Introduction Numerical chromosomal abnormalities (aneuploidies), present in approximately 30%‐50% of pediatric precursor B‐lineage acute lymphoblastic leukemia (B‐ALL) patients, are commonly identified through a laborious conventional cytogenetic (CG) technique. Flow cytometry (FCM) can identify both physical and fluorescent properties of cells together, and by using fluorescent nucleic‐acid‐binding dyes, FCM can identify variations in total nucleic‐acid content of cells. FxCycleTM Violet dye (FxCV) is a selective DNA‐binding dye which permits simultaneous multiparametric immunophenotyping and cell‐cycle/ploidy assessment in a single assay. To date, only two studies have demonstrated the feasibility of FxCV‐aided FCM‐ploidy analysis in B‐ALL patients and only one of these studies have compared their results with CG‐ploidy. Methodology Blast size‐specific FCM‐ploidy was prospectively analyzed using FxCV‐dye in 109 pediatric B‐ALL patients, and the results were compared with concurrent CG‐ploidy status. Results FCM‐ploidy categorization was feasible in 98% of samples tested and the results were 82% concordant with CG‐ploidy status. We observed significant correlation between DNA content and blast size (r = .823, P < .001) and could demonstrate size differences between diploid vs low‐hyperdiploid (P = .025), diploid vs high‐hyperdiploid (P < .001) and low‐ vs high‐hyperdiploid blasts (P = .007). Conclusion FCM‐ploidy assessment using FxCV dye is a reliable assay and the results closely concur with CG‐based ploidy stratification and risk assessment. Using blast size‐assisted DNA content analysis, the results of FCM‐ploidy analysis can be further fine‐tuned.

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BackgroundPlasma cell leukemia (PCL) is a rare and aggressive plasma cell neoplasm. In PCL, clonal plasma cells comprise ≥20% of the peripheral blood (PB) leukocytes and/or the absolute clonal PB plasma cell count is ≥2×10 9 /L. Primary PCL (PPCL) originates de novo, whereas, secondary PCL (SPCL) evolves from pre-existing multiple myeloma. MethodsClinicohematological features, immunophenotypic profile, and survival of PCL patients were analyzed retrospectively. ResultsBetween January 2007 and December 2014, ten PPCL and four SPCL patients were investigated (8 PPCLs and 3 SPCLs had complete clinical data). All were North Indians, sharing common geography and ethnicity. Our cohort showed less frequent renal failure, more frequent hepatomegaly, and non-secretory type disease. In contrast to western literature, flow cytometric immunophenotyping of our cohort revealed altered expression of CD138 (67%), CD56 (33%), and CD20 (0%). With novel therapeutic agents, these PPCL patients had a median overall survival of 15 months. ConclusionWe highlight that our PPCL patients from North India had distinct clinicohematological and immunophenotypic profiles. The significance of our findings must be tested in a larger patient cohort and must be supported by molecular and cytogenetic investigations to unmask possible significant effects on pathogenesis.

We describe an efficient procedure which could be effectively utilized in resource-limited centers and reduce the frequency of repeat BMA procedures.