Semi-automated Interphase FISH Spot Scoring in CD138-positive cells: validation study for genetic abnormalities detection in multiple myeloma
Year: 2024
Type: conference-abstract
Abstract: Objective: Genetic abnormalities detected in plasma cells are important prognostic indicators in multiple myeloma (MM), and interphase Fluorescent in Situ Hybridization (iFISH) in selected plasma cells is considered the gold standard genetic technique. However, manual iFISH analysis is time-consuming and subject to interobserver variability. Automated analysis provides an opportunity to overcome these difficulties, but clear guidelines for fluorescent spot automated analysis are lacking. This study aimed to validate the automated analysis and compare its effectiveness with standard manual analysis for iFISH in selected CD138 cells. Method: A workstation was optimized based on the manufacturer’s configurations. Six commercial probes were tested to detect genetic alterations in 20 samples (120 slides). Reference values proposed by the European Myeloma Network (10% for rearrangements and 20% for gains and losses) were used to compare the results of manual and semi-automated analyses. The time spent on the two analyses was compared in another 10 samples (60 slides). Conclusion: Automated analysis was found to be ineffective, lacking proper validation, and was excluded from further testing. Semi-automated analysis was then performed. Results showed the following abnormalities: CDKN2C/CKS1B probe detected 5 cases with abnormal signals; the RB1/DLEU1/LAMP1 probe identified 3 cases with deletion of all probes; the TP53/17cen probe detected 1 abnormal case; FGFR3/IGH probe identified 5 cases with abnormal signals; the MAF/IGH probe detected 5 cases with abnormal signals; and the CCND1/IGH probe detected 7 cases with abnormal signals. Both manual and semi-automated analyses demonstrated 100% accuracy for normal or abnormal signals patterns. Semi-automated analysis required less time compared to manual analysis for the CDKN2C/CKS1B probes (2.3 vs. 3.9 minutes, p=0.013) and for the CCND1/IGH probe (2.0 vs. 4.7, p=0.009). These findings suggest that automating iFISH analysis for MM is feasible and could become routine practice in cytogenetics laboratories.
Language: English
Subfield: Hematology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4409419006