CN-122016612-A - Device and method for classifying and counting white blood cells of patient with acute respiratory tract infection

CN122016612ACN 122016612 ACN122016612 ACN 122016612ACN-122016612-A

Abstract

The invention relates to the technical field of white blood cell counting, in particular to a white blood cell classifying and counting device and method for patients suffering from acute respiratory tract infection. The method comprises the steps of screening white blood cells according to the appearance of cell membrane fluorescence and the appearance component types to obtain white blood cells in a single cell suspension sample, dividing a plurality of cycle groups through the degree of similarity of fluorescence intensity of nuclear DNA among the white blood cells, further analyzing the scattered light characteristics of cells in a single cell group in the same cycle state to divide cell subsets, carrying out corresponding one-to-one combination on the cell subsets of the two cycle groups according to the difference between different components to obtain white blood cell classification of the single cell suspension sample, and realizing the counting of final classification according to the cell number distribution of classification results of a plurality of single cell suspension samples. The invention combines the classification and combination of the white blood cells in different growth states, improves the accuracy of the classification of the white blood cells, and thus obtains a more accurate white blood cell classification counting result for patients suffering from acute respiratory tract infection.

Inventors

LI XUEQING
TIAN ZHANHONG
LI XINRU
SONG KAI
WANG XUEQING
WANG KEXIN
QIU HUIYING

Assignees

北京怀柔医院

Dates

Publication Date: 20260512
Application Date: 20260302

Claims (10)

1. A method of differential white blood cell count for patients with acute respiratory tract infections, the method comprising: Collecting more than two single cell suspension samples from elbow vein blood of a patient suffering from acute respiratory tract infection, obtaining fluorescent intensity of different single cell components by fluorescent staining treatment, wherein the components comprise cell membranes, nuclear DNA, endoplasmic reticulum, mitochondria and Golgi apparatus; Screening white blood cells in each single cell suspension sample from all cells according to the deviation condition of the fluorescence intensity of the single cells on the cell membrane and the species number of the components showing the fluorescence intensity; Dividing the white blood cells in each single cell suspension sample based on the fluorescence intensity similarity degree of the nuclear DNA among the white blood cells to obtain periodic groups, clustering the white blood cells according to the distribution aggregation condition of all the white blood cells between forward scattering light intensity and side scattering light intensity in each periodic group to obtain a preset category number of cell subsets; the count for each cell type was determined by the distribution of cell numbers for the cell types across all single cell suspension samples.
2. The method for differential leukocyte differential count in patients with acute respiratory tract infection according to claim 1, wherein said method for obtaining leukocytes in each single cell suspension sample comprises: For any one cell, obtaining the possibility of cell membrane damage of the cell according to the deviation degree of the corresponding fluorescence intensity of the cell membrane of the cell in the single cell suspension sample; performing negative correlation mapping on the number of species of the component of which the fluorescence intensity appears in the cell, and obtaining the structural loss probability of the cell; combining the cell membrane damage probability and the structure loss probability of the cell to obtain the activity reflecting degree of the cell, and taking the cell with the activity reflecting degree larger than a preset activity threshold as a leukocyte.
3. The method for classifying and counting leukocytes for patients with acute respiratory infection according to claim 2, wherein the method for obtaining the probability of rupture of the envelope comprises: Taking the average value of the corresponding fluorescence intensities of the cell membranes of all cells in the single cell suspension sample where the cells are positioned as the integral cell membrane intensity of the sample of the cells; The difference between the fluorescence intensity of the cell at the cell membrane and the overall cell membrane intensity of the sample is used as the cell membrane breakage probability of the cell.
4. The method for classifying and counting white blood cells in patients with acute respiratory infection according to claim 1, wherein the method for acquiring the periodic group comprises: Carrying out normalization treatment on the fluorescence intensity corresponding to each leukocyte nuclear DNA to obtain the growth trend degree of each leukocyte, and carrying out negative correlation mapping on the difference of the growth trend degrees between the two leukocytes to obtain the growth approximation degree between the two leukocytes; And taking two white blood cells with the growth approximation degree larger than a preset period threshold value as a period group, wherein each white blood cell in the period group meets the condition that the growth approximation degree between at least one group of white blood cells is larger than the preset period threshold value, and taking the period group corresponding to the period group with the maximum growth approximation degree as the period group of the corresponding white blood cells when the growth approximation degree between the white blood cells and more than two period groups is larger than the preset period threshold value.
5. The method for differential leukocyte differential count in patients with acute respiratory tract infection according to claim 1, wherein said method for obtaining cell subsets comprises: the forward scattered light intensity is taken as a horizontal axis, the side scattered light intensity is taken as a vertical axis, and a two-dimensional space is established; And for any periodic group, mapping all the white blood cells in the periodic group into a two-dimensional space to obtain a scatter diagram of the periodic group, clustering the white blood cells in the scatter diagram of the periodic group into a plurality of clusters with preset category numbers through a clustering algorithm, wherein each cluster is a cell subgroup.
6. The method for classifying and counting white blood cells in patients with acute respiratory infection according to claim 1, wherein the method for obtaining cell types comprises: For any one period group, sequentially taking other period groups as groups to be combined; For any cell subgroup in the periodic group, sequentially calculating the approximate conditions of fluorescence intensity between endoplasmic reticulum, mitochondria and golgi apparatus between the cell subgroup and each cell subgroup in the group to be combined, and obtaining the similar trend degree of the cell subgroup and each cell subgroup in the group to be combined; Combining the cell subset with the cell subset corresponding to the greatest similar tendency in the to-be-combined group to form a new cell subset, and taking each cell subset obtained by combining the periodic group with all other periodic groups as each cell category.
7. The method for classifying and counting white blood cells of a patient suffering from acute respiratory infection according to claim 6, wherein the method for acquiring the similar tendencies comprises the steps of: taking the average value of fluorescence intensity of all white blood cells in the cell subgroup at each common component as the component intensity of the cell subgroup at each common component, wherein the common components comprise endoplasmic reticulum, mitochondria and golgi apparatus; and calculating the difference between the component intensities of each common component of the cell sub-population and the analysis sub-population, calculating the sum of all the differences, and carrying out negative correlation mapping to obtain the similar tendencies of the cell sub-population and the analysis sub-population.
8. The method for classifying and counting white blood cells of a patient suffering from acute respiratory infection according to claim 1, wherein the method for determining the count result of each cell type comprises: Sorting the cell types in each single cell suspension sample according to the forward scattering light intensity average value of all the white blood cells in the cell types from small to large to obtain a type sequence of each single cell suspension sample, wherein each serial number under all the type sequences is used as a final cell type; for any sequence number, according to the deviation of the cell number of the cell type of each single cell suspension sample under the sequence number and the cell number of the whole cell type under the same sequence number, obtaining the classification confidence of each single cell suspension sample under the sequence number; Taking the single cell suspension sample with the maximum classification confidence under the serial number as a trusted sample, taking the cell number of the cell class of the trusted sample under the serial number as the final number of the cell class corresponding to the serial number; The ratio of the final number of each final cell type to the total number of white blood cells was used as a count result for each final cell type.
9. The method for classifying and counting white blood cells of a patient suffering from acute respiratory infection according to claim 8, wherein the method for obtaining the classification confidence comprises the following steps: Taking the average value of the cell numbers corresponding to all the class sequences under the serial number as the number uniformity of the serial number; And carrying out negative correlation mapping and normalization treatment on the difference of the cell number and the number uniformity of the cell class of each single cell suspension sample under the serial number to obtain the classification confidence of each single cell suspension sample under the serial number.
10. A device for classifying and counting white blood cells of a patient suffering from acute respiratory tract infection, comprising a memory, a processor and a computer program stored in the memory and executable on the processor, wherein the processor performs the steps of a method for classifying and counting white blood cells of a patient suffering from acute respiratory tract infection according to any one of claims 1 to 9 when the processor executes the computer program.

Description

Device and method for classifying and counting white blood cells of patient with acute respiratory tract infection Technical Field The invention relates to the technical field of white blood cell counting, in particular to a white blood cell classifying and counting device and method for patients suffering from acute respiratory tract infection. Background Leukocyte differential count is a routine test of patients with acute respiratory infections in blood tests that generally help doctors to see if an infection or inflammation is present in the patient's body. This test evaluates the immune system status of the patient by analyzing the number of different types of leukocytes in the blood, including neutrophils, lymphocytes, monocytes, etc. The technology detects with single cell precision, and can acquire the volume, the internal particle complexity and other information of each single cell in the blood sample through the built-in laser module and the fluorescent signal module, so as to cluster multidimensional parameter information to realize the differential counting of the white blood cells. Since each leukocyte in the actual blood scene may be in different cell cycles, even if the same type of leukocyte is in different growth states, there is still a certain difference between the body type and the internal structural components, and when the cell classification is directly obtained according to the information such as cell volume, particle complexity, etc., and the influence of the cell in different growth states is not considered, the accuracy of the classification result of the cell classification may be insufficient, and the accuracy of the classification and counting of the patient's leukocyte is further insufficient. Disclosure of Invention In order to solve the technical problems that the accuracy of the classification result of the cell type is insufficient and the accuracy of the classification and counting of the white blood cells of a patient is insufficient when the influence of the different growth states of the cells is not considered in the prior art, the invention aims to provide a white blood cell classification and counting device and method for patients with acute respiratory tract infection, and the adopted technical scheme is as follows: The invention provides a leukocyte classification counting method for patients with acute respiratory tract infection, which comprises the following steps: Collecting more than two single cell suspension samples from elbow vein blood of a patient suffering from acute respiratory tract infection, obtaining fluorescent intensity of different single cell components by fluorescent staining treatment, wherein the components comprise cell membranes, nuclear DNA, endoplasmic reticulum, mitochondria and Golgi apparatus; Screening white blood cells in each single cell suspension sample from all cells according to the deviation condition of the fluorescence intensity of the single cells on the cell membrane and the species number of the components showing the fluorescence intensity; Dividing the white blood cells in each single cell suspension sample based on the fluorescence intensity similarity degree of the nuclear DNA among the white blood cells to obtain periodic groups, clustering the white blood cells according to the distribution aggregation condition of all the white blood cells between forward scattering light intensity and side scattering light intensity in each periodic group to obtain a preset category number of cell subsets; the count for each cell type was determined by the distribution of cell numbers for the cell types across all single cell suspension samples. Further, the method for obtaining the white blood cells in each single cell suspension sample comprises the following steps: For any one cell, obtaining the possibility of cell membrane damage of the cell according to the deviation degree of the corresponding fluorescence intensity of the cell membrane of the cell in the single cell suspension sample; performing negative correlation mapping on the number of species of the component of which the fluorescence intensity appears in the cell, and obtaining the structural loss probability of the cell; combining the cell membrane damage probability and the structure loss probability of the cell to obtain the activity reflecting degree of the cell, and taking the cell with the activity reflecting degree larger than a preset activity threshold as a leukocyte. Further, the method for obtaining the probability of cell membrane breakage comprises the following steps: Taking the average value of the corresponding fluorescence intensities of the cell membranes of all cells in the single cell suspension sample where the cells are positioned as the integral cell membrane intensity of the sample of the cells; The difference between the fluorescence intensity of the cell at the cell membrane and the overall cell membrane intensity of the sample is used