title: Single-Tube 10-Fluorochrome Analysis for Efficient Flow Cytometric Evaluation of Minimal Residual Disease in Plasma Cell Myeloma.
abstract: OBJECTIVE
  Widespread adoption of recent recommendations for minimal residual disease (MRD) detection in myeloma has partly been impeded by a paucity of studies detailing multiparameter flow cytometry (MPF) assay validation. In response, we have validated a novel and efficient single-tube 10-color assay for MRD detection that incorporates the recently recommended plasma cell markers.
  
  
  METHODS
  Aspirate samples from 53 patients with plasma cell disorder were analyzed using a novel single-tube 10-color method. The limit of detection, precision of measurement, and linearity of measurement of our new assay were determined using serial dilution experiments. The stability of the new antibody cocktail and the viability/specificity of stained samples were evaluated using serial time course measurements.
  
  
  RESULTS
  There was a high degree of quantitative agreement between our new 10-color method and an established eight-color method. Four positive samples detected by the 10-color assay were below or at the limit of detection of the eight-color assay, confirming its high sensitivity. In two cases, subsequent revision of the International Myeloma Working Group Uniform Response Criteria was necessary.
  
  
  CONCLUSIONS
  Adoption of our validated 10-color assay would enable clinical laboratories to satisfy current MRD recommendations without significantly increasing the demands on current workflow practices.
authors.affiliation: From the Sloan Kettering Institute.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Royston
authors.affiliation: Departments of Laboratory Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Qi
authors.initials: Q
authors.lastname: Gao
authors.affiliation: Departments of Laboratory Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Nghia
authors.initials: N
authors.lastname: Nguyen
authors.affiliation: Departments of Laboratory Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Peter
authors.initials: P
authors.lastname: Maslak
authors.affiliation: Departments of Laboratory Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Dogan
authors.affiliation: Departments of Laboratory Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY. roshalm@mskcc.org.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Roshal
mesh_terms: D000328:Adult; D000368:Aged; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D008875:Middle Aged; D009101:Multiple Myeloma / Q000175:diagnosis* / Q000473:pathology; D018365:Neoplasm, Residual / Q000175:diagnosis*
chemical_list: D005456:Fluorescent Dyes
keywords: 10-color; MRD; Minimal residual disease; Multiparameter flow cytometry; Multiple myeloma; Plasma cell disorder; Single tube; Validation