Analytical validation of platelet microparticle quantification in cats

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

doi:10.1111/vcp.12641

Analytical validation of platelet microparticle quantification in cats

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

^*Corresponding author for this work

1 Citation (Scopus)

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.

Original language	English
Journal	Veterinary Clinical Pathology
Volume	47
Issue number	3
Pages (from-to)	386-395
Number of pages	10
ISSN	0275-6382
DOIs	https://doi.org/10.1111/vcp.12641
Publication status	Published - 2018

Keywords

Annexin
cardiac
CD61
microvesicles
precision

Access to Document

10.1111/vcp.12641

Cite this

@article{e1dbbae8620c43c9a2732811b26ac594,

title = "Analytical validation of platelet microparticle quantification in cats",

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.",

keywords = "Annexin, cardiac, CD61, microvesicles, precision",

author = "Cremer, {Signe E.} and J{\o}rgen Koch and Nanna Graversen and Gravgaard, {Anne S.} and Rebecca Langhorn and Kristensen, {Annemarie T.} and Willesen, {Jakob L.} and Nielsen, {Lise N.}",

year = "2018",

doi = "10.1111/vcp.12641",

language = "English",

volume = "47",

pages = "386--395",

journal = "Veterinary Clinical Pathology",

issn = "0275-6382",

publisher = "Wiley-Blackwell",

number = "3",

}

TY - JOUR

T1 - Analytical validation of platelet microparticle quantification in cats

AU - Cremer, Signe E.

AU - Koch, Jørgen

AU - Graversen, Nanna

AU - Gravgaard, Anne S.

AU - Langhorn, Rebecca

AU - Kristensen, Annemarie T.

AU - Willesen, Jakob L.

AU - Nielsen, Lise N.

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

KW - Annexin

KW - cardiac

KW - CD61

KW - microvesicles

KW - precision

U2 - 10.1111/vcp.12641

DO - 10.1111/vcp.12641

M3 - Journal article

C2 - 30199121

AN - SCOPUS:85053008443

SN - 0275-6382

VL - 47

SP - 386

EP - 395

JO - Veterinary Clinical Pathology

JF - Veterinary Clinical Pathology

IS - 3

ER -

Analytical validation of platelet microparticle quantification in cats

Abstract

Keywords

Access to Document

Fingerprint

Cite this