Microfluidic method and system for detecting blood troponin combined with blood pressure fluctuation

Abstract

The application discloses a blood troponin microfluidic detection method and system combined with blood pressure fluctuation, relates to the field of in-vitro microfluidic biomarker detection, and the method comprises the following steps: acquiring a blood pressure waveform in a sampling period, determining a trigger moment at which a target heart beat pressure rising stage and a pressure falling stage reach a same target pressure value, and opening a micro valve; calculating an instantaneous flow and a cumulative through volume according to a pressure value and a change rate thereof, a branch outlet pressure, a blood flow resistance and a pipeline compliance, closing the micro valve when a target sample volume is reached, and obtaining blood samples in the pressure rising phase and the pressure falling phase; performing plasma separation and troponin immune detection, correcting a detection signal in combination with the cumulative through volume, a reaction residence time and immune capture parameters, obtaining a two-phase hydraulic correction concentration, calculating a geometric mean and a logarithmic absolute value of a concentration ratio, and outputting a detection result or re-sampling according to a liquid path anti-symmetry residual. Thus, the inhibition capability of troponin detection on liquid path disturbance and the result reliability are improved.

Description

Technical Field

[0001] This application relates to the field of in vitro microfluidic biomarker detection technology, and in particular to a microfluidic detection method and system for blood troponin combined with blood pressure fluctuations. Background Technology

[0002] Cardiac troponin is an important target analyte in blood sample testing, and changes in its levels can provide reference information for assessing myocardial injury-related states. Currently, in vitro detection of cardiac troponin is gradually developing from central laboratories to point-of-care and portable testing. Microfluidic detection devices can perform plasma separation, immune reactions, and signal readings on small amounts of whole blood samples collected in vitro, which helps to shorten the time required for sample processing and result reporting. In applications such as emergency resuscitation, perioperative monitoring, and intensive care, it is also necessary to test whole blood samples obtained at different sampling points separately to obtain troponin content sequence analysis data. Therefore, in addition to meeting the corresponding detection sensitivity requirements, the consistency of the metrological conditions corresponding to different in vitro samples will also affect the comparability of sequence detection data.

[0003] Current in vitro microfluidic assays typically assume that the blood sample delivered to the assay device has been stabilized, or that the sample is formed using pre-defined sampling time, driving pressure, or flow control. When different ex vivo blood samples originate from sampling points corresponding to different blood pressure states, blood pressure fluctuations associated with those sampling points may alter the instantaneous flow state of the blood sample upon entering the in vitro microfluidic device. Furthermore, the dynamic characteristics of the sampling components and the rheological state of the blood may also affect the sample measurement process. In such cases, even with the same sampling time or control parameters, differences in the injection volume, plasma separation state, and reaction residence time may still exist between different samples, causing the in vitro assay signal to be simultaneously affected by changes in the target analyte concentration and assay conditions.

[0004] Furthermore, the fluid equilibrium state may vary at different sampling points, resulting in different plasma volumes in the obtained isolated blood samples. Even when the total circulating troponin level remains unchanged, plasma dilution or concentration can still cause a decrease or increase in the detectable troponin concentration in isolated samples. Existing sequence detection methods typically compare the concentration difference between adjacent samples directly, and some methods also use indicators such as hematocrit for result correction. However, a single concentration difference or a single correction indicator cannot simultaneously eliminate the influence of sample formation conditions and changes in plasma volume. Therefore, it is difficult to obtain in vitro detection data that are based on a unified metrological foundation and can accurately characterize the changes in troponin content in different isolated blood samples. Summary of the Invention

[0005] This application provides a microfluidic detection method, system, storage medium, computer program product, and electronic device for blood troponin based on blood pressure fluctuations, which at least solves the problem in the prior art that inconsistent blood troponin detection conditions at different sampling points lead to poor comparability of sequence detection results.

[0006] In a first aspect, embodiments of this application provide a microfluidic detection method for blood troponin combined with blood pressure fluctuations. The method includes: acquiring continuous blood pressure waveform data corresponding to a predetermined sampling period of the isolated whole blood to be tested, wherein the continuous blood pressure waveform data includes pressure values ​​received through a data interface and corresponding data acquisition times; determining a target heartbeat and a target pressure value between the diastolic and systolic pressures of the target heartbeat from the continuous blood pressure waveform data, and determining the times when the pressure-boosting phase and the pressure-depressing phase of the target heartbeat reach the target pressure value as the pressure-boosting trigger time and the pressure-depressing trigger time, respectively; opening the pressure-boosting phase microvalve and the pressure-depressing phase microvalve, respectively, at the pressure-boosting trigger time and the pressure-depressing trigger time; determining the instantaneous flow rate of each phase during the sampling process based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding sampling branch, blood flow resistance parameters, and tubing compliance parameters, and performing time integration to obtain the cumulative throughput volume of each phase; and determining the cumulative throughput volume of each phase when the target sample is reached. When the volume is reached, the corresponding micro-valve is closed to form a pressurized phase blood sample and a depressurized phase blood sample. Plasma separation and troponin immunoassay are performed on the pressurized and depressurized phase blood samples respectively to obtain the troponin detection signal and reaction residence time for each phase. The immunoassay efficiency of each phase is determined based on the cumulative throughput volume, the reaction residence time of the corresponding blood sample, and preset immunoassay parameters. The corresponding troponin detection signal is corrected using the immunoassay efficiency of each phase to obtain the pressurized phase hydraulic correction concentration and the depressurized phase hydraulic correction concentration. The geometric mean of the pressurized phase hydraulic correction concentration and the depressurized phase hydraulic correction concentration is calculated to obtain the two-phase symmetry. The absolute logarithm of the ratio of the pressurized phase hydraulic correction concentration to the depressurized phase hydraulic correction concentration is calculated to obtain the antisymmetric residual of the liquid path. When the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold, the two-phase symmetry is output as the troponin detection result. When the antisymmetric residual of the liquid path is greater than the preset residual threshold, a re-injection command is generated.

[0007] Secondly, embodiments of this application provide a blood troponin microfluidic detection system incorporating blood pressure fluctuations. The system includes: a blood pressure waveform analysis unit, used to acquire continuous blood pressure waveform data corresponding to a predetermined sampling period of the extracted whole blood to be tested, wherein the predetermined sampling period is a preset time window for selecting the sampling trigger time; the continuous blood pressure waveform data includes pressure values ​​received through a data interface and the data acquisition time corresponding to each pressure value; determining a target heartbeat and its target pressure value between diastolic and systolic blood pressure from the continuous blood pressure waveform data; and determining the pressure rise phase of the target heartbeat. The moments when the target pressure values ​​are reached during the pressure increase and decrease phases are defined as the pressure increase trigger moment and pressure decrease trigger moment, respectively. A biphase quantitative injection unit is used to open the pressure increase phase microvalve and the pressure decrease phase microvalve at the pressure increase trigger moment and the pressure decrease trigger moment, respectively, allowing the tested whole blood to enter the corresponding injection branches. Based on the pressure values ​​and their rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, blood flow resistance parameters, and tubing compliance parameters, the instantaneous flow rate of each phase injection process is determined and integrated over time to obtain the cumulative throughput volume of each phase. The cumulative throughput volume of each phase reaches the target sample level. The corresponding micro-valve is closed at this volume to form a pressurized phase blood sample and a depressurized phase blood sample. A biphasic immunoassay unit is used to perform plasma separation and troponin immunoassay on the pressurized and depressurized phase blood samples respectively, obtaining the troponin detection signal and reaction residence time for each phase blood sample. A hydraulic immunocorrection unit is used to determine the immunocapture efficiency of each phase based on the cumulative throughput volume of each phase, the reaction residence time of the corresponding blood sample, and preset immunocapture parameters, and to correct the troponin detection signal for each phase blood sample using the immunocapture efficiency, obtaining the hydraulic correction concentration of the pressurized phase. The system includes a depressurization phase hydraulic correction concentration; a two-phase characteristic calculation unit, used to calculate the geometric mean of the depressurization phase hydraulic correction concentration and the pressure-increasing phase hydraulic correction concentration to obtain the two-phase symmetry, and to calculate the absolute logarithm of the ratio of the depressurization phase hydraulic correction concentration to the pressure-increasing phase hydraulic correction concentration to obtain the antisymmetric residual of the liquid path; and a detection result determination unit, used to output the two-phase symmetry as the troponin detection result corresponding to the target heartbeat when the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold; and to generate a re-injection command when the antisymmetric residual of the liquid path is greater than the preset residual threshold.

[0008] Thirdly, an electronic device is provided, comprising: at least one processor, and a memory communicatively connected to the at least one processor, wherein the memory stores instructions executable by the at least one processor, the instructions being executed by the at least one processor to enable the at least one processor to perform the steps of the blood troponin microfluidic detection method incorporating blood pressure fluctuations according to any embodiment of the present application.

[0009] Fourthly, embodiments of this application provide a storage medium storing a computer program thereon, which, when executed by a processor, implements the steps of the blood troponin microfluidic detection method incorporating blood pressure fluctuations according to any embodiment of this application.

[0010] Fifthly, embodiments of this application provide a computer program product, including a computer program / instructions, which, when executed by a processor, implement the steps of the blood troponin microfluidic detection method incorporating blood pressure fluctuations according to any embodiment of this application.

[0011] The microfluidic detection method and system for blood troponin combined with blood pressure fluctuations provided in this application can achieve at least the following technical effects:

[0012] A correspondence is established between the target pressure state in the continuous blood pressure waveform and the microfluidic injection process, and paired injections are formed by utilizing the moments when the pressure rises and falls twice within the same heartbeat, reaching the target pressure value. The two injection phases maintain correspondence in pressure amplitude but differ in the direction of pressure change, giving the injection process both a common benchmark and directional reference. By dynamically solving and integrating the instantaneous flow rate during each injection phase, and controlling the microvalve closure with the actual cumulative flow volume, the fixed opening time is avoided from being equated with a fixed injection volume. This ensures that the blood samples in the pressurizing and depressurizing phases still obtain relatively consistent target volumes under dynamic pressure conditions, and provides traceable hydraulic metrology data for subsequent comparisons.

[0013] In the result processing, instead of directly comparing the original detection signals of the two phases, the capture efficiency is determined based on the actual throughput volume, reaction residence time, and immune capture parameters of each phase. The influence of the liquid flow process and the immune reaction process on signal formation is incorporated into the correction. After correction, the concentrations of the two phases are symmetrically fused using geometric mean to reduce the impact of phase correlation bias on the final result. Furthermore, an antisymmetric residual is generated using the logarithmic absolute value of the concentration ratio to evaluate whether there are still differences between the two phases exceeding the allowable range. When this difference exceeds a threshold, potentially distorted detection values ​​are not output; instead, re-injection is triggered, thus connecting sample measurement, reaction correction, and result validity determination into a continuous quality control chain.

[0014] This technical solution does not involve simply adding an extra sample injection. Instead, it utilizes a pressurizing phase and a depressurizing phase to construct a two-phase detection system with a physically symmetrical relationship. The results from the two phases are decomposed into a symmetrical component characterizing troponin levels and an antisymmetric component characterizing inconsistencies in the liquid flow. Therefore, the detection device can obtain robust troponin detection results from the shared information of the two phases, and can also proactively identify detection processes that do not meet the output conditions based on the differences between the two phases, thereby improving the consistency, reliability, and anomaly protection capabilities of the sequence detection data. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 A flowchart is shown as an example of a microfluidic detection method for blood troponin in conjunction with blood pressure fluctuations according to an embodiment of this application;

[0017] Figure 2 A schematic diagram of the structural connection of an example microfluidic detection device according to an embodiment of this application is shown;

[0018] Figure 3 A flowchart illustrating an example of two-phase injection hydraulic metering verification in a method according to an embodiment of this application is shown.

[0019] Figure 4 A flowchart illustrating an example of troponin concentration correction under plasma volume constraints in a method according to an embodiment of this application is shown.

[0020] Figure 5 A schematic diagram illustrating the system operation mechanism of an example of a microfluidic detection method for blood troponin incorporating blood pressure fluctuations according to an embodiment of this application is shown.

[0021] Figure 6 A schematic diagram of the comparative experimental simulation results of different methods in cardiac-level hydraulic metering and fluid circuit residual evaluation is shown.

[0022] Figure 7 A schematic diagram illustrating the comparative experimental simulation results of different methods for detecting troponin sequences under conditions of fluctuating plasma volume is shown.

[0023] Figure 8 The diagram illustrates the comparative simulation results of different methods in the hematocrit-pulse pressure parameter space and an example of ablation using the complete method mechanism.

[0024] Figure 9 A structural block diagram of an example of a blood troponin microfluidic detection system incorporating blood pressure fluctuations, according to an embodiment of this application, is shown. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0026] It should be noted that, in terms of specific in vitro detection implementation, various cardiac troponin detection systems based on POCT (Point-of-Care Testing) have been developed. For example, systems using DMF (Digital Microfluidics) technology can divide pre-acquired and in vitro-placed whole blood samples into microdroplets within a microfluidic cartridge, and complete magnetic bead immunoassay, washing, and signal reading. Systems using FET (Field-Effect Transistor) biosensors can detect cTnI (cardiactroponin I) in ex vivo whole blood samples, reducing sample pretreatment operations such as centrifugation and pipetting. These systems are mainly designed around detection sensitivity, detection limit, reaction time, and sensor interface performance, and typically complete the corresponding reaction and reading processes after the ex vivo blood sample enters the detection device.

[0027] For microfluidic assays requiring plasma separation from ex vivo whole blood samples, current systems employ methods such as microchannel splitting, microstructure filtration, or capillary actuation to reduce the number of blood cells entering the detection area. Plasma purity, plasma yield, throughput, and device complexity are typically interdependent, and separation efficiency is also affected by factors such as hematocrit (Hct), inlet flow rate, and microchannel structure. At the microchannel scale, blood cell migration, sedimentation, splitting, and the Fahreus effect can cause local hematocrit to deviate from the overall state of the input whole blood sample. Therefore, the effective plasma volume reaching the detection area may vary for the same volume of ex vivo whole blood sample under different separation conditions.

[0028] In microfluidic heterogeneous immunoassay, the binding of antigens to solid-phase antibodies is influenced not only by the analyte concentration but also by sample flow rate, actual injection volume, reaction residence time, and interfacial mass transfer conditions. In continuous flow heterogeneous immunoassay systems, variations in flow conditions can alter the binding kinetics and capture efficiency of antigens and antibodies. Correspondingly, these systems typically convert the endpoint optical or electrical signal into the analyte concentration in the in vitro sample based on a pre-established standard curve; however, when the actual reaction conditions differ from the calibration conditions, the correspondence between the endpoint signal and the analyte concentration may also change.

[0029] In pressure-driven microfluidic systems within the detection device, the pressure controller, sample delivery tubing, and microfluidic chip together constitute the extracorporeal fluid delivery path. Currently, these systems typically maintain the set flow state using methods such as pressure control, flow feedback, or input compensation. However, due to the dynamic response characteristics of the sample delivery tubing and the microfluidic chip, deviations may exist between the set pressure or flow rate at the control end and the actual flow state within the chip. Furthermore, the viscosity and blood cell count of ex vivo blood samples can alter flow resistance, resulting in different flow outcomes under different sample conditions with the same control parameters.

[0030] In terms of concentration correction for ex vivo blood samples, the Dill-Costill plasma volume estimation relationship can be used to form a plasma volume change parameter characterizing the relative dilution or concentration of the sample based on the changes in hemoglobin concentration and hematocrit of ex vivo blood samples obtained at different sampling time points. This estimation relationship is based on the assumption that the red blood cell count is approximately conserved within the sample collection interval and that the measured hematocrit can characterize the overall blood composition at the corresponding sampling time point. When influencing factors such as blood transfusion, blood loss, or changes in local blood cell distribution occur within the sample collection interval, the above-mentioned basic conditions may change. When using this estimation relationship to correct the concentration of biomarkers in ex vivo whole blood, it is also necessary to distinguish the concentration change relationship between plasma or serum biomarkers and whole blood biomarkers; therefore, this type of correction parameter has corresponding applicable conditions.

[0031] Furthermore, when performing in vitro testing on ex vivo blood samples corresponding to the hemodialysis (HD) process, troponin sequence readings may vary depending on the dialysis membrane type, fluid removal method, and molecular transport conditions. These sample processing conditions differ from those for ex vivo blood samples obtained under typical blood pressure fluctuations, but they do indicate that the troponin detection concentration at different sampling points may also be affected by sample dilution or concentration, as well as in vitro processing conditions. Therefore, it is difficult to distinguish the effects of changes in target analyte content, sample dilution or concentration, and in vitro processing conditions based solely on the detection readings.

[0032] It should be understood that the above description of the relevant technologies is intended only to help the public better understand the inventive spirit and motivation of this application, and is not intended to limit this application. Furthermore, the technical solutions described in the above-mentioned relevant technologies are not prior art, and may also be undisclosed technical solutions, such as those under research or in the laboratory stage.

[0033] Figure 1 A flowchart illustrating an example of a microfluidic detection method for blood troponin incorporating blood pressure fluctuations according to an embodiment of this application is shown. Figure 2 A schematic diagram of the structural connection of an example microfluidic detection device according to an embodiment of this application is shown.

[0034] Regarding the entity responsible for implementing the method in the embodiments of this application, it may include... Figure 2 The microfluidic detection device 200 and the data processing unit 222 communicatively connected to the microfluidic detection device 200 are shown. The microfluidic detection device 200 is provided with two independent detection paths with mirrored flow channel structures for forming blood samples in the pressure-boosting phase and the pressure-depressing phase. The data processing unit 222 receives continuous blood pressure waveform data provided by an external blood pressure data source 224 through a data interface 221, controls the corresponding microvalve through a microvalve drive component 223, and receives pressure detection data and detection signals to perform flow rate determination, cumulative throughput calculation, detection signal correction, and biphasic detection result processing.

[0035] Figure 2 Solid arrows indicate the flow direction of the whole blood, separated plasma, or fluid after testing, while dashed arrows indicate the transmission relationship of continuous blood pressure waveform data, pressure detection data, detection signals, or microvalve control signals. Figure 2 The structure shown is an exemplary implementation of the microfluidic detection device 200, and does not limit the specific shape, relative position and flow channel size of each functional area.

[0036] This embodiment processes whole blood samples already acquired and located in the in vitro sample input path, and each detection step is performed after the whole blood sample enters the in vitro sample input path. Continuous blood pressure waveform data can be provided in real time by an external blood pressure data acquisition device, or it can be read from a cache module connected to the external blood pressure data acquisition device and transmitted to the data processing unit via a data interface. The continuous blood pressure waveform data and the whole blood sample injection process have a unified time reference, and the pressure values ​​in the continuous blood pressure waveform data serve as representations of the inlet pressure at the sample input end. The resulting troponin detection results are in vitro detection data obtained specifically for the whole blood sample.

[0037] like Figure 1As shown, in step S110, continuous blood pressure waveform data corresponding to the predetermined sampling period of the whole blood sample to be tested is acquired. The target heartbeat and its target pressure value between diastolic and systolic blood pressure are determined from the continuous blood pressure waveform data. The times when the pressure-raising and depressor phases of the target heartbeat reach the target pressure value are respectively determined as the pressure-raising trigger time and the pressure-depressor trigger time. The predetermined sampling period is a preset time window used to select the sampling trigger time, and the continuous blood pressure waveform data includes pressure values ​​received through a data interface and the data acquisition time corresponding to each pressure value.

[0038] like Figure 2 As shown, the external blood pressure data source 224 is connected to the data interface 221, and the data interface 221 is connected to the data processing unit 222. The external blood pressure data source 224 can be a blood pressure data acquisition device that acquires continuous blood pressure waveforms in real time, or it can be a cache module or data storage device that stores continuous blood pressure waveform data. The data interface 221 transmits the pressure value and the corresponding data acquisition time to the data processing unit 222. The data processing unit 222 determines the target heartbeat, target pressure value, pressure increase trigger time, and pressure decrease trigger time from the continuous blood pressure waveform data according to the time base corresponding to the microvalve control clock.

[0039] The data processing unit 222 is connected to the microvalve drive assembly 223. After determining the boost trigger time and the deboost trigger time, the data processing unit 222 generates corresponding microvalve control signals and sends the microvalve control signals to the microvalve drive assembly 223 so that the trigger time in the waveform data can be converted into the microvalve drive time of the corresponding injection path.

[0040] The predetermined sampling period can be pre-set based on the time when the whole blood sample is introduced into the microfluidic detection device, the preparation status of the microfluidic detection device, and the available time range of continuous blood pressure waveform data. The data processing unit establishes a correspondence between the data acquisition time corresponding to each pressure value and the microvalve control clock based on a unified time reference, so that the pressure values ​​within the predetermined sampling period can be used to determine the microvalve opening time. When the continuous blood pressure waveform data uses the data acquisition time of an external blood pressure data acquisition device, time conversion can be performed using the clock correspondence recorded by the data interface; when the data transmission process and the microvalve driving process have a calibrated response delay, the sending time of the microvalve control signal can be adjusted according to the corresponding response delay.

[0041] The data processing unit divides the continuous blood pressure waveform data within the predetermined sampling period according to the heartbeat cycle, and determines the diastolic pressure, systolic pressure, systolic-boosting phase, and depressive phase of each heartbeat based on the minimum pressure value, maximum pressure value, and direction of pressure change over time. The target heartbeat is selected from heartbeats within the predetermined sampling period that exhibit both systolic-boosting and depressive phases. The target pressure value is set between the diastolic and systolic pressures of the target heartbeat according to a preset pressure layer determination rule, thus being higher than the diastolic pressure and lower than the systolic pressure. The preset pressure layer determination rule can be pre-configured based on the sampling pressure range and heartbeat pressure range of the microfluidic detection device. Specific target heartbeat screening conditions and target pressure value determination relationships can be further defined in other embodiments.

[0042] During the pressure-boosting phase of the target heartbeat, the data processing unit searches for the moment when the pressure value reaches the target pressure value and the rate of pressure change is positive, in the direction of increasing data acquisition time, and determines the corresponding moment as the pressure-boosting trigger moment. During the pressure-deflating phase of the target heartbeat, the data processing unit searches for the moment when the pressure value reaches the target pressure value and the rate of pressure change is negative, and determines the corresponding moment as the pressure-deflating trigger moment. The pressure-boosting trigger moment and the pressure-deflating trigger moment correspond to the same target pressure value, but to opposite directions of pressure change, thus forming a biphasic trigger moment within the same target heartbeat.

[0043] In step S120, the boost phase microvalve and the depressurization phase microvalve are opened at the boost trigger time and the depressurization trigger time, respectively, so that the whole blood to be tested enters the corresponding injection branch. Based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, the blood flow resistance parameter and the pipeline compliance parameter, the instantaneous flow rate of each phase injection process is determined and integrated over time to obtain the cumulative throughput volume of each phase. When the cumulative throughput volume of each phase reaches the target sample volume, the corresponding microvalve is closed to form the boost phase blood sample and the depressurization phase blood sample.

[0044] like Figure 2 As shown, the sample input terminal 201 is connected to the shunt region 202, which is connected to both the boosting phase microvalve 203 and the depressurizing phase microvalve 204. Based on the control signal provided by the data processing unit 222, the microvalve drive assembly 223 controls the boosting phase microvalve 203 to open at the boosting trigger moment, allowing the tested whole blood sample to enter the boosting phase injection branch 205; and controls the depressurizing phase microvalve 204 to open at the depressurizing trigger moment, allowing the tested whole blood sample to enter the depressurizing phase injection branch 206. The boosting phase microvalve 203 and the depressurizing phase microvalve 204 close when their respective accumulated throughput reaches the target sample volume, enabling the two injection phases to independently complete sample measurement according to their respective pressure changes and liquid path responses.

[0045] The boosting phase injection branch 205 is sequentially connected to the boosting phase flexible metering chamber 207 and the boosting phase outlet pressure detection unit 209, while the depressurizing phase injection branch 206 is sequentially connected to the depressurizing phase flexible metering chamber 208 and the depressurizing phase outlet pressure detection unit 210. The boosting phase flexible metering chamber 207 and the depressurizing phase flexible metering chamber 208 respond to the sample loading state during the corresponding injection process, respectively. The boosting phase outlet pressure detection unit 209 and the depressurizing phase outlet pressure detection unit 210 acquire the outlet pressure of their respective injection branches and feed the outlet pressure back to the data processing unit 222. The data processing unit 222, combining the inlet pressure characterization value, the corresponding outlet pressure, blood flow resistance parameters, and tubing compliance parameters, determines the instantaneous flow rate and cumulative throughput volume of the boosting and depressurizing phases, respectively.

[0046] The instantaneous flow rate of each phase injection process is determined based on the pressure and fluid flow conditions during the microvalve opening period. The pressure conditions include the pressure values ​​in the continuous blood pressure waveform data and their rate of change with the data acquisition time. The fluid flow conditions include the corresponding injection branch outlet pressure, blood flow resistance parameters, and tubing compliance parameters. The injection branch outlet pressure characterizes the outlet pressure boundary of the injection branch; the blood flow resistance parameter characterizes the flow resistance encountered by the isolated whole blood flowing through the injection branch; and the tubing compliance parameters characterize the volumetric response of the connecting tubing and the injection branch under pressure changes.

[0047] Blood flow resistance parameters can be pre-calibrated based on the flow channel structure of the microfluidic detection device, the rheological parameters of the whole blood sample, and the ambient temperature. Alternatively, the baseline flow resistance can be corrected based on the current sample parameters of the whole blood sample. Tubing compliance parameters can be pre-determined by recording the volumetric response of the connecting tubing and the injection branch under known pressure variations. The corresponding injection branch outlet pressure can be obtained from a pressure detection unit located on the outlet side, or it can be pre-set based on the pressure boundary conditions on the outlet side.

[0048] For any injection phase, the data processing unit determines the pressure-driven flow component based on the pressure value, the corresponding injection branch outlet pressure, and blood flow resistance parameters. It also determines the flow component generated by the dynamic response of the fluid path based on the rate of change of the pressure value and the tubing compliance parameters. The data processing unit determines the instantaneous flow rate along the injection direction based on these flow components and integrates the instantaneous flow rate over time from the moment the corresponding microvalve opens until the current time, obtaining the cumulative volume passing through the corresponding injection branch from the moment the microvalve opens to the current time.

[0049] The target sample volume can be preset based on the sample volume required for troponin immunoassay and the volume of the microfluidic detection region. The data processing unit compares the cumulative throughput volume of each phase with the target sample volume. When the cumulative throughput volume of the boosting phase reaches the target sample volume, the boosting phase microvalve is closed, resulting in a boosting phase blood sample; when the cumulative throughput volume of the depressurizing phase reaches the target sample volume, the depressurizing phase microvalve is closed, resulting in a depressurizing phase blood sample. Using the cumulative throughput volume as the criterion for closing the microvalve of each phase allows the boosting and depressurizing phases to complete sample measurement based on their respective pressure changes and fluid dynamic response.

[0050] In step S130, plasma separation and troponin immunoassay are performed on the blood samples of the pressurized phase and the blood samples of the depressurized phase, respectively, to obtain the troponin detection signal and reaction residence time corresponding to each phase of the blood sample.

[0051] like Figure 2 As shown, the boosting phase outlet pressure detection unit 209 is connected to the boosting phase plasma separation region 211, and the depressurizing phase outlet pressure detection unit 210 is connected to the depressurizing phase plasma separation region 212. The boosting phase blood sample and the depressurizing phase blood sample undergo plasma separation in independent pathways to reduce fluid mixing between the two sample phases. Plasma separation can be achieved using microchannel diversion structures, microstructured filtration regions, or separation structures matched to the size of blood cells.

[0052] The output of the pressurized phase plasma separation region 211 is connected to the pressurized phase troponin immunoreaction region 213, the pressurized phase albumin reference reaction region 215, and the pressurized phase inert fluorescence reference region 217, respectively; the output of the depressurized phase plasma separation region 212 is connected to the depressurized phase troponin immunoreaction region 214, the depressurized phase albumin reference reaction region 216, and the depressurized phase inert fluorescence reference region 218, respectively. The pressurized phase detection pathway and the depressurized phase detection pathway adopt a mirror-image flow channel structure, allowing the two phases of plasma to undergo immunoreaction and reference detection under corresponding flow channel conditions.

[0053] The boosting-phase troponin immunoreaction region 213 and the depressurizing-phase troponin immunoreaction region 214 are used to generate the troponin detection signal for the corresponding injection phase; the boosting-phase albumin reference reaction region 215 and the depressurizing-phase albumin reference reaction region 216 are used to generate the albumin reference signal for the corresponding injection phase; the boosting-phase inert fluorescence reference region 217 and the depressurizing-phase inert fluorescence reference region 218 are used to generate the optical reference signal for the corresponding injection phase. The signals generated by the above reaction regions and reference regions are read by the signal reading region 219, and the fluid after detection flows into the waste liquid collection region 220.

[0054] The pressurized phase troponin immunoreaction region 213 and the depressurized phase troponin immunoreaction region 214 are each provided with an immunocapture material that matches the troponin to be tested. The immunocapture material may include capture antibodies immobilized on the surface of the corresponding immunoreaction region, or it may include immunomagnetic beads capable of controlled movement within the corresponding detection pathway. After plasma from each phase enters the corresponding troponin immunoreaction region, the troponin to be tested binds to the immunocapture material, and a troponin detection signal for the corresponding injection phase is generated through a signal readout method matching the immunocapture material.

[0055] The reaction residence time can be determined based on the moment when the corresponding phase plasma enters the troponin immune reaction zone and the moment the signal is read, or it can be determined based on the actual transit time of the corresponding phase plasma within the preset immune reaction zone. The moment can be recorded by the fluid position detection unit in the microfluidic detection device, or it can be determined based on the microvalve control moment, the flow rate of the corresponding injection process, and the channel volume between the inlet of the immune reaction zone and the signal reading position. Both the troponin detection signal and the reaction residence time carry the identifier of the corresponding injection phase, allowing the boost phase detection data and deboost phase detection data to be used separately for the capture efficiency correction of the corresponding phase.

[0056] In step S140, the immune capture efficiency of each phase is determined based on the cumulative throughput of each phase, the reaction residence time of the corresponding blood sample, and the preset immune capture parameters. The troponin detection signal corresponding to each blood sample is then corrected using the immune capture efficiency to obtain the hydraulic correction concentration of the pressurized phase and the hydraulic correction concentration of the depressurized phase.

[0057] Preset immune capture parameters are used to characterize the reaction characteristics between the troponin to be tested and the immune capture material, as well as the capture ability of the immune reaction region. These preset immune capture parameters can be obtained by calibrating quality control samples with known troponin concentrations and stored in a parameter file associated with the corresponding microfluidic detection cartridge or reagent batch.

[0058] The data processing unit can invoke a pre-established mapping relationship between cumulative throughput volume, reaction residence time, preset immune capture parameters, and immune capture efficiency to determine the immune capture efficiency of the boost phase and the immune capture efficiency of the depressurization phase, respectively. The mapping relationship can be implemented using calibration curves, lookup tables, or preset reaction-response relationships. Its input includes at least the corresponding phase cumulative throughput volume and reaction residence time, and its output is the immune capture efficiency used to correct the corresponding troponin detection signal.

[0059] exist Figure 2In the example shown, the signal reading area 219 reads the troponin detection signals formed by the boost phase troponin immune response area 213 and the depressurization phase troponin immune response area 214, and transmits the reading results to the data processing unit 222.

[0060] The signal reading area 219 can also read albumin reference signals and inert fluorescence reference signals. The albumin reference signal can be used to characterize the albumin state of the corresponding plasma sample, and the inert fluorescence reference signal can be used to characterize the optical response state of the detection pathway. These reference signals can be used in corresponding embodiments for detection signal correction or plasma volume state processing; in the main processing chain of this embodiment, the troponin detection signal, cumulative throughput volume, and reaction residence time are used to form the hydraulically corrected concentrations for the boosting phase and the depressurizing phase.

[0061] For each injection phase, the data processing unit corrects and converts the troponin detection signal based on the corresponding immunocapture efficiency and the concentration calibration relationship matched with the microfluidic detection device. After the above processing, the concentration conversion result corresponding to the pressurization phase is determined as the hydraulically corrected concentration for the pressurization phase, and the concentration conversion result corresponding to the depressurization phase is determined as the hydraulically corrected concentration for the depressurization phase. The hydraulically corrected concentration represents the troponin concentration characterization value obtained after taking into account the influence of the cumulative throughput volume and reaction residence time of the corresponding phase on the immunocapture process.

[0062] In step S150, the geometric mean of the hydraulic correction concentration of the boosting phase and the hydraulic correction concentration of the depressurizing phase is calculated to obtain the two-phase symmetry, and the absolute logarithm of the ratio of the hydraulic correction concentration of the boosting phase to the hydraulic correction concentration of the depressurizing phase is calculated to obtain the antisymmetric residual of the liquid path.

[0063] When both the hydraulic correction concentration of the pressure-raising phase and the hydraulic correction concentration of the pressure-falling phase are positive, the two-phase balancing amount is determined according to the following formula:

[0064] Equation (1)

[0065] The antisymmetric residual of the fluid path is determined according to the following formula:

[0066] Equation (2)

[0067] In the formula, For hydraulic correction concentration of the pressurized phase, For hydraulic correction concentration of the depressurized phase, For two-way symmetry, This represents the antisymmetric residual in the liquid path. The above variable form corresponds to the hydraulic correction concentration and dual relative symmetry variable forms used in subsequent sequence processing for different detection time points.

[0068] When the hydraulically corrected concentrations of the boosting and depressurizing phases are interchanged, the relative symmetry remains unchanged, the logarithmic sign of the concentration ratio changes, and the antisymmetric residual of the liquid path remains unchanged. Therefore, the relative symmetry is used to characterize the troponin concentration component shared by the boosting and depressurizing phases, while the antisymmetric residual of the liquid path is used to characterize the overall differences formed between the two injection phases during injection, plasma separation, immune response, and signal reading.

[0069] In step S160, when the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold, the dual relative symmetry is output as the troponin detection result corresponding to the target heartbeat; when the antisymmetric residual of the liquid path is greater than the preset residual threshold, a re-injection command is generated.

[0070] The preset residual threshold can be determined based on the statistical distribution of the antisymmetric residuals obtained from repeated detection of the same sample in both the boost phase and depressurization phase detection pathways. During calibration, quality control samples with different sample parameters or different troponin concentrations can be used to obtain the antisymmetric residuals under permissible detection conditions, and the preset residual threshold can be set according to preset quality control conditions. The preset residual threshold can be associated with the detection reagent batch, microfluidic detection cartridge structure, or detection signal reading method.

[0071] When the antisymmetric residual of the fluid path is less than or equal to the preset residual threshold, the data processing unit 222 outputs the dual relative symmetry as the troponin detection result corresponding to the current target heartbeat. The output data may include the troponin detection result, the time information corresponding to the target heartbeat, and the antisymmetric residual of the fluid path, so as to retain the sample time point and fluid path consistency information corresponding to the current detection result.

[0072] When the antisymmetric residual of the liquid path is greater than the preset residual threshold, the data processing unit 222 marks the current biphase symmetry as invalid and generates a re-injection command. It controls the boost phase microvalve 203 and the depressurization phase microvalve 204 to enter the next injection preparation state through the microvalve drive component 223. In subsequent heartbeats that meet the injection conditions, the data processing unit 222 re-determines the boost trigger time and the depressurization trigger time, and controls the corresponding biphase path to re-execute the injection and detection.

[0073] In this embodiment, two moments with the same pressure value but opposite directions of pressure change during the same target heartbeat are used as biphasic trigger moments. The injection is controlled to end according to the cumulative throughput volume of each phase, and the troponin detection signal is corrected based on the corresponding cumulative throughput volume and reaction residence time. Biphasic symmetry is used to form the troponin detection result of the isolated whole blood sample. The antisymmetric residual of the liquid path is used to characterize the degree of mismatch between the biphasic sampling and detection process corresponding to the detection result, and to determine whether the detection result meets the preset liquid path consistency condition. The output troponin detection result belongs to the in vitro detection data of the isolated whole blood sample.

[0074] Regarding the details of determining the target heartbeat, target pressure value, and the timing of pressure increase and pressure decrease in the method of the embodiments of this application, in some examples of the embodiments of this application, the continuous blood pressure waveform data is segmented according to the heartbeat cycle to obtain multiple candidate heartbeats arranged in chronological order, and the diastolic pressure, systolic pressure, pulse pressure, pressure increase phase, pressure decrease phase, and pressure-time integral area of ​​each candidate heartbeat are determined.

[0075] The continuous blood pressure waveform data consists of multiple pressure values ​​arranged according to the data acquisition time. Diastolic blood pressure troughs can be identified based on local minima, and the blood pressure waveform between two adjacent diastolic blood pressure troughs can be divided into a candidate heartbeat; when the external blood pressure data acquisition device provides a heartbeat cycle marker, segmentation can also be completed based on the heartbeat cycle marker.

[0076] For each candidate heartbeat, the lowest pressure value is defined as diastolic pressure, the highest pressure value as systolic pressure, and the pressure difference between the systolic and diastolic pressures is defined as pulse pressure. The waveform segment between the diastolic pressure trough and the systolic pressure peak is designated as the pressure-boosting phase, and the waveform segment between the systolic pressure peak and the next diastolic pressure trough is designated as the pressure-deflating phase. Under discrete sampling conditions, the average value of adjacent pressure values ​​can be accumulated segment by segment based on the time interval between adjacent data acquisition moments to obtain the pressure-time integral area of ​​the corresponding candidate heartbeat; each candidate heartbeat uses the same pressure benchmark and integration rule. Subsequently, based on the pulse pressure of each candidate heartbeat and a preset proportionality coefficient, a candidate pressure value is determined between the corresponding diastolic and systolic pressures.

[0077] The candidate pressure value can be determined according to the following formula:

[0078] Equation (3)

[0079] In the formula, Candidate heartbeat numbers are set in chronological order; For the first Candidate stress values ​​for each candidate heartbeat; For the first Diastolic blood pressure of each candidate heartbeat; For the first The systolic blood pressure of each candidate heartbeat; It is a unitless preset scaling factor, and satisfies The preset proportional coefficient can be pre-set based on the microvalve response time, target injection pressure layer, and blood pressure waveform calibration results. Each candidate heartbeat in the same sequence of detection uses the same preset proportional coefficient, ensuring that each candidate pressure value is formed according to the same pressure layer determination rules.

[0080] Subsequently, for each candidate heartbeat with a preceding candidate heartbeat, the candidate heartbeats that meet the corresponding preset conditions in terms of pulse pressure, data acquisition time continuity, pressure range, monotonicity of the boost phase, direction of pressure change during the depressurization phase, and pressure-time integral area difference with the preceding candidate heartbeat are determined as usable heartbeats; the earliest available heartbeat from the usable heartbeats whose heartbeat time period is within the predetermined injection time period is selected as the target heartbeat, and the corresponding candidate pressure value is determined as the target pressure value.

[0081] Availability assessment begins with a candidate heartbeat that has a preceding candidate heartbeat. For each candidate heartbeat, the pulse pressure is compared to a preset pulse pressure threshold, the maximum time interval between adjacent data acquisition moments is compared to a preset sampling interval threshold, and it is checked whether each pressure value is within the blood pressure measurement range. Monotonicity during the bolus phase is determined based on the direction of pressure change within the bolus phase; when local pressure fluctuations are within a preset monotonicity tolerance, assessment can be made based on the rate of pressure change in the neighborhood of the candidate pressure value intersection. During the depressor phase, the corresponding candidate pressure value should pass through at least once with a negative rate of pressure change.

[0082] The pressure-time integral area of ​​the current candidate heartbeat and the previous candidate heartbeat is calculated using the same integration rule, and the absolute value of the difference between the two is compared with a preset area threshold. The preset pulse pressure threshold, preset sampling interval threshold, preset monotonicity tolerance, and preset area threshold can be pre-calibrated based on the sampling parameters of the blood pressure data acquisition device, historical effective blood pressure waveforms, and microvalve response conditions. Candidate heartbeats that meet all preset conditions are added to the available heartbeat set. The data processing unit determines the target heartbeat from the available heartbeat set in chronological order and identifies the candidate pressure value corresponding to the target heartbeat as the target pressure value. .

[0083] Furthermore, the moment when the target heart rate reaches the target pressure value during the pressure rise phase under the condition that the pressure change rate is greater than zero is determined as the pressure rise trigger moment. From the candidate moments for pressure reduction when the target heart rate reaches the target pressure value during the pressure reduction phase under the condition that the pressure change rate is less than zero, the candidate moment for pressure reduction with the smallest difference between the absolute value of the pressure change rate and the absolute value of the pressure change rate at the pressure rise trigger moment is selected as the pressure reduction trigger moment. When the difference corresponding to multiple candidate moments for pressure reduction is the same, the earliest one is selected as the pressure reduction trigger moment.

[0084] The boost trigger time and the buck trigger time can be expressed by the following formula:

[0085] Equation (4)

[0086] In the formula, This refers to the moment of data acquisition. The target pressure value corresponding to the target heartbeat; This is the boost trigger moment; This is the timing of the voltage reduction trigger. For a moment The corresponding pressure value; This represents the rate of change of the pressure value relative to the time of data acquisition.

[0087] Under discrete sampling conditions, when two adjacent pressure values ​​are located on either side of the target pressure value, linear interpolation can be performed based on the two adjacent pressure values ​​and their data acquisition times. The intersection time obtained from the interpolation is taken as the corresponding trigger time. The pressure change rate can be determined based on a preset number of pressure values ​​and data acquisition times before and after the intersection time. The preset number is set according to the time range used in calculating the pressure change rate based on the sampling frequency of the blood pressure data acquisition device.

[0088] When multiple candidate pressure reduction moments are generated during the depressurization phase due to dicrotic notch or local pressure fluctuations, the difference between the absolute value of the pressure change rate at each candidate moment and the absolute value of the pressure change rate at the boost trigger moment is calculated. The depressurization trigger moments are then selected in ascending order of these differences. If the differences are the same, the earliest time is selected based on the data acquisition time. The determined boost and depressurization trigger moments correspond to the same target pressure value and, respectively, the positive and negative pressure change directions, and are used to trigger the boost and depressurization microvalves to perform two-phase injection.

[0089] Figure 3 A flowchart illustrating an example of two-phase injection hydraulic metering verification in a method according to an embodiment of this application is shown.

[0090] like Figure 3As shown, in step S310, the initial hematocrit and ambient temperature of the whole blood sample to be tested are obtained before injection. Based on the ratio between the initial hematocrit and the non-erythrocyte volume fraction determined by the initial hematocrit, the reference flow resistance at the ambient temperature is nonlinearly corrected to obtain the blood flow resistance parameter.

[0091] The initial hematocrit can be obtained from the sample test results before the injection of the whole blood sample to be tested, or it can be read from the sample test data associated with the whole blood sample to be tested. The ambient temperature is determined based on the temperature measurement data of the current microfluidic detection environment. Both the ambient temperature and the initial hematocrit are associated with the current two-phase injection process and are used for calculating the flow rates of the boost and depressurization phases.

[0092] Blood flow resistance parameters can be determined according to the following formula:

[0093] Equation (5)

[0094] In the formula, The blood flow resistance parameters of the isolated whole blood to be tested; The ambient temperature of the microfluidic detection environment; ambient temperature The reference current resistance is below, and ; The initial hematocrit of the whole blood sample to be tested before injection is given, and ; The non-erythrocyte volume fraction determined by the initial hematocrit. and The coefficients are dimensionless flow resistance coefficients, and their values ​​must be within the allowable range of the initial hematocrit. to make .

[0095] Reference flow resistance The flow resistance coefficient can be obtained by using a calibration solution with known rheological properties, pre-determining the pressure difference and steady-state flow rate of the injection branch at different calibration temperatures, and then reading or interpolating the value based on the current ambient temperature. and Quality control blood samples with different hematocrits can be used to fit the relationship between measured flow resistance and baseline flow resistance. The first and second correction terms characterize the linear and nonlinear contributions of hematocrit changes to the flow resistance of isolated whole blood, respectively.

[0096] In step S320, for each injection phase in the boosting and depressurizing phases, the pressure-driven flow rate is determined based on the non-negative pressure difference between the pressure value during the opening of the corresponding microvalve and the outlet pressure and capillary threshold pressure of the corresponding injection branch, and the blood flow resistance parameter. The compliance compensation flow rate is determined based on the pipeline compliance parameter and the rate of change of the pressure value. The non-negative instantaneous flow rate along the injection direction is determined based on the pressure-driven flow rate and the compliance compensation flow rate. Time integration is performed starting from the triggering time of the corresponding injection phase to obtain the cumulative throughput volume of the corresponding injection phase.

[0097] The non-negative instantaneous flow rate along the injection direction can be determined by the following formula:

[0098] Equation (6)

[0099] In the formula, For the identification of the injected phase, Indicates the boost phase. Indicates the pressure-reducing phase; For the injection phase At any moment Instantaneous flow rate along the injection direction; For the injection phase The corresponding outlet pressure of the injection branch; For the injection phase The corresponding capillary threshold pressure; For the injection phase The pipeline compliance parameters corresponding to the injection branch, and ; This indicates that the corresponding calculation result will be restricted to non-negative values.

[0100] The inlet pressure is the pressure value from the aforementioned continuous blood pressure waveform data. The outlet pressure of the injection branch can be obtained from the corresponding outlet pressure detection result, capillary threshold pressure. The pipeline compliance parameters can be predetermined based on the pressure difference when the calibration fluid begins to enter the corresponding injection branch. Calibration can be performed based on the branch volume response corresponding to known pressure changes. The inner non-negative constraint sets the pressure-driven flow rate to zero when the inlet pressure does not exceed the sum of the outlet pressure and the capillary threshold pressure. The outer non-negative constraint constrains the combined result of the pressure-driven flow rate and the compliance compensation flow rate, keeping the instantaneous flow rate along the injection direction.

[0101] The cumulative volume can be determined by the following formula:

[0102] Equation (7)

[0103] In the formula, To extract the sample phase The trigger time to time The cumulative volume passing through; For the injection phase The triggering time, when hour ,when hour ; This is the integration time variable. Because... Constrained to be non-negative, cumulative passing volume Since the cumulative through volume remains non-decreasing over the integration time, the moment when the cumulative through volume first reaches the target sample volume can be determined.

[0104] When continuous blood pressure waveform data is sampled discretely, the instantaneous flow rate can be determined at each data acquisition moment, and the average value of adjacent instantaneous flow rates can be accumulated segment by segment based on the time interval between adjacent data acquisition moments. When the microvalve control cycle is shorter than the blood pressure waveform data acquisition cycle, the instantaneous flow rate can be interpolated between adjacent data acquisition moments to determine the valve closing moment when the cumulative through volume first reaches the target sample volume.

[0105] In step S330, when the cumulative volume of each phase first reaches the target sample volume, the corresponding microvalve is closed, and the membrane metering volume of the corresponding phase is determined based on the membrane displacement retained by the flexible metering cavity in the corresponding injection branch after the microvalve is closed.

[0106] The cumulative volume passing through when the corresponding micro-valve is closed is denoted as... The correspondence between membrane displacement and membrane metering volume can be predetermined by injecting multiple known volumes of calibration liquid into the flexible metering chamber and recording the membrane displacement corresponding to each known volume. When the current membrane displacement is between adjacent calibration displacements, interpolation can be performed based on the corresponding known volume.

[0107] After the valve is closed, membrane displacement can be continuously acquired within a preset holding period. When the change in membrane displacement is within a preset allowable range, the membrane metering volume is determined based on the representative value of the membrane displacement during the holding period. This reduces the impact of pressure disturbances during valve closure on membrane metering results. The membrane metering volume is derived from the deformation response of the flexible metering cavity, constituting a volume characterization independent of the hydraulic integration result.

[0108] In step S340, the ratio of the absolute value of the difference between the cumulative volume of each phase and the corresponding membrane metering volume to the target sample volume is determined as the single-phase volume deviation ratio of the corresponding phase, and the ratio of the absolute value of the difference between the metering volume of the boosting phase and the metering volume of the depressurizing phase to the target sample volume is determined as the two-phase volume deviation ratio. When each volume deviation ratio is less than or equal to its corresponding preset volume deviation threshold, plasma separation and troponin immunoassay are performed on the boosting phase blood sample and the depressurizing phase blood sample. When any volume deviation ratio is greater than the corresponding preset volume deviation threshold, a re-injection command is generated.

[0109] The single-phase volume deviation ratio can be determined by the following formula:

[0110] Equation (8)

[0111] In the formula, For the injection phase The single-phase volume deviation ratio; For the injection phase The corresponding membrane volume; The target sample volume, and ; This indicates the absolute value of the difference. The single-phase volume deviation ratio is used to characterize the difference between the cumulative throughput volume and the membrane metering volume of the same injected phase.

[0112] The two-phase volume deviation ratio can be determined by the following formula:

[0113] Equation (9)

[0114] In the formula, This represents the two-phase volume deviation ratio; For measuring the volume of the boosted phase film; The volumetric volume of the depressurized phase membrane is used. The two-phase volume deviation ratio is used to characterize the difference between the volumetric volumes of the boost phase membrane and the depressurized phase membrane.

[0115] Each preset volume deviation threshold can be set based on the integral error of the cumulative passed volume, the volume calibration error of the flexible metering chamber, the microvalve shut-off delay, and the allowable injection volume variation range for immunoassay. When all volume deviation ratios are within their corresponding threshold ranges, the pressurized and depressurized blood samples are marked as having passed volume verification and proceed to subsequent plasma separation and troponin immunoassay processes. If any volume deviation ratio exceeds its corresponding threshold, a re-injection command is generated. The hydraulic integral result and membrane displacement metering result jointly participate in volume verification, restricting blood samples with intra-phase metering differences or bi-phase volume differences exceeding the corresponding thresholds from entering subsequent immunoassays.

[0116] Regarding the implementation details of optical reference correction and immune capture efficiency correction in the methods of this application's embodiments, in some examples of this application's embodiments, hematocrit detection and plasma separation are performed on blood samples of each phase. The separated pressurized phase plasma and depressurized phase plasma are respectively delivered to the troponin reaction region, albumin reference reaction region, and inert fluorescence reference region within the pressurized phase detection pathway and the depressurized phase detection pathway, respectively. The pressurized phase detection pathway and the depressurized phase detection pathway have mutually mirrored flow channel structures. Hematocrit, albumin concentration, troponin detection signal, reaction residence time, and inert fluorescence reference signal corresponding to each phase blood sample are obtained. In this embodiment, the troponin reaction region is the troponin immune reaction region.

[0117] The flow path length, cross-sectional dimensions, and reaction zone volume of the boosting phase detection pathway and the depressurizing phase detection pathway are matched. The boosting phase plasma and the depressurizing phase plasma pass through their respective reaction zones within independent flow paths. For the injection phase... The obtained hematocrit is recorded as The albumin concentration is denoted as The troponin detection signal is recorded as The reaction residence time is recorded as The inert fluorescence reference signal is denoted as Hematocrit can be obtained by optical absorption or electrical impedance detection before plasma separation; albumin concentration can be obtained based on the detection signal formed in the albumin reference reaction area and the pre-calibrated signal-concentration relationship.

[0118] An inert fluorescent reference region contains a fluorescent reference that does not participate in the troponin immune binding reaction. The amounts of fluorescent reference in the boost and deboost detection pathways correspond. The inert fluorescent reference signal is used to characterize the current response state of the light source, detector, and optical pathway. Reaction dwell time. The time interval between the moment when the corresponding plasma enters the troponin immune response region and the moment when the troponin signal is read is determined.

[0119] Subsequently, based on the proportional relationship between the inert fluorescence reference signal of each phase and the preset reference signal, the preset calibration response coefficients are adjusted to obtain the optical response coefficients of each phase.

[0120] The optical response coefficients of each phase can be determined according to the following formula:

[0121] Equation (10)

[0122] In the formula, For the injection phase The corresponding optical response coefficient; The preset calibration response coefficient is used to characterize the increase in detection signal corresponding to a unit amount of troponin under the reference optical condition. ; For the injection phase The corresponding inert fluorescent reference signal, and ; As a preset reference signal under the reference optical state, and .

[0123] Preset reference signal The calibration response coefficient can be determined by detecting a reference sample with a known fluorescence response, under the conditions of reference light source intensity, reference detector gain, and reference optical path. The correlation between the detection signal increment and troponin mass can be obtained by calibrating the detection signal increment and troponin mass using a quality control sample with known troponin mass under the reference optical conditions. The preset reference signal and preset calibration response coefficient can be stored in association with the detection cartridge batch and signal reading unit identifier. The change in the inert fluorescence reference signal relative to the preset reference signal is obtained through... This information is passed to the concentration conversion process to correct for changes in the response of the current optical state relative to the reference optical state.

[0124] For the injection phase Based on the cumulative volume Hematocrit Preset plasma separation yield Branch partition coefficients for troponin detection Determine the effective plasma volume entering the troponin immune response zone. The effective plasma volume can be determined according to the following formula:

[0125] Equation (11)

[0126] In the formula, This represents the ratio of the actual plasma volume obtained from the separation to the volume of non-erythrocytes. The ratio of the plasma volume entering the troponin detection pathway to the volume of separated plasma, and , The preset plasma separation yield and troponin detection branch allocation coefficient are pre-calibrated based on the plasma separation volume of the quality control blood sample and the flow allocation results of each parallel branch.

[0127] Subsequently, the effective reaction degree of each phase is determined based on the reaction residence time, cumulative throughput volume and preset immune capture parameters. The immune capture efficiency of each phase is determined according to the response relationship that the immune capture efficiency increases monotonically with the increase of the effective reaction degree and tends to saturate.

[0128] The degree of effective reaction in each phase is represented by a dimensionless effective reaction parameter, which can be determined according to the following formula:

[0129] Equation (12)

[0130] In the formula, For the injection phase The corresponding effective reaction parameters; The effective binding rate constant is expressed in units of volume divided by the product of the amount of substance and time. ; This represents the total number of available immune capture sites, expressed in monetary terms. ; For the injection phase The reaction time of the corresponding blood sample, and ; The effective plasma volume for entering the corresponding troponin immune response region, and Based on the measurement relationships of the above variables, This is a unitless parameter.

[0131] Effective binding rate constant The binding response can be determined by labeling quality control samples with known troponin concentrations at different reaction residence times. The total amount of available immune capture sites can be determined. The effective reaction parameters can be determined based on the amount of substance immobilized with the capture antibody within the troponin immunoreaction region and the proportion of usable sites obtained from the corresponding reagent batch calibration. Effective reaction parameters increase with increasing reaction residence time, effective binding rate constant, or total amount of usable immunoreceptor sites, and decrease with increasing effective plasma volume.

[0132] Immune capture efficiency can be determined by the following formula:

[0133] Equation (13)

[0134] In the formula, For the injection phase The corresponding immune capture efficiency. Because... ,therefore The exponential response relationship causes the immune capture efficiency to increase monotonically with increasing effective reaction parameters and tend to saturate, which is used to characterize the proportion of troponin captured by the immune system at the current sample volume and reaction residence time.

[0135] Furthermore, the signal increment of each phase troponin detection signal relative to the preset blank detection signal is determined, and the signal increment is normalized and converted to a concentration based on the corresponding optical response coefficient, cumulative throughput volume and immune capture efficiency, to obtain the hydraulic correction concentration of the pressurization phase and the hydraulic correction concentration of the depressurization phase, respectively.

[0136] The hydraulic correction concentrations for each phase can be determined according to the following formula:

[0137] Equation (14)

[0138] In the formula, For the injection phase The corresponding hydraulic correction concentration; This is a preset blank detection signal; To remove the signal increment after blank response. Preset blank detection signal. Blank samples lacking the target troponin can be pre-measured under the same reagent batch and signal reading conditions as the current test. For test data entering concentration conversion, it meets the following requirements. , , and .

[0139] Optical response coefficient Used to correct for changes in the optical response of the current detection pathway, effective plasma volume Concentration conversion based on actual plasma volume entering the troponin detection pathway; immune capture efficiency. Used to correct signal differences caused by incomplete capture. The increment of the detection signal corresponding to the unit amount of troponin protein characterized, said signal increment is... , and After conversion, it has a quantitative meaning of troponin mass concentration.

[0140] Will The concentration was determined to be the hydraulic correction concentration for the boosting phase. The hydraulic correction concentration for the depressurizing phase is determined. Only when both the pressurizing and depressurizing phases form effective hydraulic correction concentrations are the two-phase hydraulic correction concentrations used to determine the two-phase symmetry and the antisymmetric residual of the liquid path; if the corresponding injection phase does not meet the concentration conversion conditions, the injection phase is marked as invalid for detection data.

[0141] Regarding the details of constructing the feasible interval of the dual-conserved plasma volume in the method of this application embodiment, in some examples of this application embodiment, when the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold, the current detection time point is determined as the current detection time point, and the detection time point where the first antisymmetric residual of the liquid path and the ratio of each volume deviation in the sequence detection are less than or equal to their respective preset thresholds is determined as the reference detection time point; for the current detection time point and the reference detection time point, the arithmetic mean of the hematocrit of the corresponding boosting phase and depressurizing phase is determined as the hematocrit of the corresponding detection time point, and the geometric mean of the albumin concentration of the corresponding boosting phase and depressurizing phase is determined as the albumin concentration of the corresponding detection time point.

[0142] The reference detection time point is determined by the threshold judgment results of the aforementioned antisymmetric residual and volume deviation ratio of the liquid path. The hematocrit of the boosting phase and the hematocrit of the depressurizing phase are obtained from the hematocrit detection results of the corresponding phase blood sample before plasma separation. The albumin concentration of the boosting phase and the albumin concentration of the depressurizing phase are obtained from the detection results of the corresponding phase plasma in the albumin reference reaction region. When both injection phases produce effective hematocrit and effective albumin concentrations, the arithmetic mean and geometric mean are calculated respectively.

[0143] Record the current detection time as the detection time. The reference detection time point is recorded as the detection time point. Detection time point Hematocrit is recorded as The hematocrit at the reference testing time point is recorded as ; Detection time point The albumin concentration is denoted as The albumin concentration at the reference detection time point is recorded as . The arithmetic mean of biphasic hematocrit and the geometric mean of biphasic albumin concentration remain unchanged when the order of the pressurization and depressurization phases is exchanged. The geometric mean aggregates the albumin concentrations of the two phases according to a multiplicative symmetry relationship.

[0144] Detection time point It can be a reference detection time point or a valid detection time point later than the reference detection time point. When When the current detection value is the same as the reference detection value, the corresponding red blood cell conservation relative plasma volume and albumin conservation relative plasma volume are both 1, and the logarithm of both is zero, which is used as the plasma volume reference state for sequence detection.

[0145] Subsequently, the red blood cell conservation relative plasma volume is determined based on the ratio between the hematocrit at the reference detection time point and the hematocrit at the current detection time point, and the ratio between the non-red blood cell volume fraction at the current detection time point and the non-red blood cell volume fraction at the reference detection time point; the albumin conservation relative plasma volume is determined based on the ratio between the albumin concentration at the reference detection time point and the albumin concentration at the current detection time point.

[0146] When the red blood cell count conservation condition is met between the reference detection time point and the current detection time point, the red blood cell conservation relative plasma volume can be determined according to the following formula:

[0147] Equation (15)

[0148] In the formula, For the detection time point The relative plasma volume of red blood cells is conserved relative to the reference detection time point; For the detection time point The non-erythrocyte volume fraction; The non-erythrocyte volume fraction at the reference detection time point; and The red blood cell conservation relative plasma volume is a unitless ratio used to characterize the change in current plasma volume relative to a reference plasma volume when the red blood cell count remains constant.

[0149] When the albumin conservation condition is met between the reference detection time point and the current detection time point, the albumin conservation relative plasma volume can be determined according to the following formula:

[0150] Equation (16)

[0151] In the formula, For the detection time point The albumin-relative plasma volume is conserved relative to the reference testing time point, and , The same albumin detection caliber and calibration relationship are used for both the reference and current detection times. Under the condition that the albumin level remains constant, when the current albumin concentration decreases relative to the reference albumin concentration, Increase; when the current albumin concentration increases relative to the reference albumin concentration, Decrease.

[0152] Subsequently, the red blood cell conservation interval and the albumin conservation interval are constructed by expanding the intervals according to the corresponding logarithmic measurement uncertainty, with the logarithmic values ​​of the red blood cell conservation relative plasma volume and the albumin conservation relative plasma volume as the center, respectively.

[0153] The red blood cell conservation interval can be constructed according to the following formula:

[0154] Equation (17)

[0155] In the formula, For the detection time point The corresponding red blood cell conservation interval; Based on the reference detection time point and the detection time point The logarithmic domain uncertainty of the hematocrit measurement uncertainty is determined, and .

[0156] The albumin conservation zone can be constructed according to the following formula:

[0157] Equation (18)

[0158] In the formula, For the detection time point The corresponding albumin conservation interval; Based on the reference detection time point and the detection time point The logarithmic domain uncertainty of the albumin concentration measurement uncertainty is determined, and . and These are used to characterize the propagation of the corresponding measurement error to and The resulting allowable deviation range.

[0159] The logarithmic domain uncertainty can be determined based on repeated testing results of quality control blood samples with known hematocrit and known albumin concentration, and is stored in association with the batch of the test cartridge, the batch of the test reagent, or the identifier of the signal reading unit. When different calibration intervals correspond to different levels of detection dispersion, the corresponding logarithmic domain uncertainty is retrieved according to the calibration interval in which the current hematocrit or albumin concentration falls. Because... and Both are positive, unitless ratios, and both conservation intervals lie within the logarithmic relative plasma volume domain.

[0160] Subsequently, a conservation applicability state marker is obtained between the reference detection time point and the current detection time point; when the conservation applicability state marker indicates that red blood cell transfusion has occurred or the blood loss reaches a preset blood loss threshold, the red blood cell conservation interval is deactivated; when the conservation applicability state marker indicates that albumin transfusion, plasma exchange, or capillary leakage that meets preset leakage judgment conditions has occurred, the albumin conservation interval is deactivated.

[0161] The conservation applicability status marker can be provided by an external recording system through a data interface, or it can be generated based on intervention event data stored in association with the detection time point. The event occurrence time or state duration is matched with the time interval formed by the reference detection time point to the current detection time point; when the event occurrence time is within the time interval or the state duration intersects with the time interval, the corresponding status marker participates in the conservation applicability judgment at the current detection time point. Deactivated conservation intervals do not participate in the interval intersection calculation at the current detection time point.

[0162] Furthermore, when at least one conservation interval is enabled and there is a non-empty intersection of the enabled conservation intervals, the non-empty intersection is determined as a feasible interval for plasma volume; when both conservation intervals are enabled and the intersection of the red blood cell conservation interval and the albumin conservation interval is empty, a conservation conflict flag is generated; when both conservation intervals are disabled, a control flag is generated that only outputs the troponin detection result.

[0163] Detection time point The corresponding feasible range of plasma volume is denoted as ,in, This is the lower boundary of the feasible plasma volume interval within the logarithmic relative plasma volume domain. This is the upper boundary of the feasible plasma volume interval within the logarithmic relative plasma volume domain. When only one conserved interval is enabled, that conserved interval is defined as... When both conservation intervals are enabled, the larger of the lower boundaries of the two intervals is determined as... The smaller value among the upper boundaries of the two intervals is determined as .

[0164] when At that time, The feasible range for plasma volume was determined; when When a conflict occurs, a conservation conflict flag is generated, and the current detection time point correction process dependent on the feasible plasma volume interval is stopped. When both conservation intervals are in a deactivated state, the current troponin detection result is output according to the control flag. The feasible plasma volume interval is used to define the logarithmic relative plasma volume state at the current detection time point.

[0165] Figure 4 A flowchart illustrating an example of troponin concentration correction under plasma volume constraints in a method according to an embodiment of this application is shown.

[0166] like Figure 4As shown, in step S410, after the current detection time point is later than the reference detection time point and the feasible plasma volume interval is determined, the detection time point that most recently formed a non-empty plasma volume feasible interval before the current detection time point is determined as the previous available detection time point; based on the continuous blood pressure waveform data corresponding to the current detection time point and the previous available detection time point, the mean arterial pressure and pulse pressure of each of the two detection time points are determined, and the logarithmic relative plasma volume state of the previous available detection time point is obtained. The logarithmic relative plasma volume state of the reference detection time point is set to a zero-value baseline state, and the two relative weighings of the reference detection time point are determined as the pressure-neutralized troponin concentration of the reference detection time point.

[0167] The previous available detection time point is obtained by looking back from the detection time points preceding the current detection time point. Only detection time points with valid dual relative symmetry and forming a feasible interval for non-empty plasma volume are included in the plasma volume state chain. Other detection time points are allowed between two adjacent available detection time points, but these other detection time points are not used as the historical state source for the current state recursion.

[0168] The mean arterial pressure and pulse pressure at both detection time points are determined based on their respective continuous blood pressure waveform data. The mean arterial pressure at the corresponding detection time point can be obtained by averaging the pressure value within the target heartbeat over time according to the data acquisition time; the pulse pressure is determined based on the pressure difference between the systolic and diastolic pressure of the target heartbeat. The current detection time point and the previous available detection time point use the same heartbeat selection rules and parameters to determine the caliber.

[0169] In the specific implementation, the current detection time point is recorded as the detection time point. The plasma volume state chain located at the detection time point The node that is most recent and the one with the earliest time is recorded as the detection time point. The logarithmic relative plasma volume status at the reference testing time point satisfies... The corresponding pressure constraint relative to the plasma volume satisfies Therefore, the pressure-neutralized troponin concentration at the reference detection time point meets the requirements. .in, For reference, the logarithmic relative plasma volume status at the time of testing, For reference testing time points, the pressure constraint relative to plasma volume, For reference, the relative symmetry of the two sides at the time of detection. The pressure-neutralizing troponin concentration was used as a reference at the testing time point. Testing time point The logarithmic relative plasma volume state is the state result retained after plasma volume constraint processing at this detection point.

[0170] In step S420, the logarithmic relative plasma volume state of the previous available detection time point is decayed according to the state maintenance relationship that decays as the time interval between the current detection time point and the previous available detection time point increases. The mean arterial pressure change and pulse pressure change between the two detection time points are mapped to the logarithmic relative plasma volume change, and the decayed state is combined with the two mapping results to form the prior value of the current plasma volume pressure.

[0171] The prior value of the current plasma volume pressure can be determined by the following formula:

[0172] Equation (19)

[0173] In the formula, The prior value of plasma volumetric pressure at the current testing point; This represents the logarithmic relative plasma volume status at the previous available detection point; The time interval between the current detection time and the previous available detection time is denoted as , and ; Let be the state decay time constant, and ; and These are the mean arterial pressures at the current detection time and the previous available detection time, respectively; and These are the pulse pressures at the current detection time and the previous available detection time, respectively. and These are preset mapping coefficients that convert changes in mean arterial pressure and pulse pressure into logarithmic changes in relative plasma volume, with units of the reciprocal of pressure.

[0174] The state decay time constant can be pre-calibrated based on the temporal characteristics of the relative plasma volume state changing with the detection time interval in the sequence detection sample. The preset mapping coefficient can be determined based on a calibration sample with paired blood pressure data and reference plasma volume data; its sign and value are determined by the corresponding direction and proportion between blood pressure changes and logarithmic relative plasma volume changes. The historical state, after time decay, together with the current mean arterial pressure change and pulse pressure change, forms a pressure prior, which serves as the constraint value for the current plasma volume state.

[0175] In step S430, when the prior value of the current plasma volume pressure is within the feasible range of plasma volume, the prior value of the current plasma volume pressure is determined as the current logarithmic relative plasma volume state; when the prior value of the current plasma volume pressure is outside the feasible range of plasma volume, the boundary of the range closest to the prior value of the current plasma volume pressure is determined as the current logarithmic relative plasma volume state.

[0176] For example, the current logarithmic relative plasma volume state can be determined by the following formula:

[0177] Equation (20)

[0178] In the formula, This represents the logarithmic relative plasma volume at the current testing point. lie in During the internal period, retain ;when Below At that time, take ;when Higher than At that time, take Therefore, the current logarithmic relative plasma volume state retains both the prior information from the formation of the blood pressure sequence and is limited to the range allowed by the current effective conservation relations.

[0179] In step S440, the current logarithmic relative plasma volume state is exponentially converted to obtain the pressure-constrained relative plasma volume, and the pressure-constrained relative plasma volume is used to convert the dual relative weighing at the current detection time point to the reference plasma volume condition to obtain the pressure-neutralized troponin concentration.

[0180] The relative symmetry between the two sides at the current detection point can be determined by the following formula:

[0181] Equation (21)

[0182] In the formula, The relative symmetry between the two sides at the current detection point. and These are the hydraulically corrected concentrations of the boosting phase and the depressurizing phase at the current detection time. The two-phase symmetry is used in subsequent conversions when both phase hydraulically corrected concentrations meet the effective detection conditions and the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold.

[0183] The pressure constraint relative plasma volume can be determined according to the following formula:

[0184] Equation (22)

[0185] In the formula, This represents the pressure constraint relative plasma volume at the current detection time point relative to the reference detection time point. The exponential transformation converts the logarithmic relative plasma volume state to a positive relative plasma volume.

[0186] Stress-neutralizing troponin concentration can be determined using the following formula:

[0187] Equation (23)

[0188] In the formula, This represents the pressure-neutralized troponin concentration at the current detection time. The current dual relative weighing is multiplied by the pressure-constrained relative plasma volume to correct for concentration dilution or concentration caused by changes in the current plasma volume relative to the reference plasma volume, resulting in the troponin concentration using the same plasma volume diameter as the reference detection time.

[0189] In step S450, the pressure-neutralized troponin concentration is output, and the current logarithmic relative plasma volume state is used as the state input when determining the prior value of plasma volume pressure at the next detection time point.

[0190] The data processing unit will As the output of the pressure-neutralized troponin concentration at the current detection point, and The data is stored in association with the data acquisition time at the current detection time. When a feasible interval for a non-empty plasma volume is formed at the next detection time, the most recent logarithmic relative plasma volume state is retrieved from the plasma volume state chain, and new state decay and pressure prior calculations are performed according to the corresponding time intervals. Thus, historical states decay according to the detection time intervals, current blood pressure changes participate in the formation of pressure priors, the dual-conservation plasma volume feasible interval restricts the state boundary, and the resulting pressure-constrained relative plasma volume is further used to convert the current dual-relative balancing to the reference plasma volume conditions.

[0191] Regarding the details of determining the troponin release flux range in the method of this application embodiment, in some examples of this application embodiment, the pressure-neutralized troponin concentration at the previous available detection time point can also be obtained, and a preset first-order clearance constant range can be determined according to the type of troponin to be tested; wherein, the preset first-order clearance constant is used to characterize the proportion of troponin clearance per unit time.

[0192] The preset first-order clearance constant range includes a lower limit and an upper limit of the first-order clearance constant, which can be set according to the preset clearance parameters corresponding to the type of troponin to be tested. The lower limit of the first-order clearance constant is not greater than the upper limit of the first-order clearance constant, and both are non-negative values, with their units being the reciprocal of time.

[0193] Different types of troponin proteins can each correspond to a preset set of clearance parameters. These preset clearance parameter sets can be pre-established based on sequence detection calibration data or validated historical detection data for the corresponding troponin type and stored in association with the troponin type being tested. After determining the current troponin type, the lower limit and upper limit of the first-order clearance constant are read from the corresponding preset clearance parameter set. The time unit used for the clearance constant is consistent with the time unit used for the time interval between the two detection points.

[0194] The preceding available detection time point is the most recent detection time point in the aforementioned plasma volume state chain, preceding the current detection time point. The pressure neutralization and troponin concentrations at both the current and preceding available detection time points have been converted to reference plasma volume conditions, thus having the same plasma volume benchmark.

[0195] Subsequently, based on the concentration difference obtained by subtracting the pressure-neutralized troponin concentration at the previous available detection time point from the pressure-neutralized troponin concentration at the current detection time point, and the time interval between the two detection time points, the rate of change of troponin concentration per unit reference plasma volume is determined, and the arithmetic mean of the pressure-neutralized troponin concentrations at the two detection time points is determined.

[0196] The concentration difference preserves the direction of change of the current detection time point relative to the previous available detection time point. A positive concentration difference indicates an increase in pressure-neutralized troponin concentration, while a negative concentration difference indicates a decrease. Dividing the concentration difference by the time interval between the two detection time points yields the rate of change of troponin concentration per unit reference plasma volume. The arithmetic mean of the pressure-neutralized troponin concentrations at the two detection time points is used to characterize the concentration baseline participating in first-order clearance compensation within the time interval.

[0197] When performing troponin release flux interval calculations, both the current detection time point and the previous available detection time point have effective pressure-neutralized troponin concentrations, and the time interval between the two detection time points... The time interval is determined based on the data acquisition time associated with each of the two detection time points.

[0198] Subsequently, based on the lower and upper limits of the preset first-order clearance constant interval and the arithmetic mean, the lower limit of troponin clearance compensation flux and the upper limit of troponin clearance compensation flux are determined. The rate of change of troponin concentration is then added to the lower limit of troponin clearance compensation flux and the upper limit of troponin clearance compensation flux to obtain the lower limit of troponin release flux and the upper limit of troponin release flux.

[0199] The lower limit of troponin release flux can be determined by the following formula:

[0200] Equation (24)

[0201] In the formula, The lower limit of troponin release flux per unit reference plasma volume at the current detection point; The pressure-neutralized troponin concentration at the previous available detection point; To predefine the lower bound of the first-order clearing constant interval, and The first term in the formula is the rate of change of troponin concentration per unit reference plasma volume, and the second term is the lower limit of troponin clearance compensation flux determined according to the lower limit of the first-order clearance constant.

[0202] The upper limit of troponin release flux can be determined by the following formula:

[0203] Equation (25)

[0204] In the formula, The upper limit of troponin release flux per unit reference plasma volume at the current detection time; To predetermine the upper limit of the interval for the first-order clearing constant, and satisfy... The rate of change of troponin concentration is the same in both formulas, and the two clearance compensation fluxes are determined by the lower and upper limits of the first-order clearance constant, respectively.

[0205] The units for both the rate of change in troponin concentration and the troponin clearance compensation flux are concentration divided by time. Therefore, and The units are all mass divided by the product of reference plasma volume and time; when the pressure-neutralized troponin concentration is expressed in ng / L, the troponin release flux is expressed in ng / (L·time). Under the condition that the pressure-neutralized troponin concentrations at both detection time points are non-negative, their arithmetic mean is non-negative; combined with... The result .

[0206] The rate of change in troponin concentration retains both positive and negative directions. When the absolute value of the negative rate of change resulting from a decrease in concentration exceeds the corresponding clearance compensation flux, the endpoint of the corresponding troponin release flux can be negative. This endpoint is retained as a signed estimate obtained based on the concentration change and first-order clearance relationship, and is used to characterize the synthesis result of the concentration change component and the clearance compensation component under the corresponding clearance constant condition.

[0207] Then, the interval defined by the lower limit of troponin release flux and the upper limit of troponin release flux is determined as the troponin release flux interval per unit reference plasma volume at the current detection time, and the troponin release flux interval and the pressure-neutralized troponin concentration are output.

[0208] The troponin release flux range is denoted as... The interval retains the signed concentration change between two detection time points and passes the range of the first-order clearance constant to the two endpoints of the release flux. The data processing unit correlates the troponin release flux interval with the pressure-neutralized troponin concentration at the current detection time point. If any detection time point lacks a valid pressure-neutralized troponin concentration or the time interval between two detection time points is equal to zero, a data invalidation flag is generated and the calculation of the current release flux interval ends.

[0209] Figure 5 A schematic diagram illustrating the system operation mechanism of an example of a microfluidic detection method for blood troponin incorporating blood pressure fluctuations according to an embodiment of this application is shown.

[0210] like Figure 5 As shown, continuous blood pressure waveform data corresponding to a predetermined injection time period of the whole blood sample to be tested is acquired, and waveform features are extracted from the continuous blood pressure waveform data to determine the target heartbeat, target pressure value, pressure boosting trigger time, and pressure depressurization trigger time. The continuous blood pressure waveform data and the whole blood sample injection process use the same time reference, and the pressure value in the continuous blood pressure waveform data serves as the representation value of the inlet pressure at the sample input terminal. Biphasic trigger commands are generated based on the pressure boosting trigger time and the pressure depressurization trigger time, respectively controlling the opening of the pressure boosting phase microvalve and the pressure depressurization phase microvalve at the corresponding trigger times, allowing the whole blood sample to enter the corresponding injection branch. The solid arrows from left to right in the figure represent the main transmission paths of waveform features, biphasic blood samples, and multi-channel detection signals between the corresponding processing stages, while the dashed arrows labeled as biphasic trigger commands represent the transmission path of the microvalve control signals.

[0211] During the biphase triggering and hydraulic metering calibration process, the instantaneous flow rate and cumulative throughput of the boosting and depressurizing phases are determined based on the pressure value and rate of change of each microvalve during opening, the outlet pressure of the corresponding inlet branch, blood flow resistance parameters, and pipeline compliance parameters. The corresponding microvalve is then closed based on the cumulative throughput of each phase. Independent volumetric metering results are generated based on the membrane displacement of the flexible metering chamber, and these results are used to verify the volume consistency of the biphase blood samples. The boosting and depressurizing blood samples that pass the volume consistency verification enter the microfluidic cartridge. Hematocrit detection and plasma separation are performed on each phase blood sample. The separated boosting and depressurizing plasma are then simultaneously detected via multiple channels to obtain the corresponding hematocrit, albumin concentration, troponin detection signals, and inert fluorescence reference signals for each phase blood sample.

[0212] Figure 5The document also illustrates abnormal factors that may affect the validity of biphasic sampling, metrological calibration, plasma separation, or detection signals, such as air bubbles, volumetric mismatch, hemolysis, and plasma separation interference. The relevant dashed lines indicate the relationship between these abnormal factors and their corresponding processing steps. When an abnormal factor is determined to meet preset abnormality conditions based on the corresponding detection data, the affected biphasic blood sample is marked as invalid. A re-sampling instruction or an invalid detection data flag is generated based on the abnormality type to prevent the corresponding detection data from entering the subsequent calibration process.

[0213] The multi-channel detection signal is transmitted to the plasma volume constraint correction and release flux range output stage. Based on the troponin detection signal of each phase, the inert fluorescence reference signal, the cumulative throughput volume, the reaction residence time, and the preset immune capture parameters, the hydraulic correction concentrations for the pressurization phase and the depressurization phase are determined respectively. A two-phase relative balancing and a liquid path antisymmetric residual are then generated based on the two-phase hydraulic correction concentrations. When the liquid path antisymmetric residual meets a preset residual threshold, the two-phase relative balancing is determined as the troponin detection result.

[0214] For sequence detection time points that meet the detection quality control conditions, conservation intervals are constructed based on the corresponding hematocrit and albumin concentrations using the erythrocyte conservation relation and albumin conservation relation, respectively. A feasible plasma volume interval is then determined based on the conservation intervals in the enabled state. Combining the mean arterial pressure change, pulse pressure change, and historical plasma volume status represented by continuous blood pressure waveform data, a pressure-constrained relative plasma volume is determined within the feasible plasma volume interval. This pressure-constrained relative plasma volume is then used to convert the dual relative balancing to reference plasma volume conditions, yielding the pressure-neutralized troponin concentration.

[0215] For continuous available detection time points, based on the signed temporal changes in pressure-neutralized troponin concentration and the preset first-order clearance constant interval corresponding to the type of troponin to be tested, the lower limit and upper limit of troponin release flux per unit reference plasma volume are determined, forming a troponin release flux interval, and the troponin release flux interval and the corresponding pressure-neutralized troponin concentration are output.

[0216] To verify the identifiability of the method in this embodiment under non-ideal fluid circuit conditions and different combinations of physiological parameters, as well as the range of its low-error operating region, this embodiment employs simulation of digital subjects, troponin release-clearance kinetics, and the combined mechanism of microfluidic fluid circuitry. The simulation uses a fixed random seed of 20260801 to construct a validation set containing 120 digital subjects, each with 13 detection time points. The core hemodynamics and body fluid state vectors are defined as follows:

[0217] Equation (26)

[0218] In the formula, Hematocrit, Pulse pressure, Mean arterial pressure, For relative changes in plasma volume, superscript Indicates transpose; parameter ranges are set to... , mmHg, mmHg and The observation signal model includes a preset troponin baseline, delayed release, and first-order clearance process, and incorporates pipeline nonlinear compliance, microvalve response hysteresis, local plasma separation yield variations, and optical noise. All comparative methods use the same validation data, random perturbation, target sample volume, immunoreaction kinetic parameters, and baseline calibration curves. Method A uses a fixed valve opening time for injection and does not perform plasma volume correction; Method B performs active stabilization of the inlet flow rate but does not perform intracardiac pressure biphasic triggering; Method C, based on Method B, only performs plasma volume compensation according to hematocrit; and Method D uses the pressure biphasic triggering, hydraulic metering verification, antisymmetric residuals of the fluid path, double-conserved plasma volume constraints, plasma volume pressure prior projection, and pressure neutralization concentration correction processing as described in this application.

[0219] In the first set of simulation experiments, the pulse pressure was set to 20–100 mmHg, the tubing compliance to 0.02–0.12 μL / mmHg, and the microvalve response hysteresis to 2–12 ms, to evaluate the micro-injection volume error of different methods. The relative volume error was determined according to the following formula:

[0220] Equation (27)

[0221] in, and These refer to the opening and closing times of the microvalve, respectively. The instantaneous flow rate of the injection process is used as a reference in the joint mechanism simulation output. The target sample volume is defined above. The injection measurement results from methods A to D, which compare relative volume errors, are used. For method D, which generates a two-phase detection result under the same pressure, the aforementioned antisymmetric liquid path residual is also used. The mismatch state of the two-phase fluid path was evaluated. By changing the pipeline compliance, microvalve response hysteresis, and local flow resistance disturbances, the correspondence between the antisymmetric fluid path residuals and relative volume errors of method D was compared. The identification results of abnormal two-phase blood samples and the triggering results of re-injection commands were recorded to verify the ability of the antisymmetric fluid path residuals to characterize the unabsorbed fluid path anomalies in the hydraulic model. The simulation true value of the abnormal two-phase blood samples was determined by the preset abnormal disturbance labels applied to the corresponding blood samples.

[0222] In the second set of simulation experiments, implicit dilution trajectories and pseudo-concentration trajectories were constructed. The implicit dilution trajectory included sustained release of troponin and plasma volume expansion caused by fluid resuscitation, ensuring that the dilution effect from plasma volume expansion exceeded the concentration increase caused by sustained troponin release, resulting in a decreasing trend in the uncorrected troponin concentration. The pseudo-concentration trajectory did not include any new troponin release, but instead caused a decrease in plasma volume and blood pressure fluctuations through diuresis or blood loss, resulting in an increasing trend in the uncorrected troponin concentration. The sequence processing results of each method were evaluated using MAE (Mean Absolute Error) and the concentration-pressure hysteresis loop area. Both the implicit dilution trajectory and the pseudo-concentration trajectory included the process of mean arterial pressure rising from the lower boundary to the upper boundary and then returning to the lower boundary, with the pressure-raising and pressure-depressing branches connected at the same mean arterial pressure endpoint to form a closed trajectory. The concentration-pressure hysteresis loop area was determined according to the following formula:

[0223] Equation (28)

[0224] In the formula, For the detection time point The error between the troponin concentration output by each method for sequence comparison and the simulated reference concentration; This corresponds to a complete cycle of mean arterial pressure rising from the lower boundary to the upper boundary and then returning to the lower boundary. For method D, the troponin concentration used for sequence comparison is the pressure-neutralized troponin concentration. By comparing the corresponding values ​​of different methods... The effect of the direction of change in mean arterial pressure on the path dependence of troponin concentration error was evaluated; a decrease in the absolute value indicates a corresponding decrease in the difference in sequence concentration error caused by the pressor and depressor paths.

[0225] In the third set of simulation experiments, a systematic error response surface characterized by MAE was established on a two-dimensional parameter grid composed of hematocrit and pulse pressure. Connected regions with errors less than or equal to a preset error threshold were used to characterize the low-error working region of the corresponding method. The error distributions of methods A to D under different combinations of hematocrit and pulse pressure were compared. At each two-dimensional parameter grid point, the remaining simulation parameters used the same values ​​or sampling caliber to ensure the comparability of the systematic errors of different methods. Subsequently, using method D as the complete method, while keeping the other processing steps unchanged, the biphasic triggering under the same pressure, the cumulative through volume control, the albumin conservation constraint, the plasma volume pressure prior, and the flexible metrological membrane metrological calibration were removed individually to obtain the corresponding ablation method. The first ablation method was determined according to the following formula. The mean absolute error increment of each ablation method relative to the complete method:

[0226] Equation (29)

[0227] In the formula, For the first The mean absolute error of each ablation method The mean absolute error of the complete method is represented. The error increments of each ablation method are compared under corresponding fluid disturbances, body fluid changes, and blood pressure fluctuations to verify the constraint effect of each processing step on the corresponding error source and the input-output dependency between processing steps.

[0228] Figure 6 A schematic diagram of the comparative experimental simulation results of different methods in cardiac-level hydraulic metering and fluid circuit residual evaluation is shown.

[0229] like Figure 6 As shown in Figure A, the continuous arterial pressure waveform passes through the target pressure layer during the pressure-raising and pressure-depressing phases of each heartbeat, thus creating pressure-raising and pressure-depressing trigger points within each heartbeat, with the same pressure value but opposite directions of pressure change rate. Red trigger points indicate the pressure-raising phase trigger moment when the pressure change rate is greater than zero, and blue trigger points indicate the pressure-depressing phase trigger moment when the pressure change rate is less than zero. For the target heartbeat determined from the continuous heartbeats, the pressure-raising and pressure-depressing trigger points control the opening of corresponding microvalves, allowing the pressure-raising and pressure-depressing blood samples to enter their respective injection branches under the same target pressure layer.

[0230] like Figure 6 As shown in subplot B, within the simulation range of pulse pressure from 20 to 100 mmHg, Method A, using a fixed valve opening time, exhibits an injection volume relative error that increases with increasing pulse pressure, reaching approximately 10% at the upper pulse pressure boundary. Method B, employing external flow stabilization, shows a smaller variation in injection volume relative error with increasing pulse pressure compared to Method A. Method D, using dual-phase triggering at the same pressure and hydraulic metering calibration, maintains an injection volume relative error within 2% within the specified pulse pressure range. The shaded areas outside each curve represent the discrete range of results for multiple sets of digital subjects under the same simulation conditions. Method C uses the same inlet flow stabilization and injection conditions as Method B. Its hematocrit correction is applied to subsequent concentration processing; therefore, the injection volume relative error curve for Method C overlaps with that of Method B, and subplot B does not repeat the plot for Method C.

[0231] like Figure 6 As shown in the C and D subplots, to compare the two-phase mismatch states of different methods, the co-simulation platform uniformly retains the diagnostic observations of the rising and falling phases under the same target pressure layer, and determines the signed two-phase diagnostic residuals according to the following formula:

[0232] Equation (30)

[0233] In the formula, The sign of is used to characterize the direction of the concentration ratio between the two phases. Method D is used for the antisymmetric residual of the liquid path in the re-extraction gate. Methods A through C only use the signed biphasic diagnostic residuals for offline comparative evaluation, not for online sample injection control. The C subplot uses a violin plot to represent the residual probability density and distribution range of each method. After hydraulic calibration and residual gating, the residual distribution of effective biphasic blood samples in Method D is concentrated near zero. The fluid path asymmetry residuals marked on the horizontal axis of the D subplot represent the signed biphasic diagnostic residuals. The D subplot shows the relationship between the signed biphasic diagnostic residuals and the troponin concentration error. The troponin concentration error in Methods A through C is positively correlated with the signed biphasic diagnostic residuals. The effective detection results of Method D are concentrated near zero error, and the variation range of the corresponding fitted curve is limited, indicating that the residuals can be used to identify biphasic fluid path mismatches and trigger re-injection.

[0234] Figure 7 A schematic diagram illustrating the comparative experimental simulation results of different methods for detecting troponin sequences under conditions of fluctuating plasma volume is shown. Figure 7 As shown in subplot A, the gray curve represents mean arterial pressure, the black curve represents the simulated reference concentration generated by the preset troponin release-clearance kinetics, the red curve represents the original microfluidic result without plasma volume constraint correction, and the green curve represents the pressure-neutralized troponin concentration output by the complete method presented in this paper. The original microfluidic result shows a directional shift relative to the simulated reference concentration as the mean arterial pressure and plasma volume change, while the pressure-neutralized troponin concentration output by the complete method presented in this paper remains near the change path corresponding to the simulated reference concentration along the detection time.

[0235] like Figure 7 As shown in subplot B, the horizontal axis represents the mean arterial pressure during sequence detection, and the vertical axis represents the signed extraction error of the troponin concentration output by the method relative to the simulated reference concentration. The pressure increase and decrease processes shown in the figure represent the increasing and decreasing paths of mean arterial pressure during sequence detection, respectively. The original method is affected by plasma concentration or dilution, forming a direction-dependent concentration error hysteresis loop between the increasing and decreasing paths of mean arterial pressure; the concentration error of the method in this paper is distributed near the zero error line, indicating that after constraining the feasible plasma volume interval and correcting for pressure-neutralized concentration, the difference in concentration error corresponding to the direction of mean arterial pressure change is limited.

[0236] like Figure 7As shown in the C-plot, the mean absolute error of subjects (MAE) for methods A, B, C, and the complete method presented in this paper on the digital subject validation set is represented using scatter plots, box plots, and half-violin distributions. The error distributions for methods A to C decrease sequentially in value. The error data for the complete method presented in this paper are mainly distributed in the range of 0–2 ng / L, and its box plots and density concentration regions are all located in the decreasing direction of the corresponding regions for methods A to C. This is used to characterize the combined limiting effect of isobaric biphasic triggering, hydraulic calibration, dual-conservation plasma volume constraint, and pressure neutralization concentration correction on end-to-end sequence detection errors.

[0237] like Figure 7 As shown in the D-plot, the simulation results are divided into intervals of less than 3%, 3%~6%, 6%~9%, and greater than 9% based on the absolute value of the relative plasma volume change. The false sequence change alarm rates for each method are then calculated. Specifically, cases where the concentration change output by the method exceeds the preset sequence alarm threshold without setting a new release of troponin are identified as false sequence change alarms. As the interval of the absolute value of the relative plasma volume change increases, the false sequence change alarm rates for methods A to C increase accordingly. Within the interval where the absolute value of the relative plasma volume change is greater than 9%, the false sequence change alarm rates for methods A, B, C, and the complete method presented in this paper are 48%, 36%, 28%, and 8%, respectively. These results characterize the constraint effect of the complete method presented in this paper on false sequence concentration changes under the combined effects of plasma volume changes and blood pressure fluctuations.

[0238] Figure 8 The diagram illustrates the comparative experimental simulation results of different methods in the hematocrit-pulse pressure parameter space and an example of ablation using the complete method mechanism.

[0239] like Figure 8 As shown in subplots A and B, the hematocrit is... and pulse pressure Constructing a two-dimensional parametric grid, and using troponin detection results This characterizes the systematic error under different parameter combinations. Subplot A corresponds to method A, which uses a fixed valve opening time and does not perform plasma volume correction. Its low-error region is mainly distributed in the middle of the parameter space, and changes towards the parameter range boundaries as hematocrit or pulse pressure changes. The values ​​gradually increase. Subplot B corresponds to method D, which employs methods such as isostatic dual-phase triggering, hydraulic metering verification, antisymmetric residuals in the fluid path, dual-conservation plasma volume constraints, prior projection of plasma volume pressure, and pressure neutralization concentration correction. The region with an error less than or equal to 2 ng / L covers a relatively wide range of hematocrit and pulse pressure combinations. The colors and contour lines in the figure represent the corresponding parameter combinations. This characterizes the range and boundary changes of the low-error working area of ​​different methods.

[0240] like Figure 8 As shown in Figure C, taking method D as the complete method, while keeping the other processing steps and simulation conditions unchanged, the following were removed: isobaric biphasic triggering, cumulative through-volume control, albumin conservation constraint, plasma volume pressure prior, and flexible metering chamber membrane calibration. The performance of each ablation method relative to the complete method was then determined. Incremental. Delete the corresponding steps in the above processing. The increments were 3.65 ng / L, 2.82 ng / L, 1.92 ng / L, 1.28 ng / L, and 0.93 ng / L, respectively; the dots represent the corresponding... The incremental statistical value, with the horizontal error bars on both sides of the dot representing the discrete range of the results corresponding to multiple sets of numerical subjects.

[0241] like Figure 8 The comparison results of subplots A and B show that method D has a wider low-error working range when hematocrit and pulse pressure change in combination; the results of subplot C show that the various ablation methods... All of these results are higher than the complete method, and the error increment generated by deleting the same-pressure biphasic trigger or cumulative volume control is higher than the error increment generated by deleting other individual processing steps. Therefore, the response surface results and mechanism ablation results can characterize the limiting effect of the method of this application embodiment on the detection error caused by fluid disturbance, body fluid changes, and blood pressure fluctuations from both the parameter space boundary and individual processing step perspectives.

[0242] The simulation experiments described above demonstrate that the method in this application, by generating biphasic blood samples with the same pressure value and opposite directions of pressure change rate within the same target heartbeat, and combining cumulative volume control, antisymmetric residuals in the fluid path, dual-conservation plasma volume constraints, and prior projection of plasma volume pressure, can limit errors in troponin sequence detection caused by pulse pressure changes, fluid path dynamic response, and plasma volume changes. Compared to methods involving fixed valve opening time for sample injection, external flow stabilization, and compensation based solely on hematocrit, the method in this application forms a wider low-error working range within the parameter space constituted by hematocrit and pulse pressure. The mean absolute error corresponding to each ablation treatment is higher than that of the complete method, indicating that biphasic triggering under the same pressure, hydraulic metering verification, dual-conservation constraints, and prior projection of pressure respectively constrain the corresponding error sources.

[0243] The applicability of the method in this application is based on the premise that biphasic triggering, detection data quality, and corresponding conservation relationships meet preset conditions. When the cardiac cycle is insufficient to form mutually separate pressurization and depressurization triggering times, or when bubbles, hemolysis, or fluid mismatch cause the biphasic detection results to fail quality verification, subsequent correction using the corresponding biphasic blood sample is stopped, and a re-injection instruction or invalid detection data flag is generated. When red blood cell transfusion, blood loss, albumin transfusion, plasma replacement, or capillary leakage causes the corresponding conservation relationship to fail, the corresponding conservation interval is deactivated; when two conservation intervals conflict, subsequent processing at the current detection time point is prevented from entering the feasible interval dependent on plasma volume. Troponin release flux is estimated using a preset first-order clearance constant interval and output in interval form; the corresponding results are used to represent in vitro detection data between consecutive available detection time points.

[0244] It should be noted that, for the sake of simplicity, the foregoing method embodiments are all described as a series of combined actions. However, those skilled in the art should understand that this application is not limited to the described order of actions, as some steps may be performed in other orders or simultaneously according to this application. Secondly, those skilled in the art should also understand that the embodiments described in the specification are preferred embodiments, and the actions and modules involved are not necessarily essential to this application. In the above embodiments, the descriptions of each embodiment have their own emphasis; for parts not described in detail in a certain embodiment, please refer to the relevant descriptions of other embodiments.

[0245] Figure 9 A structural block diagram of an example of a blood troponin microfluidic detection system incorporating blood pressure fluctuations, according to an embodiment of this application, is shown.

[0246] like Figure 9 As shown, the blood troponin microfluidic detection system 900, which incorporates blood pressure fluctuations, includes a blood pressure waveform analysis unit 910, a biphasic quantitative injection unit 920, a biphasic immunoassay unit 930, a hydraulic immunoassay correction unit 940, a biphasic feature calculation unit 950, and a detection result determination unit 960.

[0247] The blood pressure waveform analysis unit 910 is used to acquire continuous blood pressure waveform data corresponding to a predetermined sampling period of the whole blood sample to be tested. The predetermined sampling period is a preset time window for selecting the sampling trigger time. The continuous blood pressure waveform data includes pressure values ​​received through the data interface and the data acquisition time corresponding to each pressure value. The target heartbeat and its target pressure value between diastolic and systolic pressure are determined from the continuous blood pressure waveform data. The times when the pressure rise phase and pressure fall phase of the target heartbeat reach the target pressure value are determined as the pressure rise trigger time and pressure fall trigger time, respectively.

[0248] The biphase quantitative injection unit 920 is used to open the boost phase microvalve and the depressurization phase microvalve at the boost trigger time and the depressurization trigger time, respectively, so that the whole blood to be tested enters the corresponding injection branch. Based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, the blood flow resistance parameter and the pipeline compliance parameter, the instantaneous flow rate of each phase injection process is determined and integrated over time to obtain the cumulative throughput volume of each phase. When the cumulative throughput volume of each phase reaches the target sample volume, the corresponding microvalve is closed to form the boost phase blood sample and the depressurization phase blood sample.

[0249] The biphasic immunoassay unit 930 is used to perform plasma separation and troponin immunoassay on the pressurized phase blood sample and the depressurized phase blood sample, respectively, to obtain the troponin detection signal and reaction residence time corresponding to each phase blood sample.

[0250] The hydraulic immunocorrection unit 940 is used to determine the immunocapture efficiency of each phase based on the cumulative throughput volume of each phase, the reaction residence time of the corresponding blood sample, and the preset immunocapture parameters, and to correct the troponin detection signal corresponding to each phase blood sample using the immunocapture efficiency, so as to obtain the hydraulic correction concentration of the pressurized phase and the hydraulic correction concentration of the depressurized phase.

[0251] The two-phase characteristic calculation unit 950 is used to calculate the geometric mean of the hydraulic correction concentration of the pressure-raising phase and the hydraulic correction concentration of the pressure-reducing phase to obtain the two-phase symmetry, and to calculate the absolute logarithm of the ratio of the hydraulic correction concentration of the pressure-raising phase to the hydraulic correction concentration of the pressure-reducing phase to obtain the antisymmetric residual of the liquid path.

[0252] The detection result determination unit 960 is used to output the dual relative symmetry as the troponin detection result corresponding to the target heartbeat when the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold; and to generate a re-injection command when the antisymmetric residual of the liquid path is greater than the preset residual threshold.

[0253] In some embodiments, this application provides a non-volatile computer-readable storage medium storing one or more programs including execution instructions, which can be read and executed by electronic devices (including but not limited to computers, servers, or network devices) to perform the steps of any of the above-described methods for microfluidic detection of blood troponin in conjunction with blood pressure fluctuations.

[0254] In some embodiments, this application also provides a computer program product, the computer program product including a computer program stored on a non-volatile computer-readable storage medium, the computer program including program instructions, which, when executed by a computer, cause the computer to perform the steps of any of the above-described methods for microfluidic detection of blood troponin in conjunction with blood pressure fluctuations.

[0255] In some embodiments, this application also provides an electronic device comprising: at least one processor, and a memory communicatively connected to the at least one processor, wherein the memory stores instructions executable by the at least one processor, the instructions being executed by the at least one processor to enable the at least one processor to perform steps of a microfluidic detection method for blood troponin incorporating blood pressure fluctuations.

[0256] The above-described product can perform the methods provided in the embodiments of this application, and has the corresponding functional modules and beneficial effects for performing the methods. Technical details not described in detail in this embodiment can be found in the methods provided in the embodiments of this application.

[0257] The electronic devices in this application can exist in various forms, including but not limited to: mobile communication devices, ultra-mobile personal computer devices, portable entertainment devices, or other airborne electronic devices with data interaction functions.

[0258] The device embodiments described above are merely illustrative. The units described as separate components may or may not be physically separate. The components shown as units may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs.

[0259] Through the above description of the embodiments, those skilled in the art can clearly understand that each embodiment can be implemented using software plus a general-purpose hardware platform, or of course, using hardware. Based on this understanding, the above technical solutions, in essence or the parts that contribute to the related technology, can be embodied in the form of a software product. This computer software product can be stored in a computer-readable storage medium, such as ROM / RAM, magnetic disk, optical disk, etc., and includes several instructions to cause a computer device (which may be a personal computer, server, or network device, etc.) to execute the methods described in the various embodiments or some parts of the embodiments.

[0260] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A microfluidic method for detecting blood troponin in conjunction with blood pressure fluctuations, characterized in that, The method includes: Acquire continuous blood pressure waveform data corresponding to a predetermined sampling period of the whole blood sample to be tested. The continuous blood pressure waveform data includes pressure values ​​received through a data interface and corresponding data acquisition times. Determine the target heartbeat and the target pressure value between the diastolic and systolic pressures of the target heartbeat from the continuous blood pressure waveform data. Determine the times when the target heartbeat reaches the target pressure value during the pressure-raising and pressure-depressing phases as the pressure-raising trigger time and pressure-depressing trigger time, respectively. At the pressure boost trigger time and the pressure depressurization trigger time, the boost phase microvalve and the depressurization phase microvalve are opened respectively; based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, the blood flow resistance parameter, and the tubing compliance parameter, the instantaneous flow rate of each phase injection process is determined and integrated over time to obtain the cumulative throughput volume of each phase; when the cumulative throughput volume of each phase reaches the target sample volume, the corresponding microvalve is closed, forming the boost phase blood sample and the depressurization phase blood sample; Plasma separation and troponin immunoassay were performed on the blood samples in the pressurization phase and the blood samples in the depressurization phase, respectively, to obtain the troponin detection signal and reaction residence time for each blood sample. The immune capture efficiency of each phase is determined based on the cumulative throughput of each phase, the reaction residence time of the corresponding blood sample, and the preset immune capture parameters. The corresponding troponin detection signal is then corrected using the immune capture efficiency of each phase to obtain the hydraulic correction concentration of the pressurization phase and the hydraulic correction concentration of the depressurization phase. Calculate the geometric mean of the hydraulic correction concentration of the boosting phase and the hydraulic correction concentration of the depressurizing phase to obtain the two-phase symmetry, and calculate the absolute logarithm of the ratio of the hydraulic correction concentration of the boosting phase to the hydraulic correction concentration of the depressurizing phase to obtain the antisymmetric residual of the liquid path. When the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold, the dual relative symmetry is output as the troponin detection result; when the antisymmetric residual of the liquid path is greater than the preset residual threshold, a re-injection command is generated.

2. The method according to claim 1, characterized in that, The step of determining the target heartbeat and the target pressure value between the diastolic and systolic blood pressure of the target heartbeat from the continuous blood pressure waveform data, and determining the moments when the target heartbeat reaches the target pressure value during the escalation and depressor phases as the escalation trigger moment and depressor trigger moment, respectively, includes: The continuous blood pressure waveform data is segmented according to the heartbeat cycle to obtain multiple candidate heartbeats arranged in chronological order, and the diastolic pressure, systolic pressure, pulse pressure, systolic phase, systolic phase, and pressure-time integral area of ​​each candidate heartbeat are determined. Based on the pulse pressure of each candidate heartbeat and the preset proportional coefficient, a candidate pressure value is determined between the corresponding diastolic and systolic pressures; For each candidate heartbeat that has a preceding candidate heartbeat, the candidate heartbeats that meet the corresponding preset conditions in terms of pulse pressure, data acquisition time continuity, pressure range, monotonicity of the boost phase, pressure change direction of the depressurization phase, and pressure-time integral area difference with the preceding candidate heartbeat are determined as usable heartbeats; the earliest available heartbeat is selected from the usable heartbeats whose heartbeat time period is within the predetermined injection time period as the target heartbeat, and the corresponding candidate pressure value is determined as the target pressure value; The moment when the target heart rate reaches the target pressure value during the pressure rise phase of the target heart rate under the condition that the rate of change of pressure is greater than zero is determined as the pressure rise trigger moment. From the candidate moments when the target heart rate reaches the target pressure value during the pressure fall phase under the condition that the rate of change of pressure is less than zero, the candidate moment with the smallest difference between the absolute value of the rate of change of pressure and the absolute value of the rate of change of pressure at the pressure rise trigger moment is selected as the pressure fall trigger moment. When the difference between multiple candidate moments is the same, the earliest one is selected as the pressure fall trigger moment.

3. The method according to claim 2, characterized in that, The instantaneous flow rate of each phase during injection is determined and integrated over time based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, blood flow resistance parameters, and tubing compliance parameters, to obtain the cumulative throughput volume of each phase, including: The initial hematocrit and ambient temperature of the whole blood sample to be tested are obtained before the sample is injected. Based on the ratio between the initial hematocrit and the non-erythrocyte volume fraction determined by the initial hematocrit, the reference flow resistance at the ambient temperature is nonlinearly corrected to obtain the blood flow resistance parameters. For each injection phase in the boosting and depressurizing phases, the pressure-driven flow rate is determined based on the non-negative pressure difference between the pressure value during the opening of the corresponding microvalve and the outlet pressure and capillary threshold pressure of the corresponding injection branch, as well as the blood flow resistance parameter. The compliance compensation flow rate is then determined based on the pipeline compliance parameter and the rate of change of the pressure value. A non-negative instantaneous flow rate along the injection direction is determined based on the pressure-driven flow rate and the compliance compensation flow rate. Time integration is then performed starting from the triggering moment of the corresponding injection phase to obtain the cumulative throughput volume of the corresponding injection phase. When the cumulative volume of each phase reaches the target sample volume for the first time, the corresponding microvalve is closed, and the membrane metering volume of the corresponding phase is determined based on the membrane displacement retained by the flexible metering cavity in the corresponding injection branch after the microvalve is closed. The ratio of the absolute value of the difference between the cumulative volume of each phase and the corresponding membrane measurement volume to the target sample volume is determined as the single-phase volume deviation ratio of the corresponding phase, and the ratio of the absolute value of the difference between the membrane measurement volume of the boosting phase and the membrane measurement volume of the depressurizing phase to the target sample volume is determined as the two-phase volume deviation ratio. When each volume deviation ratio is less than or equal to its corresponding preset volume deviation threshold, plasma separation and troponin immunoassay are performed on the boosting phase blood sample and the depressurizing phase blood sample. When any volume deviation ratio is greater than the corresponding preset volume deviation threshold, a re-injection command is generated.

4. The method according to claim 3, characterized in that, The process involves determining the immune capture efficiency of each phase based on the cumulative throughput volume of each phase, the reaction residence time of the corresponding blood sample, and preset immune capture parameters. The immune capture efficiency of each phase is then used to correct the corresponding troponin detection signal, resulting in the hydraulically corrected concentrations for the pressurization phase and the depressurization phase. This includes: Hematocrit and plasma separation were performed on blood samples from each phase. The separated pressurized and depressurized plasmas were then delivered to the troponin reaction region, albumin reference reaction region, and inert fluorescence reference region within the pressurized and depressurized detection pathways, respectively. The pressurized and depressurized detection pathways have mirror-image flow channel structures. Hematocrit, albumin concentration, troponin detection signal, reaction residence time, and inert fluorescence reference signal were obtained for each phase of the blood sample. Based on the proportional relationship between the inert fluorescence reference signal of each phase and the preset reference signal, the preset calibration response coefficient is adjusted to obtain the optical response coefficient of each phase; The effective reaction degree of each phase is determined based on the reaction residence time, cumulative throughput, and preset immune capture parameters. The immune capture efficiency of each phase is determined according to the response relationship that the immune capture efficiency increases monotonically and tends to saturate as the effective reaction degree increases. The signal increment of each phase troponin detection signal relative to the preset blank detection signal is determined, and the signal increment is normalized and converted to a concentration based on the corresponding optical response coefficient, cumulative throughput volume and immune capture efficiency, so as to obtain the hydraulic correction concentration of the pressurization phase and the hydraulic correction concentration of the depressurization phase.

5. The method according to claim 4, characterized in that, When the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold, the method further includes: The current detection time point is defined as the current detection time point, and the detection time point in which the first antisymmetric residual of the liquid path and the ratio of each volume deviation in the sequence detection are less than or equal to their respective preset thresholds is defined as the reference detection time point; for the current detection time point and the reference detection time point, the arithmetic mean of the hematocrit of the corresponding pressurization phase and the depressurization phase is defined as the hematocrit of the corresponding detection time point, and the geometric mean of the albumin concentration of the corresponding pressurization phase and the depressurization phase is defined as the albumin concentration of the corresponding detection time point; The red blood cell conservation relative plasma volume is determined based on the ratio between the hematocrit at the reference detection time and the hematocrit at the current detection time, and the ratio between the non-red blood cell volume fraction at the current detection time and the non-red blood cell volume fraction at the reference detection time; the albumin conservation relative plasma volume is determined based on the ratio between the albumin concentration at the reference detection time and the albumin concentration at the current detection time. Using the logarithms of the red blood cell conservation relative plasma volume and the albumin conservation relative plasma volume as the center, the intervals are expanded according to the corresponding logarithmic measurement uncertainty to construct the red blood cell conservation interval and the albumin conservation interval. Obtain the conservation applicable state marker between the reference detection time point and the current detection time point; when the conservation applicable state marker indicates that red blood cell transfusion has occurred or the blood loss reaches a preset blood loss threshold, deactivate the red blood cell conservation interval; when the conservation applicable state marker indicates that albumin transfusion, plasma exchange, or capillary leakage that meets preset leakage judgment conditions has occurred, deactivate the albumin conservation interval. When at least one conservation interval is enabled and there is a non-empty intersection among the enabled conservation intervals, the non-empty intersection is determined as a feasible interval for plasma volume; when both conservation intervals are enabled and the intersection of the red blood cell conservation interval and the albumin conservation interval is empty, a conservation conflict flag is generated; when both conservation intervals are disabled, a control flag is generated that only outputs the troponin detection result.

6. The method according to claim 5, characterized in that, After the current detection time point is later than the reference detection time point and the feasible range of plasma volume is determined, the method further includes: The detection time point that most recently formed a feasible interval of non-empty plasma volume before the current detection time point is determined as the previous available detection time point; based on the continuous blood pressure waveform data corresponding to the current detection time point and the previous available detection time point, the mean arterial pressure and pulse pressure of each of the two detection time points are determined, and the logarithmic relative plasma volume state of the previous available detection time point is obtained; wherein, the logarithmic relative plasma volume state of the reference detection time point is set as the zero-value baseline state, and the dual relative weighing of the reference detection time point is determined as the pressure neutralization troponin concentration of the reference detection time point; According to the state maintenance relationship that decays as the time interval between the current detection time point and the previous available detection time point increases, the logarithmic relative plasma volume state of the previous available detection time point is decayed, and the mean arterial pressure change and pulse pressure change between the two detection time points are mapped to the logarithmic relative plasma volume change, and the decayed state is combined with the two mapping results to form the current plasma volume pressure prior value. When the prior value of the current plasma volume pressure is within the feasible range of plasma volume, the prior value of the current plasma volume pressure is determined as the current logarithmic relative plasma volume state; when the prior value of the current plasma volume pressure is outside the feasible range of plasma volume, the boundary of the range closest to the prior value of the current plasma volume pressure is determined as the current logarithmic relative plasma volume state. The current logarithmic relative plasma volume state is exponentially transformed to obtain the pressure-constrained relative plasma volume, and the pressure-constrained relative plasma volume is used to convert the dual relative weighing at the current detection time point to the reference plasma volume condition to obtain the pressure-neutralized troponin concentration. The pressure-neutralized troponin concentration is output, and the current logarithmic relative plasma volume state is used as the state input when determining the prior value of plasma volume pressure at the next detection time point.

7. The method according to claim 6, characterized in that, The method further includes: The pressure-neutralized troponin concentration at the previous available detection time point is obtained, and a preset first-order clearance constant range is determined according to the type of troponin to be tested; wherein, the preset first-order clearance constant is used to characterize the proportion of troponin cleared per unit time. Based on the concentration difference obtained by subtracting the pressure-neutralized troponin concentration at the previous available detection time point from the pressure-neutralized troponin concentration at the current detection time point, and the time interval between the two detection time points, the rate of change of troponin concentration per unit reference plasma volume is determined, and the arithmetic mean of the pressure-neutralized troponin concentrations at the two detection time points is determined. Based on the lower and upper limits of the preset first-order clearance constant interval and the arithmetic mean, the lower limit of troponin clearance compensation flux and the upper limit of troponin clearance compensation flux are determined. The rate of change of troponin concentration is added to the lower limit of troponin clearance compensation flux and the upper limit of troponin clearance compensation flux to obtain the lower limit of troponin release flux and the upper limit of troponin release flux. The interval defined by the lower limit and the upper limit of troponin release flux is determined as the troponin release flux interval per unit reference plasma volume at the current detection time, and the troponin release flux interval and the pressure-neutralized troponin concentration are output.

8. A microfluidic detection system for blood troponin combined with blood pressure fluctuations, characterized in that, The system includes: A blood pressure waveform analysis unit is used to acquire continuous blood pressure waveform data corresponding to a predetermined sampling period of the whole blood sample to be tested. The predetermined sampling period is a preset time window for selecting the sampling trigger time. The continuous blood pressure waveform data includes pressure values ​​received through a data interface and the data acquisition time corresponding to each pressure value. The unit determines the target heartbeat and its target pressure value between diastolic and systolic blood pressure from the continuous blood pressure waveform data. The times when the pressure-raising and pressure-depressing phases of the target heartbeat reach the target pressure value are determined as the pressure-raising trigger time and the pressure-depressing trigger time, respectively. The biphase quantitative injection unit is used to open the boost phase microvalve and the depressurization phase microvalve at the boost trigger time and the depressurization trigger time, respectively, so that the whole blood sample to be tested enters the corresponding injection branch. Based on the pressure value and its rate of change during the opening of each microvalve, the outlet pressure of the corresponding injection branch, the blood flow resistance parameter and the pipeline compliance parameter, the instantaneous flow rate of each phase injection process is determined and integrated over time to obtain the cumulative throughput volume of each phase. When the cumulative throughput volume of each phase reaches the target sample volume, the corresponding microvalve is closed to form the boost phase blood sample and the depressurization phase blood sample. The biphasic immunoassay unit is used to perform plasma separation and troponin immunoassay on the pressurized phase blood sample and the depressurized phase blood sample, respectively, to obtain the troponin detection signal and reaction residence time corresponding to each phase blood sample. The hydraulic immunocorrection unit is used to determine the immunodetection efficiency of each phase based on the cumulative throughput of each phase, the reaction residence time of the corresponding blood sample, and the preset immunodetection parameters, and to correct the troponin detection signal corresponding to each phase blood sample using the immunodetection efficiency, so as to obtain the hydraulic correction concentration of the pressurization phase and the hydraulic correction concentration of the depressurization phase. The two-phase characteristic calculation unit is used to calculate the geometric mean of the hydraulic correction concentration of the pressure-raising phase and the hydraulic correction concentration of the pressure-depressurizing phase to obtain the two-phase symmetry, and to calculate the absolute logarithm of the ratio of the hydraulic correction concentration of the pressure-raising phase to the hydraulic correction concentration of the pressure-depressurizing phase to obtain the antisymmetric residual of the liquid path. The detection result determination unit is used to output the dual relative symmetry as the troponin detection result corresponding to the target heartbeat when the antisymmetric residual of the liquid path is less than or equal to a preset residual threshold; and to generate a re-injection command when the antisymmetric residual of the liquid path is greater than the preset residual threshold.

9. An electronic device, comprising: At least one processor; as well as A memory communicatively connected to the at least one processor, characterized in that the memory stores instructions executable by the at least one processor, the instructions being executed by the at least one processor to enable the at least one processor to perform the steps of the method as described in any one of claims 1-7.

10. A storage medium having a computer program stored thereon, characterized in that, When the program is executed by the processor, it implements the steps of the method as described in any one of claims 1-7.

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