阅读说明：本技术 血小板模拟粒子及其制备方法以及含该模拟粒子的质控物或校准物 (Platelet simulation particle, preparation method thereof and quality control substance or calibrator containing the simulation particle ) 是由 孙思祥 谢键 于 2018-12-25 设计创作，主要内容包括：一种制备血小板模拟粒子的方法,其中使用三氟甲基类化合物对动物红细胞进行修饰处理,以制成能结合荧光染料的血小板模拟粒子。该方法能使血小板模拟粒子与荧光染料分子简单、有效地结合。还涉及另一种制备血小板模拟粒子的方法,其中使用单宁酸和醛类对动物红细胞进行固定处理,以制成血小板模拟粒子。该方法能提高血小板模拟粒子的稳定性。此外还涉及按照该方法制备的血小板模拟粒子及含有该血小板模拟粒子的质控物或校准物。(A method for producing platelet-mimetic particles, wherein animal erythrocytes are subjected to a modification treatment with a trifluoromethyl compound to produce platelet-mimetic particles capable of binding a fluorescent dye. The method can simply and effectively combine the platelet simulation particles with the fluorescent dye molecules. Also relates to another method for preparing platelet-mimetic particles, in which animal erythrocytes are subjected to a fixation treatment using tannic acid and aldehydes to prepare platelet-mimetic particles. The method can improve stability of platelet-simulated particle. Also disclosed are platelet-simulating particles produced by the method, and a quality control material or calibrator containing the platelet-simulating particles.)

A method for producing platelet-mimetic particles, wherein animal erythrocytes are modified with a trifluoromethyl compound to produce platelet-mimetic particles capable of binding a fluorescent dye.

The method according to claim 1, wherein the trifluoromethyl-like compound is an aldehyde substituted with at least one trifluoromethyl group and/or an acid substituted with at least one trifluoromethyl group.

The method of claim 2, wherein the aldehyde substituted with at least one trifluoromethyl group has the structure of formula I:

R ₁-CHO (I)；

the structure of the acid substituted with at least one trifluoromethyl group is shown in formula II:

R ₂-COOH (II)；

wherein R is₁And R₂Are each H, C₁-C ₄Alkyl radical, C₆-C ₁₀Aryl radical, C₇-C ₁₀Phenylalkyl and C₇-C ₁₀An alkyl phenyl group.

A process according to any one of claims 1 to 3, wherein the trifluoromethyl-based compound is selected from the group consisting of p-trifluoromethylbenzaldehyde, 2, 4-bistrifluoromethylbenzaldehyde, p-trifluoromethylbenzoic acid, trifluoroacetic acid and trifluoroacetaldehyde.

The method according to any one of claims 1 to 4, wherein the animal erythrocytes are fixed with a fixing agent before or after the animal erythrocytes are modified with the trifluoromethyl compound.

The method of claim 5, wherein the step of fixing the animal red blood cells with a fixative comprises: fixing the animal erythrocytes by using aldehydes;

preferably, the aldehydes are selected from one or more of the group consisting of: paraformaldehyde, formaldehyde, acrolein, malondialdehyde, glutaraldehyde and adipaldehyde, the aldehydes including glutaraldehyde in particular;

preferably, the reaction concentration of the aldehydes is 0.0075 v/v% to 0.025 v/v%, preferably 0.01 v/v% to 0.0125 v/v%.

The method of claim 6, wherein the step of fixing the animal red blood cells with a fixative further comprises: fixing the animal red blood cells by using tannic acid;

preferably, the reaction concentration of the tannic acid is 5mg/L to 75mg/L, preferably 15mg/L to 37.5mg/L, and more preferably 15mg/L to 25 mg/L.

A method for producing platelet-mimetic particles, wherein animal erythrocytes are subjected to a fixation treatment using tannic acid and aldehydes to produce platelet-mimetic particles.

The method of claim 8, wherein the aldehydes are selected from one or more of the group consisting of: paraformaldehyde, formaldehyde, acrolein, malondialdehyde, glutaraldehyde and adipaldehyde, the aldehydes including in particular glutaraldehyde;

preferably, the reaction concentration of the aldehydes is 0.0075 v/v% to 0.025 v/v%, preferably 0.01 v/v% to 0.0125 v/v%.

The method of claim 8 or 9, wherein the reaction concentration of tannic acid is 5 to 75mg/L, preferably 15 to 37.5mg/L, more preferably 15 to 25 mg/L.

The method of any one of claims 8 to 10, wherein the step of immobilizing the animal red blood cells using tannic acid and aldehydes comprises:

firstly, tannin is used for fixing the animal red blood cells, and then aldehydes are used for fixing the animal red blood cells; or

The animal erythrocytes are fixed by aldehydes and then by tannic acid.

The method of any of claims 8 to 11, wherein the method further comprises:

before or after the fixing treatment, the animal red blood cells are subjected to a modification treatment so that the animal red blood cells can generate a fluorescent signal in a platelet detection channel.

The method of claim 12, wherein the step of modifying the animal red blood cells to produce a fluorescent signal in the platelet detection channel comprises:

modifying said animal erythrocytes with a trifluoromethyl compound such that said animal erythrocytes are capable of binding a fluorescent dye, or

And (3) dyeing the animal red blood cells by using a fluorescent dye.

The method according to claim 13, wherein the trifluoromethyl-like compound is an aldehyde substituted with at least one trifluoromethyl group and/or an acid substituted with at least one trifluoromethyl group.

The method of claim 15, wherein the aldehyde substituted with at least one trifluoromethyl group has the structure of formula I:

R ₁-CHO (I)；

the structure of the acid substituted with at least one trifluoromethyl group is shown in formula II:

R ₂-COOH (II)；

wherein R is₁And R₂Are each H, C₁-C ₄Alkyl radical, C₆-C ₁₀Aryl radical, C₇-C ₁₀Phenylalkyl and C₇-C ₁₀An alkyl phenyl group.

The method according to claim 15, wherein the trifluoromethyl-based compound is selected from the group consisting of p-trifluoromethylbenzaldehyde, 2, 4-bistrifluoromethylbenzaldehyde, p-trifluoromethylbenzoic acid, trifluoroacetic acid and trifluoroacetaldehyde.

The method according to any one of claims 1 to 16, wherein the animal red blood cells are derived from a mammal, preferably from a goat or sheep.

A platelet-mimetic particle prepared according to the method of any one of claims 1 to 17.

A quality control substance or calibrator for a blood analyzer, wherein the quality control substance or calibrator contains the platelet-mimetic particles according to claim 18.

The quality control substance or calibrator of claim 19, wherein the quality control substance or calibrator further comprises at least one of leukocyte-mimicking particles, erythrocyte-mimicking particles, and nucleated red blood cell-mimicking particles.