CN103323318A

CN103323318A - Hemocyte analyzer diluent

Info

Publication number: CN103323318A
Application number: CN2013102415688A
Authority: CN
Inventors: 尹小东; 管中来
Original assignee: NANJING PERLONG MEDICAL EQUIPMENT CO Ltd
Current assignee: NANJING PERLONG MEDICAL EQUIPMENT CO Ltd
Priority date: 2013-06-18
Filing date: 2013-06-18
Publication date: 2013-09-25

Abstract

The invention discloses a hemocyte analyzer diluent. Every 1L of diluent comprises 5-25.0g of alkali metal chloride, 0.05-5.0g of ethylenediamine tetraacetic acid or ethylenediamine tetraacetic acid derivative, 0.01-3.2g of pyridine onium salt, pH regulator for regulating the pH value to 6.0-8.0, and the balance of water. The invention solves the problem that the peak shapes of lymphocyte, monocyte and granulocyte in a leucocyte histogram of XFA6000-series and XFA6000 Intelligent-series full-automatic hemocyte analyzers produced by Nanjing Pulang Medical Equipment CO, LTD are not obvious or only the wave peaks of the lymphocyte and granulocyte can appear, and also solves a series of problems of insufficient accuracy of classification of leucocytes; and the diluent does not contain any preservative or cyanide, and thus, does not pollute the environment.

Description

A kind of blood cell analyzer dilution

Technical field

The invention belongs to medical diagnostic equipment and use the reagent field, be specifically related to a kind of blood cell analyzer dilution.

Background technology

Dilution and hemolytic agent are the most frequently used reagent of blood analyser in the medical test, are the key reagents of Measuring hemoglobin (HGB) and leucocyte (WBC) counting or classification.Hemolytic agent makes on the one hand rapidly dissolving of red blood cell (RBC) under the mating reaction of dilution, discharge haemoglobin and be combined the stable haemoglobin dervative of formation with hemolytic agent, so that quantitative determination HGB to be provided; Leucocyte is kept original form on the other hand, and keeps certain volume, for the WBC counting, and comes that according to leukocytic size leucocyte is carried out three and hives off: lymphocyte (LYM), monocyte (Mo), granulocyte (GRA).

Usually, blood cell analysis is to use the diluted blood sample, thereby then analyzing a part has diluted blood sample and has measured red blood cell and platelet parameter, again hemolytic agent is added in all the other blood samples that diluted, go out red blood cell and blood platelet, content of hemoglobin is measured, the mensuration of white blood count and differential in order to carry out.

Because red blood cell, leucocyte, blood platelet and hemoglobin concentration are to the susceptibility of chemical reagent, be difficult to find a kind of both the degraded also can close the combination that needs ground changes the chemical reagent that needs the cell characteristics analyzed and hemoglobin parameters, particularly under the condition that changes operating temperature.The physical parameter of cell can be subject to the impact of diluent ingredient and analytical approach, no matter be automatic, semi-automatic or hand-manipulated, existing dilution has all been stipulated the experimental technique that necessary usefulness is special or has been used proprietary instrument, therefore, existing dilution is can not be general or be used for kinds of experiments instrument or experimental technique.

United States Patent (USP) 4255385 has been reported method and the reagent that is used for measuring the blood sample haemoglobin, the document disclose a kind of for dissolving and dilution whole blood sample in order to carry out the reagent of cell analysis, but this reagent contains the potassium ferricyanide that is useful on Measuring hemoglobin.

United States Patent (USP) 5227304 has been reported a kind of reagent system of using of complexity when the enterprising promoting circulation of blood cell count of Cell Dyn board blood analyser and magnitude classification.This reagent system comprises thinning agent and the scaling agent that a kind of grade is oozed, and wherein thinning agent is comprised of imidazoles organic buffer liquid, antiseptic, inorganic salts and pH adjusting agent, but it is not suitable for other types.

Chinese patent 1247982 has been reported and has been used for the measurement of blood sample haemoglobin and the reagent of determination of white cells, disclose and be used for measuring the hemolytic agent that leucocyte hives off, but (see figure 1) in the leucocyte histogram that this hemolytic agent obtains after use, the crest of monocyte (Mo) is obvious not, easily causes the error of instrument software identification.

Summary of the invention

Goal of the invention: for the problems of the prior art, the purpose of this invention is to provide

Technical scheme: in order to solve the problems of the technologies described above, the invention provides a kind of blood cell analyzer dilution, in the 1L dilution:

Described alkali metal chloride is sodium chloride or potassium chloride, and certain osmotic pressure can be provided.

Described ethylenediamine tetraacetic acid derivant is disodium ethylene diamine tetraacetate or EDTAP dipotassium ethylene diamine tetraacetate.

Described pyridiniujm is pyridine copper or pyridine sodium.

A kind of in described pH buffering agent phosphate buffer, Tris – hydrochloride buffer or the Peng Suan – borate buffer solution.

To match in the hemolytic agent that uses each material composition as follows with the present invention:

Quaternary ammonium salt 5.0～40.0g/L, as: decyl trimethyl ammonium chloride, DTAC, tetradecyl trimethyl ammonium chloride, hexadecyltrimethylammonium chloride, OTAC, ten alkyl trimethyl ammonium bromides, DTAB, Tetradecyl Trimethyl Ammonium Bromide, cetyl trimethyl ammonium bromide, Cetyltrimethylammonium bromide;

Alkali metal chloride 5.0～the 25.0g/L of certain osmotic pressure can be provided; A kind of ethylenediamine tetraacetic acid or ethylenediamine tetraacetic acid derivant 0.05～5.0g/L; Mineral acid such as formic acid, acetic acid, ethane diacid, malonic acid, succinic acid, benzoic acid, maleic acid etc. 0.5～5.0 ‰; 0.5～the 8.0g/L such as pH adjusting agent example hydrochloric acid, sulfuric acid, phosphoric acid, NaOH, potassium hydroxide; Hemolytic agent pH value is between 5.0～8.0

Beneficial effect: the present invention has solved emphatically XFA6000 series that Nanjing Pu Lang medical equipment company limited produces and the serial fully automatic blood cytoanalyze of XFA6000Intelligent leucocyte histogram lymphocyte, monocyte, three peak shapes of granulocyte not obvious problems that lymphocyte and two crests of granulocyte maybe can only occur, with the not accurate enough series of problems that waits of leucocyte three classification that therefore cause, and do not contain antiseptic in the reagent, do not contain prussiate, environmentally safe.

Description of drawings

Fig. 1 is Chinese patent 1247982 leucocyte histograms.

Fig. 2 is that the present invention is at the leucocyte histogram of the enterprising promoting circulation of blood liquid cell analysis of the XFA6000 of Nanjing Pu Lang medical equipment company limited production series.Horizontal ordinate represents volume unit (FL) among the figure, and ordinate represents number of cells.

Fig. 3 is that the present invention is at the leucocyte histogram of the enterprising promoting circulation of blood liquid of the XFA6000Intelligent of Nanjing Pu Lang medical equipment company limited production series fully automatic blood cytoanalyze cell analysis.Horizontal ordinate represents volume (FL) among the figure, and ordinate represents number of cells.

Embodiment

Below in conjunction with accompanying drawing the present invention is further described.

Embodiment 1

Dilution:

Sodium chloride 8.12g/L, sodium hydrogen phosphate 11.2g/L, sodium dihydrogen phosphate 1.9g/L, EDTAP dipotassium ethylene diamine tetraacetate 0.5g/L, pyridine copper 0.5g/L, all the other are water, pH value 7.5.

Hemolytic agent:

Decyl trimethyl ammonium chloride 25.8g/L, Tetradecyl Trimethyl Ammonium Bromide 1.7g/L, sodium chloride 9.2g/L, 1M hydrochloric acid 0.5ml/L, disodium ethylene diamine tetraacetate 2.0g/L, all the other are water for NaOH 1.2g/L, pH value 6.0.

Embodiment 2:

Thinning agent:

Potassium chloride 12.44g/L, boric acid 3.2g/L, borax 1.4g/L, disodium ethylene diamine tetraacetate 1.5g/L, pyridine sodium 0.4g/L, all the other are water, pH value 7.4.

Hemolytic agent:

DTAC 22.4g/L, cetyl trimethyl ammonium bromide 2.2g/L, sodium chloride 8.6g/L, formic acid 1.0ml/L, disodium ethylene diamine tetraacetate 1.5g/L, all the other are water for NaOH 1.2g/L, pH value 5.6.

Embodiment 3

Thinning agent:

Potassium chloride 5g/L, sodium hydrogen phosphate 5.04g/L, sodium dihydrogen phosphate 38g/L, disodium ethylene diamine tetraacetate 0.05g/L, pyridine sodium 0.01g/L, all the other are water, pH value 6.0.

Hemolytic agent:

Tetradecyl trimethyl ammonium chloride 38.0g/L, Cetyltrimethylammonium bromide 2.0g/L, sodium chloride 25g/L, formic acid 5.0ml/L, disodium ethylene diamine tetraacetate 5g/L, all the other are water for NaOH 8g/L, pH value 8.0.

Embodiment 4

Sodium chloride 25g/L, sodium hydrogen phosphate 11.2g/L, sodium dihydrogen phosphate 1.9g/L, EDTAP dipotassium ethylene diamine tetraacetate 5.0g/L, pyridine copper 3.2g/L, all the other are water, pH value 7.5.

Hemolytic agent:

Hexadecyltrimethylammonium chloride 2.8g/L, Tetradecyl Trimethyl Ammonium Bromide 2.2g/L, sodium chloride 5.0g/L, 1M hydrochloric acid 2.5ml/L, disodium ethylene diamine tetraacetate 5.0g/L, all the other are water for NaOH 0.5g/L, pH value 6.0.

Embodiment 5

XFA6000 series and the XFA6000Intelligent series fully automatic blood cytoanalyze produced in Nanjing Pu Lang medical equipment company limited use the embodiment of the invention 1～4 to carry out the leukocyte analysis determination experiment, be divided into two kinds of patterns, experimental result is respectively such as Fig. 2 and Fig. 3.

A) under whole blood mode, the whole blood of 13 μ L and the dilution of 3.5mL mix, sample after the dilution of formation 1:270.

Sample after the dilution of 1:270 is divided into two parts.

Get that the dilution of sample and 3.9mL mixes after the dilution of 1:270 of 15.6 μ L, form the sample of 1:45000, be used for red blood cell and enumeration of thrombocytes and measure.

The leucocyte pond adds the mixing of 1mL dilution again with the hemolytic agent of remaining dilute sample and 0.5mL, forms the sample of 1:385, is used for leukocytic count measurement.

B) under pre-dilution mode, get the anticoagulation of 20 μ L and the dilution of 1.6mL and mix (machine dilutes) outward, sample after the dilution of formation 1:81.

Sample after the dilution of 1:81 is sucked 0.7ml, add the WBC counting chamber.Add the 1.1mL dilution to the WBC counting chamber, sample after the dilution of formation 1:263.

Sample after the dilution of 1:263 is divided into two parts.

Get that the dilution of sample and 3.9mL mixes after the dilution of 1:263 of 15.2 μ L, form the sample of 1:45000, be used for red blood cell and enumeration of thrombocytes and measure.

The hemolytic agent of sample and 0.5mL mixes and adds the 0.5mL dilution again after the remaining 1:263 dilution, forms the sample of 1:378, is used for leukocytic count measurement.

After dilution is finished for the second time, before counting, hemolytic agent is added in the WBC diluted sample with the lysed erythrocyte film, usually red blood cell is Duoed 1000 times than leucocyte, the metering leucocyte must at first be eliminated red blood cell to its interference, the common acellular nuclear of red blood cell, hemolytic agent can play the erythrocytic effect of eliminating in the leucocyte dilution.

The above only is preferred implementation of the present invention; be noted that for those skilled in the art; under the prerequisite that does not break away from the principle of the invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims

1. a blood cell analyzer dilution is characterized in that, in the 1L dilution:

2. dilution according to claim 1 is characterized in that, described alkali metal chloride is sodium chloride or potassium chloride.

3. dilution according to claim 1 is characterized in that, described ethylenediamine tetraacetic acid derivant is disodium ethylene diamine tetraacetate or EDTAP dipotassium ethylene diamine tetraacetate.

4. dilution according to claim 1 is characterized in that, described pyridiniujm is pyridine copper or pyridine sodium.

5. dilution according to claim 1 is characterized in that, a kind of in described pH buffering agent phosphate buffer, Tris – hydrochloride buffer or the Peng Suan – borate buffer solution.