Utility of a column-free cell sorting system for separation of plasma cells in multiple myeloma FISH testing in clinical laboratories.
Targeted FISH analysis is an essential component of the management of plasma cell myeloma for identification of cytogenetic abnormalities. The purpose of this study was to evaluate the column-free method, RoboSep® (RS), for sorting CD138-expressing cells in bone marrow aspirates. Comparative analysis of column-based and RS methodologies was carried out on 54 paired bone marrow aspirate validation samples from patients undergoing work-up for plasma cell dyscrasia. Abnormalities detected by FISH analysis using an IGH@/CCND1 probe set were seen in 54% with RS, and 44% with column-based. We found a statistically significant difference between the yield of abnormalities detected in paired positive cases (p = 0.0001). An additional 183 consecutive post-validation samples sorted by RS showed recurrent genetic abnormalities in 85/120 (71%) of successfully sorted samples with ≥ 1% plasma cells but in none of 63 samples in which FISH analysis was completed on samples that could not be sorted due to insufficient plasma cells upon cell sorting. The column-free method successfully sorted PC, when present in ≥ 1% of cells, for detection of abnormalities by FISH. Furthermore, our data suggest that FISH analysis should not be performed on samples with an inadequate yield at the cell selection step.
Cleveland Clinic, 9500 Euclid Avenue, Mail Code LL2-2, Cleveland, OH 44195, USA. shettys@ccf.org
Shashirekha
S
Shetty
Marion
M
Siady
Kalyan C
KC
Mallempati
Andrew
A
Wilson
Jeff
J
Poarch
Brandon
B
Chandler
Judy
J
Gray
Mohamed E
ME
Salama
D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D009101:Multiple Myeloma / Q000175:diagnosis*; D010950:Plasma Cells / Q000166:cytology* / Q000378:metabolism; D012680:Sensitivity and Specificity