Detection Capability Evaluation of Flow Cytometric Assays using a BD FACSLyric™ Flow Cytometer

Introduces the key validation parameters for flow cytometry: LOB (limit of blank), LLOD (lower limit of detection), and LLOQ (lower limit of quantification). These metrics characterize measurement accuracy and precision at the low end of assay measuring intervals.

Perform a 10-day LOB study across three BD FACSLyric flow cytometers using cell-free plasma extracted from normal peripheral blood. Stain samples in six replicates to empirically establish the highest signal obtained in the absence of the measurement of interest.

Add an abnormal specimen to hematologically normal peripheral blood and assess various target populations at different concentrations. Use the qualitative BD 1-Flow CLPD limited panel to empirically determine the lowest amount of measurement consistently detected.

Dilute whole blood using cell-free plasma to achieve four different concentrations of the population of interest. Analyze these samples using the BD MultiTest Color TBNK reagent to establish the lower limit of quantification with acceptable precision and accuracy.

Determine whether full validation or method verification is required based on the assay's intended use and regulatory status. For IVD or CE-marked assays like BD stem cell enumeration kit or BD 1-Flow CLPD panel, method verification is recommended instead of full validation.

Analyze multiple samples acquired in three to five replicates to complete full detection capability validation strategies. This ensures robust evaluation of the assay's performance across varying conditions and operator variability.