A comparison of manual counting of rabbit reticulocytes with ADVIA 2120i analyzer counting

Abstract

We compared manual counting of reticulocytes in rabbits with automatic counting using an ADVIA 2120i analyzer. Reproducibility and the influence of different anticoagulants (EDTA and Li-heparin) were also examined. Blood samples of 331 rabbits (method comparison, n = 289; reproducibility, n = 33; comparison of anticoagulants, n = 9) were tested. The reticulocyte numbers of each specimen were manually determined twice for method comparison. Passing–Bablok regressions, Bland–Altman plots, and the coefficient of variation (CV) were used to evaluate statistical significance. Good correlation (rs = 0.81) was observed between manual reticulocyte counting (groups 1–4) and the ADVIA 2120i. Quantification with the ADVIA 2120i was reproducible for relative reticulocyte numbers (EDTA, CV = 4.24%; Li-heparin, CV = 3.63%) and absolute reticulocyte numbers (EDTA, CV = 5.64%; Li-heparin, CV = 3.81%). The absolute and relative reticulocyte numbers were significantly higher in Li-heparin samples than in EDTA samples (absolute, p = 0.009; relative, p = 0.016). The ADVIA 2120i is suitable for counting reticulocytes in rabbit blood samples, but reticulocyte numbers are higher by manual counting than by ADVIA 2120i counting. Therefore, microscopic confirmation of quantifications is recommended when high numbers of reticulocytes are observed. The anticoagulant of choice is EDTA.