14-color single tube for flow cytometric characterization of CD5+ B-LPDs and high sensitivity automated minimal residual disease quantitation of CLL/SLL Journal Article
| Authors: | Goshaw, J. M.; Gao, Q.; Wardrope, J.; Dogan, A.; Roshal, M. |
| Article Title: | 14-color single tube for flow cytometric characterization of CD5+ B-LPDs and high sensitivity automated minimal residual disease quantitation of CLL/SLL |
| Abstract: | Introduction: The diagnosis of CLL/SLL relies on flow cytometric immunophenotyping. Increasing emphasis is being placed on precise detection of the minimal residual disease. Following antigen recommendations of ERIC and ESCCA's Harmonization Project, we validated a 14-color assay for the characterization CD5+ lymphoproliferative neoplasms and CLL MRD with a sensitivity of at least 10−4. Methods: The assay was designed based on ERIC/ESCCA recommended antigens with the addition of CD40 for alternate gating when CD19 expression is reduced. Lower limit of quantitation/lower limit of detection, assay procedural precision, linearity, and limit of blank were established. Then, 52 CD5+ B-cell lymphoproliferative neoplasms (41 CLL/11 non-CLL) and 29 normal samples were used for parallel evaluation. Automated cluster identification and quantitation of CLL clones in MRD setting was performed using Barned-Hutt SNE. Separation analysis between CLL and non-CLL phenotypes was performed by PCA and bh-SNE. Results: Separation ratios for each antigen exceeded ERIC/ESCCA guidelines. Precision was <20% at LLOQ (0.01%). The limit of blank was <10/500,000 cells. Concordance between the 14-color and legacy assay (Deming regression y = 1.01x, r2 =.99) was seen. All 20 samples with MRD levels 0.5%–0.006% (median 0.04%) showed an abnormal cell cluster by bh-SNE, with concordant results between manual and automated quantitation (y = x, r2 = 1). CLL cases clustered together and away from mantle cell lymphoma by bh-SNE and PCA with outlier atypical phenotype CLL cases posing diagnostic challenges by both manual and automated analysis. Conclusion: The 14-color CD5+ LPD assay provides a robust standardization platform for MRD and disease characterization using both manual and automated analysis. © 2020 International Clinical Cytometry Society |
| Keywords: | cll; mantle cell; machine learning; mrd |
| Journal Title: | Cytometry Part B—Clinical Cytometry |
| Volume: | 100 |
| Issue: | 4 |
| ISSN: | 1552-4949 |
| Publisher: | Wiley Blackwell |
| Date Published: | 2021-07-01 |
| Start Page: | 509 |
| End Page: | 518 |
| Language: | English |
| DOI: | 10.1002/cyto.b.21953 |
| PUBMED: | 32896973 |
| PROVIDER: | scopus |
| PMCID: | PMC9448444 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 August 2021 -- Source: Scopus |
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