Comparison of Flow Cytometric Methods for the Enumeration of Residual Leucocytes in Leucoreduced Blood Products: A Multicenter Study

Abstract

The BD FACSVia™ System features novel designs in hardware, software, and instrument QC. We compared the performance of the BD FACSVia System using the BD Leucocount™ kit with the BD FACSCalibur™ flow cytometer. Leucoreduced platelet (PLT, n = 252) and red blood cell (RBC, n = 278) specimens were enrolled at four sites. Each specimen was stained in four tubes using the BD Leucocount kit reagents and acquired on the two systems. BD Leucocount Control cells (high and low) were used to evaluate the inter‐site reproducibility on the BD FACSVia System at three sites over 20 days. Deming regression and Bland–Altman analysis were performed to determine the WBC absolute counts on the BD FACSVia System vs. the BD FACSCalibur system. Assay accuracy for the range of 0–350 WBCs/µl was adequate. For samples with <25 WBCs/µl, the bias with 95% limits of agreement was 0.136 (–1.897 to 2.169) WBC/µl for PLTs (n = 184) and 0.170 (–2.025 to 2.365) WBC/µl for RBCs (n = 193). For inter‐site reproducibility, the CV% was 6.46% (upper 95% CI 7.16%) for the PLT high control and 9.49% (10.52%) for the PLT low control. The CV% was 7.51% (8.32%) for the RBC high control and 10.76% (11.92%) for the RBC low control. The BD FACSVia System reported equivalent results of WBC absolute counts for leucoreduced PLT and RBC samples compared to the BD FACSCalibur system. The inter‐laboratory reproducibility of the BD FACSVia System met study specifications. © 2018 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC.

Standardization of a compensation matrix for the use in an ISHAGE‐based single‐platform for CD34+ stem cell quantitation on the FACSVia™ flow cytometer

“…The BD FACSVia™ is a new flow cytometer with which the worldwide clinical experience is still limited. Until now, BD FACSVia™ has clinically been validated for the enumeration of residual white blood cells in leukoreduced blood products and for HLA‐B27 typing . The use of BD SCE kit on BD FACSVia™ is a laboratory‐developed test.…”

“…Until now, BD FACSVia™ has clinically been validated for the enumeration of residual white blood cells in leukoreduced blood products and for HLA-B27 typing. 6,11 The use of BD SCE kit on BD FACSVia™ is a laboratory-developed test. In fact, the clinical experience with the use of BD SCE kit on BD FACSVia™ is very limited.…”

“…The beads are employed with the aim of checking the optical and fluidic performances and to adjust compensation on the BD FACSVia™. The clinical software of BD FACSVia™ verifies whether or not BD CS&T bright beads are placed within the target fluorescence channels on the instrument . When BD CS&T beads are run, the instrument QC module of the BD FACSVia™ performs an automated adjustment of fluorescence spillover and, consequently, generates “compensation settings” with the format of a color compensation matrix, namely “Correct Dye by subtracting a percentage of: Dye (%).”…”

Leukoreduction filters: Still stuck on sickle trait red cells

“…10,21,30 In order to ensure that QC requirements are met, blood centers employ testing methods that can measure WBC concentrations below the lower limit of detection for most standard automated hematology analyzers. [31][32][33] A gold standard for rWBC enumeration does not exist, and most U.S. blood centers utilize automated flow cytometry or the ADAM-rWBC (NanoEntek, Waltham, MA, USA), a portable semiautomated fluorescence analyzer that is less costly than flow cytometry and requires less staff training to operate. 31,32 Specifications for rWBC testing utilizing the ADAM-rWBC demonstrate a measuring range from 1 to 100 cells/μl, with ≥80% accuracy and a ≤20% coefficient of variation at rWBC concentrations down to 3 cells/μl.…”

“…[31][32][33] A gold standard for rWBC enumeration does not exist, and most U.S. blood centers utilize automated flow cytometry or the ADAM-rWBC (NanoEntek, Waltham, MA, USA), a portable semiautomated fluorescence analyzer that is less costly than flow cytometry and requires less staff training to operate. 31,32 Specifications for rWBC testing utilizing the ADAM-rWBC demonstrate a measuring range from 1 to 100 cells/μl, with ≥80% accuracy and a ≤20% coefficient of variation at rWBC concentrations down to 3 cells/μl. 34 This range encompasses the 15-20 cells/μl representing a rWBC content of 5 Â 10 6 in most RBC units, but is incapable of quantitating counts >3 Â 10 7 /unit.…”