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. 2022 Nov 29;11(1):1720.
doi: 10.4102/ajlm.v11i1.1720. eCollection 2022.

Commercial DURAClone panels for extending the repertoire of multicolour immunophenotypic panels in an academic flow cytometry laboratory in South Africa

Leanne Swart  1   2 , Melanie Pretorius  1   2 , Denise Lawrie  1   2 , Deborah K Glencross  1   2
Affiliations

Commercial DURAClone panels for extending the repertoire of multicolour immunophenotypic panels in an academic flow cytometry laboratory in South Africa

Leanne Swart et al. Afr J Lab Med. .

Abstract

Background: Commercial multicolour fixed immunophenotyping panels can improve flow cytometric diagnostic immunophenotyping repertoire.

Objective: This study validated the commercially available, standardised Beckman Coulter lyophilised DURAClone RE panels to discriminate specific haematolymphoid subtypes.

Methods: We compared the diagnostic capability of the DURAClone acute leukaemia B (ALB), chronic leukaemia B (CLB), and plasma cells (PC) panels to the predicate second-line panels in Charlotte Maxeke Johannesburg Academic Hospital, Johannesburg, South Africa, from April to August 2020. Clinical diagnostic concordance between the in-house second-line immunophenotyping (the predicate method) and DURAClone was established. The ALB panels tested for precursor B-cell acute lymphoblastic leukaemia (n = 11) or normal bone marrow haematogones (n = 9); CLB panels established haematolymphoid subtypes of mature B-cell lymphoproliferative disorders (B-LPD) (n = 20), while PC panels detected plasma cell dyscrasias (PCD) (n = 17). Flow cytometer setup and data interpretation to discriminate normal and aberrant immunophenotypes were per manufacturer's instructions.

Results: There was 100% clinical diagnostic concordance between the predicate and the test panels for second-line diagnostic investigation of B-ALL (with additional CD56), mature B-LPD (with additional discernment of CD81, ROR-1, CD79b and CD43) and PCD.

Conclusion: The DURAClone CLB exceeded the predicate second-line performance, offering extended second-line diagnostic discernment of mature B-LPD subtypes and discernment of CD5+ B-LPD from other non-CD5+ (or CD5-) B-LPD; likewise, the PC panels enabled discovery of PCD. While ALB testing offered no additional diagnostic advantage over existing predicate investigation, CD58 did offer additional information to discern haematogones from B-ALL.

Keywords: B-cell lymphoproliferative disorder; acute lymphoblastic leukaemia; fixed panels; flow cytometry; immunophenotyping; leukaemia; lymphoma; lyophilised reagent; multicolour; plasma cell dyscrasia.

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Conflict of interest statement

The authors declare that they have no financial or personal relationships that may have inappropriately influenced them in writing this article.

Figures

FIGURE 1
FIGURE 1
Relative expression of CD81 relative to normal T-cells and granulocytes in the CLB tube validated at the Charlotte Maxeke Johannesburg Academic Hospital flow cytometry laboratory in Johannesburg, South Africa, April 2020 to August 2020. Each plot (a, b and c) shows CD81 expression (x-axis) versus frequency (y-axis). In A, there is relative (weak) under-expression of CD81 in a patient with CLL gated on CD19+|CD5+; in B, there is relative over-expression of CD81 in a patient with follicular lymphoma gated on CD19+|CD10+ C is a control experiment of a paediatric sample with 20% haematogones showing bright expression of CD81 on normal precursor B-cells gated on weak CD45+| CD19+. Orange represents the B-cells identified with CD19; the blue population represents background granulocytes identified with CD45 and side scatter, and red represents background T-cells gated on CD5 (all markers are included in the CLB panel).
FIGURE 2
FIGURE 2
Identification of plasma cells using the DURAClone PC tube validated at the Charlotte Maxeke Johannesburg Academic Hospital flow cytometry laboratory in Johannesburg, South Africa, April 2020 to August 2020. In histogram a, negative CD45 expression of plasma cells is shown (purple), with CD45 positive (background) expression noted amongst granulocytes (blue), monocytes (green) and mature lymphocytes (red). The target plasma cell population were primary-gated on bright CD138 and CD38 to reveal aberrant CD117 expression (b) and under-expression of CD81 (c) and CD27 (d).
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