bearing device; 210 - tray; 211 - light transmission hole; 220 - rotating disc; 221 - second through hole; 230 - support component; 30 - centrifugal device; 310 - motor; 320 - motor shaft; 40 - light source; 50 - optical assembly; 60 - first control device; 70 - second control device. DETAILED DESCRIPTION

[0058] In order to make the purposes, technical solutions and advantages of the present application clearer, the present application is further described in detail below in combination with the drawings and examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application.

[0059] The present application provides a microfluidic full-automatic platelet detection method. Optionally, the microfluidic full-automatic platelet detection method is applied to a platelet analysis stirring system mentioned in the present application.

[0060] It should be noted that the platelet analysis stirring system mentioned in the following of the present application is a concept equivalent to the platelet analysis stirring device.

[0061] As shown in FIG. 1, in an embodiment of the present application, the method comprises the following S100 to S700:

[0062] S100, obtaining platelet-rich plasma, and introducing the platelet-rich plasma into a disc.

[0063] Specifically, the platelet-rich plasma is also referred to as PRP (platelet rich plasma).

[0064] S200, centrifuging the disc, and transferring the platelet-rich plasma to a detection area of the disc.

[0065] Specifically, the centrifuging can be rotating in a preset direction.

[0066] S300, introducing an inducing reagent into the disc.

[0067] Specifically, the inducing reagent can be an ADP (adenosine diphosphate) inducing reagent. When the inducing reagent is introduced into the disc, the inducing reagent is introduced into a reagent adding area of the disc.

[0068] S400, centrifuging the detection area of the disc, and transferring the inducing reagent to the detection area of the disc.

[0069] Specifically, the centrifugal speed can be 1000, the acceleration can be 3000, the deceleration can be 1000, the centrifugal time can be 5 seconds, and the rotation direction can be clockwise or counterclockwise.

[0070] S500, centrifugal shaking of the disc to mix and react the platelet rich plasma and the inducing reagent, and in the process of centrifugal shaking, the optical assembly is controlled to collect the light transmission data of the detection area.

[0071] Specifically, this step is to centrifugal shake the suspension formed by mixing and reacting the platelet rich plasma (PRP) and the inducing reagent. The optical assembly can include a light receiving tube. When S500 is performed, a light source is arranged below the disc, and an optical assembly is arranged above the disc. The light emitted by the light source irradiates the detection area of the disc to produce transmission, and the light transmitted through the detection area is directed to the light receiving tube in the optical assembly. After the light receiving tube collects the light transmitted through the detection area, light transmission data can be generated and sent to the single-chip microcomputer connected to the optical assembly. The single-chip microcomputer generates a reaction curve according to the light transmission data, and the reaction curve is used to display the detection result of the platelet rich plasma.

[0072] S600, centrifugal shaking of the disc to obtain platelet poor plasma.

[0073] Specifically, the disc is centrifuged to make the platelet particles in the suspension in the detection area settle, and platelet poor plasma (PPP) is obtained. The relatively clear plasma part after the platelet particles settle is the platelet poor plasma. The centrifugal speed in this step can be 3000, the acceleration can be 5000, the deceleration can be 3000, the centrifugal time can be 300 seconds, and the rotation direction can be clockwise or counterclockwise.

[0074] S700, centrifugal shaking of the disc, and in the process of centrifugal shaking, the optical assembly is controlled to collect the light transmission data of the detection area.

[0075] Specifically, this step is to centrifugal shake the platelet poor plasma obtained in S600. The centrifugal speed can be 120, the centrifugal time can be 10 seconds, and the rotation direction can be clockwise or counterclockwise. The detection sampling process of the platelet rich plasma and the detection sampling process of the platelet poor plasma are performed on the same platelet analysis and stirring device, without the need to set up additional cuvettes, magnetic stirring devices, and magnetic rods for stirring, and without the need for additional centrifuges to prepare platelet poor plasma. The cost is greatly reduced, and the operation is simplified for the operator.

[0076] In the embodiment, the detection and sampling process of the platelet-rich plasma and the detection and sampling process of the platelet-poor plasma are performed on the same platelet analysis and stirring device. The whole device has a small volume, does not need to additionally set a centrifuge device for preparing the platelet-poor plasma, and does not need to additionally set a magnetic stirring device for stirring. The detection steps are simplified, the cost is reduced, and the detection efficiency is improved.

[0077] In an embodiment of the present application, S100 comprises the following steps:

[0078] S110, obtaining platelet-rich plasma, and introducing the platelet-rich plasma into a sample adding area of the disc.

[0079] Specifically, the sample adding area can be a sample adding hole. In the embodiment, the platelet-rich plasma can be prepared by an external device and then added to the sample adding area.

[0080] The way of introducing the platelet-rich plasma into the sample adding area of the disc can be using a sample adding needle to add the sample. The sample adding needle can complete the sample adding by one-time sample adding, that is, single sample adding. The single sample adding amount can be in a numerical range of greater than or equal to 80 μL and less than or equal to 120 μL. Alternatively, the single sample adding amount can be 108 μL.

[0081] In the embodiment, after S110 is performed, when S200 is performed, the disc is centrifuged. The platelet-rich plasma is directly transferred from the sample adding area to the detection area of the disc. In this transfer mode, the rotation speed during centrifugation can be 1000, the acceleration can be 3000, the deceleration can be 3000, the centrifugation time can be 5 seconds, and the rotation direction can be clockwise or counterclockwise.

[0082] In an embodiment of the present application, S300 comprises:

[0083] S310, obtaining an induction reagent, and introducing the induction reagent into a reagent adding area of the disc.

[0084] Specifically, the reagent adding area can be a reagent adding hole. In the embodiment, the induction reagent can be prepared by an external device and then added to the reagent adding area.

[0085] In an embodiment of the present application, S100 comprises the following S121 to S122:

[0086] S121, obtaining a whole blood sample, and introducing the whole blood sample into a sample adding area of the disc.

[0087] S122, centrifuging the disc to separate the whole blood into platelet-rich plasma and blood cell precipitate.

[0088] Specifically, in the embodiment, the platelet-rich plasma is prepared on the disc, i.e., prepared on the platelet analysis stirring system instead of being prepared outside the system. The whole blood sample is separated into platelet-rich plasma and blood cell sediment by centrifugation of the disc by S122, so that the preparation of the platelet-rich plasma on the disc is completed.

[0089] In the embodiment, the whole blood sample is introduced into the addition area of the disc, and the whole blood sample is separated into platelet-rich plasma and blood cell sediment by centrifugation of the disc, so that the preparation of the platelet-rich plasma on the disc is completed during the detection process, without the need for pretreatment of the platelet-rich plasma, and without the need for additional preparation of the platelet-rich plasma by the centrifuge and other devices in advance, thereby saving costs.

[0090] In an embodiment of the present application, after S122 is performed, i.e., after the whole blood sample is separated into blood cell sediment and platelet-rich plasma by centrifugation of the disc, the platelet-rich plasma moves to the plasma area, and the blood cell sediment moves to the sediment area.

[0091] Specifically, in the embodiment, the disc is provided with a plasma area and a sediment area, and after the whole blood is separated, the platelet-rich plasma moves to the plasma area, and the blood cell sediment moves to the sediment area.

[0092] In the embodiment, when the whole blood sample is transferred from the sample addition area to the plasma area of the disc by centrifugation of the disc by S122, the rotation speed during centrifugation can be 2000, the acceleration can be 3000, the deceleration can be 3000, the centrifugation time can be 120 seconds, and the rotation direction can be clockwise or counterclockwise.

[0093] After S122 is performed, the disc is centrifuged by S200 to transfer the platelet-rich plasma from the plasma area to the detection area of the disc, and the rotation speed during centrifugation can be 2000, the acceleration can be 2000, the deceleration can be 500, the centrifugation time can be 10 seconds, and the rotation direction can be clockwise or counterclockwise.

[0094] In the embodiment, after the whole blood sample is separated into blood cell sediment and platelet-rich plasma, the platelet-rich plasma moves to the plasma area, and the blood cell sediment moves to the sediment area, so that the platelet-rich plasma is placed in a specific area, and the subsequent migration of the platelet-rich plasma to the detection area is facilitated.

[0095] In an embodiment of the present application, S200 includes the following S211 to S212:

[0096] S211, centrifuging the disc to transfer the platelet-rich plasma to the quantification area of the disc.

[0097] S212, centrifuging the disc to transfer the platelet-rich plasma in the quantification area to the detection area of the disc.

[0098] Specifically, the embodiment is provided with a quantification area on the disc, which is used to quantitatively transfer the platelet-rich plasma to the detection area.

[0099] In the embodiment, the platelet-rich plasma is quantitatively transferred to the detection area before being transferred to the detection area.

[0100] In an embodiment of the present application, the S200 comprises the following S221 to S222:

[0101] S221, centrifuging the disc to transfer the platelet-rich plasma in the plasma area to the quantification area of the disc.

[0102] S222, centrifuging the disc to transfer the platelet-rich plasma in the quantification area to the detection area of the disc.

[0103] Specifically, in the embodiment, the platelet-rich plasma in the plasma area is transferred to the quantification area of the disc when the disc is centrifuged in S221.

[0104] In the embodiment, since there are both a whole blood separation step and a pre-quantification step, the centrifugation parameters of each step are slightly different.

[0105] When the platelet-rich plasma is transferred from the sample addition area to the plasma area of the disc by centrifuging the disc in S122, the rotation speed during centrifugation can be 1000, the acceleration can be 3000, the deceleration can be 3000, the centrifugation time can be 120 seconds, and the rotation direction can be clockwise or counterclockwise.

[0106] When the platelet-rich plasma is transferred from the plasma area to the quantification area of the disc by centrifuging the disc in S221, the rotation speed during centrifugation can be 450, the acceleration can be 30000, the deceleration can be 1000, the centrifugation time can be 30 seconds, and the rotation direction can be clockwise or counterclockwise.

[0107] When the platelet-rich plasma is transferred from the quantification area to the detection area of the disc by centrifuging the disc in S222, the rotation speed during centrifugation can be 1000, the acceleration can be 1000, the deceleration can be 1000, the centrifugation time can be 10 seconds, and the rotation direction can be clockwise or counterclockwise.

[0108] In an embodiment of the present application, the centrifuging the disc comprises:

[0109] The disc is driven to rotate in a predetermined direction for a predetermined duration.

[0110] Specifically, in the present application, in addition to centrifugal shaking, the specific way of centrifuging the disc can be driving the disc to rotate in a preset direction for a preset duration. Driving the disc to rotate in a preset direction for a preset duration means driving the disc to rotate towards a preset direction for a preset duration. The preset direction can be clockwise or counterclockwise. The preset duration can be preset before detection.

[0111] In an embodiment of the present application, the disc is provided with a plurality of detection areas and a plurality of sample addition areas and a plurality of reagent addition areas, each sample addition area has one detection area and one reagent addition area corresponding to the sample addition area, or each sample addition area has a plurality of detection areas and a plurality of reagent addition areas corresponding to the sample addition area.

[0112] Specifically, in one embodiment, one sample addition area has one detection area and one reagent addition area corresponding to the sample addition area, and the three are one-to-one correspondence. The quantitative area provided on the disc is also one-to-one corresponding to the detection area, the reagent addition area, and the sample addition area. Each detection area, each sample addition area, each inducible reagent addition area, and each quantitative area form a detection unit. In this embodiment, the disc is provided with a plurality of detection units, and the plurality of detection units are distributed in a ring shape on the disc and are arranged in a radial manner around the positioning groove of the disc, as shown in FIG. 9, and the detection units are independent of each other, and adjacent two detection units will not affect the liquid migration of each other.

[0113] In another embodiment, one sample addition area has a plurality of detection areas and a plurality of reagent addition areas corresponding to the sample addition area. At this time, one whole blood sample (corresponding to an embodiment having a whole blood separation step to prepare platelet-rich plasma on the disc) or one platelet-rich plasma (corresponding to an embodiment of directly adding prepared platelet-rich plasma on the disc) can perform different detection items and achieve diverse detection.

[0114] The way of introducing platelet-rich plasma into the addition area of the disc can be by adding sample through a sample needle. The single sample amount is in the numerical range of greater than or equal to 80 μL and less than or equal to 120 μL. Alternatively, the single sample amount can be 108 μL.

[0115] The way of introducing inducible reagent into the reagent addition area of the disc can be by adding sample through a sample needle. The single sample amount is in the numerical range of greater than or equal to 5 μL and less than or equal to 20 μL. Alternatively, the single sample amount can be 12 μL. When the sample needle is replaced with liquid, the sample needle should be cleaned.

[0116] In the embodiment, the disc is provided with a plurality of detection areas and a plurality of adding areas, each detection area has one adding area corresponding to the detection area, so that the detection process can undertake high throughput, and a plurality of detections can be performed at the same time, and the detection efficiency is high. In addition, the detection work of multiple projects can also be compatible.

[0117] In an embodiment of the present application, S500 includes:

[0118] S510, driving the disc to rotate until a detection area reaches a position opposite to the light source, so that the light source is projected on the detection area.

[0119] S520, driving the disc to stop rotating for a preset pause time.

[0120] S530, driving the disc to rotate again until the next detection area reaches the position opposite to the light source.

[0121] Specifically, the embodiment is a specific process of centrifugal oscillation, that is, the detection and sampling process of the suspension formed after the mixing and reaction of platelet-rich plasma and inducing reagents, S510 to S520 need to be repeatedly executed, and after S510 to S520 are executed once, the sampling of one detection area is completed. Alternatively, S510 to S520 are executed for M / N seconds, N is the total number of detection units, that is, the movement and sampling of all detection areas are completed in M seconds, and the movement and sampling of each detection area use M / N seconds. The position opposite to the light source is the setting position of the light transmission hole, and the light source is always emitting light. When the disc rotates to the setting position of the light transmission hole, the light source passes through the light transmission hole and irradiates on the detection area, and then irradiates on the light-receiving tube in the optical assembly after transmitting the suspension in the detection area. N can be 16. M can be 1.

[0122] The light irradiates on the bottom of the detection area, and is emitted from the top of the detection area after transmission. The light intensity through the suspension is weakened. Every 1 second rotation is 16 times of light emission, and the light intensity of each detection area is obtained at each time node. Within 300 seconds, the reaction curve is generated with the change of the reaction. The reaction curve has multiple lines, the abscissa is time, and the ordinate is light intensity.

[0123] In this step, the rotation speed during centrifugal oscillation can be 60, the acceleration can be 15000, the total centrifugal time can be 300 seconds, and the rotation direction can be clockwise or counterclockwise.

[0124] S520, driving the disc to stop rotating for a preset pause time, which is actually the time for light transmission detection of the suspension formed after the mixing and reaction of platelet-rich plasma and inducing reagents.

[0125] After the execution of S520 once, the driving disc is rotated again until the next detection area reaches the position opposite to the light source, and the rotation mode can be various. Alternatively, one rotation mode is that one detection area is directly rotated to the next detection area, i.e. one detection area step is moved at one time. Alternatively, another rotation mode is that two detection area steps are moved at one time first, and then one detection area step is moved back. Different rotation schemes can be made according to different detection items.

[0126] In this embodiment, the mixing and stirring are realized simultaneously, and the amount of blood to be added is greatly reduced compared with the traditional off-machine magnetic rod stirring. In addition, the surface oscillation sampling is adopted instead of the vertical rotation sampling, which is a rare sampling mode on the market.

[0127] In an embodiment of the present application, when S500 is executed, i.e. when the disc is centrifugally oscillated to mix and react the platelet-rich plasma and the inducing agent, the ratio of the volume of the suspension formed after the mixing and reaction of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area is in the percentage range of greater than or equal to 50% and less than or equal to 100%.

[0128] Specifically, the ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 50%. The ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 60%. The ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 70%. The ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 80%. The ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 90%. The ratio of the volume of the suspension formed after the mixing of the platelet-rich plasma and the inducing agent in the detection area to the volume of the detection area can be 100%.

[0129] Table 1 - Detection results of the ratio of the volume of different suspensions to the volume of the detection area

[0130] The above table, i.e. Table 1, is the results of the repeatability CV and relative deviation of the ratio of the volume of five different suspensions to the volume of the detection area in five detections. The control group is the theoretical result.

[0131] The volume of the detection area is 130 μL, the volume of the suspension corresponding to the ratio of 60% is 78 μL, the volume of the suspension corresponding to the ratio of 70% is 91 μL, the volume of the suspension corresponding to the ratio of 80% is 104 μL, the volume of the suspension corresponding to the ratio of 90% is 117 μL, and the volume of the suspension corresponding to the ratio of 100% is 130 μL. 78 / 130=60%, 91 / 130=70%, 104 / 130=80%, 117 / 130=90%, and 130 / 130=100%, which correspond to the ratios of the volumes of the five different suspensions to the volume of the detection area, respectively.

[0132] As can be seen from Table 1, the relative deviations of the detection results (i.e., aggregation rates) of most of the four ratios from the aggregation rate of the control group are less than or equal to 15%, which meets the expectation.

[0133] The CV value is the repeatability CV value. The smaller the CV value, the smaller the precision, the better the detection result, or in other words, the better. The relative deviation is the aggregation rate relative deviation. The smaller the aggregation rate relative deviation, the better the detection result, or in other words, the better. To observe the CV value and the aggregation rate relative deviation at the same time, the smaller the two, the better.

[0134] The repeatability CV value and the aggregation rate relative deviation of the ratio of 100% are the worst, the repeatability CV value and the aggregation rate relative deviation of the ratio of 60%, the ratio of 70%, and the ratio of 80% are moderate, and the repeatability CV value and the aggregation rate relative deviation of the ratio of 90% are the best.

[0135] The ratio of the volume of the suspension formed after mixing the platelet-rich plasma and the inducing reagent in the detection area to the volume of the detection area is 90%, which is the optimal result. Under this ratio, the mixing effect of the platelet-rich plasma and the inducing reagent is the best.

[0136] It should be noted that the volume of the suspension can be fully mixed when the volume of the suspension is 50-100% of the total volume that can be accommodated in the detection hole. Table 1 only shows the data results of the ratios of 60%, 70%, 80%, 90%, and 100%, and does not list all the percentage conditions.

[0137] In this embodiment, the volume of the suspension can be fully mixed when the volume of the suspension is 50-100% of the total volume that can be accommodated in the detection hole.

[0138] Embodiment 1

[0139] As shown in FIG. 2, the present application also provides a microfluidic full-automatic platelet detection method, which comprises the following W100-W800.

[0140] W100, obtaining a whole blood sample, and introducing the whole blood sample into a sample adding area of a disc.

[0141] W200, centrifuging the disc to separate the whole blood into platelet-rich plasma and blood cell sediment.

[0142] W300, centrifuging the disc to transfer the platelet-rich plasma to the detection area of the disc.

[0143] W400, introducing an inducing reagent into the disc.

[0144] W500, centrifuging the disc to transfer the inducing reagent to the detection area of the disc.

[0145] W600, centrifuging and oscillating the disc to mix and react the platelet-rich plasma and the inducing reagent, and in the process of centrifuging and oscillating, controlling the optical assembly to collect the light transmission data of the detection area.

[0146] W700, centrifuging the detection area of the disc to obtain platelet-poor plasma.

[0147] W800, centrifuging the disc, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area.

[0148] Specifically, the embodiment is adapted to the disc B in FIG. 6 and the disc B1 in FIG. 11. The disc has a plasma area and a sediment area, but does not have a quantification area.

[0149] Embodiment 2

[0150] As shown in FIG. 3, the application also provides a microfluidic automatic platelet detection method, which comprises the following K100 to K900:

[0151] K100, obtaining whole blood and introducing the whole blood into the sample addition area of the disc.

[0152] K200, centrifuging the disc to separate the whole blood into platelet-rich plasma and blood cell sediment, and moving the platelet-rich plasma to the plasma area and the blood cell sediment to the sediment area.

[0153] K300, centrifuging the disc to transfer the platelet-rich plasma from the plasma area to the quantification area of the disc.

[0154] K400, centrifuging the disc to transfer the platelet-rich plasma in the quantification area to the detection area of the disc.

[0155] K500, introducing an inducing reagent into the disc.

[0156] K600, centrifuging the disc to transfer the inducing reagent to the detection area of the disc.

[0157] K700, centrifuging and oscillating the disc to mix and react the platelet-rich plasma and the inducing reagent, and in the process of centrifuging and oscillating, controlling the optical assembly to collect the light transmission data of the detection area.

[0158] K800, centrifuging the detection area of the disc to obtain platelet-poor plasma.

[0159] K900, centrifuging the disc, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area.

[0160] Specifically, the embodiment is adapted to the disc C in FIG. 7 and the disc C1 in FIG. 12. The disc has both the plasma area 140 and the sedimentation area 150, and also has the quantification area 160.

[0161] Embodiment 3

[0162] As shown in FIG. 4, the application also provides a microfluidic full-automatic platelet detection method, which comprises the following L100 to L700:

[0163] L100, obtaining platelet-rich plasma, and introducing the platelet-rich plasma into a sample addition area of a disc.

[0164] L200, centrifuging the disc, and transferring the platelet-rich plasma from the sample addition area to a detection area of the disc.

[0165] L300, introducing an inducing reagent into the disc.

[0166] L400, centrifuging the disc, and transferring the inducing reagent to the detection area of the disc.

[0167] L500, centrifuging and oscillating the disc to mix and react the platelet-rich plasma and the inducing reagent, and in the process of centrifuging and oscillating, controlling the optical assembly to collect the light transmission data of the detection area.

[0168] L600, centrifuging the detection area of the disc to obtain platelet-poor plasma.

[0169] L700, centrifuging the disc, and in the process of centrifuging, controlling the optical assembly to collect the light transmission data of the detection area.

[0170] Specifically, as shown in FIG. 5, FIG. 8 and FIG. 9, the embodiment is adapted to the disc A and the disc A1 in the figures. In the embodiment, the platelet-rich plasma is prepared outside the platelet analysis stirring system, and the prepared platelet-rich plasma is directly added into the sample addition area 110 without being prepared on the disc. Since the platelet-rich plasma is directly added into the sample addition area 110 without being prepared on the disc, the disc does not have both the plasma area 140 and the sedimentation area 150, and also does not have the quantification area 160.

[0171] The application also provides a platelet analysis stirring system.

[0172] It should be noted that, for the sake of brevity, all component structures or regions are labeled in the embodiment of the platelet analysis and stirring system, and not in the embodiment of the microfluidic automatic platelet detection method.

[0173] As shown in FIGS. 13 and 14, in an embodiment of the present application, the platelet analysis and stirring system comprises a disc 10, a bearing device 20, a centrifugal device 30, a light source 40, an optical assembly 50, a first control device 60, and a second control device 70.

[0174] The disc 10 is provided with a first through hole 12 at the center. The bearing device 20 comprises a tray 210, a rotating disc 220, and a support component 230. The rotating disc 220 is embedded in a groove on the top surface of the tray 210. The tray 210 is provided with at least one light transmission hole 211. The center of the rotating disc 220 is provided with a second through hole 221. The rotating disc 220 is further provided with a plurality of positioning holes, and the disc 10 is fixedly installed on the rotating disc 220 through the positioning holes. When installed, the first through hole 12 is aligned with the second through hole 221.

[0175] The centrifugal device 30 comprises a motor 310 and a motor shaft 320. The motor shaft 320 passes through the first through hole 12 and the second through hole 221, so that the motor shaft 320 drives the rotating disc 220 and the disc 10 to rotate synchronously when the motor shaft 320 rotates. The light source 40 is arranged inside the tray 210. The light source 40 is located below the light transmission hole 211. The optical assembly 50 is arranged above the rotating disc 220. The optical assembly 50 is arranged opposite to the light transmission hole 211. The first control device 60 is electrically connected to the motor 310. The second control device 70 is electrically connected to the optical assembly 50.

[0176] Specifically, the first control device 60 can comprise a first PCB board. The second control device 70 can comprise a second PCB board. The optical assembly 50 can comprise a light-receiving tube, which can be welded on the second PCB board.

[0177] The platelet analysis and stirring system can further comprise a single-chip microcomputer, which is electrically connected to the optical assembly 50 and used to receive the light transmission data collected by the optical assembly 50.

[0178] Please continue to refer to FIGS. 13 and 14. In an embodiment of the present application, the disc 10 comprises a positioning groove 11 and a plurality of detection units 100 arranged radially around the positioning groove 11 with the positioning groove 11 as the center.

[0179] The detection unit 100 comprises a sample adding area 110, a reagent adding area 120 and a detection area 130. The sample adding area 110 and the reagent adding area 120 are both in communication with the detection area 130. The detection area 130 comprises at least one barrier area 131, a partition 132 and a detection hole 133. The barrier area 131 is arranged close to the sample adding area 110 and the reagent adding area 120. The partition 132 is arranged close to the barrier area 131. The partition 132 is two, and the two partitions 132 are oppositely arranged. The detection hole 133 is arranged close to the partition 132.

[0180] Specifically, as shown in FIG. 9, the disc 10 comprises a positioning groove 11 and a plurality of detection units 100.

[0181] The barrier area 131 functions to prevent the suspension formed by mixing the platelet-rich plasma and the inducing reagent at the position of the detection hole 133 from flowing back when centrifugal oscillation detection is performed. The embodiment shown in FIG. 8, i.e., disc A1, shows a disc 10 with only one barrier area 131. The embodiment shown in FIG. 10, i.e., disc A2, shows a disc 10 with two barrier areas 131. The number of the barrier areas 131 is not limited, and the number of the barrier areas 131 is set to match different reaction systems, so as to ensure that the suspension does not flow back into the channel 170 when oscillation is performed. Similarly, the disc B1 in FIG. 11 and the disc C1 in FIG. 12 are both provided with the barrier area 131, which functions in the same way.

[0182] The partition 132 functions to slow down the flow back of the suspension formed by mixing the platelet-rich plasma and the inducing reagent at the position of the detection hole 133 when centrifugal oscillation detection is performed. The disc A1 in FIG. 8, the disc A2 in FIG. 10, the disc B1 in FIG. 11 and the disc C2 in FIG. 12 are all provided with the partition 132.

[0183] In an embodiment of the present application, the detection unit 100 further comprises a plasma area 140 and a sedimentation area 150. As shown in FIG. 11.

[0184] In an embodiment of the present application, the detection unit 100 further comprises a quantitative area 160 and a waste pool 180. As shown in FIG. 12.

[0185] Specifically, S221 transfers the platelet-rich plasma in the plasma area to the quantitative area 160 of the disc 10 when the disc is centrifuged, and the excess platelet-rich plasma enters the waste pool 180.

[0186] Any combination of the technical features in the above-described embodiments can be made, and the method steps are not limited in execution order. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments are described, however, as long as the combination of the technical features does not exist in contradiction, it should be considered within the scope of the present disclosure.

[0187] The above-described embodiments only express several implementation manners of the present application, and the description is relatively specific and detailed, but it should not be understood as a limitation on the patent scope of the present application. It should be noted that, for ordinary skilled persons in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the appended claims.

Claims

1. A microfluidic fully automatic platelet detection method, characterized in that: The method comprises: obtaining platelet-rich plasma and introducing the platelet-rich plasma into the disc; The disc is centrifuged to transfer the platelet-rich plasma to the testing area of ​​the disc; introducing an inducing agent into the disc; Centrifuge the disc to transfer the induction reagent to the detection area of ​​the disc; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area; The test area of ​​the disc is centrifuged to obtain platelet-poor plasma; Centrifuging the disc, and during the centrifugation process, controlling the optical component to collect light transmittance data of the detection area; The centrifuging of the disc comprises: The drive disc rotates in a preset direction and for a preset duration; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area, including: driving the disc to rotate until a detection area reaches a position opposite to the light source, so that the light source is projected onto the detection area; The drive disc stops rotating for a preset pause time; The drive disk is rotated again until the next detection area reaches the position opposite to the light source.

2. The microfluidic fully automatic platelet detection method according to claim 1, characterized in that: The method of obtaining platelet-rich plasma and introducing the platelet-rich plasma into the disc comprises: Obtain platelet-rich plasma and introduce the platelet-rich plasma into the sample addition area of ​​the disc.

3. The microfluidic fully automatic platelet detection method according to claim 1, characterized in that: The method of obtaining platelet-rich plasma and introducing the platelet-rich plasma into the disc comprises: obtaining a whole blood sample and introducing the whole blood sample into the sample addition area of ​​the disc; The disc is centrifuged to separate the whole blood sample into platelet-rich plasma and blood cell pellet.

4. The microfluidic fully automatic platelet detection method according to claim 3, characterized in that: After the disc is centrifuged to separate the whole blood sample into a blood cell pellet and platelet-rich plasma, the platelet-rich plasma moves to the plasma area and the blood cell pellet moves to the sedimentation area.

5. The microfluidic fully automatic platelet detection method according to claim 3, characterized in that: The step of centrifuging the disc to transfer the platelet-rich plasma to the detection area of ​​the disc comprises: The disc is centrifuged to transfer the platelet-rich plasma to the quantitative area of ​​the disc; The disc is centrifuged and the platelet-rich plasma in the quantification area is transferred to the detection area of ​​the disc.

6. The microfluidic fully automatic platelet detection method according to claim 4, characterized in that: The step of centrifuging the disc to transfer the platelet-rich plasma to the detection area of ​​the disc comprises: The disc is centrifuged to transfer the platelet-rich plasma in the plasma area to the quantitative area of ​​the disc; The disc is centrifuged and the platelet-rich plasma in the quantification area is transferred to the detection area of ​​the disc.

7. The microfluidic fully automatic platelet detection method according to any one of claims 1 to 6, characterized in that: The disc is provided with multiple detection areas, multiple sample addition areas and multiple reagent addition areas, each sample addition area has a detection area and a reagent addition area corresponding to the sample addition area, or each sample addition area has multiple detection areas and multiple reagent addition areas corresponding to the sample addition area.

8. The microfluidic fully automatic platelet detection method according to claim 7, characterized in that: When the disc is centrifugally shaken to allow the platelet-rich plasma and the induction reagent to mix and react, the ratio of the volume of the suspension formed after the platelet-rich plasma and the induction reagent in the detection area are mixed and reacted to the volume of the detection area is within a percentage range greater than or equal to 50% and less than or equal to 100%.

9. A microfluidic fully automatic platelet detection method, characterized in that: The method comprises: obtaining a whole blood sample and introducing the whole blood sample into the sample addition area of ​​the disc; The disc is centrifuged to separate the whole blood sample into platelet-rich plasma and blood cell pellet; The disc is centrifuged to transfer the platelet-rich plasma to the testing area of ​​the disc; introducing an inducing agent into the disc; Centrifuge the disc to transfer the induction reagent to the detection area of ​​the disc; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area; The test area of ​​the disc is centrifuged to obtain platelet-poor plasma; Centrifuging the disc, and during the centrifugation process, controlling the optical component to collect light transmission data of the detection area; the centrifuging the disc includes: The drive disc rotates in a preset direction and for a preset duration; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area, including: driving the disc to rotate until a detection area reaches a position opposite to the light source, so that the light source is projected onto the detection area; The drive disc stops rotating for a preset pause time; The drive disk is rotated again until the next detection area reaches the position opposite to the light source.

10. A microfluidic fully automatic platelet detection method, characterized in that: The method comprises: obtaining a whole blood sample and introducing the whole blood sample into the sample addition area of ​​the disc; The disc is centrifuged to separate the whole blood sample into platelet-rich plasma and blood cell sediment. The platelet-rich plasma moves to the plasma area and the blood cell sediment moves to the sedimentation area. The disc is centrifuged to transfer the platelet-rich plasma from the plasma area to the quantitative area of ​​the disc; The disc is centrifuged to transfer the platelet-rich plasma in the quantitative area to the detection area of ​​the disc; introducing an inducing agent into the disc; Centrifuge the disc to transfer the induction reagent to the detection area of ​​the disc; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area; The test area of ​​the disc is centrifuged to obtain platelet-poor plasma; Centrifuging the disc, and during the centrifugation process, controlling the optical component to collect light transmittance data of the detection area; The centrifuging of the disc comprises: The drive disc rotates in a preset direction and for a preset duration; The disc is centrifuged to allow the platelet-rich plasma and the induction reagent to mix and react, and during the centrifugal oscillation process, the optical component is controlled to collect light transmittance data of the detection area, including: driving the disc to rotate until a detection area reaches a position opposite to the light source, so that the light source is projected onto the detection area; The drive disc stops rotating for a preset pause time; The drive disk is rotated again until the next detection area reaches the position opposite to the light source.

11. A platelet analysis and homogenization system, characterized in that: Used to perform the microfluidic fully automatic platelet detection method according to any one of claims 1 to 10, the platelet analysis and homogenization system comprises: a disc, wherein a first through hole is formed in the center of the disc; The carrying device includes a tray, a rotating disk, and a supporting member. The rotating disk is embedded in a groove on the top surface of the tray. The tray is provided with at least one light-through hole. A second through hole is opened at the center of the rotating disk. The rotating disk is also provided with a plurality of positioning holes. The disk is fixedly mounted on the rotating disk through the positioning holes. During installation, the first through hole is aligned with the second through hole. A centrifugal device comprising a motor and a motor shaft, wherein the motor shaft passes through the first through hole and the second through hole simultaneously, so that when the motor shaft rotates, the rotating disk and the disc rotate in coordination; The light source is arranged inside the tray and below the light hole; The optical component is arranged above the rotating disk, and the optical component is arranged opposite to the light hole; a first control device electrically connected to the motor; The second control device is electrically connected to the optical component.

12. The platelet analysis and homogenization system according to claim 11, characterized in that: The disc includes a positioning groove and a plurality of detection units arranged radially around the positioning groove with the positioning groove as the center; The detection unit comprises: Sample addition area; Reagent addition area; The detection area, the sample addition area and the reagent addition area are all connected to the detection area; The detection area includes: at least one barrier area disposed adjacent to the sample addition area and the reagent addition area; A partition is provided near the blocking area; there are two partitions, and the two partitions are provided opposite to each other; The detection hole is arranged close to the partition.

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