Maria Arroz1, Neil Came2, Pei Lin3, Weina Chen4, Constance Yuan5, Anand Lagoo6, Mariela Monreal7, Ruth de Tute8, Jo-Anne Vergilio9, Andy C Rawstron8, Bruno Paiva10. 1. Department of Clinical Pathology, Cytometry Laboratory, CHLO, Hospital S. Francisco Xavier, Lisbon, Portugal. 2. Pathology Department, Peter MacCallum Cancer Centre, Melbourne, Australia. 3. Department of Hematopathology, MD Anderson Cancer Center, Houston, Texas, USA. 4. Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. 5. Laboratory of Pathology, NCI, NIH, Bethesda, Maryland, USA. 6. Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA. 7. Interpflow Corporation, Miami, Florida, USA. 8. HMDS, Department of Haematology, St. James's Institute of Oncology, Leeds, United Kingdom. 9. University Of Michigan Medical Center Hematology Oncology Laboratory, Ann Arbor, Michigan, USA. 10. Clinica Universidad de Navarra, Centro de Investigación Médica Aplicada, University of Navarra, Pamplona, Spain.
Abstract
BACKGROUND: Major heterogeneity between laboratories in flow cytometry (FC) minimal residual disease (MRD) testing in multiple myeloma (MM) must be overcome. Cytometry societies such as the International Clinical Cytometry Society and the European Society for Clinical Cell Analysis recognize a strong need to establish minimally acceptable requirements and recommendations to perform such complex testing. METHODS: A group of 11 flow cytometrists currently performing FC testing in MM using different instrumentation, panel designs (≥ 6-color) and analysis software compared the procedures between their respective laboratories and reviewed the literature to propose a consensus guideline on flow-MRD analysis and reporting in MM. RESULTS/ CONCLUSION: Consensus guidelines support i) the use of minimum of five initial gating parameters (CD38, CD138, CD45, forward, and sideward light scatter) within the same aliquot for accurate identification of the total plasma cell compartment; ii) the analysis of potentially aberrant phenotypic markers and to report the antigen expression pattern on neoplastic plasma cells as being reduced, normal or increased, when compared to a normal reference plasma cell immunophenotype (obtained using the same instrument and parameters); and iii) the percentage of total bone marrow plasma cells plus the percentages of both normal and neoplastic plasma cells within the total bone marrow plasma cell compartment, and over total bone marrow cells. Consensus guidelines on minimal current and future MRD analyses should target a lower limit of detection of 0.001%, and ideally a limit of quantification of 0.001%, which requires at least 3 × 10(6) and 5 × 10(6) bone marrow cells to be measured, respectively.
BACKGROUND: Major heterogeneity between laboratories in flow cytometry (FC) minimal residual disease (MRD) testing in multiple myeloma (MM) must be overcome. Cytometry societies such as the International Clinical Cytometry Society and the European Society for Clinical Cell Analysis recognize a strong need to establish minimally acceptable requirements and recommendations to perform such complex testing. METHODS: A group of 11 flow cytometrists currently performing FC testing in MM using different instrumentation, panel designs (≥ 6-color) and analysis software compared the procedures between their respective laboratories and reviewed the literature to propose a consensus guideline on flow-MRD analysis and reporting in MM. RESULTS/ CONCLUSION: Consensus guidelines support i) the use of minimum of five initial gating parameters (CD38, CD138, CD45, forward, and sideward light scatter) within the same aliquot for accurate identification of the total plasma cell compartment; ii) the analysis of potentially aberrant phenotypic markers and to report the antigen expression pattern on neoplastic plasma cells as being reduced, normal or increased, when compared to a normal reference plasma cell immunophenotype (obtained using the same instrument and parameters); and iii) the percentage of total bone marrow plasma cells plus the percentages of both normal and neoplastic plasma cells within the total bone marrow plasma cell compartment, and over total bone marrow cells. Consensus guidelines on minimal current and future MRD analyses should target a lower limit of detection of 0.001%, and ideally a limit of quantification of 0.001%, which requires at least 3 × 10(6) and 5 × 10(6) bone marrow cells to be measured, respectively.
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