Blood analyzer and blood analyzing method

• US 8,524,505 B2
• Filed: 06/30/2010
• Issued: 09/03/2013
• Est. Priority Date: 07/03/2009
• Status: Active Grant

• Current Assignee: Sysmex Corporation
• Original Assignee: Sysmex Corporation
• Inventors: Kono, Mari, Takagi, Yuri, Asada, Shoichiro
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US13/381,052
• Publication Number:
  US 20120115159A1
• Time in Patent Office: 1,161 Days
• Field of Search: 435/6, 435/372.2, 435/372.3, 435/287.2, 435/7.24, 435/287.1, 436/546, 436/10, 436/64, 436/164, 436/171, 436/172, 436/800, 436/56, 422/73, 422/82.05, 422/82.08, 422/82.09
• US Class Current: 436/164
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/0069 with lysing, e.g. of erythrocyts
• CPC Class Codes: G01N 2015/008 White cells
• 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 that contains a cationic surfactant and a nonionic surfactant, 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 each of cells contained in the measurement sample irradiated with light by the light source to output fluorescence intensity of each of the cells;

a second detector for detecting scattered light emitted from each of the cells contained in the measurement sample irradiated with light by the light source to output scattered light intensity of each of the cells;

an information processing portion configured to;

a) distinguish lymphocytes from leucocytes contained in the cells based on fluorescence intensities outputted from the first detector and scattered light intensities outputted from the second detector;

b) obtain an indicator selected from the group consisting of a measure of central tendency of fluorescence intensities of the distinguished lymphocytes, a dispersion of fluorescence intensities of the distinguished lymphocytes, a kurtosis of a particle distribution of fluorescence intensities of the distinguished lymphocytes and a skewness of the particle distribution of fluorescence intensities of the distinguished lymphocytes; and

c) obtain a ratio between a B-lymphocyte count and a T-lymphocyte count based on the obtained indicator in b) and a predetermined relationship value between B-lymphocyte and T-lymphocyte, wherein the relationship value directs one-to-one relationship between the indicator and the ratio; and

an output portion for outputting the ratio obtained by the information processing portion.

2. The blood analyzer according to claim 1, wherein the ratio includes at least one of a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count and;

a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count.

3. The blood analyzer according to claim 1, wherein

the information processing portion is configured to compare the indicator with a predetermined threshold value to determine whether the ratio is abnormal or not, and

the output portion is configured to, in the case where the ratio is determined to be abnormal by the information processing portion, output that the ratio is abnormal.

4. The blood analyzer according to claim 1, wherein

the information processing portion is configured to:

obtain a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count based on the indicator and the predetermined relationship value; and

compare the obtained ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count with a predetermined threshold value to determine whether the ratio between the B-lymphocyte count and the T-lymphocyte count is abnormal or not, and

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

5. The blood analyzer according to claim 1, wherein

the information processing portion is configured to:

obtain a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count based on the indicator and the predetermined relationship value; and

compare the obtained ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count with a predetermined threshold value to determine whether the ratio between the B-lymphocyte count and the T-lymphocyte count is abnormal or not, and

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

6. The blood analyzer according to claim 1, wherein

the information processing portion is configured to distinguish lymphocytes from leucocytes contained in the cells by classifying the leucocytes, and

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

7. The blood analyzer according to claim 6, wherein

the information processing portion is configured to classify the leucocytes contained in the cells into at least four groups including one group of lymphocytes.

8. 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 containing a cationic surfactant and a nonionic surfactant, and a fluorescent dye that stains nucleic acid;

irradiating light onto the prepared measurement sample;

detecting fluorescence and scattered light emitted from each of cells contained in the measurement sample irradiated with light to output fluorescence intensity and scattered light intensity of each of the cells;

distinguishing lymphocytes from leucocytes contained in the cells based on outputted fluorescence intensities and outputted scattered light intensities;

obtaining an indicator selected from the group consisting of a measure of central tendency of fluorescence intensities of the distinguished lymphocytes, a dispersion of fluorescence intensities of the distinguished lymphocytes, a kurtosis of the particle distribution of fluorescence intensities of the distinguished lymphocytes and a skewness of the particle distribution of fluorescence intensities of the distinguished lymphocytes;

obtaining a ratio between a B-lymphocyte count and a T-lymphocyte count based on the obtained indicator and a predetermined relationship value between B-lymphocyte and T-lymphocyte, wherein the relationship information directs one-to-one relationship between the indicator and the ratio; and

outputting the obtained ratio.

9. The blood analyzing method according to claim 8, wherein the ratio between the B-lymphocyte count and the T-lymphocyte count includes at least one of a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count and a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count.

10. 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 that contains a cationic surfactant and a nonionic surfactant, 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 each of cells contained in the measurement sample irradiated with light by the light source to output fluorescence intensity of each of the cells;

a second detector for detecting scattered light emitted from each of cells contained in the measurement sample irradiated with light by the light source to output scattered light intensity of each of the cells;

an information processing portion configured to;

a) distinguish lymphocytes from leucocytes contained in the cells based on fluorescence intensities outputted from the first detector and scattered light intensities outputted from the second detector;

b) obtain an indicator selected from the group consisting of a measure of central tendency of fluorescence intensities of the distinguished lymphocytes, a dispersion of fluorescence intensities of the distinguished lymphocytes, a kurtosis of a particle distribution of fluorescence intensities of the distinguished lymphocytes and a skewness of the particle distribution of fluorescence intensities of the distinguished lymphocytes;

c) obtain a ratio between a B-lymphocyte count and a T-lymphocyte count based on the obtained indicator and a predetermined relationship value between B-lymphocyte and T-lymphocyte, wherein the relationship value directs one-to-one relationship between the indicator and the ratio; and

d) compare the indicator with a predetermined threshold value to determine whether the ratio is abnormal or not; and

an output portion for outputting that the ratio is abnormal, in the case where the ratio is determined to be abnormal by the information processing portion.

11. The blood analyzer according to claim 10, wherein the ratio includes at least one of a ratio of a B-lymphocyte count to a total B-lymphocyte and T-lymphocyte count, and a ratio of a T-lymphocyte count to a total B-lymphocyte and T-lymphocyte count.

12. The blood analyzer according to claim 10, wherein the information processing portion is configured to distinguish lymphocytes from leucocytes contained in the cells by classifying the leucocytes, and

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

13. The blood analyzer according to claim 10, wherein

the information processing portion is configured to classify the leucocytes contained in the cells into at least four groups including one group of lymphocytes.