Enhanced sensitivity of flow cytometry for routine assessment of minimal residual disease

Domingo, Esther; Moreno, Cristina; Sánchez-Ibarrola, A; Panizo, Carlos; Páramo, José A.; Merino, Juana

doi:10.3324/haematol.2009.018911

Cited by 16 publications

(14 citation statements)

References 10 publications

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“…The percentage of MM cases where a robust and sensitive discrimination between normal and myelomatous PCs may be achieved increases significantly when additional aberrant phenotypes are simultaneously investigated (e.g., abnormally low expression levels of CD27 and CD81, overexpression of CD117 and CD200) (Table ); simultaneous assessment of the CyIgκ/CyIgλ ratio within small populations of suspicious PCs, as defined with the previous marker combination(s), will further strengthen the robustness of a highly sensitive flow‐MRD assay down to 10 −5 , whenever enough cells (i.e., >5 × 10 6 cells) are investigated (Fig. ) . This reinforces the ultimate relevance of the precise antibody combinations and panels used (e.g., the way specific antibodies are combined among them in a given antibody combination in a panel) over the simple list of reagents they include, as well as the need for acquiring enough cells (several millions) per sample.…”

Section: Aberrant Plasma Cell Phenotypesmentioning

confidence: 99%

Immunophenotype of normal vs. myeloma plasma cells: Toward antibody panel specifications forMRDdetection in multiple myeloma

Flores‐Montero

Tute²,

Paiva

et al. 2015

Cytometry Part B Clinical

203

186

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In recent years, several studies on large series of multiple myeloma (MM) patients have demonstrated the clinical utility of flow cytometry monitoring of minimal residual disease (flow-MRD) in bone marrow (BM), for improved assessment of response to therapy and prognostication. However, disturbing levels of variability exist regarding the specific protocols and antibody panels used in individual laboratories. Overall, consensus exists about the utility of combined assessment of CD38 and CD138 for the identification of BM plasma cells (PC); in contrast, more heterogeneous lists of markers are used to further distinguish between normal/reactive PCs and myeloma PCs in the MRD settings. Among the later markers, CD19, CD45, CD27, and CD81, together with CD56, CD117, CD200, and CD307, have emerged as particularly informative; however, no single marker provides enough specificity for clear discrimination between clonal PCs and normal PCs. Accordingly, multivariate analyses of single PCs from large series of normal/reactive vs. myeloma BM samples have shown that combined assessment of CD138 and CD38, together with CD45, CD19, CD56, CD27, CD81, and CD117 would be ideally suited for MRD monitoring in virtually every MM patient. However, the specific antibody clones, fluorochrome conjugates and sources of the individual markers determines its optimal (vs. suboptimal or poor) performance in an eight-color staining. Assessment of clonality, via additional cytoplasmic immunoglobulin (CyIg) j vs. CyIgk evaluation, may contribute to further establish the normal/reactive vs. clonal nature of small suspicious PC populations at high sensitivity levels, provided that enough cells are evaluated.

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Section: Aberrant Plasma Cell Phenotypesmentioning

confidence: 99%

Immunophenotype of normal vs. myeloma plasma cells: Toward antibody panel specifications forMRDdetection in multiple myeloma

Flores‐Montero

Tute²,

Paiva

et al. 2015

Cytometry Part B Clinical

203

186

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“…In MM as in other diseases, it is almost certain that as treatment regimens continue to improve, the threshold of 0.01%/10 24 is likely to become less relevant. The consensus is that current and future MRD analyses should target a lower LOD preferable of < 0.001%/10 25 (18,22), which requires acquisition of at least 3 3 10 6 bone marrow cells; ideally a limit of quantification of 0.001% should be targeted in the new MM flow-MRD approaches which requires that a minimum of 5 3 10 6 BM cells are acquired. If it is required that a categorical assessment (MRD-positive or MRD-negative) is provided, it is important to state the threshold used and ideally reference the threshold appropriately, e.g.…”

Section: Reporting a Sample As Mrd Positive Mrd Negative Or Unsuitamentioning

confidence: 99%

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

Arroz

Came

Lin

et al. 2015

Cytometry Part B Clinical

148

176

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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 3 10 6 and 5 3 10 6 bone marrow cells to be measured, respectively. V C 2015 International Clinical Cytometry Society

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“…The minimum cluster size was 50 events over a total of 5 million analyzed reaching an expected 10 −5 sensitivity with a coefficient of variation according to Poisson distribution (100/√ n ) of 15% .…”

Section: The Concordance/discordance Is Referred To In Qualitative Termsmentioning

confidence: 98%

A reproducible strategy for analysis of minimal residual disease measured by Standardized multiparametric flow cytometry in b acute lymphoblastic leukemia

Palao

Tarín

Martirena

et al. 2018

Cytometry Part B Clinical

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Enhanced sensitivity of flow cytometry for routine assessment of minimal residual disease

Cited by 16 publications

References 10 publications

Immunophenotype of normal vs. myeloma plasma cells: Toward antibody panel specifications forMRDdetection in multiple myeloma

Immunophenotype of normal vs. myeloma plasma cells: Toward antibody panel specifications forMRDdetection in multiple myeloma

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

A reproducible strategy for analysis of minimal residual disease measured by Standardized multiparametric flow cytometry in b acute lymphoblastic leukemia

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