11. Validation and implementation of BioDot instruments for semi-automated FISH slide processing

      FISH set-up is often a manual and labor-intensive process that includes slide preparation, probe application and hybridization, post-hybridization washing and counter-stain application. We have evaluated the BioDot CellWriter system for normalization of cell concentration, spotting of cells, dispensing FISH probes, denaturation and hybridization, and application of DAPI and coverslips and compared these results with manual processing. Probe signal intensity and background quality were assessed using a numerical scoring criteria and quality of the BioDot method was comparable to the manual process. To evaluate the accuracy of the BioDot process, cells from 65 samples were dropped into a total of 260 BioDot wells and tested using probes for CML, AML, CLL and plasma cell disease, as appropriate to clinical indication. Nine of these 260 samples (3.5%) showed differences compared with their manually processed counterparts. Eight showed a low-level discordant signal pattern with <10% difference in abnormal cell proportion between preparations. The remaining sample showed >20% difference in abnormal signal patterns, although both results were abnormal. Additional clinical validation was performed on 75 blood and bone marrow samples representing a total of 220 BioDot wells. Samples were selected to include both normal and abnormal results for each of the FISH probes included in our plasma cell myeloma, myeloproliferative disease, adult ALL, pediatric ALL, MDS and aggressive lymphoma panels. Twelve of these 220 (5.5%) sample wells showed discrepancies with the manual method, all of which showed low-level discordance with abnormal signal patterns differing by <10% or concordant results with signal counts differing by >20%. The BioDot process has been implemented for clinical use and preliminary data suggest reduced tech time for slide processing, decreased volume of probe use and improved efficiency in analysis.
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