Analytical errors in nucleated red blood cell enumeration.

Abstract

BACKGROUND: Enumeration of nucleated red blood cells (NRBCs) in peripheral blood of dogs and cats is performed by manual counting during blood film evaluation. Automated methods have increased precision and accuracy; however, most analyzers cannot distinguish leukocytes and NRBCs. The Sysmex XN-V Series may distinguish NRBCs and leukocytes; however, analytical errors occur. OBJECTIVES: We aimed to investigate cases with discrepant automated and manual NRBC counts, and to evaluate reasons for analytical errors. METHODS: Data from samples with increased NRBCs were collected retrospectively and compared with manual counts performed on blood films using Spearman's correlation, Passing-Bablok agreement analysis, and Bland-Altman comparisons. Precision of the automated method and interobserver agreement of manual counts were evaluated. Cases with discrepant results were investigated. RESULTS: Agreement between the methods was good at ≤1NRBC ×10/L in dogs and cats, and inadequate at ≥1NRBC ×10/L. The automated method demonstrated a negative proportional difference to the manual method. Precision was good for the automated method (overall CV 7.1%) and interobserver agreement for the manual method was poor overall (mean CV 27.3%, range 0%-106.1%). Inaccuracies in NRBC enumeration by the automated method occurred with high hematocrits, the mergence of the cell fragments and leukocyte clouds, and the presence of earlier erythroid precursors. CONCLUSIONS: NRBC enumeration by the WNR channel on the Sysmex XN-1000 V is precise and has good agreement with manual counts in canine and feline blood samples at ≤1NRBC ×10/L. Manual film review is indicated for samples with ≥1NRBC ×10/L, earlier erythroid precursors, samples from greyhounds and dehydrated patients, and if gating errors are noted.