Flow cytometry analysis method, device, computer device and storage medium

Abstract

The application provides a flow lymphocyte analysis method, including: obtaining to-be-analyzed data; performing density distribution analysis on the to-be-analyzed data to obtain a density distribution result of the to-be-analyzed data; determining, based on a FlowDentity function, whether a first threshold value of a flow lymphocyte group and a non-flow lymphocyte group exists in a preset density distribution range; if the first threshold value does not exist in the preset density distribution range, determining, based on the FlowDentity function, an initial region containing the flow lymphocyte group from the density distribution result; adjusting a region boundary of the initial region by adjusting a function parameter of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and taking the adjusted region boundary as the first threshold value; determining the flow lymphocyte group in the to-be-analyzed data based on the first threshold value; determining a ratio of cells expressing a specific fluorescence signal in the flow lymphocyte group to cells in the flow lymphocyte group; and outputting a flow lymphocyte analysis result according to the ratio.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of flow cytometry, and in particular to a flow lymphocyte analysis method and device, a computer device and a storage medium. BACKGROUND

[0002] Lymphocytes are the main cell population that constitutes the immune system of the body and performs immune functions, and participate in cellular immunity and humoral immunity of the body. Lymphocyte subgroup detection is an important examination for evaluating the immune state of the body in clinical practice, which often reflects the functional state of various immune cells in the body in terms of quantity and proportion, and is widely used in the diagnosis, treatment and prognosis of tumors, AIDS and autoimmune diseases. Flow cytometry (FCM) is one of the conventional means for lymphocyte immunophenotyping analysis in clinical practice. FCM is a technology that analyzes the signal intensity of scattered light and fluorescence generated by single cells (biological microparticles) that are fluorescently labeled and flow at high speed under high-energy laser irradiation, thereby classifying and counting the cells, and realizing the analysis or sorting of target cells. At present, although various supervised and unsupervised FCM data analysis algorithms have been developed, their analysis efficiency and accuracy are not ideal under various conditions such as large number of sample cells and non-fixed number of cell categories. SUMMARY

[0003] The present application provides a flow lymphocyte analysis method, device, computer device and storage medium to solve the technical problem that the analysis efficiency and accuracy are not ideal in the existing flow lymphocyte analysis technology.

[0004] In a first aspect, a flow lymphocyte analysis method is provided, which comprises:

[0005] acquiring to-be-analyzed data;

[0006] performing density distribution analysis on the to-be-analyzed data to obtain a density distribution result of the to-be-analyzed data;

[0007] determining whether a first threshold value of a flow lymphocyte group and a non-flow lymphocyte group exists in a preset density distribution range based on a FlowDentity function; the first threshold value is a trough between two peaks in the density distribution result;

[0008] if the first threshold value does not exist in the preset density distribution range, determining an initial region containing the flow lymphocyte group from the density distribution result based on the FlowDentity function;

[0009] adjusting a region boundary of the initial region by adjusting a function parameter of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and taking the adjusted region boundary as the first threshold value;

[0010] determining the flow lymphocyte group in the to-be-analyzed data based on the first threshold value;

[0011] determining a ratio of cells expressing a specific fluorescence signal in the flow lymphocyte group to cells in the flow lymphocyte group;

[0012] outputting a flow lymphocyte analysis result according to the ratio.

[0013] With reference to the first aspect, in a possible implementation manner, the obtaining of the to-be-analyzed data comprises: performing data cleaning on original data obtained through clinical detection to obtain cleaned data, and taking the cleaned data as the to-be-analyzed data.

[0014] With reference to the first aspect, in a possible implementation manner, the determining of the flow lymphocyte group based on the first threshold value comprises: adopting the FlowDentity function to define a boundary of the flow lymphocyte group in the first threshold value range by using an elliptical gate, so as to determine the flow lymphocyte group.

[0015] With reference to the first aspect, in a possible implementation manner, the specific fluorescence signal comprises CD3, CD4, CD8, CD19, and CD16+56; and the determining of the ratio of cells expressing the specific fluorescence signal in the flow lymphocyte group to cells in the flow lymphocyte group comprises: determining a first ratio of CD3-positive expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; determining a second ratio of CD3-positive expression and CD4-positive expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; determining a third ratio of CD3-positive expression and CD4-negative expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; determining a fourth ratio of CD3-positive expression and CD8-positive expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; determining a fifth ratio of CD3-positive expression and CD8-negative expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; determining a sixth ratio of CD3-negative expression and CD19-positive expression cells in the flow lymphocyte group to cells in the flow lymphocyte group; and determining a seventh ratio of CD3-negative expression and CD16+56-positive expression cells in the flow lymphocyte group to cells in the flow lymphocyte group.

[0016] In a possible implementation manner of the first aspect, the outputting the flow cytometry analysis result according to the ratio includes: calculating a sum of the second ratio and the fourth ratio to obtain a first analysis result; calculating a sum of the first ratio, a sixth ratio and a seventh ratio to obtain a second analysis result; and outputting the first analysis result and the second analysis result if the first analysis result is not greater than the first ratio, the second analysis result is in a first preset ratio range, a difference between the second ratio and a fifth ratio is not more than a second preset ratio range, and a difference between the fourth ratio and a third ratio is not more than a third preset ratio range.

[0017] In a possible implementation manner of the first aspect, the determining the ratio of the cells with the specific fluorescence signal expression in the flow cytometry cell population to the cells in the flow cytometry cell population further includes: determining an eighth ratio of the cells with CD3 negative expression and CD4 positive expression in the flow cytometry cell population to the cells in the flow cytometry cell population.

[0018] In a possible implementation manner of the first aspect, the outputting the flow cytometry analysis result according to the ratio further includes: determining whether the eighth ratio is in a fourth preset ratio range; if the eighth ratio is in the fourth preset ratio range, performing the step of outputting the flow cytometry analysis result according to the ratio; and if the eighth ratio is not in the fourth preset ratio range, adjusting the first threshold according to the eighth ratio until the eighth ratio is in the fourth preset ratio range.

[0019] The second aspect provides a flow cytometry analysis device, and the device includes:

[0020] A data acquisition module is configured to acquire to-be-analyzed data.

[0021] A density distribution analysis module is configured to perform density distribution analysis on the to-be-analyzed data to obtain a density distribution result of the to-be-analyzed data.

[0022] A first determination module is configured to determine, based on a FlowDentity function, whether a first threshold of a flow cytometry cell population and a non-flow cytometry cell population exists in a preset density distribution range.

[0023] A second determination module is configured to determine, based on the FlowDentity function, an initial region containing the flow cytometry cell population from the density distribution result if the first threshold does not exist in the preset density distribution range.

[0024] a first threshold value determination module, configured to adjust a region boundary of the initial region by adjusting a function parameter of a FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and take the adjusted region boundary as the first threshold value;

[0025] a flow cytometry lymphocyte population determination module, configured to determine a flow cytometry lymphocyte population in the to-be-analyzed data based on the first threshold value;

[0026] a ratio determination module, configured to determine a ratio of cells expressing a specific fluorescence signal in the flow cytometry lymphocyte population to cells in the flow cytometry lymphocyte population;

[0027] a data output module, configured to output a flow cytometry lymphocyte analysis result according to the ratio.

[0028] With reference to the second aspect, in a possible design, the data acquisition module is specifically configured to perform data cleaning on original data obtained through clinical detection to obtain cleaned data, and take the cleaned data as the to-be-analyzed data.

[0029] With reference to the second aspect, in a possible design, the flow cytometry lymphocyte population determination module is specifically configured to adopt a FlowDentity function to define a boundary of a flow cytometry lymphocyte population in the first threshold value range by using an elliptical gate, to determine the flow cytometry lymphocyte population.

[0030] With reference to the second aspect, in a possible design, the specific fluorescence signal includes CD3, CD4, CD8, CD19, and CD16+56; and the ratio determination module is specifically configured to determine a first ratio of cells positively expressing CD3 to cells in the flow cytometry lymphocyte population, determine a second ratio of cells positively expressing CD3 and CD4 to cells in the flow cytometry lymphocyte population, determine a third ratio of cells positively expressing CD3 and negatively expressing CD4 to cells in the flow cytometry lymphocyte population, determine a fourth ratio of cells positively expressing CD3 and CD8 to cells in the flow cytometry lymphocyte population, determine a fifth ratio of cells positively expressing CD3 and negatively expressing CD8 to cells in the flow cytometry lymphocyte population, determine a sixth ratio of cells negatively expressing CD3 and positively expressing CD19 to cells in the flow cytometry lymphocyte population, and determine a seventh ratio of cells negatively expressing CD3 and positively expressing CD16+56 to cells in the flow cytometry lymphocyte population.

[0031] With reference to the second aspect, in a possible design, the data output module is specifically configured to: calculate a sum of the second ratio and the fourth ratio to obtain a first analysis result; calculate a sum of the first ratio, a sixth ratio and a seventh ratio to obtain a second analysis result; and output the first analysis result and the second analysis result if the first analysis result is not greater than the first ratio, the second analysis result is within a first preset ratio range, a difference between the second ratio and a fifth ratio is not greater than a second preset ratio range, and a difference between the fourth ratio and a third ratio is not greater than a third preset ratio range.

[0032] With reference to the second aspect, in a possible design, the ratio determination module is further specifically configured to: determine an eighth ratio of cells that express CD3 negatively and express CD4 positively in the flow cytometry lymphocyte population to cells in the flow cytometry lymphocyte population.

[0033] With reference to the second aspect, in a possible design, the data output module is further specifically configured to: determine whether the eighth ratio is within a fourth preset ratio range; and perform the step of outputting the flow cytometry lymphocyte analysis result according to the ratio if the eighth ratio is within the fourth preset ratio range; and adjust the first threshold according to the eighth ratio until the eighth ratio is within the fourth preset ratio range if the eighth ratio is not within the fourth preset ratio range.

[0034] According to a third aspect, a computer device is provided, including a memory and a processor, the memory storing a computer program, and the computer program, when executed by the processor, causes the processor to implement the flow cytometry lymphocyte analysis method.

[0035] According to a fourth aspect, a computer readable storage medium is provided, storing a computer program, and the computer program, when executed by a processor, causes the processor to implement the flow cytometry lymphocyte analysis method.

[0036] The present application can achieve the following beneficial effects: the present application determines the first threshold value based on the FlowDentity function, and determines an initial region containing the flow cytometry lymphocyte population based on the FlowDentity function when the density distribution result is not within the preset density distribution range; adjusts the region boundary of the initial region by adjusting the function parameter of the FlowDentity function, so that the adjusted region boundary matches the preset density distribution range, and the adjusted region boundary is used as the first threshold value; and finally determines the flow cytometry lymphocyte population and the analysis result based on the first threshold value; the analysis result is not affected by the large number of sample cells and the unfixed number of cell categories, and the efficiency and accuracy of the analysis can be effectively improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 A schematic diagram of a flow cytometry lymphocyte analysis method provided in an embodiment of the present application;

[0038] Figure 2 A schematic diagram of analysis results obtained based on a flow cytometry lymphocyte analysis method provided in an embodiment of the present application;

[0039] Figure 3 A schematic structural diagram of a flow cytometry lymphocyte analysis device provided in an embodiment of the present application;

[0040] Figure 4 A schematic diagram of the structure of a computer device provided in an embodiment of the present application. DETAILED DESCRIPTION

[0041] The technical solutions in the embodiments of the present application will be described below in conjunction with the drawings in the embodiments of the present application.

[0042] The technical solution of the present application is applicable to various scenarios of flow lymphocyte analysis. Specifically, the technical solution of the present application is applicable to various scenarios of flow lymphocyte analysis using the FlowDentity function to perform flow lymphocyte analysis. In practical applications, the FlowDentity function can be used to identify flow lymphocytes in the scenario to determine the analysis results.

[0043] To facilitate understanding of the technical solution of the present application, the flow lymphocytes involved in the present application are first introduced. Flow lymphocytes refer to lymphocytes sorted by flow cytometry. Flow cytometry (FCM) is a technology that uses high-energy laser irradiation to detect the signal intensity of scattered light and fluorescence generated by fluorescently labeled single cells (biological particles) in a high-speed flow state, thereby qualitatively classifying and quantitatively counting cells to achieve analysis or sorting of target cells. The main immune cells in the human body are lymphocytes, including T cells, B cells and NK cells. T lymphocytes reflect cellular immune function, and immunodeficiency and autoimmune diseases are closely related to the content of T lymphocytes; B lymphocytes exert humoral immune function by producing various autoantibodies, anti-bacterial antibodies and pathogenic antibodies; NK cells can directly kill target cells, and have anti-tumor, anti-infection, immune regulation and other functions, thereby exercising immune surveillance and playing an important role in tumor suppression and anti-tumor metastasis. Different types of lymphocytes have different cell surface antigens. FCM uses a variety of different fluorescently labeled monoclonal antibody reagents to label human peripheral blood cells and classify lymphocyte subsets.

[0044] In one embodiment,Figure 1 as shown, Figure 1 A flowchart of a flow cytometry analysis method provided by an embodiment of the present application is shown. The method comprises the following steps:

[0045] In step 101, data to be analyzed is obtained.

[0046] The data to be analyzed can be real-time data or historical data obtained from a database.

[0047] In one embodiment, the step of obtaining data to be analyzed comprises data cleaning of raw data obtained by clinical detection to obtain cleaned data, and the cleaned data is used as the data to be analyzed.

[0048] Specifically, data cleaning mainly includes reading data, converting parameter names, and standardizing data scales. First, the function read.FCS() in the R package flowCore is used to read flow raw data in FCS or LMD format. Second, for signal parameters FSC.A, SSC.A, CD45, CD3, CD4, CD8, CD19, and CD16+56 used for lymphocyte subtype analysis, different laboratories may have different naming methods. To facilitate subsequent code construction, the present application unifies different parameter names into fixed names. The implementation method is to pre-establish a data table including fixed names corresponding to each parameter name and description information corresponding to each parameter name. When cleaning the raw data obtained by clinical detection, the parameter names are matched with the fixed names according to the description information. Finally, since flow raw data has multiple formats such as FCS2.0, FCS3.0, and FCS3.1, and the training data used for algorithm construction in the present application is mainly in FCS2.0 format, it is necessary to convert non-FCS2.0 data to FCS2.0 format in the algorithm. The conversion method is to construct a conversion function according to the correspondence between non-FCS2.0 format data and FCS2.0 format data.

[0049] In step 102, density distribution analysis is performed on the data to be analyzed to obtain a density distribution result of the data to be analyzed.

[0050] The density distribution algorithm is used to count the fluorescence signals of cell populations and predict the probability density distribution of each cell population. Specifically, the density distribution algorithm can be a Gaussian kernel density algorithm.

[0051] In step 103, it is determined whether the density distribution result has a first threshold value of flow cytometry cell populations and non-flow cytometry cell populations within a preset density distribution range based on a FlowDentity function. The first threshold value is a trough between two peaks in the density distribution result.

[0052] The preset density distribution range is a normal density distribution range of lymphocytes obtained based on the density distribution of lymphocytes obtained from a large number of samples. This range can be expressed as a predetermined percentage range.

[0053] A gatedFD_lym() function for automatically identifying lymphocyte populations was constructed based on multiple functions of the existing FlowDentity algorithm (R package). Specifically, the deGate() function in flowDensity was used to find all inflection points in the lymphocyte density distribution based on the density distribution. It was then determined whether an inflection point existed within a preset density distribution range. If an inflection point existed, the inflection point within the predetermined range could be directly used as the first threshold between the lymphocyte population and the non-lymphocyte population.

[0054] In one embodiment, taking the SSC.A signal parameter as an example: based on the distribution of lymphocytes on SSC.A obtained from a large number of samples, the normal distribution range of lymphocytes on SSC.A data can be obtained and used as the preset density distribution range; using the deGate() function in flowDensity, all inflection points of the SSC.A data density distribution are found based on the density distribution, and it is determined whether there is an inflection point within the preset density distribution range. If there is an inflection point, then the inflection point within the predetermined range can be directly used as the first threshold between the lymphocyte population and the non-lymphocyte population.

[0055] Specifically, when finding the SSC.A threshold, only lymphocytes and monocytes are present within the predetermined range. Typically, there is only one inflection point within the predetermined range, which marks the boundary between lymphocytes and non-lymphocytes. If there are multiple inflection points within the predetermined range, it is generally because monocytes are distributed in more than one cluster, forming inflection points within the monocyte population. Since lymphocyte SSC.A values ​​are smaller than those of monocytes, the smallest inflection point is used.

[0056] In one embodiment, taking the CD45 signal parameter as an example: based on the distribution of lymphocytes on CD45 obtained from a large number of samples, the normal distribution range of lymphocytes on CD45 data can be obtained and used as the preset density distribution range; using the deGate() function in flowDensity, all inflection points of the CD45 data density distribution are found based on the density distribution, and it is determined whether there is an inflection point within the preset density distribution range. If there is an inflection point, then the inflection point within the predetermined range can be directly used as the first threshold between the lymphocyte population and the non-lymphocyte population.

[0057] Specifically, using the results of SSC.A axis division, take the cells less than the SSC.A threshold, this part generally only contains fragmented cells and lymphocytes, occasionally contains a small amount of mononuclear cells, the composition of the cells is not as complex as the total amount of cells, so only using this part of the cells can be more simple to find the threshold of CD45, the division of CD45 axis is also based on density. If the density distribution is bimodal, there is only one inflection point, at this time the fragmented cells and lymphocytes are clustered respectively, so the position of the inflection point can be used for division to distinguish the two cell populations; if the density distribution is unimodal, at this time the fragmented cells are not clustered, and there is only a cluster of lymphocytes. At this time, the front end of the density wave peak is used for division; if the density distribution exceeds two peaks, since the lymphocytes are located at the rightmost peak of the density distribution, the last density inflection point is used as the boundary position.

[0058] Step 104, if the first threshold does not exist in the preset density distribution range, determining an initial region containing a flow lymphocyte population from the density distribution result based on a FlowDentity function;

[0059] If the preset density distribution range does not contain an inflection point, that is, there is no first threshold, the flowDensity() function can be used to first circumscribe a region containing a lymphocyte population from the density distribution result, and the region is compared with the preset density distribution range, so as to determine the first threshold, that is, to determine the boundary between the cell populations.

[0060] Step 105, adjusting the region boundary of the initial region by adjusting the function parameters of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and the adjusted region boundary is taken as the first threshold;

[0061] If the preset density distribution range does not contain an inflection point, that is, there is no first threshold, the flowDensity() function can be used to first circumscribe a region containing a lymphocyte population from the density distribution result, and the region is compared with the preset density distribution range, so as to determine the first threshold, that is, to determine the boundary between the cell populations.

[0062] Step 106, determining the flow lymphocyte population in the data to be analyzed based on the first threshold;

[0063] Wherein, after determining the first threshold, the flowDensity() function is used to define the boundary of the lymphocyte group within the first threshold range (in the form of coordinates) with the elliptical gate. And the polygonGate() and Subset() functions are used to extract the lymphocyte data from the original data.

[0064] Step 107, determining the ratio of the cells expressing the specific fluorescence signal in the flow lymphocyte group to the cells in the flow lymphocyte group;

[0065] Wherein, the specific fluorescence signal includes CD3, CD4, CD8, CD19, CD16+56. After determining the lymphocytes, the cells expressing the specific fluorescence signal can be determined from the lymphocyte group, so as to determine the ratio of the cells expressing the specific fluorescence signal in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0066] Specifically, the cells expressing the specific fluorescence signal are determined from the lymphocyte group by automatic gating. The automatic gating refers to drawing the threshold dividing line of the fluorescence signal positive and negative expression in the two-dimension scatter diagram composed of two groups based on the CD3, CD4, CD8, CD19, CD16+56 fluorescence signal data of the lymphocytes, and circling the cell group expressing the specific fluorescence signal, for example, circling the cell group expressing CD3 positive and CD4 positive. The flowDensity() and deGate() functions are used in the present application to find the threshold of the lymphocytes between the CD3 negative and positive expression according to the density distribution of the CD3 fluorescence signal. Generally, the density distribution of the lymphocyte group on CD3 only has one inflection point, which is the dividing line (i.e. the threshold) between the CD3 negative and positive expression. In some special cases, the density distribution of CD3 has multiple inflection points, in which case the inflection point within the preset density distribution range is selected as the threshold of the CD3 negative and positive expression. For the density distribution of the lymphocytes on CD4, CD8, CD19 and CD16+56, the flowDensity() function can be used to select the inflection point after the highest peak of the fluorescence signal density distribution as the threshold. If the threshold of CD4 (or CD8) falls outside the preset density distribution range, the cell group expressing CD3 positive is used for analysis, and the inflection point of the density distribution of the CD3 positive cell group on CD4 (or CD8) is selected as the threshold. If the threshold of CD19 or CD16+56 falls outside the preset density distribution range, the cell group expressing CD3 negative is used for analysis, and the inflection point of the density distribution of the CD3 negative cell group on CD19 (or CD16+56) signal is selected as the threshold. After obtaining the threshold, the flowDensity() function is used to obtain the cell group expressing the specific fluorescence and the ratio of the cell group to the lymphocytes.

[0067] In one embodiment, the determining the ratio of the cells expressing the specific fluorescent signal in the flow lymphocyte group to the cells in the flow lymphocyte group comprises: determining a first ratio of the cells positively expressing CD3 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a second ratio of the cells positively expressing CD3 and positively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a third ratio of the cells positively expressing CD3 and negatively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fourth ratio of the cells positively expressing CD3 and positively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fifth ratio of the cells positively expressing CD3 and negatively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a sixth ratio of the cells negatively expressing CD3 and positively expressing CD19 in the flow lymphocyte group to the cells in the flow lymphocyte group; and determining a seventh ratio of the cells negatively expressing CD3 and positively expressing CD16+56 in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0068] Specifically, the first ratio of the cells positively expressing CD3 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the first ratio of the T lymphocytes to the cells in the flow lymphocyte group is determined; the second ratio of the cells positively expressing CD3 and positively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the second ratio of the Th cells, a subgroup of the T lymphocytes, to the cells in the flow lymphocyte group is determined; the third ratio of the cells positively expressing CD3 and negatively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the third ratio of the cells positively expressing CD3 and negatively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group is taken as one of the judgment indexes; the fourth ratio of the cells positively expressing CD3 and positively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the fourth ratio of the Ts cells, a subgroup of the T lymphocytes, to the cells in the flow lymphocyte group is determined; the fifth ratio of the cells positively expressing CD3 and negatively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the fifth ratio of the cells positively expressing CD3 and negatively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group is taken as one of the judgment indexes; the sixth ratio of the cells negatively expressing CD3 and positively expressing CD19 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the sixth ratio of the B lymphocytes to the cells in the flow lymphocyte group is determined; and the seventh ratio of the cells negatively expressing CD3 and positively expressing CD16+56 in the flow lymphocyte group to the cells in the flow lymphocyte group is determined, i.e. the seventh ratio of the NK lymphocytes to the cells in the flow lymphocyte group is determined.

[0069] In one embodiment, the determining the ratio of the cells expressing the specific fluorescent signal in the flow lymphocyte group to the cells in the flow lymphocyte group further comprises: determining an eighth ratio of the cells expressing CD3 negatively and CD4 positively in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0070] Specifically, the eighth ratio of the cells expressing CD3 negatively and CD4 positively in the flow lymphocyte group to the cells in the flow lymphocyte group is taken as one of the judgment indexes.

[0071] In step 108, the flow lymphocyte analysis result is output according to the ratio.

[0072] After the ratio of the cells expressing the specific fluorescent signal in the flow lymphocyte group to the cells in the flow lymphocyte group is obtained, the abnormal data can be identified based on the ratio of the cells expressing the specific fluorescent signal to the cells in the flow lymphocyte group, and the flow lymphocyte analysis result is output according to the non-abnormal data.

[0073] In one embodiment, the outputting the flow lymphocyte analysis result according to the ratio comprises: calculating a sum of the second ratio and the fourth ratio to obtain a first analysis result; calculating a sum of the first ratio, the sixth ratio and the seventh ratio to obtain a second analysis result; and outputting the first analysis result and the second analysis result if the first analysis result is not greater than the first ratio, the second analysis result is within a first preset ratio range, a difference between the second ratio and the fifth ratio is not more than a second preset ratio range, and a difference between the fourth ratio and the third ratio is not more than a third preset ratio range.

[0074] Wherein, first, the number of lymphocyte cells is obtained, then the ratio of the five groups of cells, i.e. CD3 positive cell group (i.e. T lymphocytes), CD3 positive and CD4 positive cell group (i.e. Th cell subgroup of T lymphocytes), CD3 positive and CD8 positive cell group (i.e. Ts cell subgroup of T lymphocytes), CD3 negative and CD19 positive cell group (i.e. B lymphocytes), and CD3 negative and CD16+56 positive cell group (i.e. NK lymphocytes) in the lymphocytes is obtained. Then, the sum of the second ratio and the fourth ratio is calculated to obtain the first analysis result, i.e. the sum of the proportion of CD3 positive and CD4 positive cell group (i.e. Th cell subgroup of T lymphocytes) and the proportion of CD3 positive and CD8 positive cell group (i.e. Ts cell subgroup of T lymphocytes) is calculated. Finally, the sum of the first ratio, the sixth ratio and the seventh ratio is calculated to obtain the second analysis result, i.e. the sum of the proportion of CD3 positive cell group (i.e. T lymphocytes), the proportion of CD3 negative and CD19 positive cell group (i.e. B lymphocytes), and the proportion of CD3 negative and CD16+56 positive cell group (i.e. NK lymphocytes) in the lymphocytes is calculated. The above results are stored in the form of a data table in a csv file.

[0075] Wherein, the statistical results of the above calculation are logically judged to identify the abnormal analysis results. The judgment conditions include:

[0076] 1) The second analysis result is in a first preset ratio range, i.e. the sum of the proportion of T lymphocytes, B lymphocytes and NK lymphocytes in the lymphocytes is in the range of 95%-100%;

[0077] 2) The first analysis result is not greater than the first ratio, i.e. the sum of the proportion of CD3 positive and CD4 positive cell group and the proportion of CD3 positive and CD8 positive cell group does not exceed the proportion of T lymphocytes;

[0078] 3) The difference between the second ratio and the fifth ratio does not exceed a second preset ratio range, i.e. the difference between the proportion of CD3 positive and CD4 positive cell group and the proportion of CD3 positive and CD8 negative cell group does not exceed 20%;

[0079] 4) The difference between the fourth ratio and the third ratio does not exceed a third preset ratio range, i.e. the difference between the proportion of CD3 positive and CD4 negative cell group and the proportion of CD3 positive and CD8 positive cell group does not exceed 20%.

[0080] In an implementation method, the non-abnormal data also needs to satisfy: 1) the number of lymphocytes exceeds 2000; 2) the proportion of mononuclear cells does not exceed 5%; 3) the proportion of B lymphocytes does not exceed 50%.

[0081] If the statistical results do not meet the above rules, the algorithm analysis results for the sample are considered abnormal. The abnormal information is stored as a separate column in the CSV file. If the statistical results meet the above rules, the algorithm analysis results for the sample are considered normal, and the normal information is stored in the CSV file.

[0082] In one embodiment, the outputting of the flow lymphocyte analysis result according to the ratio further includes: determining whether the eighth ratio is within a fourth preset ratio range; if the eighth ratio is within the fourth preset ratio range, executing the step of outputting the flow lymphocyte analysis result according to the ratio; if the eighth ratio is not within the fourth preset ratio range, adjusting the first threshold according to the eighth ratio until the eighth ratio is within the fourth preset ratio range.

[0083] Among them, since the FlowDensity algorithm is used to identify lymphocytes, it is easy to identify adjacent monocytes as lymphocytes. Therefore, the threshold of the lymphocyte population on SSC.A can be adjusted inversely according to the proportion of monocytes (i.e., CD3 negative CD4 positive cell population) (eighth ratio). According to large sample analysis, when the analysis results are normal, the normal range of monocyte proportion is 0%-5%. When the proportion of monocytes in the algorithm analysis results exceeds 5%, it indicates that the threshold of lymphocytes and non-lymphocytes in the SSC.A signal is too high, and a large number of monocytes are included. The analysis result can be classified as abnormal. Therefore, in this application, when the algorithm recognizes that the proportion of monocytes in the analysis results exceeds 5%, the threshold of lymphocytes and non-lymphocytes in the SSC.A data is adjusted. Specifically, when the proportion of monocytes exceeds 5% but does not exceed 8%, the SSC.A threshold is multiplied by 0.95; when the proportion of monocytes exceeds 8%, the SSC.A threshold is multiplied by 0.90. The adjusted threshold is then used again for monocyte identification, gating, and abnormality identification. This process is repeated until the proportion of monocytes does not exceed 5%.

[0084] In one embodiment, after the analysis is completed, the analysis result image, statistical results and warning information will be output. After each sample is analyzed, the corresponding analysis result image will be output immediately (see example Figure 2 ).like Figure 2 As shown, Figure 2An analysis result schematic diagram obtained by a flow cytometry analysis method provided by an embodiment of the present application, which contains nine two-dimensional scatter plots. Among them, green scatter points are CD3 positive cells (i.e. T lymphocytes), and red scatter points are CD3 negative cells (i.e. B lymphocytes, NK lymphocytes and possibly existing monocytes). In the fourth to ninth scatter plots, the horizontal (vertical) line represents the threshold value for distinguishing the positive and negative expressions of the fluorescence signals, and the threshold value divides the four quadrants representing the fluorescence signal expressions of different cell groups (for example, the cell group in the first quadrant is double negative expression of the two fluorescence signals), and the numbers marked in each phenomenon represent the ratio of the number of cells in the quadrant to the lymphocytes. After analyzing a batch of samples, the statistical results and warning information of all samples in the batch are integrated into a data table and output to a CSV file. The data table contains the sample names, statistical results and warning information of all samples in the batch. The content of the warning information includes which judgment conditions are not met by the analysis results of the sample, and the warning information is empty when the analysis results are normal.

[0085] The present application proposes a flow lymphocyte analysis method, which includes: obtaining data to be analyzed; performing density distribution analysis on the data to be analyzed to obtain a density distribution result of the data to be analyzed; judging whether the density distribution result has a first threshold value of a flow lymphocyte population and a non-flow lymphocyte population within a preset density distribution range based on a FlowDentity function; the first threshold value is a trough between two peaks in the density distribution result; if the first threshold value does not exist within the preset density distribution range, determining an initial region containing a flow lymphocyte population from the density distribution result based on the FlowDentity function; adjusting the region boundary of the initial region by adjusting the function parameters of the FlowDentity function so that the adjusted region boundary is within the preset density distribution range, and using the adjusted region boundary as the first threshold value; determining the flow lymphocyte population in the data to be analyzed based on the first threshold value; determining the ratio of cells expressing a specific fluorescent signal in the flow lymphocyte population to the cells in the flow lymphocyte population; and outputting the flow lymphocyte analysis result according to the ratio. The present application determines whether the density distribution result has a first threshold of a flow lymphocyte population and a non-flow lymphocyte population within a preset density distribution range based on the FlowDentity function; and when the first threshold does not exist within the preset density distribution range, determines an initial region containing a flow lymphocyte population based on the FlowDentity function; and adjusts the region boundary of the initial region by adjusting the function parameters of the FlowDentity function so that the adjusted region boundary matches the preset density distribution range, and uses the adjusted region boundary as the first threshold; finally, determines the flow lymphocyte population and the analysis result based on the first threshold; the analysis result is not affected by the large number of sample cells and the non-fixed number of cell categories, and can effectively improve the efficiency and accuracy of the analysis.

[0086] like Figure 3 As shown, in one embodiment, the present invention provides a flow lymphocyte analysis device, the device comprising:

[0087] Data acquisition module 301, used to acquire data to be analyzed;

[0088] Density distribution analysis module 302, configured to perform density distribution analysis on the data to be analyzed to obtain density distribution results of the data to be analyzed;

[0089] A first determination module 303 is configured to determine, based on a FlowDentity function, whether the density distribution result contains a first threshold of a flow lymphocyte population and a non-flow lymphocyte population within a preset density distribution range; the first threshold is a trough between two peaks in the density distribution result;

[0090] The second determining module 304 is configured to determine an initial region containing the flow lymphocyte group from the density distribution result based on the FlowDentity function if the first threshold value does not exist in the preset density distribution range.

[0091] The first threshold value determining module 305 is configured to adjust the region boundary of the initial region by adjusting the function parameter of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and take the adjusted region boundary as the first threshold value.

[0092] The flow lymphocyte group determining module 306 is configured to determine the flow lymphocyte group in the data to be analyzed based on the first threshold value.

[0093] The ratio determining module 307 is configured to determine the ratio of the cells expressing the specific fluorescence signal in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0094] The data output module 308 is configured to output the flow lymphocyte analysis result according to the ratio.

[0095] As shown in FIG. 1, Figure 4 In an embodiment, the computer device is an internal structure diagram. The computer device can be a flow lymphocyte analysis device, or a terminal or server connected with a flow lymphocyte analysis device. As shown in FIG. 2, Figure 4 The computer device includes a processor, a memory, and a network interface connected through a system bus. The memory includes a non-volatile storage medium and an internal memory. The non-volatile storage medium of the computer device stores an operating system, and can also store a computer program. When the computer program is executed by the processor, the processor can implement a flow lymphocyte analysis method. The internal memory can also store a computer program. When the computer program is executed by the processor, the processor can execute a flow lymphocyte analysis method. The network interface is used for communication with an external device. Those skilled in the art can understand that Figure 4 The structure shown in FIG. 2 is only a block diagram of part of the structure related to the scheme of the present application, and does not constitute a limitation on the computer device to which the scheme of the present application is applied. The specific computer device can include more or fewer components than those shown in the figure, or combine certain components, or have a different component arrangement.

[0096] In an embodiment, the flow lymphocyte analysis method provided by the present application can be implemented in the form of a computer program. The computer program can be executed on a computer device such as Figure 4The computer device shown is running. The memory of the computer device can store various program templates constituting the lymphocyte analysis device. For example, the data acquisition module 301, the density distribution analysis module 302, the first judgment module 303, the second judgment module 304, the first threshold determination module 305, the lymphocyte population determination module 306, the ratio determination module 307, and the data output module 308.

[0097] A computer device includes a memory and a processor, the memory storing a computer program, the computer program being executed by the processor to cause the processor to perform the following steps: acquiring data to be analyzed; performing density distribution analysis on the data to be analyzed to obtain a density distribution result of the data to be analyzed; determining, based on a FlowDentity function, whether a first threshold value of a lymphocyte population and a non-lymphocyte population exists in a preset density distribution range of the density distribution result; the first threshold value being a valley between two peaks in the density distribution result; if the first threshold value does not exist in the preset density distribution range, determining an initial region containing a lymphocyte population from the density distribution result based on the FlowDentity function; adjusting the region boundary of the initial region by adjusting the function parameters of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and taking the adjusted region boundary as the first threshold value; determining a lymphocyte population in the data to be analyzed based on the first threshold value; determining a ratio of cells expressing a specific fluorescent signal in the lymphocyte population to cells in the lymphocyte population; and outputting a lymphocyte analysis result according to the ratio.

[0098] In one embodiment, the acquiring data to be analyzed includes: performing data cleaning on original data obtained by clinical detection to obtain cleaned data, and taking the cleaned data as the data to be analyzed.

[0099] In one embodiment, the determining a lymphocyte population based on the first threshold value includes: using the FlowDentity function to define the boundary of the lymphocyte population in the first threshold value range with an elliptical gate to determine the lymphocyte population.

[0100] In one embodiment, the specific fluorescent signals include CD3, CD4, CD8, CD19, CD16+56; and the determining the ratio of the cells expressing the specific fluorescent signals in the flow lymphocyte group to the cells in the flow lymphocyte group includes: determining a first ratio of the cells positively expressing CD3 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a second ratio of the cells positively expressing CD3 and CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a third ratio of the cells positively expressing CD3 and negatively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fourth ratio of the cells positively expressing CD3 and CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fifth ratio of the cells positively expressing CD3 and negatively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a sixth ratio of the cells negatively expressing CD3 and positively expressing CD19 in the flow lymphocyte group to the cells in the flow lymphocyte group; and determining a seventh ratio of the cells negatively expressing CD3 and positively expressing CD16+56 in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0101] In one embodiment, the outputting the flow lymphocyte analysis result according to the ratios includes: calculating a first analysis result by summing the second ratio and the fourth ratio; calculating a second analysis result by summing the first ratio, the sixth ratio and the seventh ratio; and outputting the first analysis result and the second analysis result if the first analysis result is not greater than the first ratio, the second analysis result is within a first preset ratio range, a difference between the second ratio and the fifth ratio is not greater than a second preset ratio range, and a difference between the fourth ratio and the third ratio is not greater than a third preset ratio range.

[0102] In one embodiment, the determining the ratio of the cells expressing the specific fluorescent signals in the flow lymphocyte group to the cells in the flow lymphocyte group further includes: determining an eighth ratio of the cells negatively expressing CD3 and positively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0103] In one embodiment, the outputting the flow lymphocyte analysis result according to the ratios further includes: judging whether the eighth ratio is within a fourth preset ratio range; if the eighth ratio is within the fourth preset ratio range, performing the step of outputting the flow lymphocyte analysis result according to the ratios; and if the eighth ratio is not within the fourth preset ratio range, adjusting the first threshold according to the eighth ratio until the eighth ratio is within the fourth preset ratio range.

[0104] A computer readable storage medium stores a computer program, the computer program is executed by a processor to make the processor execute the following steps: obtaining to-be-analyzed data; performing density distribution analysis on the to-be-analyzed data to obtain a density distribution result of the to-be-analyzed data; determining, based on a FlowDentity function, whether a first threshold value of a flow lymphocyte group and a non-flow lymphocyte group exists in a preset density distribution range of the density distribution result; the first threshold value is a trough between two peaks in the density distribution result; if the first threshold value does not exist in the preset density distribution range, determining, based on the FlowDentity function, an initial region containing the flow lymphocyte group from the density distribution result; adjusting a region boundary of the initial region by adjusting a function parameter of the FlowDentity function, so that the adjusted region boundary is within the preset density distribution range, and taking the adjusted region boundary as the first threshold value; determining the flow lymphocyte group in the to-be-analyzed data based on the first threshold value; determining a ratio of cells expressing a specific fluorescence signal in the flow lymphocyte group to cells in the flow lymphocyte group; and outputting a flow lymphocyte analysis result according to the ratio.

[0105] In one embodiment, the obtaining to-be-analyzed data comprises: performing data cleaning on original data obtained by clinical detection to obtain cleaned data, and taking the cleaned data as the to-be-analyzed data.

[0106] In one embodiment, the determining the flow lymphocyte group based on the first threshold value comprises: using the FlowDentity function to define a boundary of the flow lymphocyte group in the first threshold value range by using an elliptical gate to determine the flow lymphocyte group.

[0107] In one embodiment, the specific fluorescent signals include CD3, CD4, CD8, CD19, CD16+56; and the determining the ratio of the cells expressing the specific fluorescent signals in the flow lymphocyte group to the cells in the flow lymphocyte group includes: determining a first ratio of the cells positively expressing CD3 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a second ratio of the cells positively expressing CD3 and CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a third ratio of the cells positively expressing CD3 and negatively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fourth ratio of the cells positively expressing CD3 and CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a fifth ratio of the cells positively expressing CD3 and negatively expressing CD8 in the flow lymphocyte group to the cells in the flow lymphocyte group; determining a sixth ratio of the cells negatively expressing CD3 and positively expressing CD19 in the flow lymphocyte group to the cells in the flow lymphocyte group; and determining a seventh ratio of the cells negatively expressing CD3 and positively expressing CD16+56 in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0108] In one embodiment, the outputting the flow lymphocyte analysis result according to the ratios includes: calculating a first analysis result by summing the second ratio and the fourth ratio; calculating a second analysis result by summing the first ratio, the sixth ratio and the seventh ratio; and outputting the first analysis result and the second analysis result if the first analysis result is not greater than the first ratio, the second analysis result is within a first preset ratio range, a difference between the second ratio and the fifth ratio is not greater than a second preset ratio range, and a difference between the fourth ratio and the third ratio is not greater than a third preset ratio range.

[0109] In one embodiment, the determining the ratio of the cells expressing the specific fluorescent signals in the flow lymphocyte group to the cells in the flow lymphocyte group further includes: determining an eighth ratio of the cells negatively expressing CD3 and positively expressing CD4 in the flow lymphocyte group to the cells in the flow lymphocyte group.

[0110] In one embodiment, the outputting the flow lymphocyte analysis result according to the ratios further includes: judging whether the eighth ratio is within a fourth preset ratio range; if the eighth ratio is within the fourth preset ratio range, performing the step of outputting the flow lymphocyte analysis result according to the ratios; and if the eighth ratio is not within the fourth preset ratio range, adjusting the first threshold according to the eighth ratio until the eighth ratio is within the fourth preset ratio range.

[0111] Those skilled in the art can understand that all or part of the processes in the above-mentioned embodiment methods can be completed by a computer program instructing related hardware. The program can be stored in a computer readable storage medium, and when the program is executed, the processes of the above-mentioned embodiment methods can be included. The storage medium can be a magnetic disc, an optical disc, a read-only memory (ROM), a random access memory (RAM), or the like.

[0112] The above disclosure is only the preferred embodiment of the present application, and of course cannot limit the scope of the right of the present application, so the equivalent changes made according to the claims of the present application still belong to the scope covered by the present application.

Claims

1. A flow cytometry lymphocyte analysis method, characterized in that: The method comprises: Obtain the data to be analyzed; Performing density distribution analysis on the data to be analyzed to obtain a density distribution result of the data to be analyzed; Determining, based on the FlowDentity function, whether the density distribution result includes a flow lymphocyte population and a non-flow lymphocyte population within a preset density distribution range; the first threshold is a trough between two peaks in the density distribution result; If the first threshold does not exist within the preset density distribution range, determining an initial region containing a flow cytometry lymphocyte population from the density distribution result based on a FlowDentity function; Adjusting the region boundary of the initial region by adjusting the function parameters of the FlowDentity function so that the adjusted region boundary is within the preset density distribution range, and using the adjusted region boundary as the first threshold; determining a flow cytometry lymphocyte population in the data to be analyzed based on the first threshold; Determining the ratio of cells expressing a specific fluorescent signal in the flow cytometry lymphocyte population; outputting flow lymphocyte analysis results according to the ratio; Wherein, determining the flow cytometry lymphocyte population based on the first threshold comprises: Using the FlowDentity function, elliptical gating is used to delineate the boundaries of the flow lymphocyte population within the first threshold range to determine the flow lymphocyte population; The specific fluorescent signals include: CD3, CD4, CD8, CD19, and CD16+56; and determining the ratio of cells expressing the specific fluorescent signals in the flow lymphocyte population to the cells in the flow lymphocyte population includes: determining a first ratio of cells expressing CD3 positively in a flow cytometry lymphocyte population to cells in the flow cytometry lymphocyte population; determining a second ratio of cells in the flow cytometry lymphocyte population that are CD3-positive and CD4-positive; and determining a third ratio of cells in the flow lymphocyte population that are CD3 positive and CD4 negative in the flow lymphocyte population; determining a fourth ratio of cells in the flow cytometry lymphocyte population that are both CD3-positive and CD8-positive; and determining a fifth ratio of cells in a flow cytometry lymphocyte population that are CD3 positive and CD8 negative in the flow cytometry lymphocyte population; determining a sixth ratio of cells in the flow lymphocyte population that are CD3 negative and CD19 positive in the flow lymphocyte population; The seventh ratio of cells in the flow lymphocyte population that are CD3 negative and CD16+56 positive is determined.

2. The method according to claim 1, characterized in that The obtaining of the data to be analyzed includes: The raw data obtained from the clinical test are cleaned to obtain cleaned data, and the cleaned data are used as the data to be analyzed.

3. The method according to claim 1, characterized in that Outputting the flow lymphocyte analysis result according to the ratio includes: Calculating the sum of the second ratio and the fourth ratio to obtain a first analysis result; Calculating the sum of the first ratio, the sixth ratio, and the seventh ratio to obtain a second analysis result; If the first analysis result is not greater than the first ratio, the second analysis result is within the first preset ratio range, the difference between the second ratio and the fifth ratio does not exceed the second preset ratio range, and the difference between the fourth ratio and the third ratio does not exceed the third preset ratio range, then the first analysis result and the second analysis result are output.

4. The method according to claim 1, wherein The determining of the ratio of cells expressing a specific fluorescent signal in the flow lymphocyte population to the cells in the flow lymphocyte population further includes: The eighth ratio of cells in the flow lymphocyte population that are CD3 negative and CD4 positive is determined.

5. The method according to claim 4, characterized in that Outputting the flow lymphocyte analysis result according to the ratio further includes: determining whether the eighth ratio is within a fourth preset ratio range; If the eighth ratio is within the fourth preset ratio range, performing the step of outputting the flow lymphocyte analysis result according to the ratio; If the eighth ratio is not within the fourth preset ratio range, the first threshold is adjusted according to the eighth ratio until the eighth ratio is within the fourth preset ratio range.

6. A flow cytometry lymphocyte analysis device, characterized in that: The device comprises: A data acquisition module is used to obtain data to be analyzed; A density distribution analysis module is used to perform density distribution analysis on the data to be analyzed to obtain a density distribution result of the data to be analyzed; A first judgment module is configured to judge, based on a FlowDentity function, whether the density distribution result contains a first threshold value of a flow lymphocyte population and a non-flow lymphocyte population within a preset density distribution range; the first threshold value is a trough between two peaks in the density distribution result; a second judgment module, configured to determine an initial region containing a flow cytometry lymphocyte population from the density distribution result based on a FlowDentity function if the first threshold does not exist within the preset density distribution range; a first threshold determination module, configured to adjust a region boundary of the initial region by adjusting a function parameter of a FlowDentity function so that the adjusted region boundary is within the preset density distribution range, and use the adjusted region boundary as the first threshold; a flow lymphocyte population determination module, configured to determine a flow lymphocyte population in the data to be analyzed based on the first threshold; wherein the flow lymphocyte population determination module is specifically configured to use a FlowDentity function to use elliptical gating to delineate the boundaries of the flow lymphocyte population within the first threshold range to determine the flow lymphocyte population; A ratio determination module is used to determine the ratio of cells expressing a specific fluorescent signal in a flow lymphocyte population to the cells in the flow lymphocyte population; wherein the specific fluorescent signals include: CD3, CD4, CD8, CD19, CD16+56; the ratio determination module is specifically used to: determine a first ratio of cells expressing CD3 positively in a flow lymphocyte population to the cells in the flow lymphocyte population; determine a second ratio of cells expressing CD3 positively and CD4 positively in a flow lymphocyte population to the cells in the flow lymphocyte population; determine a second ratio of cells expressing CD3 positively and CD4 negatively in a flow lymphocyte population determining a third ratio of cells in the flow lymphocyte population that are CD3 positively expressed and CD8 positively expressed in the flow lymphocyte population; determining a fourth ratio of cells in the flow lymphocyte population that are CD3 positively expressed and CD8 negatively expressed in the flow lymphocyte population; determining a fifth ratio of cells in the flow lymphocyte population that are CD3 negatively expressed and CD19 positively expressed in the flow lymphocyte population; determining a seventh ratio of cells in the flow lymphocyte population that are CD3 negatively expressed and CD16+56 positively expressed in the flow lymphocyte population; A data output module is used to output the flow lymphocyte analysis results according to the ratio.

7. A computer device comprising a memory and a processor, wherein the memory stores a computer program, and when the computer program is executed by the processor, the processor performs the steps of the method according to claims 1 to 5.

8. A computer-readable storage medium storing a computer program, wherein when the computer program is executed by a processor, the processor is caused to perform the steps of the method according to claims 1 to 5.

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