Blood immunity in-vitro diagnosis equipment and diagnosis method thereof
By integrating microfluidic design and electrochemical precision excitation, combined with intelligent algorithm processing, the problems of cumbersome operation, insufficient accuracy and high noise of traditional blood immunoassay in vitro diagnostic equipment have been solved, achieving efficient and accurate detection results.
Patent Information
- Application Number
- CN202511118055.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-11
- Publication Date
- 2025-11-14
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Traditional blood immunoassay in vitro diagnostic equipment suffers from problems such as cumbersome operation, insufficient accuracy in detecting trace samples, high background noise, and poor stability of electrochemiluminescence.
Employing microfluidic integrated design, precise electrochemical excitation, and intelligent algorithm processing, this system integrates a sample loading module, microfluidic chip, detection core module, and human-machine interaction module. Combined with a three-dimensional spiral flow channel, photomultiplier tube, potentiostat, and embedded processor, it achieves fully automated detection, eliminates human operation errors, and improves signal-to-noise ratio and detection accuracy.
It achieves efficient and accurate blood immune detection, reduces operational steps, lowers sample requirements, improves detection accuracy and environmental adaptability, and enhances signal-to-noise ratio and luminescence stability.
Smart Images
Figure CN120948780A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of medical device technology, and in particular to a blood immunology in vitro diagnostic device and its diagnostic method. Background Technology
[0002] Blood immunoassay in vitro diagnostic equipment refers to medical testing equipment that analyzes and tests blood samples, mainly serum or plasma samples, outside the human body using immunological principles and technologies to diagnose diseases, assess health status, or monitor treatment effectiveness. It is one of the core pillars of modern medical testing. The analysis is performed in an in vitro laboratory by drawing blood and using the principle of specific binding of antigens and antibodies to detect various immune-related markers in the blood, serving a variety of clinical needs such as disease diagnosis, efficacy evaluation, health screening, and transfusion safety.
[0003] Traditional ELISA methods for detecting blood immune biomarkers require multiple manual operations, which are time-consuming. While chemiluminescence analyzers offer high sensitivity, they are bulky and expensive. Although microfluidic technology can achieve integration, existing devices suffer from problems such as unoptimized flow channel design leading to sample residue, signal crosstalk during parallel detection of multiple biomarkers, and a lack of low-noise coupling schemes with electrochemiluminescence. To address these issues, we propose an in vitro diagnostic device and method for blood immune biomarkers. Through microfluidic integrated design, precise electrochemical excitation, and intelligent algorithm processing, this system systematically solves the technical problems of cumbersome operation, large sample requirements, insufficient accuracy, and poor environmental adaptability in blood immune biomarkers. Summary of the Invention
[0004] This invention provides a blood immunoassay in vitro diagnostic device and method, which solves the problems of cumbersome traditional immunoassay procedures, insufficient accuracy of micro-sample detection, high background noise, and poor stability of electrochemiluminescence.
[0005] The present invention provides the following solution to the above-mentioned technical problems: a blood immunoassay in vitro diagnostic device, comprising a sample loading module, a microfluidic chip, a detection core module, a control system, and a human-computer interaction module. The sample loading module is provided with an inlet with an anti-backflow valve and a capillary quantitative cavity. The microfluidic chip includes a substrate layer, a reaction layer, and an electrode layer. The substrate layer is provided with a three-dimensional spiral flow channel. The detection core module includes a photomultiplier tube and a potentiostat. The control system includes an embedded processor, a temperature controller, and a negative pressure pump. The human-computer interaction module includes a touch screen and a Bluetooth module.
[0006] The diagnostic method includes the following steps:
[0007] S1, Equipment initialization: After powering on, the device performs a self-test, scans the RFID tags on the sample tubes, and preheats.
[0008] S2, Sample loading and pretreatment: After centrifuging anticoagulated whole blood to separate plasma, 50 μL is quantitatively added to the sample inlet of the device through the capillary quantitative cavity;
[0009] S3, Immunological reaction and signal excitation: The sample is driven by a negative pressure pump to flow through the spiral channel of the basal layer to the reaction layer. The sample is incubated with coated antibody magnetic beads in the reaction layer for 10 min. A potentiostat applies a voltage of +1.2V to the electrode layer and injects luminol solution to excite the luminescence reaction.
[0010] S4, signal acquisition and processing, uses a photomultiplier tube to detect the number of photons at a wavelength of 483nm and converts them into electrical signals;
[0011] S5, Concentration Calculation and Output: Calculates biomarker concentration; human-computer interaction module outputs test report.
[0012] Based on the above technical solution, the present invention can be further improved as follows.
[0013] Furthermore, the capillary quantitative cavity has an accuracy of ±0.1 μL. The base layer is made of PDMS material with etched three-dimensional spiral channels. The reaction layer has an antibody-coated magnetic nanobead immobilization region and streptavidin magnetic beads coated with anti-IL-6 antibody. The electrode layer is an integrated three-electrode system on ITO conductive glass, which precisely controls the plasma sample injection volume, eliminates human operation errors, and ensures the consistency of the immune reaction system. The PDMS material provides biocompatibility and gas permeability, maintaining a stable reaction environment. The antibody is directionally immobilized through the high affinity binding of streptavidin and biotin.
[0014] Furthermore, the three-dimensional spiral channel has a width of 200 μm and a depth of 100 μm. The three-electrode system consists of a working electrode, a reference electrode, and a counter electrode. The spiral channel design extends the sample path and enhances the antigen-antibody contact efficiency. The three-electrode system constructs a closed-loop electrochemical circuit to precisely control the excitation voltage.
[0015] Furthermore, the photomultiplier tube has a wavelength range of 400-700nm and a signal-to-noise ratio >100:1. The potentiostat applies a scanning voltage of -1.5V to +1.5V, the embedded processor implements timing control, the temperature controller achieves a constant temperature reaction of 37±0.5℃, captures the characteristic light emission signal of 483nm, suppresses ambient light interference, the potentiostat dynamically optimizes the excitation voltage to adapt to different marker energy levels, and the embedded processor synchronously coordinates the fluid drive, reaction incubation, and signal excitation process.
[0016] Furthermore, the touchscreen displays the concentration curve and transmits data via Bluetooth. The touchscreen visualizes the original signal and the fitted curve, and supports wireless report export.
[0017] Further, in step S1, after powering on, a self-test program is executed to verify the electrode impedance and the dark current of the photomultiplier tube. The normal range of electrode impedance is 1-5 kΩ, and the threshold of the dark current of the photomultiplier tube is <0.1 nanoamp. The RFID tag of the sample tube is scanned, the detection parameters (such as IL-6 detection mode) are loaded, the temperature controller is preheated to 37°C, the photomultiplier tube gain is set to 1000 volts, and the hardware status is verified upon power-on to ensure the stability of the detection baseline.
[0018] Furthermore, in step S2, the anticoagulated whole blood sample is centrifuged at 3000 rpm, and plasma is separated in 10 minutes. 50 μL of plasma is drawn and injected into the inlet of the device. The negative pressure pump drives the sample at a pressure of -30 kPa. The plasma is transferred to the reaction layer through the spiral channel at a flow rate of 2 μL / sec. The inner wall of the channel is treated with plasma hydrophilic treatment (contact angle ≤15°) to ensure zero residue and achieve standardized plasma separation and zero-residue transfer.
[0019] Further, in step S3, the sample is incubated with antibody-coated magnetic beads at 37°C for 10 minutes in the reaction layer to complete the antigen-antibody specific binding. Phosphate-buffered saline (PBS, pH 7.4) is injected to wash away unbound material, the flow rate is increased to 5 μL / s, an electrochemiluminescent reagent containing 0.1 mmol luminol and 0.5 mmol hydrogen peroxide is injected, a step voltage of +1.2 volts (relative to the reference electrode) is applied, and chemiluminescence is excited for 5 seconds to efficiently remove non-specific binders and trigger directional luminescence.
[0020] Furthermore, in step S4, the electrical signal is decomposed into three levels using the Daubechies4 wavelet basis function. The three levels of decomposition are wavelet decomposition, high-frequency coefficient thresholding, and wavelet reconstruction, which separate the effective signal from high-frequency noise and baseline drift.
[0021] Furthermore, in step S5, an abnormal handling mechanism is set up. If I_max < 1000 nanoamps, an "insufficient signal" warning is triggered, suggesting retesting or increasing the sample volume to 100 microliters. A negative pressure sensor is set to monitor the pressure in real time. If the pressure > -10 kPa, a backflushing procedure is automatically executed, injecting 50 microliters of deionized water and increasing the pressure to +50 kPa. This intelligently addresses the risk of detection failure, ensuring the robustness of the equipment. A fault-tolerant design is adopted, automatically suggesting additional samples when the signal is insufficient to avoid invalid report output. An anti-clogging design is adopted, and pressure monitoring and backflushing reduce the flow channel failure rate to < 1 time / thousandth of tests.
[0022] The beneficial effects of this invention are as follows: This invention provides a blood immunoassay in vitro diagnostic device and method, which have the following advantages:
[0023] 1. This solution solves the problem of cumbersome traditional immunoassay processes. Traditional immunoassay processes involve many manual steps, such as centrifugation, sample loading, incubation, and washing, which are time-consuming. This solution achieves fully automated one-stop testing by integrating a microfluidic chip to achieve "sample in - result out", thereby effectively improving testing efficiency.
[0024] 2. This solution solves the problem of insufficient accuracy in detecting trace samples in traditional immunoassay equipment. Existing equipment requires 200μL of plasma and has an error >5%. This solution, with its capillary quantitative cavity with an accuracy of ±0.1μL and a plasma hydrophilic flow channel, can achieve low-error quantification with only 50μL of sample.
[0025] 3. This solution addresses the problem of severe non-specific binding interference in traditional immunoassay devices. Unbound impurities in traditional diagnostic devices result in high background noise. This solution reduces non-specific signals and effectively improves the signal-to-noise ratio by using 5μL / s pressurized rinsing and PBS buffer.
[0026] 4. This solution solves the problem of poor electrochemiluminescence stability in traditional immunoassay detection equipment. Traditional voltage fluctuations easily lead to signal drift. This solution uses a three-electrode system to make the potential drift < ±5mV. Combined with a +1.2V step voltage for precise excitation, it can effectively improve the luminescence efficiency.
[0027] 5. This blood immunoassay in vitro diagnostic device and its diagnostic method systematically solve the technical problems of cumbersome operation, large sample requirements, insufficient accuracy, and poor environmental adaptability in blood immunoassay through microfluidic integrated design, precise electrochemical excitation, intelligent algorithm processing, and triple innovation.
[0028] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it according to the contents of the specification, the preferred embodiments of the present invention are described in detail below with reference to the accompanying drawings. Specific embodiments of the present invention are given in detail below with reference to the accompanying drawings. Attached Figure Description
[0029] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and, together with their description, serve to explain the invention and do not constitute an undue limitation thereof. In the drawings:
[0030] Figure 1 This is a flowchart illustrating a blood immunoassay in vitro diagnostic device and diagnostic method according to an embodiment of the present invention.
[0031] Figure 2 This is a system architecture diagram of a blood immunological in vitro diagnostic device and diagnostic method provided in an embodiment of the present invention. Detailed Implementation
[0032] The following is in conjunction with the appendix Figure 1-2 The principles and features of the present invention are described below. The examples given are for illustrative purposes only and are not intended to limit the scope of the invention. The invention is described more specifically in the following paragraphs by way of example with reference to the accompanying drawings. The advantages and features of the invention will become clearer from the following description. It should be noted that the drawings are in a very simplified form and use non-precise proportions, and are only used to facilitate and clarify the illustration of the embodiments of the invention.
[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.
[0034] like Figure 1-2 As shown, this invention provides a blood immunoassay in vitro diagnostic device, including a sample loading module, a microfluidic chip, a detection core module, a control system, and a human-computer interaction module. The sample loading module is equipped with an inlet with an anti-backflow valve and a capillary quantitative cavity. The microfluidic chip includes a base layer, a reaction layer, and an electrode layer. The base layer is provided with a three-dimensional spiral flow channel. The detection core module includes a photomultiplier tube and a potentiostat. The control system includes an embedded processor, a temperature controller, and a negative pressure pump. The human-computer interaction module includes a touch screen and a Bluetooth module.
[0035] Preferably, the capillary quantitative cavity has an accuracy of ±0.1μL, the base layer is made of PDMS material with etched three-dimensional spiral flow channels, the reaction layer is provided with an antibody-coated magnetic nanobead immobilization area, the reaction layer is provided with streptavidin magnetic beads coated with anti-IL-6 antibody, and the electrode layer is an integrated three-electrode system on ITO conductive glass, which precisely controls the plasma sample injection volume, eliminates human operation errors, and ensures the consistency of the immune reaction system. The PDMS material provides biocompatibility and gas permeability, maintains a stable reaction environment, and the antibody is directionally immobilized through the high affinity binding of streptavidin and biotin.
[0036] Preferably, the three-dimensional spiral channel has a width of 200μm and a depth of 100μm. The three-electrode system consists of a working electrode, a reference electrode, and a counter electrode. The spiral channel design extends the sample path and enhances the antigen-antibody contact efficiency. The three-electrode system constructs a closed-loop electrochemical circuit and precisely controls the excitation voltage.
[0037] Preferably, the photomultiplier tube has a wavelength range of 400-700nm and a signal-to-noise ratio of >100:1. A potentiostat applies a scanning voltage of -1.5V to +1.5V, an embedded processor implements timing control, a temperature controller achieves a constant temperature reaction of 37±0.5℃, captures the characteristic light emission signal of 483nm, suppresses ambient light interference, the potentiostat dynamically optimizes the excitation voltage to adapt to different marker energy levels, and the embedded processor synchronously coordinates the fluid drive, reaction incubation, and signal excitation process.
[0038] Preferably, the touchscreen displays the concentration curve and transmits data via Bluetooth. The touchscreen visualizes the original signal and the fitted curve, and supports wireless report export.
[0039] The specific working principle and usage method of this invention are as follows:
[0040] S1, Equipment initialization: After powering on, the device performs a self-test, scans the RFID tags on the sample tubes, and preheats.
[0041] After powering on, execute the self-test program to verify the electrode impedance (normal range 1-5 kΩ) and the dark current of the photomultiplier tube (threshold < 0.1 mAh), scan the RFID tag of the sample tube, load the test parameters (such as IL-6 test mode), preheat the temperature controller to 37°C, and set the photomultiplier tube gain to 1000 volts.
[0042] S2, Sample loading and pretreatment: After centrifuging anticoagulated whole blood to separate plasma, 50 μL is quantitatively added to the inlet of the device through the capillary quantitative cavity;
[0043] Take whole blood samples anticoagulated with EDTA, centrifuge at 3000 rpm for 10 minutes to separate the plasma, and inject 50 μL of plasma into the inlet of the device. The negative pressure pump drives the sample at a pressure of -30 kPa. The plasma is transferred to the reaction layer through the spiral channel at a flow rate of 2 μL / sec. The inner wall of the channel is treated with plasma hydrophilic treatment (contact angle ≤15°) to ensure zero residue.
[0044] S3, Immunological reaction and signal excitation: The sample is driven through the spiral channel by a negative pressure pump, and incubated with the coated antibody magnetic beads in the reaction layer for 10 min. A voltage of +1.2V is applied to the electrode layer, and luminol solution is injected to excite the luminescence reaction.
[0045] The sample was incubated with antibody-coated magnetic beads in the reaction layer at 37°C for 10 minutes to complete the antigen-antibody specific binding. Unbound material was washed off with phosphate-buffered saline (PBS, pH 7.4), the flow rate was increased to 5 μL / s, and an electrochemiluminescent reagent containing 0.1 mmol luminol and 0.5 mmol hydrogen peroxide was injected. A step voltage of +1.2 volts (relative to the reference electrode) was applied and chemiluminescence was excited for 5 seconds.
[0046] S4, signal acquisition and processing, uses a photomultiplier tube to detect the number of photons at a wavelength of 483nm and converts them into electrical signals;
[0047] Optical signal acquisition: The photomultiplier tube integrates at a wavelength of 483 nm for 200 milliseconds to collect the number of photons and convert them into a current signal (unit: nanoampere);
[0048] Signal denoising algorithm: Three-level decomposition using Daubechies4 wavelet basis functions;
[0049] The original signal is sequentially processed by wavelet decomposition, high-frequency coefficient thresholding (Sureshrink method), and wavelet reconstruction, and the signal-to-noise ratio is improved to 120:1 after noise reduction.
[0050] Baseline correction algorithm: Asymmetric Least Squares (ALS) is used.
[0051] Objective function: minΣ{w_i*(y_i-b_i)^2+λ*Σ(Δ 2 b_i)^2};
[0052] The weight w_i is 0.01 at the signal peak and 1 in the baseline region, and the smoothing parameter λ = 1 × 10 5 ;
[0053] Peak Extraction: Calculate the first derivative of the corrected signal. The extreme values that cross zero and have a half-width greater than 5 sampling points are determined as valid peaks, and the peak current intensity I_max is recorded.
[0054] S5, Concentration Calculation and Output: Calculates biomarker concentration, and outputs test report via human-computer interaction module;
[0055] Standard curve fitting was performed using a four-parameter logistic regression (4PL) model.
[0056] I = D + (AD) / [1 + (C / x)^B]
[0057] Parameter definition,
[0058] A: Minimum asymptote (zero concentration signal value, typically 15.2 nanoamps)
[0059] D: Maximum asymptote (saturation signal value, typical value 9820 nanoamperes)
[0060] C: Half-maximal effect concentration (EC5) 50 (Typical value 25.3 picograms / mL)
[0061] B: Slope factor (typical value -1.08);
[0062] Concentration inverse algorithm:
[0063] Concentration x is calculated from the detection signal value I_max:
[0064] x=C*exp{ln[(AD) / (I_max-D)-1] / B};
[0065] Results output: The touchscreen displays "IL-6 concentration: XX.X picograms / mL" and generates a test report containing the original signal curve;
[0066] Key algorithm validation was performed by establishing a 4PL model through calibration at 5 concentration points (0 / 5 / 20 / 50 / 100 picograms / mL);
[0067] Linear correlation coefficient R 2 =0.998 (n=10)
[0068] Limit of detection (LOD) = 0.1 picograms / mL (signal-to-noise ratio S / N ≥ 3)
[0069] Repeatability test: 20 picograms / mL sample tested 10 times consecutively, relative standard deviation (RSD) = 2.8%;
[0070] Exception handling mechanism
[0071] Low signal alarm: If I_max < 1000 nanoamps, an "insufficient signal" warning is triggered. It is recommended to re-examine or increase the sample size to 100 microliters.
[0072] Channel blockage handling: The negative pressure sensor monitors the pressure in real time. If the pressure is >-10 kPa, the reverse flushing procedure is automatically executed (50 μL of deionized water is injected at +50 kPa).
[0073] Temperature drift compensation:
[0074] The compensated signal I_comp = I_raw * [1 - 0.015 * (T_actual - 37)];
[0075] The value of 0.015 is the temperature compensation coefficient (% / ℃), which is determined by accelerated aging experiments.
[0076] It should be noted that the operation is limited to isolated plasma samples, and the output result is the IL-6 protein concentration value, in picograms per milliliter. It does not contain any disease diagnosis conclusions. The contents not described in detail in this specification are existing technologies known to those skilled in the art.
[0077] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Those skilled in the art can readily implement the present invention based on the accompanying drawings and the above description. However, any modifications, alterations, or variations made by those skilled in the art without departing from the scope of the present invention, utilizing the disclosed technical content, are equivalent embodiments of the present invention. Furthermore, any modifications, alterations, or variations made to the above embodiments based on the essential technology of the present invention are still within the protection scope of the present invention.
Claims
1. A blood immunoassay in vitro diagnostic device, comprising a sample loading module, a microfluidic chip, a detection core module, a control system, and a human-computer interaction module, characterized in that: The sample loading module is equipped with an inlet with an anti-backflow valve and a capillary quantitative cavity. The microfluidic chip includes a substrate layer, a reaction layer, and an electrode layer. The substrate layer is provided with a three-dimensional spiral flow channel. The detection core module includes a photomultiplier tube and a potentiostat. The control system includes an embedded processor, a temperature controller, and a negative pressure pump. The human-machine interaction module includes a touch screen and a Bluetooth module. The diagnostic method includes the following steps: S1, Equipment initialization: After powering on, the device performs a self-test, scans the RFID tags on the sample tubes, and preheats. S2, Sample loading and pretreatment: After centrifuging anticoagulated whole blood to separate plasma, 50 μL is quantitatively added to the sample inlet of the device through the capillary quantitative chamber. S3, Immunological reaction and signal excitation: The sample is driven by a negative pressure pump to flow through the spiral channel of the basal layer to the reaction layer. The sample is incubated with coated antibody magnetic beads in the reaction layer for 10 min. A potentiostat applies a voltage of +1.2V to the electrode layer and injects luminol solution to excite the luminescence reaction. S4, signal acquisition and processing, uses a photomultiplier tube to detect the number of photons at a wavelength of 483nm and converts them into electrical signals; S5, Concentration Calculation and Output: Calculates biomarker concentration; human-computer interaction module outputs test report.
2. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, The capillary quantitative cavity has an accuracy of ±0.1μL. The substrate layer is made of PDMS material and has a three-dimensional spiral flow channel etched on it. The reaction layer is provided with a region for immobilization of antibody-coated magnetic nanobeads. The reaction layer is provided with streptavidin magnetic beads coated with anti-IL-6 antibody. The electrode layer is a three-electrode system integrated on ITO conductive glass.
3. The blood immunoassay in vitro diagnostic device according to claim 2, characterized in that, The three-dimensional spiral channel is 200μm wide and 100μm deep, and the three electrode systems are the working, reference, and counter electrode systems.
4. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, The photomultiplier tube has a wavelength range of 400-700nm and a signal-to-noise ratio of >100:
1. A potentiostat applies a scanning voltage of -1.5V to +1.5V. The embedded processor implements timing control, and the temperature controller achieves a constant temperature response of 37±0.5℃.
5. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, The touchscreen displays the concentration curve and transmits data via Bluetooth.
6. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, In step S1, after powering on, a self-test program is executed to verify the electrode impedance and the dark current of the photomultiplier tube. The normal range of electrode impedance is 1-5 kΩ, and the threshold of the dark current of the photomultiplier tube is <0.1 nanoamp. The RFID tag of the sample tube is scanned, the test parameters are loaded, the temperature controller is preheated to 37°C, and the gain of the photomultiplier tube is set to 1000 volts.
7. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, In step S2, the anticoagulated whole blood sample is centrifuged at 3000 rpm, and plasma is separated after 10 minutes. 50 μL of plasma is aspirated and injected into the inlet of the device. The negative pressure pump drives the sample at a pressure of -30 kPa. The plasma is transferred to the reaction layer through the spiral channel at a flow rate of 2 μL / sec. The inner wall of the channel is treated with plasma hydrophilic treatment to ensure zero residue.
8. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, In step S3, the sample is incubated with antibody-coated magnetic beads in the reaction layer at 37°C for 10 minutes to complete the antigen-antibody specific binding. Phosphate buffer is injected to wash away unbound material, the flow rate is increased to 5 μL / s, an electrochemiluminescent reagent containing 0.1 mmol luminol and 0.5 mmol hydrogen peroxide is injected, a +1.2 volt step voltage is applied, and chemiluminescence is excited for 5 seconds.
9. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, In step S4, the electrical signal is decomposed into three levels using wavelet basis functions: wavelet decomposition, high-frequency coefficient thresholding, and wavelet reconstruction.
10. The blood immunoassay in vitro diagnostic device according to claim 1, characterized in that, In step S5, an abnormal handling mechanism is set up. If I_max < 1000 nanoamps, an "insufficient signal" warning is triggered, and it is recommended to retest or increase the sample volume to 100 microliters. The negative pressure sensor monitors the pressure in real time. If the pressure > -10 kPa, the reverse flushing procedure is automatically executed.