Accurate And Reproducible Enumeration Of T-, B-, And Nk Lymphocytes Using The Bd Facslyric 10-Color System: A Multisite Clinical Evaluation

Clinical flow cytometry is a reliable methodology for whole blood cell phenotyping for different applications. The BD FACSLyric (TM) system comprises a flow cytometer available in different optical configurations, BD FACSuite (TM) Clinical software, and optional BD FACS (TM) Universal Loader. BD FACSuite Clinical software used with BD (TM) FC Beads and BD CS&T Beads enable universal setup for performance QC, instrument control, data acquisition/storage, online/offline data analysis, and instrument standardization. BD Biosciences sponsored the clinical evaluation of the BD FACSLyric 10-color configuration at seven clinical sites using delinked and de-identified blood specimens from HIV-infected and uninfected subjects to enumerate T-, B-, and NK-lymphocytes with the BD Multitest (TM) reagents (BD Multitest IMK kit and BD Multitest 6-color TBNK). Samples were analyzed on the BD FACSLyric system with BD FACSuite Clinical software, and on the BD FACSCanto (TM) II system with BD FACSCanto clinical software and BD FACS 7-Color Setup beads. For equivalency between methods, data (n = 362) were analyzed with Deming regression for absolute count and percentage of lymphocytes. Results gave R-2 >= 0.98, with slope values > 0.96, and slope ranges between 0.90-1.05. The percent (%) bias values were <10% for T- and NK cells and < 15% for B-cells. The between-site (n = 4) total precision was tested for 5 days (2 runs/day), and gave % coefficient of variation below 10% for absolute cell counts. The stability claims were confirmed (n = 186) for the two BD Multitest reagents. The reference intervals were re-established in male and female adults (n = 134). The analysis by gender showed statistically significant differences for CD3(+) and CD4(+) T-cell counts and % CD4. In summary, the BD FACSLyric and the BD FACSCanto II systems generated comparable measurements of T-, B-, and NK- cells using BD Multitest assays.