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A cyanide-free lytic reagent compositiona nd method for hemoglobin and cell analysis

A technology for lysing reagents and hemoglobin, which is applied in biological testing, analytical materials, biological material analysis, etc., and can solve problems such as counting white blood cells without instructions

Inactive Publication Date: 2000-02-09
COULTER INTERNATIONAL CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, Kim does not state the ability to count white blood cells or distinguish white blood cell subpopulations
[0013] None of the cyanide-free hemoglobin assays described above provide monocyte and whole granulocyte counts at the same time as hemoglobin; however, both parameters are valuable tools for clinical diagnosis of various diseases

Method used

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  • A cyanide-free lytic reagent compositiona nd method for hemoglobin and cell analysis
  • A cyanide-free lytic reagent compositiona nd method for hemoglobin and cell analysis
  • A cyanide-free lytic reagent compositiona nd method for hemoglobin and cell analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Reagents of the following compositions were prepared. Formulation 1a tetrazole 5.0g tetradecyltrimethylammonium bromide 15.0g distilled water to adjust to 1 liter pH 2.77 Formulation 1b triazole 10.0g dodecyltrimethylammonium chloride (50% solution) 35.0ml tetradecyltrimethylammonium Alkyltrimethylammonium bromide 3.5g distilled water to adjust to 1 liter pH 6.19 recipe 1c quinalcinic acid 5.0g dodecyltrimethylammonium chloride (50% solution) 50.0ml distilled water to adjust to 1 liter pH 2.55 recipe 1d Melamine 5.0g Tetradecyltrimethylammonium Bromide 5.0g Dodecyltrimethylammonium Chloride (50% solution) 36.0ml Distilled water Adjust to 1 liter pH 4.55 Formulation 1e Tetrazole 5.0g Chemfac NB-104 (complex phosphate ester manufactured by Chemax Corporation) 30.0 g of distilled water adjusted to 1 liter of pH 7.11

[0071] With 2500 μl of ISOTON  II Dilute 11.6 μl of whole blood sample, then manually mix 403 μl of one of the above lysing reagent compositions with the ...

Embodiment 2

[0073] Reagents of the following compositions were prepared. Tetrazole 5.0g dodecyltrimethylammonium chloride (50% solution) 36.0ml tetradecylammonium bromide 3.6g distilled water 1 liter pH 2.73

[0074] 11.6 μl of whole blood samples were diluted with 2500 μl of blood thinner, then 403 μl of the above lysing reagent composition were manually mixed with the pre-diluted samples. Absorption spectra of the samples were immediately measured on a BeekmanDU 7500 spectrophotometer. Five commercial blood thinners were used for hemoglobin determination, the COULTER  ISOTON  III.COULTER  ISOTON  II. Technicon H TM System RBC DIL (commercial from Technicon Instruments), standard phosphate buffered saline and standard sodium chloride saline. The chromogen formed has an absorption maximum at about 540 nm and a shoulder at about 565 nm. When different diluents are used for the determination of hemoglobin, the maximum absorption of the chromogen is only slightly different, that i...

Embodiment 3

[0076] Reagents of the following compositions were prepared. Recipe 3a Tetrazole 5.0g Tetradecyltrimethylammonium bromide 15.0g Distilled water Adjust to 1 liter pH 12.06 Recipe 3b Aniline-2-sulfonic acid 5.0g Tetradecyltrimethylammonium bromide 10.0g Hexadecane Trimethylammonium bromide 4.0g distilled water 1 liter pH 1.67 Recipe 3c quinaldixic acid 5.0g trimethylammonium lauryl chloride (50% solution) 40.0ml distilled water 1 liter pH 2.58

[0077] About 70 blood samples (with a hemoglobin concentration of 6-17g / dL and a white blood cell count of 1,000 / μL-40,000 / μL) were collected on a calibrated COULTER  On the STKS instrument under standard instrument settings (except for lysing reagents, LYSE S  III diff was replaced by the above preparation) analysis. For each formulation, approximately half of the samples were clinical samples for various diseases. In the case of formulation 3a, 70% of the samples were clinical samples. Figure 5 shows that by using the reference r...

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Abstract

The present invention relates to a cyanide-free lytic reagent composition and method for measuring the total hemoglobin concentration in a blood sample, for counting the number of leukocytes and for differential counting of three leukocyte subpopulations including lymphocytes, monocytes, and granulocytes. The cyanide-free lytic reagent composition comprises a hemolytic surfactant selected from the group consisting of quaternary ammonium salts, pyridinium salts, organic phosphate esters, and alkyl sulfonates to lyse erythrocytes and release hemoglobin, and an organic ligand selected from the group consisting of triazole and its derivatives, tetrazole and its derivatives, alkaline metal salts of oxonic acid, melamine, aniline-2-sulfonic acid, quinaldic acid, 2-amino-1,3,4-thiadiazole, triazine and its derivatives, urazole, DL-pipecolinic acid, isonicotinamide, anthranilonitrile, 6-aza-2-thiothymine, adenine, 3-(2-thienyl)acrylic acid, benzoic acid and alkali metal and ammonium salts of benzoic acid, and pyrazine and its derivatives to form a stable chromogen with hemoglobin. Alternatively, the organic ligands can be added to a suitable blood diluent for hemoglobin and leukocyte analysis.

Description

Background of the invention [0001] The technical field of the invention [0002] The present invention relates to lysing reagent compositions, diluents and methods for determining the concentration of total hemoglobin in a blood sample (manually or automatically) and for simultaneously performing leukocyte counts or differential counts of leukocyte subpopulations. [0003] discussion of prior art [0004] The measurement of total hemoglobin is an indicator of the oxygen-carrying capacity of whole blood. Examination of clinically symptomatic patients and electrophoretic studies of clinically normal populations have revealed more than 300 abnormal hemoglobins. Many of these abnormalities lead to clinical pathology of altered hemoglobin levels or altered ability of hemoglobin to bind oxygen. These diseases are sickle cell anemia, alpha and beta-thalassemia, and hemoglobin M, among others. [0005] The ability to measure hemoglobin (Hgb) in blood samples is a very important pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/49G01N33/50G01N33/569G01N33/72
CPCG01N33/56972Y10T436/101666Y10T436/10Y10T436/105831G01N33/5094G01N33/721Y10T436/108331Y10T436/107497Y10T436/25125
Inventor 李毅C·永
Owner COULTER INTERNATIONAL CORPORATION
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