CN112673088A - Method for detecting white blood cells, blood cell analyzer and storage medium - Google Patents
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Abstract
A method for detecting leukocytes, a blood cell analyzer and a storage medium. A method of detecting leukocytes for use in a blood cell analyzer, comprising: obtaining first optical signal information of particles of a first test sample, wherein the first test sample is prepared by subjecting a blood sample to a hemolysis process and a fluorescent staining process (101); classifying and counting particles in the blood sample according to the first optical signal information to obtain a first leukocyte particle group (102); determining whether a leukocyte accumulation is present in the first test sample based on pulse width information of the first optical signal information of the first leukocyte population (103); and performing a retest (104) of the blood sample when it is determined that there is leukocyte aggregation in the first test sample.
Description
The invention relates to the field of blood sample detection, in particular to the detection of leucocytes.
When the blood cell analyzer detects white blood cells, firstly, red blood cells in a blood sample are ruptured by a hemolytic agent, the white blood cells are treated, then the generated blood sample is enabled to pass through a detection area to classify particles in the sample according to different optical signals generated by the ruptured red blood cell fragments and the white blood cells passing through the detection area, the red blood cell fragment particles (ghost) and the white blood cell particle regions can be separated, and further, the optical signal information of the white blood cell particles is further analyzed to obtain the required detection result. There are several detection methodologies for leukocytes, such as flow cytometry, in which leukocytes in a blood sample are sequentially queued through a detection zone by the action of a sheath fluid as the leukocytes are measured. As each leukocyte passes through the detection zone, an optical pulse is generated in the corresponding detection channel. The number and classification of the white blood cells can be obtained by counting and analyzing the optical pulse.
For normal samples, the vast majority of leukocytes are able to pass through the detection zone independently as single cells, i.e., one leukocyte may produce one pulse. However, it has been found that during the collection and preparation of a blood sample, there may be instances where multiple leukocytes pass through the detection zone in overlapping relationship within the sample. In this case, a plurality of aggregated leukocytes form only one allotype pulse, so that a plurality of leukocytes are recognized as one leukocyte, resulting in inaccurate leukocyte count.
This is very rare in normal samples, but for some abnormal samples, it is likely that more severe white blood cells will accumulate. It was also noted in "analysis of decrease in leukocyte pseudoreduction in routine blood tests due to leukocyte aggregation" in 50 cases (Yangkai, Huishui county, Guizhou, Jilin medicine 2014, volume 35, 20): "EDTA anticoagulation can cause aggregation between leukocytes or between leukocytes and platelets, infectious mononucleosis and weakening of repulsive force between leukocytes caused by acute bacterial infection can cause aggregation of leukocytes, and the pseudoreduction of leukocytes is caused. The leucocyte aggregation usually causes the pseudo-reduction of leucocytes, influences the correctness of diagnosis, and needs to improve the understanding, cannot completely depend on the result of cytometric detection, and avoids misleading caused by the leucocyte reduction caused by various reasons as much as possible. "in another example, in the case where blood contains a tumor marker of malignant pleural mesothelioma, leukocytes are accumulated in a large amount. Multiple leukocytes caused by different reasons are overlapped together to pass through the detection area, so that the detection result of the leukocyte count is low, and the judgment of a clinician is influenced.
Disclosure of Invention
The invention aims to provide a leukocyte detection method, a blood cell analyzer and a computer storage medium storing the program of the method, which can judge the occurrence of leukocyte aggregation in a sample and further eliminate the aggregation, aiming at the problem that the leukocyte aggregation possibly exists in the leukocyte detection and further influences the accuracy of the leukocyte counting.
To this end, according to a first aspect of the present invention, there is provided a method of detecting leukocytes in a blood sample of a subject for use in a blood cell analyzer, the method comprising:
obtaining first optical signal information of particles in a first test sample, wherein the first test sample is prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process;
classifying and counting particles in the first test sample according to the first optical signal information to obtain a first leukocyte particle group;
judging whether leukocyte aggregation exists in the first test sample according to pulse width information of first optical signal information of the first leukocyte particle group; and
when it is determined that there is leukocyte aggregation in the first test sample,
acquiring second optical signal information of particles in a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; and
classifying and counting particles in the second test sample according to the second optical signal information to obtain a second leukocyte particle group.
According to one embodiment of the method, the step of determining whether there is leukocyte aggregation in the first test sample based on the pulse width information of the first optical signal information of the first leukocyte particles may comprise:
counting the total number of first leukocyte particle groups according to the first optical signal information and counting the number of abnormal leukocyte particles with pulse widths larger than a first preset threshold value according to the pulse width information;
determining whether the ratio of the number of abnormal leukocyte particles to the total number of the first leukocyte particle population is greater than a second predetermined threshold; and
determining that leukocyte aggregation is present in the first test sample when the ratio is greater than the second predetermined threshold.
According to one embodiment, the first predetermined threshold may be determined from pulse width information in the light signal information of leukocyte particles in a normal blood sample.
According to another embodiment, the first predetermined threshold value may be determined from an average value of pulse widths in the first optical signal information of the first leukocyte population.
In the method of the present invention, the first and/or second optical signal information may include a fluorescent signal and a forward scattered light signal. Furthermore, the first and/or second optical signal information may further comprise side scattered light signals.
According to a particular embodiment, when it is determined that there is a leukocyte accumulation in the first test sample, the method may further comprise: and alarming the leucocyte aggregation.
According to a further specific embodiment, when it is determined that there is leukocyte aggregation in the first test sample, the method may further comprise: recording and storing information that the subject's blood sample is a leukocyte aggregate sample in association with the characteristic information of the subject.
According to an embodiment, the method may further comprise: and outputting the classification and counting results of the first leukocyte particle group and/or the second leukocyte particle group.
There is also provided, in accordance with a second embodiment of the first aspect of the present invention, a method of detecting leukocytes in a blood sample of a subject, for use in a blood cell analyzer, the method comprising:
obtaining information of the subject;
matching the obtained information of the subject with stored information of the subject in which the presence of the leukocyte accumulation has been detected; and
detecting leukocytes in a blood sample of the subject according to the method of the first embodiment when no matching information is found, and
when matching information is found, the following steps are performed:
acquiring second optical signal information of particles of a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; and
classifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
After the matching information is found and the above steps are performed, the method may further include: and outputting the classification and counting results of the second leukocyte particle group.
In a second aspect of the present invention, there is provided a blood cell analyzer according to a third embodiment. The blood cell analyzer includes:
a sampling device for aspirating a blood sample of a subject;
a pretreatment device for pretreating the blood sample to prepare a pretreated test sample;
optical detection means for passing particles in the pretreated test sample through detection zones one by one to detect and output optical signal information of the particles in the test sample; and
a processor for performing the steps of:
obtaining first optical signal information of particles of a first test sample, wherein the first test sample is prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process;
classifying and counting particles in the blood sample according to the first optical signal information to obtain a first leukocyte particle group;
judging whether leukocyte aggregation exists in the first test sample according to pulse width information of first optical signal information of the first leukocyte particle group; and
when it is determined that there is leukocyte aggregation in the first test sample,
acquiring second optical signal information of particles of a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; and
classifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
According to one embodiment, the processor, when performing the step of determining whether a leukocyte accumulation is present in the first test sample from pulse width information of the first optical signal information of the first leukocyte population, may perform the steps of:
counting the total number of first leukocyte particle groups according to the first optical signal information and counting the number of abnormal leukocyte particles with pulse widths larger than a first preset threshold value according to the pulse width information;
determining whether the ratio of the number of abnormal leukocyte particles to the total number of the first leukocyte particle population is greater than a second predetermined threshold; and
determining that leukocyte aggregation is present in the first test sample when the ratio is greater than the second predetermined threshold.
According to an embodiment of the blood cell analyzer of the present invention, the first predetermined threshold value may be determined based on pulse width information in the optical signal information of leukocyte particles in a normal blood sample.
In accordance with another embodiment of the blood cell analyzer of the present invention, the first predetermined threshold value may be determined based on an average value of pulse widths in the first optical signal information of the first leukocyte particle group.
In the blood cell analyzer of the present invention, the first and/or second optical signal information may include fluorescence signal information and forward scattered light information. Further, the first and/or second optical signal information may further comprise side scattered light information.
According to a specific embodiment, the processor may be further configured to output an alarm signal of the leukocyte accumulation when it is determined that the leukocyte accumulation is present in the first test sample.
According to another specific embodiment, wherein the processor is further operable to record and store information that the blood sample of the subject is a leukocyte aggregation sample in association with the characteristic information of the subject when it is determined that leukocyte aggregation is present in the first test sample.
The blood cell analyzer of the present invention may further include a display device for displaying the results of classification and counting of the first leukocyte particle group and/or the second leukocyte particle group output by the processor.
According to an embodiment of the present invention, the pretreatment device of the blood cell analyzer may have a diluent addition port for performing a dilution process on the blood sample.
In a second aspect of the present invention, there is also provided according to a fourth embodiment a blood cell analyzer comprising:
a sampling device for drawing a blood sample from a subject;
a pretreatment device for pretreating the blood sample to prepare a pretreated test sample;
optical detection means for passing particles in the pretreated test sample through detection zones one by one to detect and output optical signal information of the particles in the test sample;
a processor for performing the steps of:
obtaining information of the subject;
matching the acquired information of the subject with stored information of the subject in which the presence of aggregation of leukocytes has been detected, detecting leukocytes in a blood sample of the subject according to the method defined in the aforementioned first embodiment when no matching information is found, and performing the following steps when matching information is found:
acquiring second optical signal information of particles in a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; and
classifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
A third aspect of the present invention provides an assay device for detecting leukocytes in a blood sample of a subject, comprising:
a memory configured to store executable instructions;
a processor configured to execute the executable instructions stored in the memory to perform the method for detecting leukocytes in a blood sample of a subject as defined in the first and second embodiments of the aforementioned first aspect.
A fourth aspect of the present invention provides a computer-readable storage medium having stored thereon executable instructions configured to cause a processor to perform the method for detecting leukocytes in a blood sample of a subject as defined in the first and second embodiments of the aforementioned first aspect, when the executable instructions are executed.
The method and the blood cell analyzer of the invention simply solve the problem of inaccurate white blood cell counting caused by white blood cell aggregation in a sample by adding an in-machine dilution step during rechecking when judging that the white blood cell aggregation phenomenon occurs. In addition, by correlating characteristic information of the subject, such as ID information and disease information, with information that a blood sample is primarily detected as a leukocyte aggregation sample, a dilution step for the blood sample can be directly added when the subject next goes to examine the blood sample to obtain accurate leukocyte classification and counting results.
FIG. 1 is a schematic flow chart of a first embodiment of a method of detecting leukocytes in a blood sample of a subject according to the invention;
FIG. 2 is a schematic view of a pretreatment device including a reaction cell in a conventional blood cell analyzer, in which a sampling needle of the blood cell analyzer is also shown;
FIG. 3 is a pulse waveform (A) of an optical signal of a leukocyte particle passing through an optical detection device and a pulse shape distribution scattergram (B) of forward scattered light signals of all leukocyte particles in a normal blood sample;
FIG. 4 is a pulse profile of an aggregated leukocyte particle of a blood sample with leukocyte aggregation (A) and a pulse shape distribution scattergram of forward scattered light signals of all leukocyte particles (B);
FIG. 5 is a schematic view of a pretreatment device including a reaction cell in the blood cell analyzer of the present invention, in which a sampling needle of the blood cell analyzer is also shown;
FIG. 6 is a pulse shape distribution scattergram of forward scattered light signals of all leukocyte granules obtained after a blood sample in which leukocyte aggregation is present is retested according to the method for detecting leukocytes in a blood sample of a subject of the present invention;
FIG. 7 is a schematic flow chart of a variant embodiment of the first embodiment of the method for detecting leukocytes in a blood sample of a subject according to the invention; and
FIG. 8 is a schematic flow chart of a second embodiment of a method of detecting leukocytes in a blood sample of a subject according to the invention.
FIG. 9 is a schematic configuration diagram of one embodiment of a blood cell analyzer according to the present invention.
FIG. 10 is a schematic view of the structure of an analysis device for detecting leukocytes in a blood sample of a subject according to the present invention.
The present invention will be described in further detail below with reference to the accompanying drawings and examples. It should be understood that the examples provided herein are merely illustrative of the present invention and are not intended to limit the present invention. In addition, the following embodiments are provided as partial embodiments for implementing the present invention, not all embodiments for implementing the present invention, and the technical solutions described in the embodiments of the present invention may be implemented in any combination without conflict.
It should be noted that, in the embodiments of the present invention, the terms "comprises", "comprising" or any other variation thereof are intended to cover a non-exclusive inclusion, so that a method or apparatus including a series of elements includes not only the explicitly recited elements but also other elements not explicitly listed or inherent to the method or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other related elements in a method or apparatus including the element (e.g., steps in a method or elements in an apparatus, such as a part of a circuit, a part of a processor, a part of a program or software, etc.).
It should be noted that the terms "first \ second \ third" related to the embodiments of the present invention only distinguish similar objects, and do not represent a specific ordering for the objects, and it should be understood that "first \ second \ third" may exchange a specific order or sequence when allowed. It should be understood that the terms first, second, and third, as used herein, are interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in other sequences than those illustrated or otherwise described herein.
The invention provides a method for detecting leucocytes in a blood sample of a subject, which is applied to a blood cell analyzer. Fig. 1 shows a first embodiment of the present invention.
The term "subject" as used herein, unless otherwise indicated, refers to a mammal, particularly a human.
Referring to fig. 1, step 101: first optical signal information of particles of a first test sample prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process is acquired while passing through an optical detection device of the blood cell analyzer.
The blood sample in the present invention generally refers to a whole blood sample unless otherwise specified.
The first optical signal information includes forward scattered light information and fluorescent signal information, and further may include side scattered light information.
In this step 101, a first test sample is obtained by subjecting a blood sample to conventional hemolysis and fluorescent staining. After treatment, the red blood cells in the sample are ruptured. The particles in the first test sample include at least ghost particles formed by ruptured red blood cells and include white blood cells.
Specifically, referring to fig. 5, there is shown a pretreatment device of a blood cell analyzer according to the present invention including at least a reaction cell 10. In the detector, firstly, a hemolytic agent is added into the reaction cell 10 through the hemolytic agent addition port 40, and then a blood sample is added into a hemolytic agent bottom liquid in the reaction cell 10 through the needle port 3 of the sampling needle 2, and is uniformly mixed. Followed by the addition of a fluorescent dye through the fluorescent dye addition port 50. Wherein the blood sample, the hemolytic agent, and the fluorescent dye are added to the reaction cell 10 at a conventional ratio (e.g., conventionally 20. mu.L: 1000. mu.L: 20. mu.L) through the sampling needle 2, the hemolytic agent addition port 40, and the fluorescent dye addition port 50, respectively, to prepare a first test sample. After a period of incubation, the first test sample is transferred to the optical detection device through the reaction solution extraction port 60 for detection, so as to obtain first optical signal information of the particles in the first test sample.
In step 102, the particles in the first test sample are classified and counted according to the first optical signal information to obtain a first leukocyte particle group.
In step 101, a first test sample is comprised primarily of ghost particles and white blood cells, wherein the ghost particles are substantially smaller in size than the white blood cells. Thus, in step 102, the ghost particles and the white blood particles may be distinguished based on the forward scattered light information reflecting the particle size in the first optical signal information, or based on at least two of the forward scattered light, the side scattered light, and the fluorescence information. First optical signal information of the leukocyte particles in the first test sample is thereby obtained.
The vast majority of leukocytes in a normal blood sample can pass through the detection zone as single cells, i.e., a single leukocyte can generate a pulse. Referring to FIG. 3, FIG. 3A shows the forward scattered light signal pulse shape of a normal leukocyte particle. FIG. 3B shows a scatter plot of the forward scattered light signal pulse shape distribution of all leukocyte granules from a normal blood sample. In the forward scattered light signal pulse shape distribution scattergram of fig. 3B, the horizontal axis represents the width of the forward scattered light signal pulse of the white blood cell, and the vertical axis represents the height of the forward scattered light signal pulse of the white blood cell. As shown in the leukocyte region of a normal blood sample, a large number of leukocyte pulses are characterized by varying pulse heights, but close pulse widths, forming a narrow, diffuse spot shape.
With further reference to FIG. 4, the results of testing an abnormal blood sample containing a large number of aggregated leukocytes are shown. FIG. 4A shows a waveform of an abnormal pulse in which a plurality of white blood cells are gathered together to pass through the optical detection region, and the pulse width of the abnormal pulse is significantly wider than that of a normal pulse in which a single white blood cell in the normal blood sample passes through the optical detection region as shown in FIG. 3A. The forward scattered light signal pulse shape distribution scattergram reflected in all leukocyte granules of the abnormal blood sample, see fig. 4B, forms an abnormal pulse region with a large pulse width generated by aggregated leukocytes and a normal pulse region generated by a single leukocyte.
Based on the above phenomenon, in step 103, it is determined whether or not there is leukocyte aggregation in the detected blood sample, particularly based on the pulse width information in the forward scattered light information of leukocytes.
According to an embodiment of the present invention, the total number of the first leukocyte populations is first counted according to the first optical signal information and the number of abnormal leukocyte populations with pulse widths larger than a first predetermined threshold is counted according to the pulse width information. Then, it is determined whether the ratio of the number of abnormal leukocyte particles to the total number of the first leukocyte particle population is greater than a second predetermined threshold. Determining that there is no leukocyte aggregation in the first test sample when the ratio is less than the second predetermined threshold (as shown in FIG. 3B); and determining that there is leukocyte aggregation in the first test sample when the ratio is greater than the second predetermined threshold (as shown in FIG. 4B).
The first predetermined threshold and the second predetermined threshold may both be detection experience values.
Further, the first predetermined threshold may also be determined based on pulse width information in the optical signal information of leukocyte particles in the normal blood sample. For example, in this manner, the first predetermined threshold may be 1.2us or 1.4 us. Alternatively, the first predetermined threshold may be determined based on an average value of pulse widths in the first optical signal information of the leukocyte particles in the detected blood sample. It should be understood that the pulse widths of the particles in the measurement systems of different manufacturers are not exactly the same, and the pulse widths of the particles are related to the characteristics of the optical path, the characteristics of the fluid, etc. in the optical measurement system, so that the first predetermined threshold value can be adjusted according to actual conditions.
The second predetermined threshold is determined according to a different first predetermined threshold determination such that the presence of white blood cell aggregates in the sample is determined when the number of aggregated white blood cells will affect the accuracy of the detection. The second predetermined threshold may be, for example, 5%.
When it is determined that there is no leukocyte accumulation in the first test sample (in the case shown in fig. 3), step 106 is performed to directly output the classification and counting results of the first leukocyte population.
When it is determined that there is leukocyte aggregation in the first test sample (in the case shown in fig. 4), the blood sample is retested by the blood cell analyzer. That is, the blood sample is retested.
In step 104, when it is determined that there is leukocyte aggregation in the first test sample, second optical signal information is acquired of particles of a second test sample that is prepared by subjecting the blood sample to a hemolysis process, a fluorescence staining process, and a dilution process while passing through the optical detection apparatus.
This step 104 differs from the previous step 101 in that the blood sample is subjected to a dilution process.
Referring again to fig. 5, there is shown a pretreatment device including a reaction cell 10 of the blood cell analyzer of the present invention. In the analyzer, first, a small amount of diluent is added to the reaction cell 10 through the diluent addition port 70, then a blood sample is added to the diluent bottom liquid in the cell through the needle port 3 of the sampling needle 2, and after being uniformly mixed, a hemolytic agent and a fluorescent dye are added to the reaction cell 10 through the hemolytic agent addition port 40 and the fluorescent dye addition port 50, thereby preparing a second test sample. Herein, the order of adding the diluent, the hemolytic agent and the fluorescent dye is not limited, and the ratio of the blood sample to the diluent may be a conventional ratio or may be empirically obtained, for example, 1: 10. The ratio of the amount of the blood sample, the hemolytic agent and the fluorescent dye is a conventional ratio, and thus, the ratio of the amount of the blood sample, the diluent, the hemolytic agent and the fluorescent dye may be, for example, 20. mu.L to 200. mu.L to 1000. mu.L to 20. mu.L. After a period of incubation, the second test sample is transferred to the optical detection device through the reaction solution extraction port 60 for detection, so as to obtain second optical signal information of the particles in the second test sample.
The blood sample with leukocyte aggregates shown in fig. 4 was retested to obtain a pulse shape distribution scattergram of the forward scattered light signal of leukocytes from the second test sample as shown in fig. 6. As can be seen in fig. 6, the abnormal pulse on the forward scattered light signal pulse shape scattergram of the second test sample has disappeared. The phenomenon of leukocyte aggregation in the second test sample obtained by the pre-dilution treatment is eliminated, so that a normal pulse morphology scatter diagram can be obtained, and a more accurate leukocyte count can be obtained.
Then, in step 106, the classification and counting results of the first leukocyte population and/or the second leukocyte population of the blood sample subjected to the retest before/after the retest are output. The classification and counting results of the first leukocyte particle group and the second leukocyte particle group may be output, or only the classification and counting results of the second leukocyte particle group subjected to the retest may be output.
Fig. 7 shows a variant of the first embodiment of the invention. As shown in FIG. 7, steps 201 to 206 in this modified embodiment are the same as steps 101 to 106 in the first embodiment, respectively. Except that when it is determined that there is leukocyte aggregation in the first test sample, step 207 of alarming for leukocyte aggregation is performed, and step 208 of recording and storing information that the blood sample of the subject is a leukocyte aggregation sample in association with the characteristic information of the subject is performed.
According to this mode, when it is determined that there is leukocyte aggregation in the first test sample, an alarm is given. The manner of the alarm is not particularly limited, and includes but is not limited to: visual alarms (e.g., flashing lights, displaying text, displaying warning icons, displaying warning colors, etc.), audible alarms (e.g., beeps, voice alarms, other alarm tones, etc.), tactile alarms (e.g., vibrations, etc.), and the like. The alarm may be one of the above-described modes, or may be a combination of two or more modes.
Further, when it is determined that there is leukocyte aggregation in the first test sample, information that the blood sample of the subject is a leukocyte aggregation sample is recorded and stored in association with the characteristic information of the subject. The characteristic information of the subject may include, but is not limited to, the identity information of the subject (e.g., ID number, age (or date of birth), sex, name, etc.), physical condition information of the subject (e.g., current illness, medication, etc.), medical history of the subject, family history of the subject, etc. A plurality of information of the characteristic information of the subject can be recorded, at least including information for determining the identity of the subject and disease information related to the generation of leukocyte aggregates. This embodiment is particularly advantageous when a blood sample of a subject is subject to leukocyte aggregation due to a disease.
The present invention also includes other modified embodiments based on the specific embodiment shown in fig. 7. In one embodiment, only an alarm is provided when it is determined that there is a leukocyte accumulation in the first test sample. In another embodiment, the information of the subject is recorded and stored in association with only characteristic information of the subject when it is determined that there is leukocyte aggregation in the first test sample.
In addition, fig. 7 only shows a manner in which steps 207 and 208 are sequentially performed before step 204, but the embodiment of the present invention is not limited thereto. For example, steps 207 and/or 208 may be performed sequentially, in reverse order, or simultaneously. Further, step 207/208 is not limited to being performed before step 204, for example, either or both may be performed between steps 205 and 206, or after step 206, etc.
With further reference to fig. 8, there is shown a second embodiment of the method of the invention for detecting leukocytes in a blood sample of a subject.
As shown in fig. 8, the second embodiment of the present invention includes: step 310, obtaining characteristic information of the subject; step 320, matching the acquired characteristic information of the subject with stored characteristic information of the subject, wherein the leucocyte aggregation condition is detected; and when no matching information is found, performing a detecting step 330, and when matching information is found, performing a detecting step 340.
In this embodiment, when a blood sample of a subject is to be tested, first, characteristic information of the subject is acquired (step 310). The characteristic information of the subject is as defined above. The identity information of the subject and the necessary disease information can be obtained, such as by inputting, or by reading code information or a chip.
Next, the obtained subject profile is matched with already stored subject profiles in which the presence of leukocyte aggregation was detected (step 320). In particular, matching the subject's identity information with stored subject identity information and matching the subject's disease information, particularly disease information associated with leukocyte aggregation, with stored subject disease information may be included.
When no match is found, the same steps as those in the first embodiment of the present invention described above are performed (step 330).
In step 330, the following steps may be specifically included:
step 331 of obtaining first optical signal information of particles of a first test sample passing through an optical detection device of the blood cell analyzer, wherein the first test sample is prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process.
When it is determined that there is no leukocyte aggregation in the first test sample, step 336 is performed to output a leukocyte classification result of the first test sample.
When it is determined that there is a leukocyte accumulation in the first test sample, the blood sample is retested. That is, in step 334, second optical signal information of particles of the second test sample passing through the optical detection device is obtained, wherein the second test sample is prepared by subjecting the blood sample to hemolysis treatment, fluorescence staining treatment and dilution treatment; step 335, classifying and counting the particles in the second test sample according to the second optical signal information to obtain a second leukocyte particle group; and a step 336 of outputting the results of classification and counting of the first and second populations of cell particles, or only the second population of leukocyte particles.
The steps 331 to 336 are substantially the same as the steps 101 to 106 in the first embodiment, and are not described herein again.
In addition, similarly, the detecting step 330 may further include a step (not shown) of alarming and/or recording and storing information that the blood sample of the subject is a leukocyte aggregation sample in association with the characteristic information of the subject.
When the stored past information of the subject is found to have information matching the information of the subject, it can be determined that there is a possibility of leukocyte aggregation in the blood sample of the subject, and the detecting step 340 is performed, that is, the detecting step of diluting the blood sample of the subject is directly performed.
Step 340 specifically includes steps 344, 345 and 346. Steps 344, 345 and 346 are the same as steps 334, 335 and 336, respectively, and are not described in detail herein.
The invention further provides a blood cell analyzer. The above-described method for detecting leukocytes in a blood sample of a subject can be applied to the blood cell analyzer of the present invention.
According to one embodiment, referring to FIG. 9, a blood cell analyzer 500 of the present invention includes a sampling device, such as a sampling needle (not shown); a pretreatment device 510 comprising at least one reaction cell 511; an optical detection device 520; and a processor 530.
The sampling device is used to draw a blood sample. After the sampling device has aspirated the blood sample, it is transported by a transport device (i.e., tubing) to the pretreatment device 510.
The pre-treatment device is used for pre-treating the blood sample so as to prepare a pre-treated test sample. In the present embodiment, the pretreatment device includes at least one reaction cell 511 for providing a reaction site for the blood sample and the treatment reagent. An example of a pretreatment device used in the present invention is shown in FIG. 5.
Referring again to fig. 5, the pretreatment device shown therein includes at least a reaction cell 10. The sampling needle 2 can inject the sucked blood sample into the reaction cell 10 through the hole provided in the needle head. In the analyzer of the present invention, a plurality of openings are arranged on the lower side wall of the reaction cell 10, and the plurality of openings are respectively communicated with the reservoirs of different sample processing reagents through the pipes.
Among them, the opening 70 is a diluent addition port for adding a diluent to the reaction cell 10 when the test sample (i.e., the second test sample in the foregoing embodiments) is prepared by the blood cell analyzer 500 when it is determined that there is a white blood cell aggregation in the blood sample. The openings 40 and 50 are a hemolytic agent inlet port and a fluorescent dye inlet port, respectively, for adding a suitable hemolytic agent and fluorescent dye to perform a hemolytic treatment and a fluorescent staining treatment on the sample. The opening 60 is a reaction solution extraction port, and the prepared test sample is transferred to the optical detection device 520 for detection.
The reaction tank 10 of the pretreatment apparatus of the present invention is different from that of the conventional pretreatment apparatus. Fig. 2 shows a reaction cell 1 of a conventional pretreatment apparatus. Referring to FIG. 2, the lower side wall of the reaction cell 1 is also provided with a plurality of openings, which are respectively communicated with reservoirs of different sample processing reagents through pipes. Wherein, the openings 4 and 5 are a hemolytic agent inlet and a fluorescent dye inlet respectively, for adding appropriate hemolytic agent and fluorescent dye to perform hemolytic treatment and fluorescent staining treatment on the sample. The opening 6 is a reaction solution extraction port, and the prepared test sample is transferred to an optical detection device for detection.
As can be seen from a comparison of fig. 2 and 5, the reaction cell 10 of the present invention further has a diluent introduction port 70, unlike the conventional reaction cell 1, so that the dilution of the sample is performed in-machine to obtain an accurate result of the leukocyte measurement when it is determined that the sample has the leukocyte accumulation.
The optical detection device 520 is used to make the particles in the pretreated test sample pass through the detection area one by one, so as to detect and output the optical signal information of the particles in the test sample. Specifically, the optical detection device 520 is used to irradiate the blood sample treated by the treatment reagent, i.e., the test sample, with light, collect optical signals generated by each particle in the sample, and convert the optical signals into electrical signals to output optical signal information. The optical signal may be a forward scattered light signal (FSC), a side scattered light signal (SSC), or a fluorescence signal (FL). The optical detection device 520 generally includes a light source 521 and a sheath flow chamber 522 having an aperture 5221, etc., particles in the blood sample can flow in the sheath flow chamber 522 and pass through the aperture 5221 one by one, and light emitted by the light source 521 can irradiate the particles in the aperture 5221 and generate a scattered light signal and/or a fluorescence signal accordingly. The optical detection device 520 may further include a lens group 523, a photo sensor 524 (e.g., a photodiode, a photomultiplier tube, etc.), and an a/D converter respectively disposed in front of and laterally to the aperture, where the a/D converter may be disposed in the processor 530 or separately form a component, so that the lens group 523 may capture corresponding scattered light signals and fluorescence signals, the photo sensor 524 may convert the captured light signals (e.g., scattered light signals and fluorescence signals) into electrical signals, and the electrical signals are a/D converted by the a/D converter to obtain digital signals, which may be output as optical signal information.
And a processor 530 for receiving and processing the optical signal information outputted from the optical detection device 520 to obtain the cellular parameters of the blood sample. Processor 530 is configured to perform the steps of the method of the present invention for detecting leukocytes in a blood sample of a subject. According to a specific embodiment, the processor 530 is configured to perform each step of the first and second embodiments described above. And will not be described in detail herein.
The blood cell analyzer of the present invention further includes a memory (not shown). The memory may be implemented by any type of volatile or non-volatile storage device, or combination thereof. Various types of data may be stored in the memory to support the operation of the blood cell analyzer 500. Examples of such data include, but are not limited to, obtaining optical/electrical signal information, subject information, detection results obtained after processing by processor 520, and/or threshold or normal value ranges for various types of parameters for comparison with the detection results/optical/electrical signal information, and the like.
The blood cell analyzer of the present invention may further comprise a display device (not shown) for displaying the detection result output by the processor.
The present invention further provides an analyzer for detecting leukocytes in a blood sample of a subject.
Fig. 10 is a schematic structural diagram of an analysis apparatus 600 according to an embodiment of the present application, where the analysis apparatus 600 includes at least one processor 601 and a memory 602, and the memory 602 stores instructions executable by the at least one processor 601, and the instructions, when executed by the at least one processor 601, implement the above method for detecting leukocytes in a blood sample of a subject.
Furthermore, the analysis apparatus 600 may further comprise at least one network interface 604 and a user interface 603. The various components in the control device 600 are coupled together by a bus system 605. It is understood that the bus system 605 is used to enable communications among the components. The bus system 605 includes a power bus, a control bus, and a status signal bus in addition to a data bus. For clarity of illustration, however, the various buses are labeled as bus system 605 in fig. 10.
The user interface 603 may include, among other things, a display, a keyboard, a mouse, a trackball, a click wheel, a key, a button, a touch pad, or a touch screen.
It will be appreciated that the memory 602 can be either volatile memory or nonvolatile memory, and can include both volatile and nonvolatile memory. Among them, the nonvolatile Memory may be a Read Only Memory (ROM), a Programmable Read Only Memory (PROM), an Erasable Programmable Read-Only Memory (EPROM), an Electrically Erasable Programmable Read-Only Memory (EEPROM), a magnetic random access Memory (FRAM), a Flash Memory (Flash Memory), a magnetic surface Memory, an optical disk, or a Compact Disc Read-Only Memory (CD-ROM); the magnetic surface storage may be disk storage or tape storage. Volatile Memory can be Random Access Memory (RAM), which acts as external cache Memory. By way of illustration and not limitation, many forms of RAM are available, such as Static Random Access Memory (SRAM), Synchronous Static Random Access Memory (SSRAM), Dynamic Random Access Memory (DRAM), Synchronous Dynamic Random Access Memory (SDRAM), Double Data Rate Synchronous Dynamic Random Access Memory (DDRSDRAM), Enhanced Synchronous Dynamic Random Access Memory (ESDRAM), Enhanced Synchronous Dynamic Random Access Memory (Enhanced DRAM), Synchronous Dynamic Random Access Memory (SLDRAM), Direct Memory (DRmb Access), and Random Access Memory (DRAM). The memory 602 described in embodiments herein is intended to comprise these and any other suitable types of memory.
The memory 602 includes, but is not limited to: the ternary content addressable memory, static random access memory, and the like, are capable of storing a wide variety of data, such as received sensor signals, to support the operation of the analysis device 600.
The Processor 601 may be a Central Processing Unit (CPU, or other general purpose Processor), a Digital Signal Processor (DSP), an Application Specific Integrated Circuit (ASIC), a Field-Programmable Gate Array (FPGA) or other Programmable logic device, discrete Gate or transistor logic, discrete hardware components, etc.
Furthermore, the present invention further provides a computer-readable storage medium. The computer readable storage medium has stored thereon executable instructions that, when executed by the processor 601, perform the steps of the method for detecting leukocytes in a blood sample of a subject as previously described. The computer readable storage medium may be the aforementioned memory or a component thereof, in which the computer program is stored and executed by the processor 601 of the blood cell analyzer to perform the aforementioned method steps. The computer readable storage medium may be FRAM, ROM, PROM, EPROM, EEPROM, Flash Memory, magnetic surface Memory, optical disk or CD-ROM, etc., or may be various devices including one or any combination of the above storage media.
The features mentioned above can be combined with one another as desired, insofar as they are within the scope of the invention. The advantages and features described for the method according to the invention apply in a corresponding manner to the blood cell analyzer according to the invention. The above description is only for the embodiments of the present invention and is not intended to limit the scope of the present invention, and all equivalent structural changes made by using the contents of the present specification and the drawings or directly/indirectly applied to other related technical fields under the inventive concept of the present invention are included in the scope of the present invention.
Claims (23)
- A method of detecting leukocytes in a blood sample of a subject for use in a blood cell analyzer, the method comprising:obtaining first optical signal information of particles in a first test sample, wherein the first test sample is prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process;classifying and counting particles in the first test sample according to the first optical signal information to obtain a first leukocyte particle group;judging whether leukocyte aggregation exists in the first test sample according to pulse width information of first optical signal information of the first leukocyte particle group; andwhen it is determined that there is leukocyte aggregation in the first test sample,acquiring second optical signal information of particles in a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; andclassifying and counting particles in the second test sample according to the second optical signal information to obtain a second leukocyte particle group.
- The method of claim 1, wherein the step of determining whether leukocyte aggregation is present in the first test sample based on pulse width information of the first light signal information of the first leukocyte particle comprises:counting the total number of first leukocyte particle groups according to the first optical signal information and counting the number of abnormal leukocyte particles with pulse widths larger than a first preset threshold value according to the pulse width information;determining whether the ratio of the number of abnormal leukocyte particles to the total number of the first leukocyte particle population is greater than a second predetermined threshold;determining that leukocyte aggregation is present in the first test sample when the ratio is greater than the second predetermined threshold.
- The method of claim 2, wherein the first predetermined threshold is determined based on pulse width information in the light signal information of leukocyte particles in the normal blood sample.
- The method according to claim 2, wherein the first predetermined threshold is determined from a pulse width average in first optical signal information of the first leukocyte population.
- The method of any one of claims 1 to 4, wherein the first and/or second optical signal information comprises a fluorescent signal and a forward scattered light signal.
- The method of claim 5, wherein the first and/or second optical signal information further comprises side scatter optical signals.
- The method of any one of claims 1 to 6, wherein when it is determined that there is leukocyte aggregation in the first test sample, the method further comprises: and alarming the leucocyte aggregation.
- The method of any one of claims 1 to 7, wherein when it is determined that there is leukocyte aggregation in the first test sample, the method further comprises: recording and storing information that the subject's blood sample is a leukocyte aggregate sample in association with the characteristic information of the subject.
- The method of any of claims 1 to 8, wherein the method further comprises: and outputting the classification and counting results of the first leukocyte particle group and/or the second leukocyte particle group.
- A method of detecting leukocytes in a blood sample of a subject for use in a blood cell analyzer, the method comprising:obtaining information of the subject;matching the obtained information of the subject with stored information of the subject in which the presence of the leukocyte accumulation has been detected; anddetecting leukocytes in a blood sample of the subject according to the method of any one of claims 1 to 9 when no matching information is found, andwhen matching information is found, the following steps are performed:acquiring second optical signal information of particles of a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; andclassifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
- A blood cell analyzer, comprising:a sampling device for aspirating a blood sample of a subject;a pretreatment device for pretreating the blood sample to prepare a pretreated test sample;optical detection means for passing particles in the pretreated test sample through detection zones one by one to detect and output optical signal information of the particles in the test sample; anda processor for performing the steps of:obtaining first optical signal information of particles of a first test sample, wherein the first test sample is prepared by subjecting the blood sample to a hemolysis process and a fluorescent staining process;classifying and counting particles in the blood sample according to the first optical signal information to obtain a first leukocyte particle group;judging whether leukocyte aggregation exists in the first test sample according to pulse width information of first optical signal information of the first leukocyte particle group; andwhen it is determined that there is leukocyte aggregation in the first test sample,acquiring second optical signal information of particles of a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; andclassifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
- The blood cell analyzer of claim 11, wherein the processor in performing the step of determining whether there is a white blood cell aggregate in the first test sample based on pulse width information of the first optical signal information of the first population of white blood cells performs the steps of:counting the total number of first leukocyte particle groups according to the first optical signal information and counting the number of abnormal leukocyte particles with pulse widths larger than a first preset threshold value according to the pulse width information;determining whether the ratio of the number of abnormal leukocyte particles to the total number of the first leukocyte particle population is greater than a second predetermined threshold;determining that leukocyte aggregation is present in the first test sample when the ratio is greater than the second predetermined threshold.
- The blood cell analyzer of claim 12, wherein the first predetermined threshold value is determined based on pulse width information in the optical signal information of leukocyte particles in a normal blood sample.
- The blood cell analyzer of claim 12, wherein the first predetermined threshold is determined according to a pulse width average value in the first optical signal information of the first leukocyte population.
- The blood cell analyzer of any one of claims 11 to 14, wherein the first and/or second optical signal information comprises fluorescent signal information and forward scattered light information.
- The hematology analyzer of claim 15, wherein the first and/or second optical signal information further includes side scattered light information.
- The hematology analyzer of any one of claims 11-16, wherein the processor is further configured to output a white blood cell aggregation alarm signal when it is determined that a white blood cell aggregation is present in the first test sample.
- The blood cell analyzer of any one of claims 11 to 17, wherein the processor is further configured to record and store information that the blood sample of the subject is a leukocyte aggregation sample in association with the characteristic information of the subject when it is determined that leukocyte aggregation is present in the first test sample.
- The blood cell analyzer of any one of claims 11 to 18, further comprising a display device for displaying the classification and counting results of the first and/or second leukocyte particle populations output by the processor.
- The blood cell analyzer of any one of claims 11 to 19, wherein the pretreatment device has a diluent addition port for performing a dilution process on the blood sample.
- A blood cell analyzer, comprising:a sampling device for drawing a blood sample from a subject;a pretreatment device for pretreating the blood sample to prepare a pretreated test sample;optical detection means for passing particles in the pretreated test sample through detection zones one by one to detect and output optical signal information of the particles in the test sample;a processor for performing the steps of:obtaining information of the subject;matching the obtained information of the subject with stored information of the subject from which the presence of leukocyte aggregates has been detected, detecting leukocytes in a blood sample of the subject according to the method of any one of claims 1 to 9 when no matching information is found, and performing the following steps when matching information is found:acquiring second optical signal information of particles in a second test sample, wherein the second test sample is prepared by subjecting the blood sample to a hemolysis process, a fluorescent staining process, and a dilution process; andclassifying and counting particles in the blood sample according to the second optical signal information to obtain a second leukocyte particle group.
- An assay device for detecting leukocytes in a blood sample of a subject, comprising:a memory configured to store executable instructions;a processor configured to execute the memory stored executable instructions to perform the method of detecting leukocytes in a blood sample of a subject of any of claims 1-10.
- A computer readable storage medium storing executable instructions, wherein the computer readable storage medium is configured to cause a processor to implement the method of detecting leukocytes in a blood sample of a subject of any of claims 1-10 when the executable instructions are executed.
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| PCT/CN2018/119614 WO2020113527A1 (en) | 2018-12-06 | 2018-12-06 | Method for detecting white blood cells, blood cell analyzer, and storage medium |
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