Detection and retrieval of multiple myeloma cells with single cell molecular confirmation from peripheral blood by multi-parameter immunofluorescence, automated scanning and image analysis
Multiple myeloma is assessed by bone marrow aspirate, an invasive procedure that limits how often the disease can be monitored; this AACR 2018 poster reports a 5-parameter immunofluorescence assay that detected, retrieved and molecularly confirmed myeloma cells in a patient's peripheral blood.
Presented by RareCyte and All4Cure.
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- A 5-parameter immunofluorescence assay identified myeloma cells in peripheral blood at 55 cells per mL. In a pre-treatment patient sample, cells with a CD138+, CD56+, CD38- and CD45- phenotype were counted at 55 per mL of whole blood; buffy coats were separated from 7.5 mL draws with AccuCyte, stained for CD138, CD38, CD56, CD45 and a nuclear dye, and scanned on CyteFinder.
- Retrieved single cells carried the patient's own tumor mutations, confirming their identity. Cells picked individually with the CytePicker module were whole-genome amplified and Sanger sequenced against four genes taken from the patient's bone marrow genomic profile (IDH1, NRAS, PHLDA1, RPH3AL); mutations were found in 9 of 15 myeloma cells and in 0 of 13 white blood cell controls.
- The same panel also identified normal plasma cells in healthy donors, at 5 per mL. Plasma cells co-expressing CD138 and CD38 were identified at 5 per mL of whole blood in healthy volunteer samples; RPMI8226 myeloma cells spiked into normal donor blood provided the model system used to develop the assay, and patient samples were run under an IRB-approved protocol.
