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Blast size‐specific flowcytometric ploidy assessment using FxCycle TM Violet dye and its correlation with conventional cytogenetic ploidy in pediatric precursor B‐lineage acute lymphoblastic leukemia patients
Author(s) -
Bommannan Karthik,
Arumugam Jhansi Rani,
Koshy Teena,
Radhakrishnan Venkatraman,
Sagar Tenali Gnana,
Sundersingh Shirley
Publication year - 2021
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/ijlh.13436
Subject(s) - ploidy , flow cytometry , biology , immunophenotyping , aneuploidy , nucleic acid , microbiology and biotechnology , cytometry , enumeration , genetics , gene , chromosome , combinatorics , mathematics
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. FxCycle TM 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.