BLOOD ANALYZER AND BLOOD ANALYZING METHOD

• US 20120115159A1
• Filed: 06/30/2010
• Published: 05/10/2012
• Est. Priority Date: 07/03/2009
• Status: Active Grant

• Current Assignee: Sysmex Corporation
• Original Assignee: Sysmex Corporation
• Inventors: Kono, Mari, Takagi, Yuri, Asada, Shoichiro
• Granted Patent: US 8,524,505 B2
• Time in Patent Office: Days
• US Class Current: 435/6.19
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/011 with lysing, e.g. of erythrocytes
• CPC Class Codes: G01N 2015/016 White blood cells
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2021/6493 by alternating fluorescence/transmission or fluorescence/reflection
• CPC Class Codes: G01N 21/51 inside a container, e.g. in an ampoule G01N21/53 takes precedence
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

The present invention provides a blood analyzer and a blood analyzing method capable of obtaining information regarding B lymphocytes and T lymphocytes without using a fluorescence-labeled antibody. The blood analyzer of the present invention includes a blood specimen supplying portion, a sample preparation portion that prepares a measurement sample without using a fluorescence-labeled antibody by mixing a blood specimen supplied from the blood specimen supplying portion, a hemolyzing agent, and a fluorescent dye that stains nucleic acid, a light source, a first detector that detects fluorescence, a second detector that detects scattered light, and information processing portion that classifies lymphocytes based on the intensity of fluorescence and scattered light, and based on the fluorescence intensity of the classified lymphocytes, obtains information regarding B-lymphocytes and T-lymphocytes.

Claims

1. A blood analyzer comprising:

a blood specimen supplying portion for supplying a blood specimen;

a sample preparation portion for preparing a measurement sample without using a fluorescence-labeled antibody by mixing the blood specimen supplied from the blood specimen supplying portion, a hemolyzing agent, and a fluorescent dye that stains nucleic acid;

a light source for irradiating light onto the measurement sample prepared by the sample preparation portion;

a first detector for detecting fluorescence emitted from the measurement sample irradiated with light by the light source;

a second detector for detecting scattered light emitted from the measurement sample irradiated with light by the light source;

an information processing portion for obtaining an indicator representing a particle size distribution of fluorescence intensity of lymphocytes based on intensity of fluorescence detected by the first detector and intensity of scattered light detected by the second detector, and obtaining information regarding a ratio of a B-lymphocyte count to a T-lymphocyte count based on the obtained indicator representing the particle size distribution of fluorescence intensity of lymphocytes; and

an output portion for outputting the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtained by the information processing portion.

2. The blood analyzer according to claim 1, wherein the information processing portion is configured to obtain, as the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count, a value representing a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count;

a value representing of a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count;

information indicating that a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count is in a predetermined range;

information indicating that a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count is in a predetermined range;

or information indicating the presence or absence of an abnormal ratio of a T-lymphocyte count to a B-lymphocyte count.

3. The blood analyzer according to claim 1, wherein the information processing portion is configured to obtain, as the indicator representing the particle size distribution of fluorescence intensity of lymphocytes, a measure of central tendency of fluorescence intensity of lymphocytes, and based on the obtained measure of central tendency, obtain the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

4. The blood analyzer according to claim 1, wherein the information processing portion is configured to obtain, as the indicator representing the particle size distribution of fluorescence intensity of lymphocytes, a dispersion of fluorescence intensity of lymphocytes, and based on the obtained dispersion, obtain the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

5. The blood analyzer according to claim 1, wherein the information processing portion is configured to obtain, as the indicator representing the particle size distribution of fluorescence intensity of lymphocytes, a value representing a bias in a distribution of fluorescence intensity of lymphocytes, and based on the value representing the bias in the distribution, obtain the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

6. The blood analyzer according to claim 5, wherein the information processing portion is configured to obtain, as the value representing the bias in the distribution, kurtosis or skewness of the distribution of fluorescence intensity of lymphocytes, and based on the obtained kurtosis or skewness, obtain the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

7. The blood analyzer according to claim 1, wherein

the information processing portion is configured to compare the indicator representing the particle size distribution of fluorescence intensity of lymphocytes with a predetermined threshold value to determine whether the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal or not, and

the output portion is configured to, in the case where the ratio of the B-lymphocyte count to the T-lymphocyte count is determined to be abnormal by the information processing portion, output information indicating that the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal as the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

8. The blood analyzer according to claim 1, wherein

the information processing portion is configured to obtain a value representing a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count based on the indicator representing the particle size distribution of fluorescence intensity of lymphocytes, and compare the obtained value with a predetermined threshold value to determine whether the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal or not, and

the output portion is configured to, in the case where the ratio of the B-lymphocyte count to the T-lymphocyte count is determined to be abnormal by the information processing portion, output information indicating that the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal as the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

9. The blood analyzer according to claim 1, wherein

the information processing portion is configured to obtain a value representing a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count based on the indicator representing the particle size distribution of fluorescence intensity of lymphocytes, and compare the obtained value with a predetermined threshold value to determine whether the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal or not, and

the output portion is configured to, in the case where the B-lymphocyte count to T-lymphocyte count ratio is determined to be abnormal by the information processing portion, output information indicating that the ratio of the B-lymphocyte count to the T-lymphocyte count is abnormal as the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

10. The blood analyzer according to claim 1, wherein the information processing portion is configured to distinguish lymphocytes contained in the measurement sample from other leucocytes based on the intensity of fluorescence detected by the first detector and the intensity of scattered light detected by the second detector, and obtain the indicator representing the particle size distribution of fluorescence intensity of lymphocytes.

11. The blood analyzer according to claim 10, wherein

the information processing portion is configured to distinguish lymphocytes contained in the measurement sample from other leucocytes to classify leucocytes, and

the output portion is configured to output a result of leucocyte classification performed by the information processing portion.

12. The blood analyzer according to claim 11, wherein

the information processing portion is configured to classify leucocytes contained in the measurement sample into at least four groups including lymphocytes.

13. The blood analyzer according to claim 1, wherein the sample preparation portion is configured to prepare the measurement sample by mixing a blood specimen supplied from the blood specimen supplying portion, a hemolyzing agent containing a cationic surfactant and a nonionic surfactant, and a fluorescent dye that stains nucleic acid.

14. A blood analyzing method comprising the steps of:

preparing a measurement sample without using a fluorescence-labeled antibody by mixing a blood specimen, a hemolyzing agent, and a fluorescent dye that stains nucleic acid;

irradiating light onto the prepared measurement sample;

detecting both fluorescece and scattered light emitted from the measurement sample irradiated with light;

obtaining an indicator representing a particle size distribution of fluorescence intensity of lymphocytes based on intensity of fluorescence and intensity of scattered light detected;

obtaining information regarding a ratio of a B-lymphocyte count to a T-lymphocyte count based on the obtained indicator representing the particle size distribution of fluorescence intensity of lymphocytes; and

outputting the obtained information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count.

15. The blood analyzing method according to claim 14, wherein the step of obtaining the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtaines, as the information, a value representing a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count;

a value representing of a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count;

information indicating that a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count is in a predetermined range;

information indicating that a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count is in a predetermined range;

or information indicating the presence or absence of an abnormal ratio of a T-lymphocyte count to a B-lymphocyte count.

16. The blood analyzing method according to claim 14,

wherein the step of obtaining the indicator representing the particle size distribution of fluorescence intensity of lymphocytes obtaines, as the indicator, a measure of central tendency of fluorescence intensity of lymphocytes, and

wherein the step of obtaining the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtaines the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count based on the obtained measure of central tendency.

17. The blood analyzing method according to claim 14,

wherein the step of obtaining the indicator representing the particle size distribution of fluorescence intensity of lymphocytes obtaines, as the indicator, a dispersion of fluorescence intensity of lymphocytes, and

wherein the step of obtaining the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtaines the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count based on the obtained dispersion.

18. The blood analyzing method according to claim 14,

wherein the step of obtaining the indicator representing the particle size distribution of fluorescence intensity of lymphocytes obtaines, as the indicator, a value representing a bias in a distribution of fluorescence intensity of lymphocytes, and

wherein the step of obtaining the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtaines the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count based on the value representing the bias in the distribution.

19. The blood analyzing method according to claim 18,

wherein the step of obtaining the indicator representing the particle size distribution of fluorescence intensity of lymphocytes obtaines, as the value representing the bias in the distribution, kurtosis or skewness of the distribution of fluorescence intensity of lymphocytes, and

wherein the step of obtaining the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count obtaines the information regarding the ratio of the B-lymphocyte count to the T-lymphocyte count based on the obtained kurtosis or skewness.

20. The blood analyzing method according to claim 14, wherein the step of obtaining the indicator representing the particle size distribution of fluorescence intensity of lymphocytes distinguishes lymphocytes contained in the measurement sample from other leucocytes based on the detected intensity of fluorescence and the detected intensity of scattered light, and obtaines the indicator.