Semi-Automated Interphase FISH (iFISH) Spot Scoring in CD138-Positive Cells: Validation Study for Genetic Abnormalities Detection in Multiple Myeloma

Microsc Res Tech. 2025 Jul;88(7):2001-2008. doi: 10.1002/jemt.24844. Epub 2025 Mar 3.

Abstract

We conducted a study to validate the automated scoring of Fluorescent in Situ Hybridization (FISH) in a routine cytogenetics laboratory using selected CD138-positive cells in samples from patients with multiple myeloma. A workstation was optimized based on the manufacturer's configurations. Six commercial probes (CDKN2C/CKS1B, RB1/DLEU1/LAMP1, TP53/CEN17, FGFR3::IGH, CCND1::IGH, and IGH::MAF) were examined to detect gains, losses, and rearrangements of genes across a total of 180 slides. We used reference values proposed by the European Myeloma Network (10% for rearrangements and 20% for gains and losses) to compare the accuracy of manual, automated, and semi-automated (automated coupled with manual revision) scoring. The time spent by the biologist to perform semi-automated and manual scoring was compared. Automated scoring was not effective and lacked validation. In contrast, semi-automated scoring proved to be efficient and highly accurate for all probes and offered time-saving benefits for deletion/gain probes. These findings suggest that semi-automated iFISH scoring for multiple myeloma is feasible and could become a routine practice in cytogenetics laboratories.

Keywords: CD138‐positive cells; automation iFISH; cytogenetics laboratory; multiple myeloma; validation.

Publication types

  • Validation Study

MeSH terms

  • Chromosome Aberrations*
  • Humans
  • In Situ Hybridization, Fluorescence* / methods
  • Interphase
  • Multiple Myeloma* / diagnosis
  • Multiple Myeloma* / genetics
  • Multiple Myeloma* / pathology
  • Syndecan-1* / analysis
  • Syndecan-1* / genetics

Substances

  • Syndecan-1
  • SDC1 protein, human