Blood testing device and method of testing thereof

Abstract

The present disclosure belongs to the technical field of in-vitro diagnostic medical devices, and particularly relates to a blood detection device and a detection method thereof. The blood detection device comprises a sample loading assembly, a control processing assembly, an optical detection assembly, a driving assembly and a display assembly. The present disclosure directly inserts a standard blood collection tube as a sampling piece into a containing portion to complete detection, avoids sample contamination and platelet activation, and thus realizes tube detection. The differential signal calibration tube wall interference and the influence of individual plasma background difference on the detection result are eliminated. By setting the driving assembly, the driving assembly drives the optical detection assembly to move away from the layered interface and accurately lock the detection area of the sampling piece to perform positioning detection on the sampling piece in the containing portion, thereby avoiding the interference of adjacent layers.

Description

Technical Field

[0001] This disclosure belongs to the field of in vitro diagnostic medical device technology, and in particular relates to a blood testing device and its testing method. Background Technology

[0002] Platelet-rich plasma (PRP) is a blood product obtained by centrifuging autologous whole blood, resulting in a platelet concentration higher than baseline. PRP contains high concentrations of platelets, growth factors (such as PDGF, TGF-β, VEGF, EGF, IGF, etc.), and fibrin, among other bioactive components, and has broad clinical applications in tissue repair, regenerative medicine, dental implants, wound healing, cosmetic surgery, and sports medicine.

[0003] The clinical efficacy of PRP is directly related to its platelet concentration. PRP concentrations reach 3-5 times the baseline whole blood concentration (approximately 600-1000 × 10⁻⁶). 9 When the platelet concentration of PRP reaches a certain level (e.g., 1 / L), the release of growth factors can reach the therapeutic threshold. Therefore, rapid and accurate detection and evaluation of PRP platelet concentration before clinical application is of great significance for ensuring treatment efficacy and standardizing clinical procedures.

[0004] PRP is usually prepared by centrifugation. After preparation, the detection of platelet concentration in PRP becomes a key step in quality control.

[0005] Currently, the commonly used platelet counting methods in clinical practice mainly include the following: (1) Electrical impedance method (blood cell analyzer): This method counts platelets by utilizing the change in resistance generated when they pass through tiny pores. This method is highly automated, but it requires samples, regular calibration, and the equipment is large and expensive, making it unsuitable for rapid point-of-care testing.

[0006] (2) Flow cytometry: This method uses fluorescently labeled platelet-specific antibodies (such as CD41 and CD61) for counting and is considered a reference method for platelet counting. However, it is complex to operate, requires professional personnel and expensive equipment, and has a long testing cycle, making it unsuitable for routine PRP quality control.

[0007] (3) Optical microscopy counting method: The hemocytometer is used to count the cells manually under a microscope. This method has simple equipment, but it is cumbersome to operate, highly subjective, has poor repeatability, and requires sample transfer, which poses a risk of contamination.

[0008] (4) Light Transmission Aggregometry (LTA): This method utilizes the principle of changes in light transmittance during platelet aggregation and is considered the "gold standard" for platelet function testing. The traditional LTA method requires transferring PRP to a special cuvette, using platelet-poor plasma (PPP) as a 100% transmittance reference, and assessing platelet aggregation function by adding an inducing agent and detecting changes in transmittance.

[0009] (5) Spectrophotometry: In recent years, Toyoda, T., Isobe, K., Tsujino, T. et al. Directactivation of platelets by addition of CaCl2 leads coagulation of platelet-rich plasma. Int J Implant Dent 4, 23 (2018) reported a method for estimating platelet concentration by measuring PRP absorbance at 615 nm using a spectrophotometer. This method showed good linearity (R0). 2 =0.995), but the sample needs to be transferred to a micro-volume cuvette, making it impossible to perform in-tube detection.

[0010] Therefore, there is an urgent need for a device and method that can rapidly and accurately perform semi-quantitative detection of PRP platelet concentration under standard blood collection tube conditions, in order to overcome the shortcomings of the existing technology. Summary of the Invention

[0011] This disclosure provides a blood testing device and method to solve technical problems in the prior art, such as sample transfer, interference from plasma background color, tube wall differences, and interference from layered interfaces, and to achieve rapid semi-quantitative detection of platelet count in platelet-rich plasma under the original state of a standard blood collection tube.

[0012] In a first aspect, embodiments of this disclosure provide a blood testing device, comprising: A sample loading assembly, the sample loading assembly including a receiving portion, wherein a positioning detection element is disposed within the receiving portion, the positioning detection element being configured to generate a positioning signal; A control processing component is connected to the positioning detection element, and the control processing component is configured to receive the positioning signal; An optical detection component is movable relative to the sample loading component. The optical detection component includes a light source module and an optical path system. The light source module is connected to the control and processing component, and the light source module includes multiple light sources. The control and processing component is configured to control the multiple light sources to switch and alternately light up in a time sequence. The optical path system detects light signals during the alternating lighting of the multiple light sources and transmits the detected light signals to the control and processing component. A driving component, connected to the control processing component, is configured to drive the optical detection component to move when the control processing component receives the positioning signal, so as to perform positioning detection on the sampler within the accommodating portion; A display component is connected to the control processing component, and the display component is configured to display the processing result output by the control processing component.

[0013] In some embodiments, the sample loading assembly further includes an identification element connected to the control processing assembly, the identification element being configured to identify the sample located in the receiving portion and generate an identification signal; and / or, The sample loading assembly also includes a clamping member disposed on the outer periphery of the receiving portion, and the clamping member has a preset clamping stroke.

[0014] In some embodiments, the sample loading assembly further includes a support member, and the receiving portion is disposed on the support member.

[0015] In some embodiments, the identification signal includes at least one of the specifications of the sample and the type of anticoagulation.

[0016] In some embodiments, the light source module includes a first light source and a second light source, wherein the first light source has a wavelength range of 650-670nm and an optical power range of 30-70mW, and the second light source has a wavelength range of 795-815nm and an optical power range of 30-70mW; and / or, The first light source and the second light source are alternately lit using a time-division multiplexing method, with the alternation switching frequency ranging from 0.5 to 2 kHz.

[0017] In some embodiments, the optical path system includes: An incident light path is configured to guide incident light emitted by the light source in the light source module to the detection area of ​​the sampling element; The outgoing light path and the incoming light path are respectively located on opposite sides of the sample loading assembly; The detector module is located on the side of the outgoing optical path away from the sample loading assembly; The outgoing optical path is configured to guide the transmitted light passing through the sampling element to the detector module, and the detector module is configured to convert the received optical signal into a photocurrent signal.

[0018] In some embodiments, the incident light path includes: a collimating lens and a bandpass filter; the collimating lens is located between the light source module and the sample loading assembly; the bandpass filter is located between the collimating lens and the sample loading assembly; and / or, The incident light path further includes a beam combiner disposed between the bandpass filter and the sample loading assembly. The beam combiner is configured to combine the light paths of multiple light sources at the intersection point to share the same outgoing light path.

[0019] In some embodiments, the driving component includes: A driving component, wherein the driving end of the driving component is connected to the optical detection assembly to drive the optical detection assembly to move; A monitoring element, installed on the optical detection assembly, is configured to monitor the liquid level and stratification interface within the sampling element; A positioning element is installed on the optical detection assembly, and the positioning element is configured to locate the liquid level position within the sampling element.

[0020] In some embodiments, the monitoring device includes a liquid level sensor array, which includes 4-8 liquid level sensors, and the spacing between two adjacent liquid level sensors is in the range of 8-12 mm; And / or, the positioning element includes a positioning probe.

[0021] In some embodiments, the driving component includes a stepper motor with a step angle of 1.8°, microsteps subdivided into 1 / 8-1 / 32, a positioning accuracy of ±0.5mm, and a repeatability of ±0.1mm.

[0022] In some embodiments, the control processing component includes: The controller is configured to control the timing switching of multiple light sources in the light source module, the movement of the optical detection component, the movement and positioning of the driving component, the positioning signal, the identification signal, and the acquisition and processing of the optical signal; An analog-to-digital converter is configured to convert the photocurrent signal output by the detector module into a digital signal; The memory is configured to store factory baseline calibration parameters and detection thresholds; The communication interface is configured to transmit data with external devices.

[0023] In some embodiments, the blood testing device further includes: a housing, wherein the sample loading component, the control processing component, the optical detection component, and the driving component are all disposed within the housing, the housing has a first opening corresponding to the sample loading component for inserting the sample into the receiving portion, and / or, the housing has a second opening, the second opening including at least one of a heat dissipation hole, a power interface, and a communication interface.

[0024] In some embodiments, the display component includes: an indicator light array disposed on the outer surface of the housing, the indicator light array being configured to display semi-quantitative detection results at different levels; and / or, A display screen is disposed on the outer surface of the housing and on the same side as the indicator array. The display screen is configured to display specific detection parameter results.

[0025] Secondly, embodiments of this disclosure provide a detection method for a blood detection device as described in any embodiment of the first aspect, comprising the following steps: Preprocessing of samples containing venous blood to obtain samples with a detection layer; The pre-treated sample is placed on the sample loading component of the blood testing device, and a position signal is received. In response to the arrival signal, the driving component is activated to perform positioning detection of the detection layer of the sample; Control the alternating illumination of multiple light sources in the optical detection assembly; The optical path system receives the optical signals detected during the alternating illumination of the multiple light sources; The optical signal is processed, and the processed result is output to the display component.

[0026] In some embodiments, the preprocessing of the sample containing venous blood to obtain a sample with a detection layer includes: Samples containing venous blood are centrifuged to obtain samples with a detection layer. The centrifugal force is 50-200×g, and the centrifugation time is 5-10 minutes.

[0027] In some embodiments, controlling the alternating illumination of multiple light sources in the optical detection assembly includes: The two light sources in the optical detection component are controlled to be lit alternately. The two light sources are a light source with a first preset wavelength and a light source with a second preset wavelength, and the first preset wavelength and the second preset wavelength are different. Calculate the difference absorbance ΔA between the first preset wavelength and the second preset wavelength.

[0028] In some embodiments, the processed result includes multiple semi-quantitative levels, which include a first concentration, ΔA < 0.50; a second concentration, 0.50 ≤ ΔA < 1.00; a third concentration, 1.00 ≤ ΔA < 1.50; a fourth concentration, 1.50 ≤ ΔA < 2.00; and a fifth concentration, ΔA ≥ 2.00.

[0029] In some embodiments, the display component includes an LED indicator array, where a first color represents a first concentration; a second color represents a second concentration; a third color represents a third concentration; a fourth color represents a fourth concentration; and a fifth color represents a fifth concentration, with a 10% overlap buffer between adjacent levels.

[0030] In some embodiments, the detection method further includes: Interference is corrected using factory baseline calibration parameters, which include: blank calibration values, pipe wall calibration values ​​of the sampled component, and standard curve parameters. The standard curve is in the range of 50-800×10⁻⁶. 9 It is linear within the range of / L, R 2 ≥0.995.

[0031] In some embodiments, in response to the positioning signal, the driving component is activated to perform positioning detection in the detection layer of the sampling element, wherein the lower middle part of the detection layer is the detection area.

[0032] The blood testing device and method provided in this disclosure, by setting up a sample loading component including a receiving part, allows standard blood collection tubes to be directly used as testing containers. Without opening the cap or transferring the sample, the standard blood collection tubes can be directly inserted into the receiving part as sampling components to complete the test, avoiding sample contamination and platelet activation, thereby realizing tube-based testing.

[0033] By setting up an optical detection component including a light source module and an optical path system, the light source module includes multiple light sources that are switched and lit alternately in sequence. The optical path system detects the light signal during the alternating lighting of the multiple light sources, and uses differential signal calibration to eliminate tube wall interference and the influence of individual plasma background color differences on the detection results.

[0034] By setting up a driving component, when the control processing component receives the positioning signal, the driving component drives the optical detection component to move, avoiding the layer interface, accurately locking the detection area of ​​the sample to perform positioning detection on the sample in the accommodating part, and avoiding interference from adjacent layers. Attached Figure Description

[0035] To more clearly illustrate the technical solutions of the embodiments of this disclosure, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this disclosure. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0036] Figure 1 This is a schematic diagram of the structure of a blood testing device provided in an embodiment of this disclosure; Figure 2 This is a schematic diagram of the dual-wavelength differential optical path design of an optical detection component provided in one embodiment of the present disclosure; Figure 3 This is a schematic diagram of the structure of the drive component in a blood testing device provided in an embodiment of this disclosure; Figure 4 This is a flowchart of a blood testing method provided by an embodiment of the present disclosure.

[0037] Explanation of reference numerals in the attached figures: 100 - Sample loading assembly; 101 - Receiving part; 103 - Identification component; 104 - Clamping component; 105 - Support component; 200 - Control processing component; 300 - Optical detection assembly; 301 - Light source module; 3011 - First light source; 3012 - Second light source; 302 - Optical path system; 3021 - Incident light path; 303 - Collimating lens; 304 - Bandpass filter; 305 - Beam combiner; 3022 - Outgoing light path; 3023 - Detector module; 400 - Drive assembly; 401 - Drive component; 402 - Monitoring component; 403 - Positioning component; 500 - Display Components; 600 - Housing. Detailed Implementation

[0038] To enable those skilled in the art to better understand the technical solutions in this disclosure, the technical solutions in the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. Based on the embodiments in this disclosure, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this disclosure.

[0039] It should be noted that many specific details are set forth in the following description in order to provide a full understanding of this disclosure; however, the intermediate means described herein are indirect contacts.

[0040] In this disclosure, unless otherwise expressly specified and limited, the terms "connected," "linked," and "fixed," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral unit; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. However, specifying a direct connection indicates that the two entities connected are not linked by an intermediate structure, but are simply connected to form a whole. For those skilled in the art, the specific meaning of the above terms in this disclosure can be understood according to the specific circumstances.

[0041] In this disclosure, the terms "first," "second," etc., are used for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of those features.

[0042] The detection of platelet concentration in PRP using related technologies suffers from the following common limitations, restricting its application in rapid quality control at the PRP preparation site: (1) Sample transfer requirements: Existing detection methods all require the transfer of PRP from blood collection tubes to special detection containers such as cuvettes, cuvettes, and flow cytometers, which is cumbersome, increases the risk of sample contamination and platelet activation, and prolongs the detection time.

[0043] (2) Interference from individual plasma background color: There are differences in plasma color among different individuals, such as lipemia, hemolysis, jaundice, etc. These differences will affect the absorbance measurement results. Existing methods lack an effective plasma background color correction mechanism.

[0044] (3) Differences in tube wall and tube type: There are differences in material, wall thickness and light transmittance of blood collection tubes from different manufacturers and of different specifications. These differences will introduce detection errors, and the existing methods do not consider the correction of tube wall factors.

[0045] (4) Interference at the layer interface: In the blood collection tube after centrifugation, the PRP layer is located between the plasma layer and the red blood cell layer. If it is not accurately located during detection, it is easily affected by the interference of the adjacent layer, which affects the accuracy of the detection.

[0046] Therefore, there is an urgent need for a device and method that can rapidly and accurately perform semi-quantitative detection of PRP platelet concentration under standard blood collection tube conditions, in order to overcome the shortcomings of the existing technology.

[0047] To this end, in a first aspect, this disclosure provides a blood testing device.

[0048] Figure 1 This is a schematic diagram of the structure of a blood testing device provided in one embodiment of the present disclosure. Figure 1The overall architecture of the blood testing device was demonstrated.

[0049] like Figure 1 As shown, the blood testing device may include: a sample loading component 100.

[0050] The sample loading assembly 100 includes a receiving portion 101, in which a positioning detection element is disposed, and the positioning detection element is configured to generate a positioning signal.

[0051] The receiving part 101 can be a slotted structure, which can be adapted to the sampling device. The sampling device can be a standard vacuum blood collection tube of Φ13×75mm or Φ13×100mm, supporting sodium citrate, dipotassium ethylenediaminetetraacetate, EDTA dipotassium salt (EDTA-K2), and heparin anticoagulant tubes.

[0052] After the sampling component is inserted into the receiving part 101, it will trigger the positioning detection component to generate a positioning signal. The positioning detection component can be a micro switch.

[0053] In some embodiments, such as Figure 1 As shown, the sample loading assembly 100 may also include an identification element 103 connected to the control processing assembly 200. The identification element 103 is configured to identify the sample located in the receiving portion 101 and generate an identification signal.

[0054] The identification element 103 can be a tubular identification sensor, such as a capacitive proximity sensor. The tubular identification sensor can automatically identify the specifications and anti-coagulation type of the sampling element within the housing 101.

[0055] In some embodiments, the identification signal includes at least one of the specifications of the sampling tube and the type of anticoagulation. The specifications of the sampling tube may be a standard vacuum blood collection tube of Φ13×75mm or Φ13×100mm, and the type of anticoagulation may be sodium citrate, EDTA-K2, or heparin anticoagulation tubing, which is not limited in this embodiment.

[0056] For example, a tube type identification sensor, model OmronE2K-X4ME1, can be installed at the bottom of the receiving part 101. This capacitive proximity sensor automatically identifies the specifications and anticoagulant type of the sampling component by detecting the shape of its bottom (e.g., flat bottom, round bottom), the height of the tube, and the color of its cap. The identification signal is transmitted to the control processing component 200 to retrieve the corresponding factory baseline calibration parameters.

[0057] In this embodiment of the disclosure, the standard blood collection tube is directly used as the detection container through the sample loading component. There is no need to open the cap or transfer the sample. The standard blood collection tube is directly inserted into the receiving part as a sampling component to complete the detection, avoiding sample contamination and platelet activation, thereby realizing the detection of the original tube.

[0058] In some embodiments, such as Figure 1 As shown, the sample loading assembly 100 may also include a clamping member 104, which is disposed on the outer periphery of the receiving portion 101 and has a preset clamping stroke.

[0059] The clamping member 104 is used to clamp and fix the sampling member in the fixed accommodating part 101 to ensure that the position of the sampling member can be relatively fixed when it is detected, thereby ensuring the alignment accuracy of the optical path and the detection layer during the detection process.

[0060] Exemplarily, the clamping member 104 can be installed on the periphery of the receiving portion 101. The clamping member 104 can be a purely mechanical spring clip, requiring no electrical connection. When the sampling piece is inserted into the receiving portion 101, the spring clip automatically tightens, clamping the sampling piece with a fixing force of 2-3N to ensure that the sampling piece does not shift, shake, or tilt during the testing process. The spring clip can be made of medical-grade 304SS stainless steel with a polished surface to avoid scratching the sample tube wall. The preset clamping stroke of the clamping member 104 can be 12-14mm, which can be adapted to a standard Φ13mm blood collection tube.

[0061] In some embodiments, such as Figure 1 As shown, the sample loading assembly 100 also includes a support member 105, and a receiving portion 101 is disposed on the support member 105.

[0062] It is understandable that the support 105 can be a testing seat or a testing frame, and the clamping component 104 and the support 105 can be an integrated structural design. Of course, the support 105 can also be an independent modular design. It is made of transparent acrylic, specifically polymethyl methacrylate (PMMA).

[0063] In this embodiment, the transparent material support 105 facilitates observation of the insertion state and liquid level of the sampling component. Precision light-transmitting holes, for example, 3mm in diameter with a tolerance of ±0.05mm, can be opened at corresponding positions in the incident and exit light paths to allow the detection beam to pass through, thus realizing a lateral transmission light path system. Furthermore, to prevent ambient light from entering the light path from the side, light-shielding sleeves can be installed on both sides of the light-transmitting holes.

[0064] In some embodiments, the inner diameter of the support 105 can be set to 13.2 ± 0.1 mm as a radial positioning reference for the sampling element.

[0065] In some embodiments, such as Figure 1 As shown, the blood testing device may include: a control processing component 200.

[0066] The control processing component 200 is connected to the position detection element and is configured to receive the position signal.

[0067] Once the sampling component is inserted into the receiving section 101, the positioning detection component will trigger to generate a positioning signal. This positioning signal is input to the controller (MotorControlUnit, MCU) via a general-purpose input / output (GPIO) port.

[0068] For example, the control processing component 200 may include: a controller MCU, an analog-to-digital converter ADC, an Analog-to-Digital Converter, a memory, and a communication interface.

[0069] The controller MCU is configured to control the timing switching of multiple light sources in the light source module 301, the movement of the optical detection component 300, the movement and positioning of the drive component 400, and the acquisition and processing of position signals, tube-shaped signals, and optical signals.

[0070] The analog-to-digital converter (ADC) is configured to convert the photocurrent signal output by the detector module 3023 into a digital signal.

[0071] The memory is configured to store factory baseline calibration parameters and detection thresholds.

[0072] The communication interface is configured for data transmission with external devices. The communication interface is the data interface between the controller MCU and external devices, supporting USB wired or Bluetooth wireless connections. The communication interface is used to communicate with devices such as personal... l It enables data transmission with external devices such as computers, PCs, smartphones, hospital laboratory information systems, and LIS systems. Two methods are supported: a USB wired connection, using the CH340E USB-UART converter chip to achieve serial communication between the controller MCU and the PC; and a Bluetooth wireless connection, using a Bluetooth Low Energy (BLE) module (model HC-42) to achieve wireless data transmission with smartphones. The communication interface outputs information such as test results, quality control data, and equipment status to external devices.

[0073] In some embodiments, such as Figure 1 As shown, the blood testing device may include an optical detection component 300. The optical detection component 300 is movable relative to the sample loading component 100.

[0074] The optical detection component 300 may include a light source module 301 and an optical path system 302. The light source module 301 is connected to the control processing component 200 and includes multiple light sources. The control processing component 200 is configured to control the multiple light sources to alternately light up in a time sequence. The optical path system 302 detects the optical signal during the alternating lighting of the multiple light sources and transmits the detected optical signal to the control processing component 200.

[0075] It should be noted that the optical detection principle is based on the light scattering theory of turbidity measurement. PRP is a platelet suspension in which platelet particles scatter and absorb incident light, resulting in a decrease in the intensity of transmitted light. According to an extended form of Lambert-Beer's law, the absorbance of the suspension is positively correlated with the concentration of suspended particles.

[0076] However, absorbance measurements at a single wavelength are affected by a variety of factors: individual differences in plasma background color, such as lipemia (milky white and turbid), hemolysis (red), and jaundice (yellow); differences in blood collection tube material, wall thickness, and translucency; and ambient light interference.

[0077] In this embodiment of the disclosure, by setting the light source module 301 in the optical detection component 300 to include multiple light sources, the multiple light sources are switched and lit alternately in sequence, and the optical path system 302 detects the optical signal during the alternating lighting of multiple light sources, and the tube wall interference is calibrated by differential signal calibration and the influence of individual plasma background color differences on the detection results is eliminated.

[0078] Therefore, this embodiment introduces an 805nm reference wavelength and utilizes dual-wavelength differential technology ΔA = A660 - A805 to eliminate the aforementioned interference factors. 805nm is located in the near-infrared region, where platelet scattering signals are weak, while the contributions from plasma background color and vessel wall factors are similar to those at the 660nm wavelength. Therefore, differential operations can effectively cancel out these interferences. For example, as... Figure 1 As shown, the light source module 301 includes a first light source 3011 and a second light source 3012. For example, the first light source 3011 can be a 660nm main detection LED light source, and the second light source 3012 can be an 805nm reference correction LED light source.

[0079] It should be noted that platelets exhibit significant light scattering at a wavelength of 660 nm. This wavelength is also located far from the hemoglobin absorption peaks of approximately 415 nm, 540 nm, and 576 nm, effectively reducing interference from residual red blood cells in the detection. Within the 570-660 nm range, the absorbance of PRP shows a good linear relationship with platelet concentration.

[0080] Therefore, the 660nm main detection LED light source is located in the platelet scattering sensitive region, namely the 570-660nm range, and is used to detect the scattering absorbance of platelets in PRP. The 805nm reference correction LED light source is used to correct baseline drift caused by individual plasma background color, tube wall transmittance, and factors such as lipemia / hemolysis. The wavelength range of the first light source 3011 is 650-670nm, and the optical power range is 30-70mW. The wavelength range of the second light source 3012 is 795-815nm, and the optical power range is 30-70mW.

[0081] The first light source 3011 and the second light source 3012 are alternately lit using a time-division multiplexing method, with the alternation switching frequency ranging from 0.5 to 2 kHz.

[0082] For example, the first light source 3011 is a 660nm LED light source, which can be a Thorlabs M660F1, 50mW; the second light source 3012 is an 805nm LED light source, which can be a Thorlabs M805F1, 50mW. The first light source 3011 and the second light source 3012 are mounted on a printed circuit board, and the luminous intensity stability is controlled by a constant current drive circuit to be better than ±1%. The two light sources are alternately lit using a time-division multiplexing method, with a switching frequency of 1kHz, to avoid mutual interference between the two optical signals.

[0083] Through the integrated circuit bus, Inter-Integrated Circuit, I 2 The C-bus communicates with the controller MCU. The MCU retrieves the corresponding factory baseline calibration parameters from memory based on the identified tube type and specifications, and then sends them to the light source driver circuit via the Serial Peripheral Interface (SPI) bus to adjust the LED drive current. The LED light source is powered by a constant current drive circuit, which receives the Pulse Width Modulation (PWM) control signal from the MCU. The 660nm and 805nm LEDs are alternately illuminated using time-division multiplexing with a 1kHz switching frequency. After passing through collimating lenses L1 / L2 and bandpass filters BPF660 / BPF805, the light is combined into a coaxial optical path at the combiner BS and directed towards the blood collection tube detection area.

[0084] Each LED light source is fitted with a collimating lens, for example, with a focal length of 10mm, a numerical aperture (NA) of 0.3, and a bandpass filter, for example, a passband full width at half maximum (FWHM) of 660nm ± 10nm or 805nm ± 10nm FWHM, to ensure the monochromaticity and collimation of the incident light.

[0085] In this embodiment of the disclosure, the influence of individual differences in plasma background color on the detection results is eliminated by employing 660nm / 805nm dual-wavelength differential detection technology.

[0086] Among these steps, factory baseline calibration is a crucial step in eliminating interference from tube wall and plasma background color. The calibration process is as follows: Blank calibration: Using a standard blood collection tube filled with distilled water as a blank reference, the transmitted light intensity I0(660) and I0(805) were measured at wavelengths of 660nm and 805nm, respectively, and recorded in the device memory.

[0087] Tube wall calibration: Use standard blood collection tubes containing platelet-rich plasma (PPP), for example, platelet concentration <10 × 10⁻⁶. 9 / L, measure the transmitted light intensity, calculate the baseline absorbance A_baseline(660) and A_baseline(805) of the tube wall and plasma background color, and record them in the memory.

[0088] Standard curve establishment: A series of PRP standard concentration gradients were determined using flow cytometry to determine platelet concentrations: 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800 × 10⁻⁶. 9 / L, A660 and A805 were measured separately, ΔA was calculated, and a standard curve of ΔA versus platelet concentration (PLT) was established. In this embodiment, the standard curve equation is: ΔA = 0.002627 × [PLT] - 0.022, and the correlation coefficient R is 0.022. 2 =0.9999.

[0089] Sample testing: For the sample to be tested, after measuring A660 and A805, calculate ΔA=(A660-A_baseline(660))-(A805-A_baseline(805)), and use the standard curve to calculate the platelet concentration.

[0090] In some embodiments, such as Figures 1-2 As shown, the optical path system 302 may include: an incident optical path 3021, an outgoing optical path 3022, and a detector module 3023.

[0091] The incident light path 3021 is configured to guide the incident light emitted by the light source in the light source module 301 to the detection area of ​​the sampling device.

[0092] The photodetector (PD) outputs a photocurrent that is converted into a voltage signal by a built-in transimpedance amplifier (TIA). The TIA has an adjustable gain range of 10. 3 -106 V / A. The preamplifier transimpedance amplifier (TIA) can be integrated within the same photodetector module. Through a printed circuit board (PCB), copper traces connect to the input of the analog-to-digital converter (ADC). The ADC converts the analog voltage into a digital signal, which is then transmitted to the controller MCU for data processing via the SPI bus.

[0093] Figure 2 This is a schematic diagram of the dual-wavelength differential optical path design of an optical detection component provided in one embodiment of the present disclosure. Figure 2 The layout of the 660nm main detection optical path and the 805nm reference correction optical path is shown, as well as the optical path routing for beam splitting / combining, bandpass filtering, and lateral transmission detection.

[0094] For example, such as Figure 2 As shown, the incident light path 3021 includes a collimating lens 303 and a bandpass filter 304. The collimating lens 303 can be a first collimating lens L1 and a second collimating lens L2, which are located between the light source module 301 and the sample loading assembly 100. The bandpass filter 304 can be a first bandpass filter BPF660 and a second bandpass filter BPF805, with the first bandpass filter BPF660 located between the first collimating lens L1 and the sample loading assembly 100, and the second bandpass filter BPF805 located between the second collimating lens L2 and the sample loading assembly 100, to ensure the monochromaticity and parallelism of the incident light.

[0095] The bandpass filter is a bandpass filter (BPF). In this embodiment, two bandpass filters are used: a first bandpass filter (BPF660) and a second bandpass filter (BPF805), located in the 660nm and 805nm optical paths, respectively. Relevant parameters of the first bandpass filter (BPF660) and the second bandpass filter (BPF805) are shown in Table 1.

[0096] Table 1. Parameter settings for the first bandpass filter BPF660 and the second bandpass filter BPF805

[0097] The core function of a bandpass filter (BPF) is to filter out stray light of non-target wavelengths, ensuring that the light beam reaching the detection area is monochromatic. The functions of a BPF include: Wavelength selection: The BPF660 allows only light in the 650-670nm range to pass through, filtering out other wavelength components emitted by the LED and ambient light. The BPF805 allows only light in the 795-815nm range to pass through. Noise reduction: It prevents non-target wavelength light from reaching the photodetector, reducing detection noise and improving the signal-to-noise ratio. Crosstalk prevention: It avoids wavelength crosstalk caused by combining the 660nm and 805nm light beams, ensuring the accuracy of dual-wavelength detection.

[0098] For example, such as Figure 2 As shown, the incident light path 3021 also includes a beam combiner 305. The beam combiner, BS, is positioned between the bandpass filter 304 and the sample loading assembly 100. The beam combiner 305 is configured to combine the light paths of multiple light sources at the intersection point to share the same outgoing light path 3022. For example, the first bandpass filter BPF660 is mounted after the first collimating lens L1 and before the beam combiner 305, centered along the optical axis of the 660nm light path. The second bandpass filter BPF805 is mounted after the second collimating lens L2 and before the beam combiner 305, centered along the optical axis of the 805nm light path. The beam combiner 305 is mounted at a 45° angle at the intersection point of the 660nm and 805nm light paths. For example, the beam combiner 305 can be designed using a dichroic mirror, model Edmund Optics#49-654, with specific parameters shown in Table 2.

[0099] Table 2 Parameter settings for the combiner BS

[0100] The core function of the beam combiner 305 is to merge two independent optical paths, 660nm and 805nm, into a single coaxial optical path, including: Optical beam combining: Utilizing the wavelength-selective reflection / transmission characteristics of dichroic mirrors, 660nm and 805nm beams from different directions are combined onto the same optical axis, allowing the two beams to share the same detection optical path and the same detector, simplifying the optical path structure and reducing the size and cost of the device.

[0101] Wavelength isolation: While combining the beams, wavelength isolation is maintained between the two beams to avoid optical crosstalk. 660nm light is reflected, and 805nm light is transmitted, so the two beams do not interfere with each other.

[0102] Coaxial detection: The two beams after beam combining pass through the PRP sample completely coaxially, ensuring that the detection area is consistent, so that the differential calculation of ΔA=A660-A805 has physical meaning, that is, the two beams pass through the same optical path and sample volume.

[0103] In some embodiments, such as Figure 2 As shown, the incident light path 3021 and the exit light path 3022 are located on opposite sides of the sample loading assembly 100. The exit light path 3022 receives the transmitted light that passes through the sampling element and the PRP layer.

[0104] The detector module 3023 is located on the side of the outgoing light path 3022 away from the sample loading assembly 100. The outgoing light path 3022 is configured to guide the transmitted light passing through the sample to the detector module 3023, and the detector module 3023 is configured to convert the received optical signal into a photocurrent signal.

[0105] The photodetector can be a photodiode, such as a silicon photodiode, Thorlabs PDA100A2, with a spectral response range of 320-1100 nm.

[0106] The outgoing light path 3022 and the incoming light path 3021 are arranged 180° opposite each other, located on opposite sides of the sampling device, such as a blood collection tube. The incident light enters from one side, passes through the tube wall and the PRP detection layer, and then exits from the other side (the outgoing side) and is received by the photodetector. The outgoing light path 3022 may include a focusing lens to focus the outgoing light beam onto the photosensitive surface of the photodetector, thereby improving light collection efficiency. Of course, the outgoing light path 3022 does not contain any active optical elements such as a light source or filters; its function is to efficiently transmit the transmitted light carrying PRP platelet concentration information to the photodetector.

[0107] The optical path of the 660nm LED is as follows: 660nm LED light source → First collimating lens L1 → First bandpass filter BPF660 → Beam combiner BS, reflection path → blood collection tube detection area.

[0108] The 660nm optical path is connected as follows: first bandpass filter BPF660 → reflection from the upper surface of the beam combiner BS at a reflection angle of 90° → outgoing optical path → blood collection tube detection area. The 660nm beam arrives at the upper surface of the beam combiner at an incident angle of 45°, is reflected 90° by the dichroic film layer, and then shines horizontally onto the blood collection tube.

[0109] The optical path of an 805nm LED is as follows: 805nm LED light source → second collimating lens L2 → second bandpass filter BPF805 → beam combiner BS, transmission path → blood collection tube detection area.

[0110] The 805nm optical path is connected as follows: second bandpass filter BPF805 → beam combiner BS transmission, transmission direction remains unchanged → outgoing optical path → blood collection tube detection area. The 805nm beam arrives at the BS beam combiner at an incident angle of 45°, penetrates the dichroic film layer, and coincides with the reflected 660nm beam to form a coaxial beam combiner.

[0111] The combined optical path is: beam combiner BS → blood collection tube, PRP detection layer → photodetector PD. The two beams share the same outgoing optical path after being combined, pass through the PRP layer in the blood collection tube, and are received by the photodetector on the opposite side.

[0112] In some embodiments, such as Figure 1 As shown, the blood testing device also includes a drive assembly 400. The drive assembly 400 is connected to the control processing assembly 200 and is configured to drive the optical detection assembly 300 to move when the control processing assembly 200 receives a positioning signal, so as to perform positioning detection on the sample within the receiving portion 101.

[0113] After receiving the positioning signal, the controller MCU starts the mechanical positioning program, and the drive component 400 drives the optical detection component 300 to move in order to perform positioning detection on the sampling component in the accommodating part 101.

[0114] Figure 3 This is a schematic diagram of the structure of the driving component in a blood testing device provided in an embodiment of this disclosure. Figure 3 The relative positions of the monitoring and positioning components driven by the drive assembly are shown, as well as the distribution of the plasma layer, PRP layer, white membrane layer, and red blood cell layer in the sampling component after centrifugation.

[0115] In some embodiments, such as Figure 3 As shown, the drive component 400 may include: a drive element 401.

[0116] The driving end of the driving component 401 is connected to the optical detection component 300 to drive the optical detection component 300 to move.

[0117] For example, the driver 401 may include a stepper motor. The stepper motor driver receives control pulses from the controller MCU via the DirectionSignal, DIR / STEP signal lines. The stepper motor has a step angle of 1.8°, microsteps ranging from 1 / 8 to 1 / 32, a positioning accuracy of ±0.5 mm, and a repeatability of ±0.1 mm. Using a stepper motor driven by a precision leadscrew effectively avoids interference from sampling components such as the layering interface of blood collection tubes, ensuring that the detection layer is located in the optimal detection area of ​​the PRP layer.

[0118] In some embodiments, such as Figure 3 As shown, the drive component 400 may include: a monitoring component 402.

[0119] The monitoring element 402 is installed in the optical detection assembly 300 and is configured to monitor the liquid level and stratification interface within the sampling element.

[0120] For example, the monitoring device 402 includes a liquid level sensor array S1-S6, which comprises 4-8 liquid level sensors, with a spacing between adjacent liquid level sensors ranging from 8-12 mm. For instance, the liquid level sensors can be infrared photoelectric sensors, and the liquid level sensor array S1-S6 monitors the position of each interface layer within the sampling device in real time.

[0121] For example, the liquid level sensor array consists of 6 infrared photoelectric sensors with a spacing of 10 mm between adjacent infrared photoelectric sensors, used to detect the position of the liquid level and the layering interface inside the sampling piece.

[0122] The analog output signals from the infrared photoelectric sensors in the liquid level sensor array S1-S6 are multiplexed (MUX) and then acquired by the analog-to-digital converter (ADC) before being input to the controller MCU. After processing by the MCU, the liquid level sensor output signals automatically identify the plasma-PRP interface and the PRP-erythrocyte interface using a gradient algorithm, calculating the midpoint position of the PRP layer. The drive detection window is aligned with the lower part of the PRP layer, for example, at a distance of 2-3 mm from the PRP-erythrocyte interface.

[0123] In some embodiments, such as Figure 3 As shown, the drive assembly 400 may include a positioning element 403. The positioning element 403 is mounted on the optical detection assembly 300 and is configured to position the liquid level within the sampling element.

[0124] For example, the positioning element 403 includes a positioning probe, and the driving element 401 drives the positioning probe to move along the sampling element, for example, a stepper motor drives the positioning probe to move up and down along the axis of the blood collection tube.

[0125] In this embodiment of the disclosure, based on the detection of the liquid level and the layer interface within the sampling member, the positioning member 403 further positions the area to be detected within the sampling member, so as to accurately target the lower part of the PRP layer.

[0126] The positioning algorithm of driver component 400 is as follows: Initialization: After the sample is inserted into the sample loading assembly 100, the optical detection assembly 300 is located at the initial position at the bottom of the sample, for example, 5 mm from the bottom of the sample.

[0127] Liquid level scanning: The drive unit 401 moves upward at a speed of 2 mm / s, while the infrared sensor array of the monitoring unit 402 continuously acquires signals. When the signal intensity changes abruptly, the position is recorded as the liquid level position.

[0128] Interface recognition: Continuing the upward scan, the infrared sensor array's sensor signal shows two distinct gradient change points at the plasma-PRP interface and the PRP-erythrocyte interface. The controller MCU automatically identifies these two interface locations using a differential algorithm.

[0129] Location calculation: Calculate the PRP layer thickness h = h2 - h1, where h1 is the height of the plasma-PRP interface, h2 is the height of the PRP-erythrocyte interface, and the target detection location is h1 + h / 3, which is the lower part of the PRP layer about 2-3 mm away from the PRP-erythrocyte interface.

[0130] Precise positioning: The optical detection component 300 moves quickly to the target position, and the positioning accuracy is finely adjusted to ±0.1mm by the positioning component 403, and then the position is locked for optical detection.

[0131] In some embodiments, such as Figure 1 As shown, the blood testing device may further include a display component 500. The display component 500 is connected to the control processing component 200 and is configured to display the processing results output by the control processing component 200.

[0132] In some embodiments, the display component 500 may include an indicator light array disposed on the outer surface of the housing 600, the indicator light array being configured to display semi-quantitative detection results of different levels.

[0133] Understandably, the five-level semi-quantitative grading system based on a wide threshold design—low, low-medium, medium, medium-high, and high—accommodates minor fluctuations in platelet status, such as slight aggregation or temperature changes, and provides grading results that can be directly used clinically for PRP quality assessment.

[0134] The LED indicator array can be 5 WS2812BRGB LEDs, or other numbers of LED indicator arrays. The controller MCU connects via I... 2 C-bus drives the LED indicator array.

[0135] In this embodiment of the disclosure, the indicator light array can be red-yellow-green-blue-violet corresponding to low-medium-low-medium-medium-high-high concentrations, respectively, to distinguish the detection structure. Of course, it can also be an indicator light array of other colors, and this disclosure does not limit it.

[0136] In some embodiments, such as Figure 1 As shown, the blood testing device also includes: a housing 600.

[0137] The sample loading component 100, the control processing component 200, the optical detection component 300, and the drive component 400 are all disposed within the housing 600. The housing 600 has a first opening corresponding to the sample loading component 100 for inserting the sample into the receiving portion 101, and / or, the housing 600 has a second opening, which includes at least one of a heat dissipation hole, a power interface, and a communication interface.

[0138] The first opening can be a Φ14mm circular hole, and the second opening can be a circular hole or a square hole; the embodiments disclosed herein do not limit this.

[0139] The housing 600 can be injection molded from acrylonitrile-butadiene-styrene copolymer, ABS engineering plastic.

[0140] In this embodiment, the housing 600 serves both as a structural support, housing all internal functional modules, and as an optical shield to block ambient light from entering the detection area and prevent stray light interference. It also provides safety protection, preventing operators from contacting the internal optical detection component 300 and drive component 400. Furthermore, electromagnetic shielding is achieved by coating the inner wall of the housing 600 with conductive paint, reducing the impact of electromagnetic interference on photoelectric detection.

[0141] The support 105 and the housing 600 can be detachably connected by a snap fastener to facilitate cleaning and replacement of the support 105.

[0142] In this embodiment, the blood testing device is small in size, lightweight, and low in power consumption. It can be integrated into a portable in vitro diagnostic (IVD) sample loading device, making it suitable for rapid PRP quality control in operating rooms, outpatient clinics, bedside settings, and other similar environments.

[0143] In some embodiments, the display component 500 may include a display screen. The display screen is disposed on the outer surface of the housing 600 and on the same side as the indicator light array, and the display screen is configured to display specific detection parameter results.

[0144] The display screen can be an LCD screen, which communicates via SPI / I 2 The controller MCU communicates via the C-bus. The LED indicator array can be positioned facing the operator, with the display screen located above it. The controller MCU and display screen are connected via on-board wiring. The controller MCU communicates via I-bus. 2The C bus, consisting of two signal lines (SCL / SDA), is traced on the PCB using copper foil and directly drives the LED indicator array and the LCD screen. This connection is a short-range wired communication within the circuit board and does not involve wireless transmission. The LED indicator uses WS2812 BRGB LEDs and is controlled via a single-wire serial protocol; the LCD screen is controlled via I... 2 C / SPI protocol communication.

[0145] In this embodiment of the disclosure, the liquid crystal display screen shows specific absorbance values ​​and estimated platelet concentration ranges, which can be directly used for the grading results of PRP quality assessment.

[0146] Figure 4 This is a flowchart of a blood testing method provided by an embodiment of the present disclosure. Figure 4 It demonstrates the complete testing process from blood collection, centrifugation, sample loading, positioning, detection, calculation to result output.

[0147] Secondly, this disclosure provides a detection method using a blood testing device according to any embodiment of the first aspect described above. The complete detection time from blood collection, centrifugation, sample loading, positioning, detection, calculation to result output is approximately 10-12 minutes, including the centrifugation operation; excluding the centrifugation operation, it is approximately 1 minute. For example... Figure 4 As shown, the detection method includes the following steps S100-S600:

[0148] S100: Preprocess the sample containing venous blood to obtain a sample with a detection layer.

[0149] Step S100 includes: Samples containing venous blood are centrifuged to obtain samples with a detection layer. The centrifugal force is 50-200×g and the centrifugation time is 5-10 minutes.

[0150] The sample after step S100 consists of, from top to bottom, a plasma layer, a detection target layer (i.e., platelet-rich plasma layer), a white membrane layer, and a red blood cell layer.

[0151] S200: Place the pre-processed sample onto the sample loading assembly of the blood testing device and receive a position signal.

[0152] S300: In response to the positioning signal, the drive component is activated to perform the detection layer of the positioning detection sample.

[0153] The detection zone is located in the lower middle part of the detection layer. For example, it can be located 2-3 mm from the PRP-erythrocyte interface.

[0154] S400: Controls multiple light sources in the optical detection assembly to be lit alternately.

[0155] Step S400 includes: The two light sources in the optical detection component are controlled to be lit alternately. The two light sources are a light source with a first preset wavelength and a light source with a second preset wavelength, and the first preset wavelength and the second preset wavelength are different.

[0156] Calculate the difference absorbance ΔA between the first preset wavelength and the second preset wavelength.

[0157] S500: Receives optical signals detected by the optical path system during the alternating illumination of multiple light sources.

[0158] S600: Processes the optical signal and outputs the processed result to the display component.

[0159] The processed results in step S600 include multiple semi-quantitative levels, which include: first concentration, ΔA < 0.50; second concentration, 0.50 ≤ ΔA < 1.00; third concentration, 1.00 ≤ ΔA < 1.50; fourth concentration, 1.50 ≤ ΔA < 2.00; and fifth concentration, ΔA ≥ 2.00.

[0160] The display component includes an array of LED indicators, with the first color representing the first concentration, the second color representing the second concentration, the third color representing the third concentration, the fourth color representing the fourth concentration, and the fifth color representing the fifth concentration, with a 10% overlap buffer between adjacent levels.

[0161] A five-level semi-quantitative grading system was adopted, based on the corrected dual-wavelength differential absorbance ΔA. Table 3 shows the five-level semi-quantitative grading standard for PRP platelets.

[0162] Table 3. PRP Platelet-Rich Classification Standards (5 Grades)

[0163] The grading thresholds employ a wide threshold design, with a 10% overlap buffer between adjacent grades to tolerate measurement fluctuations caused by factors such as slight platelet aggregation and temperature fluctuations. When a measurement falls within the overlap area, the device displays a combined indication of the two adjacent grades (e.g., "low to medium concentration"), prompting the operator to make a judgment based on clinical needs.

[0164] The testing method also includes: correcting for interference using factory baseline calibration parameters, the calibration process of which is as follows: Blank calibration: Using a standard blood collection tube filled with distilled water as a blank reference, the transmitted light intensity I0(660) and I0(805) were measured at wavelengths of 660nm and 805nm, respectively, and recorded in the device memory.

[0165] Tube wall calibration: Use standard blood collection tubes containing platelet-rich plasma (PPP), for example, platelet concentration <10 × 10⁻⁶. 9 / L, measure the transmitted light intensity, calculate the baseline absorbance A_baseline(660) and A_baseline(805) of the tube wall and plasma background color, and record them in the memory.

[0166] Standard curve establishment: A series of PRP standard concentration gradients were determined using flow cytometry to determine platelet concentrations: 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800 × 10⁻⁶. 9 / L, A660 and A805 were measured separately, ΔA was calculated, and a standard curve of ΔA versus platelet concentration was established. In this embodiment, the standard curve equation is: ΔA = 0.002627 × [PLT] - 0.022, and the correlation coefficient R is 1 / L. 2 =0.9999.

[0167] Sample testing: For the sample to be tested, after measuring A660 and A805, calculate ΔA=(A660-A_baseline(660))-(A805-A_baseline(805)), and use the standard curve to calculate the platelet concentration.

[0168] The factory baseline calibration parameters include: blank calibration values, tube wall calibration values ​​of the sampled parts, and standard curve parameters. The standard curve is in the range of 50-800×10⁻⁶. 9 It is linear within the range of / L, R 2 ≥0.995.

[0169] To verify the performance of the blood testing device disclosed herein, the following embodiments and comparative examples were designed and implemented. All embodiments and comparative examples were carried out in accordance with the requirements of the "Regulations for the Registration and Management of In Vitro Diagnostic Reagents" and the "Good Clinical Practice for Medical Devices".

[0170] Experimental materials: (1) Sample source: Thirty healthy volunteers (16 males and 14 females, aged 22-55 years, BMI 18.5-28.0 kg / m²) were recruited, and venous blood was collected after obtaining informed consent. Basic information of the samples and whole blood platelet count can be found in Table 4.

[0171] Table 4. Basic information of the samples and whole blood platelet count

[0172] (2) Blood collection tubes: Blood samples for PRP preparation were collected using BDVacutainer sodium citrate anticoagulation vacuum blood collection tubes (product number 363083, batch number 5234021, 3.2% sodium citrate, 2.7 mL); blood samples for whole blood platelet count were collected using BDVacutainer EDTA-K2 anticoagulation vacuum blood collection tubes (product number 367844, batch number 8239012).

[0173] (3) Instruments and equipment: Eppendorf 5804R low-speed horizontal centrifuge (item number 5804R, batch number EP20240156); Sysmex XN-550 fully automated hematology analyzer (batch number SN55078901) for whole blood platelet counting; BDFACS CantoII flow cytometer (batch number FC20230512) as a reference method for platelet counting; UshioPi COSCOPE micro spectrophotometer (batch number PS20240501) for methodological comparison; Thorlabs M660F1 and M805F1 LED light sources and PDA100A2 photodiode detector.

[0174] (4) Reagents: Sigma-Aldrich ADP inducer (catalog number A2754, batch number SLBT4567); GibcoPBS buffer (catalog number 10010023, batch number 2456789); BDBiosciences CD41-PE and CD61-FITC antibodies; Sysmexe-Check quality control products (low value L20240601, medium value M20240601, high value H20240601).

[0175] For a detailed list of experimental materials and reagents, please refer to Table 5.

[0176] Table 5 List of Experimental Materials and Reagents

[0177] PRP preparation method: A two-step centrifugation method is used. Step 1: Centrifuge sodium citrate anticoagulated whole blood at 50-200×g (depending on the experimental group) for 5-10 minutes; Step 2: Aspirate the supernatant plasma (containing the PRP layer), transfer it to a new centrifuge tube, and further concentrate it as needed.

[0178] Platelet concentrations in 30 PRP samples were simultaneously determined using the blood testing apparatus disclosed herein and flow cytometry (reference method). Flow cytometry was performed using the CD41 / CD61 dual labeling method, following the protocol recommended by the International Committee for Standardization in Hematology (ICSH).

[0179] The verification results compared with the gold standard of flow cytometry are shown in Table 6.

[0180] Table 6. Comparison and verification results with the gold standard of flow cytometry

[0181] The comparison results show that the Pearson correlation coefficient r = 0.9979 between the blood testing device disclosed herein and the flow cytometry reference method, and the coefficient of determination R0.05 2 =0.9958, indicating a very high degree of consistency between the two. The validation results of the grading thresholds for 30 clinical samples are shown in Table 7. In the 30 samples, the semi-quantitative grading concordance rate was 100%, and the grading results for all samples were completely consistent with the flow cytometry validation results. The mean absolute deviation was (10.2±6.8)×10⁻⁶. 9 / L, with an average relative deviation of (2.8±1.9)%, meeting the accuracy requirements for clinical semi-quantitative detection.

[0182] Table 7. Validation results of grading thresholds in 30 clinical samples.

[0183] Establishing a standard curve

[0184] A series of PRP standards (concentration gradients: 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800 × 10⁻⁶) were used to determine platelet concentration by flow cytometry. 9 / L, a total of 14 concentration points), and the dual-wavelength differential absorbance ΔA was measured using the blood detection device disclosed herein.

[0185] The calibration data for the dual-wavelength standard curve (flow cytometry calibration) are shown in Table 8.

[0186] Table 8. Calibration data for dual-wavelength standard curves (flow cytometry calibration)

[0187] The standard curve results showed that ΔA was related to platelet concentration in the range of 50-800×10⁻⁶. 9 The system exhibits a good linear relationship within the range of / L, with the linear regression equation being ΔA = 0.002627 × [PLT] - 0.022 and the correlation coefficient R0. 2 =0.9999, sensitivity (slope) is 0.002627 A / (×10 9 The detection limit was 5.7 × 10⁻⁶ (L / L). 9 / L (3σ method). The linear range was determined based on multi-point dilution experiments, and no significant nonlinear deviation was found within the test range.

[0188] Precision and repeatability experiments

[0189] Intra-batch precision: Selected concentration is approximately 400 × 10⁻⁶. 9 A PRP sample of / L was repeatedly measured 20 times consecutively by the same operator using the same equipment on the same day. The results showed that the coefficient of variation (CV) of A660 was 0.42%, the CV of ΔA was 0.55%, and the CV of calculated PLT was 2.1%, all of which were less than the 5% precision requirement.

[0190] Inter-batch precision: The same PRP sample was measured twice daily (once in the morning and once in the afternoon) for 10 consecutive days, and the inter-day precision was calculated. The results showed that the total mean PLT was 401 × 10⁻⁶. 9 / L, with a standard deviation of 8.5×10 9 / L, CV is 2.12%, and the maximum inter-day deviation is 18×10 9 / L, with an average diurnal deviation of 9.2×10 9 / L, meeting the batch-to-batch precision requirements.

[0191] Precision and repeatability experimental data are shown in Tables 9 and 10.

[0192] Table 9. Intra-batch precision and repeatability test data

[0193] Table 10. Data on inter-batch precision and repeatability experiments

[0194] Quality control product verification

[0195] Use Sysmexe-Check low value (50×10) 9 / L), median (200×10 9 / L), high value (500×10 9 Each of the / L quality control samples was measured 10 times to evaluate the accuracy and stability of the testing system. The validation results of the quality control samples are shown in Table 11.

[0196] Table 11 Validation Results of Quality Control Products

[0197] The results showed that the measured mean value of the low-value quality control sample was 52×10. 9 / L, deviation +4.0%, CV=3.2%; the measured mean of the median quality control sample is 198×10 9 / L, deviation -1.0%, CV=2.8%; the measured mean of high-value quality control products is 512×10 9 / L, deviation +2.4%, CV=2.5%. All three concentration levels of quality control samples passed validation, demonstrating that the detection system of this invention has good accuracy and stability.

[0198] Interference experiment

[0199] To evaluate the impact of common interference factors on the detection results, the following interference experiments were conducted: (1) Lipid interference: Different concentrations of triglycerides (0, 2, 5, 10 mmol / L) were added to PRP samples to simulate lipemia. The results showed that when the triglyceride concentration was below 5 mmol / L, the effect on the detection results was <5%; when it was 10 mmol / L, the effect was 8.2%, but within the tolerance range of the semi-quantitative grading threshold.

[0200] (2) Hemolysis interference: Different concentrations of free hemoglobin (0, 0.5, 1, 2 g / L) were added to the PRP sample to simulate hemolysis. The results showed that, due to the use of a 660 nm detection wavelength (far from the hemoglobin absorption peak) and dual-wavelength differential correction in this invention, the influence of hemolysis on the detection results was <3%, which is much lower than the 15-20% of the single-wavelength method.

[0201] (3) Jaundice interference: Different concentrations of total bilirubin (0, 50, 100, 200 μmol / L) were added to the PRP sample to simulate jaundice. After dual-wavelength differential correction, the effect of jaundice on the test results was <4%, which meets the clinical testing requirements.

[0202] The above interference experiment results show that the dual-wavelength differential calibration technology disclosed herein can effectively resist the influence of common interference factors and ensure the accuracy and reliability of the detection results.

[0203] Clinical sample validation

[0204] Clinical sample validation was conducted at the Department of Laboratory Medicine of XX Hospital. Fifty clinical PRP samples (covering departments such as orthopedics, dentistry, and plastic surgery) were collected. The blood testing device disclosed herein was used for semi-quantitative grading, and flow cytometry was used to determine platelet concentration as a reference.

[0205] The verification results showed that the overall concordance rate between the blood testing device disclosed herein and flow cytometry grading was 96% (48 / 50), with the ΔA values ​​of two inconsistent samples falling precisely at the grading threshold boundary and within the overlap buffer. The Kappa consistency coefficient was 0.95, indicating extremely high consistency. Operator satisfaction with the device was rated 4.6 / 5.0, indicating that it was easy to operate, provided intuitive results, and was suitable for clinical use.

[0206] It is readily understood that those skilled in the art can combine, split, or reorganize the embodiments provided in this disclosure to obtain other embodiments, none of which exceed the protection scope of this disclosure.

[0207] The above detailed embodiments further illustrate the purpose, technical solutions, and beneficial effects of the embodiments of this disclosure. It should be understood that the above are merely specific embodiments of the embodiments of this disclosure and are not intended to limit the protection scope of the embodiments of this disclosure. Any modifications, equivalent substitutions, improvements, etc., made on the basis of the technical solutions of the embodiments of this disclosure should be included within the protection scope of the embodiments of this disclosure.

Claims

1. A blood detection device, characterized in that, include: The sample loading assembly (100) includes a receiving portion (101) and a positioning detection element is disposed therein, the positioning detection element being configured to generate a positioning signal; A control processing component (200) is connected to the positioning detection element, and the control processing component (200) is configured to receive the positioning signal; An optical detection component (300) is movable relative to the sample loading component (100). The optical detection component (300) includes a light source module (301) and an optical path system (302). The light source module (301) is connected to the control processing component (200), and the light source module (301) includes multiple light sources. The control processing component (200) is configured to control the multiple light sources to switch and alternately light up in a time sequence. The optical path system (302) detects light signals during the alternating lighting of the multiple light sources and transmits the detected light signals to the control processing component (200). A drive assembly (400) is connected to the control processing assembly (200), and the drive assembly (400) is configured to drive the optical detection assembly (300) to move when the control processing assembly (200) receives the positioning signal, so as to perform positioning detection on the sampler in the accommodating portion (101); A display component (500) is connected to the control processing component (200), and the display component (500) is configured to display the processing results output by the control processing component (200).

2. The blood detection device according to claim 1, characterized in that, The sample loading assembly (100) further includes an identification element (103) connected to the control processing assembly (200), the identification element (103) being configured to identify a sample located in the receiving portion (101) and generate an identification signal; and / or, The sample loading assembly (100) further includes a clamping member (104), which is disposed on the outer periphery of the receiving portion (101) and has a preset clamping stroke; and / or, The sample loading assembly (100) also includes a support member (105), and the receiving portion (101) is disposed on the support member (105).

3. The blood detection device according to claim 1, characterized in that, The light source module (301) includes a first light source (3011) and a second light source (3012). The first light source (3011) has a wavelength range of 650-670nm and an optical power range of 30-70mW. The second light source (3012) has a wavelength range of 795-815nm and an optical power range of 30-70mW; and / or, The first light source (3011) and the second light source (3012) are alternately lit using a time-division multiplexing method, with the alternation switching frequency ranging from 0.5 to 2 kHz.

4. The blood detection device according to claim 2, characterized in that, The optical path system (302) includes: An incident light path (3021) is configured to guide incident light emitted from the light source in the light source module (301) to the detection area of ​​the sampling element; The outgoing light path (3022) and the incoming light path (3021) are respectively located on opposite sides of the sample loading assembly (100); The detector module (3023) is disposed on the side of the outgoing optical path (3022) away from the sample loading assembly (100); The outgoing optical path (3022) is configured to guide the transmitted light passing through the sampling element to the detector module (3023), and the detector module (3023) is configured to convert the received optical signal into a photocurrent signal.

5. The blood detection device according to claim 4, characterized in that, The incident light path (3021) includes a collimating lens (303) and a bandpass filter (304). The collimating lens (303) is located between the light source module (301) and the sample loading assembly (100). The bandpass filter (304) is located between the collimating lens (303) and the sample loading assembly (100), and / or, The incident light path (3021) further includes a beam combiner (305) disposed between the bandpass filter (304) and the sample loading assembly (100), wherein the beam combiner (305) is configured to combine the light paths of the multiple light sources at the intersection point to share the same outgoing light path (3022).

6. The blood detection device according to claim 1, characterized in that, The drive component (400) includes: A driving element (401) is provided, the driving end of which is connected to the optical detection assembly (300) to drive the optical detection assembly (300) to move. A monitoring element (402) is installed on the optical detection assembly (300), the monitoring element (402) being configured to monitor the liquid level and stratification interface within the sampling element; A positioning element (403) is installed on the optical detection assembly (300), and the positioning element (403) is configured to position the liquid level within the sampling element.

7. The blood detection device according to claim 6, characterized in that, The monitoring element (402) includes a liquid level sensor array, which comprises 4-8 liquid level sensors, with a spacing of 8-12 mm between adjacent liquid level sensors; and / or, The positioning element (403) includes a positioning probe; and / or, The driving component (401) includes a stepper motor with a step angle of 1.8°, microstep subdivision of 1 / 8-1 / 32, positioning accuracy of ±0.5mm, and repeatability of ±0.1mm.

8. The blood detection device according to claim 4, characterized in that, The control processing component (200) includes: The controller is configured to control the timing switching of multiple light sources of the light source module (301), the movement of the optical detection component (300), the movement and positioning of the drive component (400), the positioning signal, the identification signal, and the acquisition and processing of the light signal; An analog-to-digital converter is configured to convert the photocurrent signal output by the detector module (3023) into a digital signal; The memory is configured to store factory baseline calibration parameters and detection thresholds; The communication interface is configured to transmit data with external devices.

9. The blood detection device according to claim 1, characterized in that, Also includes: The housing (600), the sample loading component (100), the control processing component (200), the optical detection component (300), and the driving component (400) are all disposed within the housing (600). The housing (600) has a first opening corresponding to the sample loading component (100) for inserting the sample into the receiving portion (101), and / or, the housing (600) has a second opening, the second opening including at least one of a heat dissipation hole, a power interface, and a communication interface.

10. The blood detection device according to claim 9, characterized in that, The display component (500) includes: an indicator light array disposed on the outer surface of the housing (600), the indicator light array being configured to display semi-quantitative detection results of different levels; and / or, A display screen is disposed on the outer surface of the housing (600) and on the same side as the indicator array, and the display screen is configured to display specific detection parameter results.

11. A detection method using the blood detection device according to any one of claims 1-10, characterized in that, Includes the following steps: Preprocessing of samples containing venous blood to obtain samples with a detection layer; The pre-treated sample is placed on the sample loading component of the blood testing device, and a position signal is received. In response to the arrival signal, the driving component is activated to perform positioning detection of the detection layer of the sample; Control the alternating illumination of multiple light sources in the optical detection assembly; The optical path system receives the optical signals detected during the alternating illumination of the multiple light sources; The optical signal is processed, and the processed result is output to the display component.

12. The detection method of the blood detection device according to claim 11, characterized in that, The control of multiple light sources in the optical detection component to be lit alternately includes: The two light sources in the optical detection component are controlled to be lit alternately. The two light sources are a light source with a first preset wavelength and a light source with a second preset wavelength, and the first preset wavelength and the second preset wavelength are different. Calculate the difference absorbance ΔA between the first preset wavelength and the second preset wavelength.

13. The detection method of the blood detection device according to claim 11, characterized in that, The processed results include multiple semi-quantitative levels, which include a first concentration, ΔA < 0.50; a second concentration, 0.50 ≤ ΔA < 1.00; a third concentration, 1.00 ≤ ΔA < 1.50; a fourth concentration, 1.50 ≤ ΔA < 2.00; and a fifth concentration, ΔA ≥ 2.

00.

14. The detection method of the blood detection device according to claim 13, characterized in that, The display component includes an LED indicator array, where the first color represents the first concentration; the second color represents the second concentration; the third color represents the third concentration; the fourth color represents the fourth concentration; and the fifth color represents the fifth concentration. A 10% overlap buffer is provided between adjacent levels.

15. The detection method of the blood detection device according to claim 11, characterized in that, The detection method further includes: Interferences are corrected using factory baseline calibration parameters, wherein the factory baseline calibration parameters include a blank calibration value, a tube wall calibration value for the sampling piece, and standard curve parameters, the standard curve being linear over a range of 50-800 x 10 9 / L with R 2 ≥0.995.

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