Evaluation of a novel moving threshold gating strategy for assessment of reticulated platelets in dogs using the ADVIA 2120 analyzer

Anja Kuhn; Kristina Weiler; Graham Gibbs; Gottfried Prechtl; Natali Bauer; Andreas Moritz

doi:10.1111/vcp.13168

Evaluation of a novel moving threshold gating strategy for assessment of reticulated platelets in dogs using the ADVIA 2120 analyzer

Vet Clin Pathol. 2023 Mar;52(1):11-21. doi: 10.1111/vcp.13168. Epub 2022 Oct 25.

Authors

Anja Kuhn¹, Kristina Weiler², Graham Gibbs³, Gottfried Prechtl⁴, Natali Bauer², Andreas Moritz^{1

2}

Affiliations

¹ Clinic for Small Animals, Internal Medicine, Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine, Justus Liebig University, Giessen, Germany.
² Clinical Pathophysiology and Clinical Pathology, Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine, Justus Liebig University, Giessen, Germany.
³ Siemens Healthcare Laboratory Diagnostics, Sudbury, UK.
⁴ Siemens Healthcare Diagnostics GmbH, Vienna, Austria.

PMID: 36284074
DOI: 10.1111/vcp.13168

Abstract

Background: A novel method using a moving threshold (r-PLTmt) to determine canine r-PLTs (reticulated platelets) has been introduced for ADVIA 2120 software v6.11.7.

Objectives: We aimed to evaluate absolute (ar-PLTmt) and percent (%r-PLTmt) prior to and after visual inspection of scattergrams (ar-PLTmtv, %rPLTmtv) compared with flow cytometry (flow) and to determine reference intervals (RIs) in 120 dogs.

Methods: For method comparison, 42 blood samples of healthy and thrombocytopenic dogs were included. Calculation of Spearman's rho, Bland-Altman, and Passing-Bablok analysis was performed. Coefficients of variation (CVs) were determined for three concentration levels.

Results: Moderate correlations between %r-PLTmt and %r-PLTmtv (r_s 0.75-0.76) were seen compared with flow cytometry. The CV for medium %r-PLTs counts assessed with flow cytometry was 12.9%. Comparable CVs were obtained for ar-PLTmt (14.4%) and %r-PLTmt (15.7%), and ar-PLTmtv and %r-PLTmtv (10.9% and 12.9%, respectively). At low and high concentration levels, CVs for % and absolute r-PLTmt/rPLTmtv ranged between 23%-30% and 15%-20%. In patients with microcytic hypochromic erythrocytes, CVs for ar-PLTmt and %r-PLTmt were 36%-66%. Visual inspection of scattergrams resulted in a marked decrease in CV ranging between 15% and 20%. A proportional bias of 10.8% between %r-PLTmt and flow cytometry became lower (9.7%) after visual validation of scattergrams. Passing-Bablok analysis showed proportional and constant error. RIs for r-PLTmt and r-PLTmtv were 0.2%-3.8% and 0.6-10.2 × 10⁹ /L and 0.3%-4.5% and 1.1-10.3 × 10⁹ /L, respectively. Median values for %r-PLTmtv were higher in young adults (≤2 years) than in older dogs (P = 0.03).

Conclusions: r-PLTmt and r-PLTmtv were moderately correlated with flow cytometry. Visual inspection of scattergrams is recommended.

Keywords: automated measurement; canine; flow cytometry; hematology; immature platelets; scattergram.

MeSH terms

Animals
Blood Platelets*
Dogs
Flow Cytometry / veterinary