Quality control method for blood cell testing in medical testing

Abstract

The present invention provides a method for quality control of blood cell testing in medical testing. [Solution] The method includes: obtaining the initial number of platelets in a measurement sample using an electrical resistance method; obtaining a two-dimensional scattering scatter diagram of the measurement sample using a scattered light method; determining a total light intensity set based on the sum of side scattered light intensities and the sum of forward scattered light intensities in the two-dimensional scattering scatter diagram; and performing linear or nonlinear correction on the initial number of platelets according to a preset correction rule. The present invention establishes a correction rule appropriate for the platelet count based on experimental data from a blood sample set, determines the corresponding light intensity range for the initial number of platelets obtained from a conventional blood test and the two-dimensional scattering scatter diagram, and applies the appropriate correction rule to correct the initial number of platelets and ensure the accuracy of the platelet measurement results.

Description

[Technical Field]

[0001] The present invention relates to the field of blood cell medical testing technology, and more particularly to a quality control method for blood cell testing in medical testing. [Background technology]

[0002] When measuring the number of platelets in blood, conventional blood cell analyzers use the Coulter principle (electrical resistance method) for detection. The basic principle is to measure the volume and number of particles by utilizing the change in resistance that occurs when particles in a blood sample pass through a detection hole. However, in actual detection, the volume of small white blood cell fragments is very similar to the volume of platelets. This similarity in volume makes it difficult to distinguish the range of resistance changes, resulting in counting errors. In particular, in samples with a high concentration of white blood cell fragments, the number of white blood cell fragments is incorrectly determined as the number of platelets, resulting in a significant decrease in the accuracy of platelet counting.

[0003] To solve this problem, some techniques have introduced the scattered light method. In this method, the forward-scattered light (FSC) signal is used to represent the volumetric size of particles, while the side-scattered light (SSC) signal reflects the internal complexity of particles. However, the scattered light method still has obvious limitations in practical applications. The forward-scattered light signal of small-volume leukocyte debris is very similar to that of platelets, making it impossible to effectively distinguish the two using the FSC signal. On the other hand, while side-scattered light can represent the internal complexity of particles, the complexity of small-volume leukocyte debris is low and there is significant overlap with the SSC signal of platelets, limiting the effectiveness of differentiation using two-dimensional scatter plots (FSC-SSC diagrams).

[0004] These two methods still cannot completely eliminate the interference of white blood cell debris on platelet counting. Especially in large-scale detection scenarios, the limitations of traditional methods become even more pronounced due to the diversity of samples and fluctuations in the debris ratio, making it impossible to meet the requirements for accuracy and consistency in platelet counting. Summary of the Invention [Problem to be solved by the invention]

[0005] 1) Technical issues to be resolved The present invention aims to provide a quality control method for blood cell testing in medical testing, and to solve the problem that small volumes of white blood cell debris are easily misidentified as platelets in conventional platelet counting tests, which results in an impact on the accuracy of platelet counting. [Means for solving the problem]

[0006] 2) Technical plan In order to achieve the above object, the present invention provides the following technical solution: A method for quality control of blood cell testing in medical testing, Obtaining an initial number of platelets in the sample to be measured using an electrical resistance method; Obtaining a two-dimensional scattering scatter diagram of the measurement target sample based on a scattered light method, wherein the horizontal axis of the two-dimensional scattering scatter diagram represents side scattered light intensity and the vertical axis represents forward scattered light intensity; determining a total light intensity set based on a sum of side scattered light intensities and a sum of forward scattered light intensities in the two-dimensional scattering scatter plot; According to a preset correction rule, when a total light intensity set within a set region in the two-dimensional scattering scatter diagram is located within a preset first light intensity interval, linearly correcting the initial number of platelets; performing nonlinear correction on the initial number of platelets when the total light intensity set within the set region is located in a predetermined second light intensity range; The correction rule construction step includes: obtaining blood sample sets, including a patient sample set and a healthy sample set; determining the first light intensity interval based on a total light intensity set of the set area in the two-dimensional scattering scatter plot of the healthy sample set; and determining a second light intensity interval based on a total light intensity set of the set area in the two-dimensional scattering scatter plot of the patient sample set; performing fluorescent staining on white blood cell debris in the blood sample set to determine the number of white blood cell debris in the blood sample set; constructing a linear correction function based on the number of white blood cell debris in the healthy sample set; and constructing a non-linear correction function based on the number of white blood cell debris in the patient sample set.

[0007] Furthermore, the fluorescent staining of the leukocyte debris specifically refers to staining the nucleic acids in the leukocyte debris.

[0008] Furthermore, the dilution concentration of each blood sample in the blood sample set is the same, and after obtaining the two-dimensional scattering scatter diagram for each blood sample using the light scattering method, the white blood cell debris is stained so that it has a fluorescent signal that can be distinguished from platelets, and the number of white blood cell debris in each blood sample is calculated.

[0009] Furthermore, in the two-dimensional scattering scatter plot, the total light intensity set within the set region is TIFF0007743138000002.tif9168, where: TIFF0007743138000003.tif8168 is the sum of the side scattered light intensity of white blood cell debris and platelets within the set area. TIFF0007743138000004.tif7168 is the sum of the forward scattered light intensity of white blood cell debris and platelets within the set area.

[0010] Furthermore, the step of determining the set region includes: obtaining a two-dimensional scatter plot of each blood sample in the healthy sample set; determining an initial region based on the intersection of platelet and leukocyte debris distribution regions in each two-dimensional scattering scattergram; The initial region in each two-dimensional scatter plot Total light intensity set Calculate and statistically analyze TIFF0007743138000005.tif9168. and determining the initial region where the total light intensity set has the highest consistency based on a stability analysis method as the set region.

[0011] Furthermore, in the healthy sample set, a total light intensity set of the set region in the two-dimensional scattering scatter plot of each of the blood samples is calculated, and from there, Maximum total light intensity set TIFF0007743138000006.tif9168 and Minimum total light intensity set TIFF0007743138000007.tif9168 is extracted, and the first light intensity interval is:

number

[0012] Furthermore, in the patient sample set, a total light intensity set of the set region in the two-dimensional scattering scatter plot of each of the blood samples is calculated, and from there, Maximum total light intensity set TIFF0007743138000010.tif10168 and Minimum total light intensity set TIFF0007743138000011.tif9168 is extracted, and the second light intensity interval is:

number

[0013] 3) Beneficial Effects Compared with the prior art, the present invention has the following beneficial effects:

[0014] The present invention establishes a correction rule suitable for platelet count based on experimental data of a blood sample set, in which the experimental data of a group of healthy samples and patient samples is used to fluorescently label white blood cell debris in the sample set, calculate the number of white blood cell debris, and then establish linear and nonlinear correction functions, respectively. For blood samples before white blood cell debris staining, a first light intensity range to which linear correction is applied is determined based on the total light intensity set of a set area in the two-dimensional scattering scatter diagram of the healthy sample set, and a second light intensity range to which nonlinear correction is applied is determined based on the total light intensity set of a set area in the two-dimensional scattering scatter diagram of the patient sample set.

[0015] A normal blood test is performed on the blood sample to be measured, and the obtained initial platelet count and two-dimensional scattering scatter plot are used to calculate the total light intensity set in a set area in the two-dimensional scattering scatter plot. The corresponding light intensity range is determined and an appropriate correction rule is applied, thereby correcting the initial platelet count and ensuring the accuracy of the platelet measurement results. [Brief explanation of the drawings]

[0016] [Figure 1] 1 is a schematic flow diagram of a quality control method for blood cell testing in medical testing provided by an embodiment of the present invention. [Figure 2] 1 is a schematic diagram of a flow chart for acquiring a preset correction rule in a quality control method for blood cell testing in medical testing provided by an embodiment of the present invention. [Figure 3] 1 is a schematic diagram of a flow chart for defining a set area in a quality control method for blood cell testing in medical testing provided by an embodiment of the present invention; [Figure 4] 1 is a schematic diagram showing the distribution of each type of blood cell in a two-dimensional scattering scatter diagram obtained in a quality control method for blood cell testing in medical testing provided by an embodiment of the present invention. [Figure 5] 1 is a schematic diagram illustrating a setting area A of platelets and white blood cell debris in a two-dimensional scattering scatter diagram in a quality control method for blood cell testing in medical testing provided by an embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0017] The following clearly and completely describes the technical solutions in the embodiments of the present invention with reference to the drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments that can be obtained by those skilled in the art without creative work are included in the protection scope of the present invention.

[0018] In describing the present invention, terms relating to orientation or positional relationships, such as "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inside," and "outside," are based on the orientations or positional relationships shown in the drawings and are intended to facilitate description and understanding of the present invention, but do not suggest or imply that the subject devices or elements must be configured or operated in a particular orientation. Therefore, they should not be construed as limitations on the present invention.

[0019] Furthermore, terms such as "first," "second," etc. are merely descriptive and should not be understood as indicating or implying relative importance.

[0020] It should be noted that, unless there is a contradiction, the features in the embodiments of the present invention can be combined with each other.

[0021] A diluted blood sample is introduced into a chamber with a tiny detection hole, and the platelets in the blood are counted using electrical impedance (Coulter principle).When cells in the blood pass through the detection hole, they change the impedance value, and the volume and number of cells are detected based on this change.

[0022] When measuring platelets in blood cells using electrical impedance, the inventors discovered that white blood cell debris inevitably exists in diluted blood samples. The volume ranges of white blood cell debris and platelets are very similar, typically between 2 μm and 4 μm. Because electrical impedance mainly counts particles based on their volume, small volumes of white blood cell debris may be mistaken for platelets by the system, resulting in an inflated platelet count.

[0023] When the number of leukocyte debris in a blood sample is low, it has little effect on the overall platelet count results, but when the sample has a high leukocyte count or is affected by sample preparation factors such as vigorous shaking or hemolysis, the number of leukocyte debris increases significantly, exacerbating counting errors.

[0024] To solve this problem, conventional techniques have introduced scattered light methods, combining optical detection with electrical resistance. In scattered light measurement of blood cells, the forward scattered light (FSC) signal reflects the volumetric size of the particle, with larger particles typically producing stronger forward scattered light. Platelets and white blood cell debris have similar volumes, but with minor differences. The side scattered light (SSC) signal reflects the internal structure and complexity of the particle, such as the granular structure of the cell and the ratio of nucleus to cytoplasm. In practice, small volumes of white blood cell debris and platelets are typically very similar in volume, resulting in nearly identical FSC signals. Due to this similarity, the FSC axis (forward scattered light intensity) cannot effectively distinguish white blood cell debris from platelets in two-dimensional scatter plots. White blood cells, especially white blood cell debris, have a highly complex internal structure, but their small volume results in weaker scattered light signals and lower complexity. Platelets typically lack an internal nucleus, resulting in a relatively weak SSC signal. However, there is a certain overlap between the SSC signals of white blood cell debris and platelets, and the scattered light intensity of small-volume debris is similar to that of platelets, making it difficult to distinguish between the two.

[0025] Considering the above issues, when a sample contains a large amount of leukocyte debris, it is clear that the volumetric similarity between leukocyte debris and platelets makes it difficult to directly distinguish between the two using the Coulter principle based on electrical resistance changes or optical methods based on forward and side scattered light intensity. In actual measurements, this volumetric approximation often results in erroneous results that are difficult to eliminate using simple calibration or filtering algorithms. In particular, samples with a high number of leukocyte debris, such as blood cells from patients with inflammation or disease, contain a high content of leukocytes and leukocyte debris, so platelet counting using the existing electrical resistance method is subject to significant errors due to leukocyte debris.

[0026] Therefore, in order to address the above-mentioned problems, an embodiment of the present invention provides a quality control method for blood cell testing in medical testing. Specifically, referring to Figure 1, Figure 1 is a workflow diagram of the quality control method for blood cell testing in medical testing provided by the present invention.

[0027] First, perform S1: obtain the initial number of platelets in the sample to be measured using the electrical resistance method. In some embodiments of the present invention, the electrical resistance signal is obtained based on the resistance method, which is detected after the blood sample passes through the small hole.

[0028] First, an anticoagulated blood sample, such as a blood sample containing EDTA anticoagulant, is collected, and then the blood sample is diluted at a certain ratio (usually 1:1000 to 1:5000). A low-conductivity electrolyte, such as sodium chloride solution, is used to prevent cell clumping and overlapping, thereby reducing cell density and ensuring that cells pass through the micropores as single particles, improving detection accuracy.

[0029] Based on the Coulter principle, cells are passed one by one through a micropore (detection hole). The diameter of the micropore is typically in the range of 50 μm to 100 μm (a range optimized for platelet counting). Highly sensitive electrodes are placed on both sides of the micropore, and a stable DC electric field is applied. The cells act as insulators, and as they move through the micropore, they momentarily change the resistance within the micropore, causing a change in the electrical signal. High-frequency sampling captures the instantaneous signal of the resistance change.

[0030] Perform S2: Based on the scattered light method, obtain a two-dimensional scattering scatter diagram of the sample to be measured, where the horizontal axis of the two-dimensional scattering scatter diagram is the side scattered light intensity and the vertical axis is the forward scattered light intensity.

[0031] In some embodiments of the present invention, a light scattering system in a hematology analyzer is used to obtain a two-dimensional scattering scattergram of a blood sample. Specifically, a laser light source (typically a laser diode) in the light scatterometer emits a laser beam and irradiates the flowing blood sample. When the laser beam irradiates different cells in the blood sample (e.g., platelets, white blood cells, red blood cells, etc.), the cells scatter the light, generating scattered light signals at different angles. Forward-scattered light (FSC) refers to the light signal scattered by cells in the direction of laser beam propagation after the laser beam passes through the blood sample. The FSC signal mainly reflects the volume size of the cell particles. Large cells or particles scatter a strong forward-scattered light signal. Side-scattered light (SSC) refers to the scattering signal generated in a direction perpendicular to the laser beam propagation direction when the laser beam interacts with cells in the blood sample. The SSC signal reflects the internal complexity of the particles (e.g., cell structure, particle density, etc.). Cells with complex structures (e.g., white blood cells) scatter a strong side-scattered light signal.

[0032] Light scatterometers use photoelectron detectors (e.g., photodiodes) to collect scattered light signals, which are converted into electrical signals that are then converted into digital signals and compared to a reference standard to obtain a scattered light intensity value.

[0033] In the two-dimensional scattering scatter plot obtained in S2, the horizontal axis represents side-scattered light intensity (SSC), which is related to the complexity of the cell particle and indicates the internal structure and complexity of the cell particle. Among them, complex cells such as white blood cells exhibit greater scattered light intensity in this dimension. The vertical axis represents forward-scattered light intensity (FSC) and particle volume size. Particles with larger volumes (e.g., large white blood cells) generate stronger forward-scattered signals; i.e., large-volume blood cells are distributed in the larger FSC region in the two-dimensional scattering scatter plot. On the other hand, small particles (e.g., platelets) generate weaker signals; i.e., small-volume blood cells are distributed in the smaller FSC region in the two-dimensional scattering scatter plot.

[0034] Based on these signals, a two-dimensional scatter plot (FSC-SSC plot) is generated. Referring to Figure 4, this scatter plot shows how various cells (platelets, white blood cells, red blood cells, etc.) in a blood sample are scattered on the two-dimensional FSC and SSC.

[0035] Step S3 is performed: In the two-dimensional scattering scatter diagram, the total light intensity set is determined based on the sum of the side scattered light intensity and the sum of the forward scattered light intensity. Here, in the two-dimensional scattering scatter diagram, the total light intensity set within the set region is TIFF0007743138000014.tif10168, where: TIFF0007743138000015.tif9168 is the sum of the side scattered light intensity of white blood cell debris and platelets within the set area, TIFF0007743138000016.tif8168 is the sum of the forward scattered light intensity of white blood cell debris and platelets within the set area.

[0036] It should be understood that decomposing the total intensity value into two parts, lateral total light intensity and forward total light intensity, allows the independent characteristics of each direction to be preserved rather than directly integrating them into a single value, which helps to analyze the lateral and forward contributions separately. Furthermore, because laboratory analysis of two-dimensional scatter plots is essentially a computational logic based on two-dimensional coordinates, expressing the total light intensity in a two-dimensional format naturally fits this analysis mode better, and the two-dimensional total intensity format can more precisely describe the overlap characteristics of platelets and white blood cell debris.

[0037] Perform S4: According to a preset correction rule, if the total light intensity set within a set region in the two-dimensional scattering scatter plot is located within a preset first light intensity interval, a linear correction is performed on the initial number of platelets.

[0038] If the total light intensity set within the set area in the two-dimensional scattering scatter plot does not belong to the first light intensity interval, perform S5: if the total light intensity set within the set area is located in the predetermined second light intensity interval, perform nonlinear correction on the initial number of platelets.

[0039] In the above two steps, referring to the flow diagram of obtaining preset correction rules shown in FIG. 2, the specific operation flow for establishing the preset rules is as follows:

[0040] First, perform S41: obtain a blood sample set, which includes a patient sample set and a healthy sample set. The experimental data required for constructing the correction rule include a healthy sample set and a patient sample set. Both have different characteristics and are used to establish linear and nonlinear correction rules, respectively.

[0041] The healthy sample set is composed of blood samples from healthy individuals, characterized by a relatively stable ratio relationship between platelets and white blood cell debris, which typically exhibits linear distribution characteristics within a set range. Healthy individuals without infection, inflammation, or other diseases that affect blood characteristics can be selected. The patient sample set is collected from individuals with specific diseases (such as inflammation, infection, or blood disorders) and is characterized by the possibility of a significantly increased number of white blood cell debris compared to the healthy sample set. Furthermore, platelets and white blood cell debris may exhibit nonlinear distribution characteristics within a set range. It is important to note that the dilution concentrations of each blood sample in the blood sample set are identical.

[0042] Perform S42: Determine a first light intensity interval based on the total light intensity set of a set area in the two-dimensional scattering scatter plot of the healthy sample set, and determine a second light intensity interval based on the total light intensity set of a set area in the two-dimensional scattering scatter plot of the patient sample set.

[0043] In some embodiments of the present invention, and with reference to the flow diagram shown in FIG. 3, defining the platelet and white blood cell debris set area includes the following steps:

[0044] First, perform S421: Obtain a two-dimensional scatter plot of each blood sample in the healthy sample set.

[0045] Perform step S422: For each two-dimensional scattering scattergram, determine an initial region based on the intersection of the distribution regions of platelets and white blood cell debris. For each two-dimensional scattering scattergram, determine the overlapping distribution range of platelets and white blood cell debris based on existing experimental experience or characteristic curves in the literature, and define the initial region.

[0046] Perform S423: Initial area of ​​each 2D scatterplot Total light intensity set Calculate and statistic TIFF0007743138000017.tif10168. That is, for each two-dimensional scattering scatter plot, statisticize all the scattered point data within the initial region and calculate the total light intensity set on the horizontal axis (side light intensity) and vertical axis (forward light intensity).

[0047] Perform step S424: Based on the stability analysis method, the initial region with the most consistent total light intensity set is selected as the set region. By analyzing the stability of the light intensity distribution of the scatter points within the initial region in the two-dimensional scattering scatter plot of the healthy sample set, we can determine the region with the highest statistical consistency as the final set region, i.e., region A in Figure 5, and use it for subsequent set region correction for platelets and white blood cell debris. Here, the most consistent total light intensity set refers to the initial region with the most concentrated and stable light intensity distribution. Selecting such a region as the set region through stability analysis (e.g., calculating the mean and standard deviation) can significantly improve the reliability and accuracy of the correction method.

[0048] Returning to step S42, in the healthy sample set, the platelet count and white blood cell debris count show linear distribution characteristics within the physiological range, and the light intensity distribution of these two components is relatively stable within the set region in the two-dimensional scatter plot. Because healthy samples are not affected by pathological factors, the total light intensity set in the set region has smaller variability and higher consistency. Due to this consistency, the light intensity range calculated for the healthy sample set (i.e., the first light intensity interval) can be used as the basis for subsequent analysis and correction.

[0049] It should be noted that the healthy sample set usually covers normal individuals of different genders, ages, and constitutions, and the statistical results are more universal and representative. The determined first light intensity interval can adapt to the normal blood characteristics of the majority of individuals and provide a standardized reference for subsequent analysis.

[0050] In patient sample sets, the number of white blood cell debris may increase significantly due to pathological conditions (e.g., inflammation, immune response, etc.), so platelets and white blood cell debris exhibit nonlinear characteristics in their distribution within the set area. By calculating the total light intensity set of the set area using the scatter plot of the patient sample set, the light intensity distribution in pathological conditions can be actually reflected, providing a basis for correcting abnormal samples.

[0051] The advantage of selecting a healthy sample set to determine the first light intensity interval is that the data is stable, providing a universal standard and helping to define normal distribution characteristics. The advantage of selecting a patient sample set to determine the second light intensity interval is that it dynamically reflects pathological characteristics, adapts to complex distributions, and provides an optimization direction for the correction model. By combining the two, the standard and dynamic characteristics complement each other, allowing for the construction of more accurate and adaptable correction rules.

[0052] More specifically, in the healthy sample set, the total light intensity set of a set region in the two-dimensional scattering scatter plot of each blood sample is calculated, and from that, Maximum total light intensity set TIFF0007743138000018.tif9168 and Minimum total light intensity set Extract TIFF0007743138000019.tif9168, and the first light intensity interval is:

number

[0053] Set margin For TIFF0007743138000024.tif8168, some embodiments of the present invention introduce the calculation of the mean value and standard deviation, i.e., statistically calculate the mean value and standard deviation of each dimension value in the total light intensity set, thereby defining a light intensity interval based on data distribution, which can better reflect the distribution characteristics of the data. When the data has large fluctuations or uneven distributions, the statistically based interval can more effectively capture the actual fluctuation range of the data and improve the accuracy of the correction rule.

[0054] Similarly, for the patient sample set, we calculate the total light intensity set for a set region in the two-dimensional scattering scatter plot for each blood sample, and then select Maximum total light intensity set TIFF0007743138000025.tif11168 and Minimum total light intensity set Extract TIFF0007743138000026.tif10168, and the second light intensity interval is:

number

[0055] Specifically, the light intensity intervals are used to define two different correction strategies. If the total light intensity set in the set area is within a first light intensity interval, it can be determined that the impact of white blood cell debris in the sample on platelets is small. In this case, correcting the platelet count obtained from this blood sample is relatively simple. Due to the small interference of white blood cell debris, in some possible implementations of the present invention, when the impact of white blood cell debris on platelets is small, a linear model or linear function is used to correct the platelet count obtained. This is because the platelet count and the interference of white blood cell debris exhibit a relatively simple linear relationship, and linear correction can quickly and effectively adjust the platelet count while maintaining the simplicity and efficiency of the correction process. Furthermore, the linear correction method is simple to calculate and has a fast processing speed, is suitable for large-scale data processing, and does not cause excessive adjustments to the platelet count. For situations where detection results are demanding, linear correction can provide a good balance between accuracy and efficiency.

[0056] When the total light intensity set of the set area is in the second light intensity interval, the influence of white blood cell debris shows a complex nonlinear relationship, so a more complex correction function needs to be adopted. In some possible embodiments of the present invention, a nonlinear model or function is used to correct the obtained platelet count. This correction method can more accurately adjust the platelet count and avoid the shortcomings of linear correction.

[0057] The above method effectively defines the light intensity ranges of the first light intensity section and the second light intensity section, and different correction strategies can be applied to the areas of different influence, thereby ensuring the accuracy and reliability of the platelet counting.

[0058] Regarding how to perform linear or nonlinear correction for the platelet count, some possible examples of the present invention perform S43: perform fluorescent staining for white blood cell debris in the blood sample set, determine the number of white blood cell debris in the blood sample set, construct a linear correction function based on the number of white blood cell debris in the healthy sample set, and construct a nonlinear correction function based on the number of white blood cell debris in the patient sample set.

[0059] In some embodiments of the present invention, fluorescent staining of leukocyte debris specifically refers to staining of nucleic acids in leukocyte debris. When performing specific nucleic acid staining of leukocytes, the selection of a fluorescent dye is key. It is necessary to select a dye that is highly specific to leukocyte debris and does not interfere with platelets. In some possible embodiments of the present invention, leukocyte debris is labeled by conjugating a CD45 antibody with a fluorescent dye. CD45 is a marker molecule for leukocytes, and almost all leukocytes express CD45. Using a fluorescent dye conjugated to a CD45 antibody (e.g., FITC, PE, or APC dye) can specifically label leukocytes and their debris. In other possible embodiments of the present invention, Hoechst 33342 or DAPI dye is used. These dyes can bind to nuclear DNA and are typically used to label all leukocyte debris or nucleic acids, helping to identify leukocyte debris containing nucleic acid material.

[0060] For diluted blood samples, a specific fluorescent dye (e.g., FITC dye labeled with CD45 antibody) is added to the blood sample, and the fluorescent signal is acquired using an apparatus such as a flow cytometer or a fluorescent microscope.

[0061] It can be seen that white blood cell debris emits fluorescence at a specific wavelength after staining, while platelets do not have a fluorescent signal. After staining white blood cell debris, the number of white blood cell debris can be calculated using a flow cytometer, imaging flow cytometer, fully automated hematology analyzer, etc.

[0062] Based on the statistical data of white blood cell debris fluorescence staining, linear and nonlinear correction functions are constructed respectively. Specifically, the blood sample set is divided into a healthy sample set and a patient sample set. The number of white blood cell debris obtained by fluorescent staining is respectively TIFF0007743138000031.tif10168The initial number of platelets TIFF0007743138000032.tif11168. In a two-dimensional scatter plot, the set areas for white blood cell debris and platelets cannot directly distinguish between their numbers, so a correction function must be used to estimate the numbers of white blood cell debris and platelets within the set areas.

[0063] A linear correction function is constructed based on the healthy sample set, specifically, each blood sample in the healthy sample set is subjected to fluorescent staining, Number of white blood cell debris TIFF0007743138000033.tif11168 and Initial platelet count TIFF0007743138000034.tif9168. In the blood samples from the healthy sample set, the white blood cell debris and platelets show a linear proportional relationship within the set range:

number

[0064] Based on the total light intensity signal of a set region in the two-dimensional scattering scatter plot, distribution statistics are used to determine the ratio of the number of white blood cell debris to platelets, and then k1 is obtained by fitting the data of the healthy sample set.

[0065] Therefore, the linear correlation function between platelet count and white blood cell debris count is:

number

[0066] It should be noted that the initial number of platelets is the initial number of platelets in the sample to be measured obtained by the electrical resistance method.

[0067] A nonlinear correction function is constructed based on the patient sample set, specifically, by performing fluorescent staining on each blood sample in the patient sample set, Number of white blood cell debris TIFF0007743138000039.tif12168 and Initial platelet count TIFF0007743138000040.tif11168 respectively. In some laboratory examples, in patient sample sets, white blood cell debris and platelets show an exponential relationship within a set range.

[0068] Through experimental data analysis, the numbers of white blood cell debris and platelets within the set area in the patient sample satisfy the following relationship:

number

[0069] The correction function for the exponential function is:

number

[0070] Furthermore, some embodiments of the present invention also consider that if the total light intensity set of a set area in the measurement sample exceeds the value of the second light intensity interval, the white blood cell debris in the measurement sample will have too great an effect on the platelet count, making conventional platelet counting methods unable to provide accurate results. This situation may indicate that the number of white blood cell debris in the blood sample is large, or that the characteristics of the debris in the scattering spectrum overlap highly with those of platelets, making them ineffectively distinguishable by conventional correction methods. Therefore, re-detection measures should be taken for such samples to ensure the accuracy of platelet counting.

[0071] The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention. The patent protection scope of the present invention is governed by the claims, and equivalent structural changes made based on the content of the specification and drawings of the present invention should also be included in the protection scope of the present invention.

Claims

1. A quality control method for blood cell testing in medical testing, comprising: Obtaining an initial number of platelets in the sample to be measured using an electrical resistance method; Obtaining a two-dimensional scattering scatter diagram of the measurement target sample based on a scattered light method, wherein the horizontal axis of the two-dimensional scattering scatter diagram represents side scattered light intensity and the vertical axis represents forward scattered light intensity; determining a total light intensity set based on a sum of side scattered light intensities and a sum of forward scattered light intensities in the two-dimensional scattering scatter plot; According to a preset correction rule, when a total light intensity set within a set region in the two-dimensional scattering scatter diagram is located within a preset first light intensity interval, linearly correcting the initial number of platelets; performing nonlinear correction on the initial number of platelets when the total light intensity set within the set region is located in a predetermined second light intensity range; The correction rule construction step includes: obtaining blood sample sets, including a patient sample set and a healthy sample set; determining the first light intensity interval based on a total light intensity set of the set area in the two-dimensional scattering scatter plot of the healthy sample set; and determining a second light intensity interval based on a total light intensity set of the set area in the two-dimensional scattering scatter plot of the patient sample set; A quality control method for blood cell testing in medical testing, comprising: performing fluorescent staining on white blood cell debris in the blood sample set, determining the number of white blood cell debris in the blood sample set, constructing a linear correction function based on the number of white blood cell debris in the healthy sample set, and constructing a non-linear correction function based on the number of white blood cell debris in the patient sample set.

2. 2. The quality control method for blood cell testing in medical testing according to claim 1, wherein the fluorescent staining of the white blood cell debris specifically refers to staining nucleic acids in the white blood cell debris.

3. 2. The quality control method for blood cell testing in medical testing according to claim 1, wherein the dilution concentration of each blood sample in the blood sample set is the same, and the two-dimensional scattering scatter diagram is obtained for each blood sample by a light scattering method, and then the white blood cell debris is stained so that the white blood cell debris has a fluorescent signal that can be distinguished from platelets, and the number of white blood cell debris in each blood sample is calculated.

4. In the two-dimensional scattering scatter plot, the total light intensity set within the set region is where: is the sum of the side scattered light intensity of white blood cell debris and platelets within the set area, 2. The quality control method for blood cell testing in medical testing according to claim 1, wherein ≡(f) is the sum of the forward scattered light intensity of white blood cell debris and platelets within the set region.

5. The step of determining the set region includes: obtaining a two-dimensional scatter plot of each blood sample in the healthy sample set; determining an initial region based on the intersection of platelet and leukocyte debris distribution regions in each two-dimensional scattering scattergram; The total light intensity set of the initial region in each two-dimensional scattering scatter plot Calculating and statistically 5. The quality control method for blood cell testing in medical testing according to claim 4, further comprising: determining the initial region in which the total light intensity set has the highest consistency based on a stability analysis method as the set region.

6. In the healthy sample set, a total light intensity set of the set area in the two-dimensional scattering scatter plot of each of the blood samples is calculated, and the maximum total light intensity set is calculated from the set area. 【number】 and the minimum total light intensity set 【number】 The first light intensity interval is: [Equation 1] where: 【number】 6. The quality control method for blood cell testing in medical testing according to claim 5, wherein the difference is a set margin.

7. In the patient sample set, a total light intensity set of the set area in the two-dimensional scattering scatter plot of each of the blood samples is calculated, and the maximum total light intensity set is calculated from the set area. 【number】 and the minimum total light intensity set 【number】 and the second light intensity interval is: [Equation 2] where: 【number】 6. The quality control method for blood cell testing in medical testing according to claim 5, wherein the difference is a set margin.

Images