猫血小板微粒定量的分析验证

Analytical validation of platelet microparticle quantification in cats.

作者信息

Cremer Signe E, Koch Jørgen, Graversen Nanna, Gravgaard Anne S, Langhorn Rebecca, Kristensen Annemarie T, Willesen Jakob L, Nielsen Lise N

机构信息

Department of Veterinary Clinical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

出版信息

Vet Clin Pathol. 2018 Sep;47(3):386-395. doi: 10.1111/vcp.12641.

DOI:10.1111/vcp.12641

PMID:30199121

Abstract

BACKGROUND

Cardiogenic embolism (CE) in cats is a devastating condition primarily associated with hypertrophic cardiomyopathy (HCM). Hypercoagulability may pose a risk for thrombus formation; however, no single test can predict CE development. Platelet microparticles (PMPs) released from platelet membranes are associated with thrombosis in humans.

OBJECTIVES

The aims were to validate flow cytometric PMP quantification in cats analytically and, in a pilot study, evaluate the procoagulant annexin V (AnV) positive PMP concentration in healthy cats and cats with asymptomatic HCM.

METHODS

With CD61 as a platelet marker, CD61 AnV PMPs (0.3-1.0 μm) were quantified in citrated whole blood (WB) and platelet-poor plasma (PPP) using flow cytometry. Analyses were performed in 6 healthy cats and 5 cats with asymptomatic HCM. The coefficient of variation (CV) for duplicate (intra-assay) and parallel (inter-assay) analyses were calculated.

RESULTS

PMP concentrations were quantified with acceptable intra-assay CV for WB (CD61 /AnV ; 2.4%, 0.2%-8.4% (median, range), CD61 /AnV ; 3.8%, 0.1%-12.5%) and PPP (CD61 /AnV ; 5.0%, 0.7%-12.8%, CD61 /AnV ; 7.4%, 0.5%-15.3%), and acceptable inter-assay CV for WB in 10/11 cats (CD61 /AnV ; 6.2%, 1.4%-13.3%, CD61 /AnV ; 6.4%, 0.7%-17.2%), but unacceptable for PPP (CD61 /AnV ; 15.6%, 5.8%-42.7%, CD61 /AnV ; 27.8%, 8.4%-77.1%). For WB PMP concentrations, the pilot data demonstrated no differences between healthy cats and cats with asymptomatic HCM (4/5 with left ventricular outflow obstruction) for either the CD61 /AnV or the CD61 /AnV PMPs.

CONCLUSIONS

Only WB PMP concentrations could be quantified reliably in cats in a clinical setting. PMP concentrations did not differ between healthy and asymptomatic HCM cats in this pilot study.

摘要

背景

猫的心源性栓塞（CE）是一种严重疾病，主要与肥厚型心肌病（HCM）相关。高凝状态可能会增加血栓形成的风险；然而，没有单一的检测方法能够预测CE的发生。从血小板膜释放的血小板微粒（PMPs）与人类的血栓形成有关。

目的

本研究旨在通过分析验证流式细胞术对猫PMP的定量检测，并在一项初步研究中，评估健康猫和无症状HCM猫中促凝血的膜联蛋白V（AnV）阳性PMP的浓度。

方法

以CD61作为血小板标志物，采用流式细胞术对枸橼酸盐抗凝血（WB）和少血小板血浆（PPP）中的CD61 AnV PMPs（0.3 - 1.0μm）进行定量。对6只健康猫和5只无症状HCM猫进行分析。计算重复分析（批内）和平行分析（批间）的变异系数（CV）。

结果

WB中PMP浓度的批内CV可接受（CD61 /AnV ；2.4%，0.2% - 8.4%（中位数，范围），CD61 /AnV ；3.8%，0.1% - 12.5%），PPP中也可接受（CD61 /AnV ；5.0%，0.7% - 12.8%，CD61 /AnV ；7.4%，0.5% - 15.3%）；WB批间CV在10/11只猫中可接受（CD61 /AnV ；6.2%，1.4% - 13.3%，CD61 /AnV ；6.4%，0.7% - 17.2%），但PPP不可接受（CD61 /AnV ；15.6%，5.8% - 42.7%，CD61 /AnV ；27.8%，8.4% - 77.1%）。对于WB中的PMP浓度，初步数据表明，无论是CD61 /AnV还是CD61 /AnV PMPs，健康猫和无症状HCM猫（4/5伴有左心室流出道梗阻）之间均无差异。