• Document: Single Tube, Six-Color Flow Cytometric Analysis Is a Sensitive and Cost-Effective Technique for Assaying Clonal Plasma Cells
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Hematopathology / Flow Cytometric Analysis of Clonal Plasma Cells Single Tube, Six-Color Flow Cytometric Analysis Is a Sensitive and Cost-Effective Technique for Assaying Clonal Plasma Cells Derek K. Marsee, MD, PhD, Betty Li, MS, MT(ASCP), and David M. Dorfman, MD, PhD Key Words: Plasma cell myeloma; Multiple myeloma; Flow cytometry DOI: 10.1309/AJCPKKNPMLWX9ZXB Abstract Plasma cell neoplasms are defined by a neoplastic clonal Bone marrow flow cytometric analysis is a population of mature plasma cells. According to criteria powerful and rapid tool for evaluating plasma cell from the International Myeloma Working Group and the myeloma. By using a noncontrolled patient population 2008 World Health Organization Classification of Tumours in various stages of diagnosis and treatment, we of Hematopoietic and Lymphoid Tissues, these entities are compared 6-color (single-tube) and 4-color (multiple- subdivided into monoclonal gammopathy of undetermined tube) flow cytometric immunophenotyping protocols. significance, plasma cell myeloma, extraosseous plasmacy- Prospective comparison in 52 cases demonstrated toma, and monoclonal immunoglobulin deposition diseases improved ability to detect clonal plasma cells or based on several criteria, including the presence of a serum or identical diagnoses in 100% of the cases using 6-color, urine M spike, organ or tissue damage, and demonstration of single-tube analysis. In cases in which 6-color flow a clonal plasma cell population.1-3 A minimal percentage of cytometric analysis improved detection of a clonal abnormal plasma cells is not strictly required for a diagnosis population, concurrent biopsy showed less than 5% of symptomatic plasma cell myeloma. involvement by plasma cell myeloma, suggesting Bone marrow flow cytometric analysis is a powerful and that 6-color flow cytometry has an advantage in rapid tool for evaluating plasma cell clonality. Nonneoplastic patients with a low disease burden. In addition, the plasma cells are characterized by high levels of surface expres- simplification of the procedure resulted in substantial sion of CD19, CD38, and CD138 and dim expression of savings in technologist time and reagent costs. Taken CD45.2,4 In contrast, neoplastic plasma cells typically show loss together, this study demonstrates that 6-color flow of CD19 and aberrant expression of markers such as CD56 and cytometry is an excellent, cost-effective means to assay CD117.2,4,5 In addition, immunoglobulin heavy and light chain for clonal plasma cells in a noncontrolled patient staining can also be assayed to assess for plasma cell clonality. population. The assessment of light chain clonality is complicated by the cytoplasmic localization of immunoglobulins in plasma cells, requiring that they be permeabilized before determining the immunoglobulin profile. In 4-color analysis, the combina- tion of numerous surface and cytoplasmic markers requires separate protocols and multiple tubes for analysis. In addition to increasing the chances of error, the presence of multiple tubes and protocols results in increased cost of the assay in terms of reagents and technician hours. With the advent of 6- and 8-color analysis, the use of single-tube flow cytometric analysis for plasma cell myeloma 694 Am J Clin Pathol 2010;133:694-699 © American Socie

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