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Reagent and method for differential determination of leukocytes in blood

A white blood cell and red blood cell technology, applied in the field of dissolving agents, can solve problems such as prolonging reaction time, raising temperature, and reducing the working efficiency of automatic analyzers

Inactive Publication Date: 2005-02-09
COULTER INTERNATIONAL CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This elevated temperature requires significantly more complex instrumentation, since the temperature of the reaction must be controlled
In addition, prolonging the reaction time will directly reduce the working efficiency of the automatic analyzer

Method used

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  • Reagent and method for differential determination of leukocytes in blood
  • Reagent and method for differential determination of leukocytes in blood
  • Reagent and method for differential determination of leukocytes in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0057] Solvent compositions were formulated with the following components:

[0058] 1. T-mulz (Harcros Chemical, Inc., 50% potassium salt of organophosphate) 5.0 g

[0059] 2. Sodium lauryl sulfate 1.0g

[0060] 3. Hetoxol STA30 (Heterene, ethoxylated stearyl alcohol) 10.0g

[0061] 4. Pluronic 10R5 (BASF) 15.0g

[0062] 5. Formic acid 0.2ml

[0063] 6. Sodium sulfate 9.0g

[0064] 7. Potassium chloride 6.0g

[0065] 8. BHT pre-dissolved in ethanol 0.05g

[0066] 9. proclin300 (Rohm & Haas Co.) 0.5ml

[0067] 10. PH (adjusted by 4M HCl) 2.2

[0068] 11. Adjust the deionized water to 1L

[0069] To 28ml of EDTA anticoagulated normal whole blood sample, add 1800ml of lysing agent composition, and at room temperature (about 212 ℃), mix the mixture gently by shaking for 9 seconds, after adding the lysing agent composition, about At 13 seconds ready for differential analysis. The blood mixture was maintained under acidic conditions (pH approximately 2.2) and an osmolality...

Embodiment II

[0071] A lyser composition was formulated with the following components and used in a leukocyte subpopulation differentiation assay. To 28 μl of EDTA anticoagulated normal whole blood sample, add 1600 μl of lysing agent composition, and at room temperature (approximately 212° C.), mix the mixture gently by shaking for 8 seconds, about 12 seconds after adding the lysing agent composition Seconds are ready for differential analysis. Figure 3a and 3b Two plots of DC versus turbidity are shown. Figure 3a Corresponding to normal blood samples, Figure 3b Corresponds to a blood sample with elevated eosinophils (manual differential count of 12%).

[0072] 1. Burco TME (Burlington Chemical Co. Inc., 1.5 g of ethoxylated dodecylmercaptan, HLB=13.0)

[0073] 2. Sodium lauryl sulfate 1.0g

[0074] 3. Hetoxol STA30 (Heterene, ethoxylated octadecyl alcohol 8.0g)

[0075] 4. Pluronic L35 (BASF) 10.0g

[0076] 5. Citric acid 17.0ml

[0077] 6. Sodium sulfate 9.0g

[0078] 7. Potas...

Embodiment III

[0082] A lyser composition was formulated with the following components and used in a leukocyte subpopulation differentiation assay. In 28 μl of EDTA anticoagulated normal whole blood sample, add about 1880 μl of lysing agent composition, and at room temperature (about 212 ℃), mix the mixture gently by shaking for 11 seconds, after adding the lysing agent composition about At 15 seconds ready for differential analysis. Samples were then analyzed on an experimental hematology analyzer.

[0083] 1. Tergitol NP-13 (Union Carbide, ethoxylated nonylphenol) 1.0g

[0084] 2. Sodium Tetradecyl Sulfate 1.0g

[0085] 3. Hetoxol STA30 (Heterene) 10.0g

[0086] 4. Formic acid 0.2g

[0087] 5. Sodium sulfate 9.0ml

[0088] 6. Potassium chloride 5.0g

[0089] 7. Pluronic 10R5 15.0g

[0090] 8. Adjust the deionized water to 1L

[0091] 9. PH 2.2

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Abstract

This invention relates to a lytic reagent and a method of using the lytic reagent for automatically determining leukocyte subpopulations in blood. More specifically, the new lytic reagent lyses red blood cells and affects the eosinophils which enables the differentiation of at least one subpopulation of leukocytes. The lytic reagent is an acidic, hypertonic aqueous solution comprising an alkali metal salt of alkyl sulfate, an eosinolytic agent, a nonionic surfactant and a physiological salt. When used in combination with a second lytic reagent system, one is able to obtain at least a five part differential of leukocytes using DC and RF measurements.

Description

field of invention [0001] The present invention relates to a lysing agent and a method for differentiating leukocyte subgroups in a blood sample by suitable electronic instrumentation. Background of the invention [0002] Analysis of white blood cell populations from blood samples is a necessary and fundamental part of the methods for diagnosing many diseases. Measuring eosinophils of white blood cell subsets is important for diagnosing certain diseases. For example, increased numbers of eosinophils are found in Hodgkin's disease, parasitic and allergic diseases. [0003] Traditional blood analysis involves smearing a blood sample on a microscope slide and then viewing and analyzing the slide. This method is extremely time-consuming and there are individual differences in the interpretation of slide analysis. Therefore, a method for automatic analysis of leukocytes using mobile leukocyte counts was developed. An essential step in the method of analyzing white blood cells...

Claims

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Application Information

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IPC IPC(8): G01N33/49
Inventor 李毅C・尤恩格S・C・维兰斯
Owner COULTER INTERNATIONAL CORPORATION