Aims Cytogenetic abnormalities involving the IGH gene are seen in up to 55% of patients with multiple myeloma. Current testing is performed manually by fluorescence in situ hybridisation (FISH) on purified plasma cells. We aimed to assess whether an automated imaging flow cytometric method that uses immunophenotypic cell identification, and does not require cell isolation, can identify IGH abnormalities.
Methods Aspirated bone marrow from 10 patients with multiple myeloma were studied. Plasma cells were identified by CD38 and CD138 coexpression and assessed with FISH probes for numerical or structural abnormalities of IGH. Thousands of cells were acquired on an imaging flow cytometer and numerical data and digital images were analysed.
Results Up to 30 000 cells were acquired and IGH chromosomal abnormalities were detected in 5 of the 10 marrow samples. FISH signal patterns seen included fused IGH signals for IGH/FGFR3 and IGH/MYEOV, indicating t(4;14) and t(11;14), respectively. In addition, three IGH signals were identified, indicating trisomy 14 or translocation with an alternate chromosome. The lowest limit of detection of an IGH abnormality was in 0.05% of all cells.
Conclusions This automated high-throughput immuno-flowFISH method was able to identify translocations and trisomy involving the IGH gene in plasma cells in multiple myeloma. Thousands of cells were analysed and without prior cell isolation. The inclusion of positive plasma cell identification based on immunophenotype led to a lowest detection level of 0.05% marrow cells. This imaging flow cytometric FISH method offers the prospect of increased precision of detection of critical genetic lesions involving IGH and other chromosomal defects in multiple myeloma.
- Multiple Myeloma
- Image Cytometry
Data availability statement
No data are available.
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Handling editor Tahir S Pillay.
Contributors Conceptualisation: HH, KF and WNE. Methodology and validation: HH, KF, JLE, RF, GS, YK, SEC. Formal analysis: HH, JLE and YK. Investigation: HH, JLE, YK and TFN. Resources: KF, WNE, RF, GP and KYl. Data curation: HH, TFN, KF and WE. Writing, review and editing: KF, WNE, SEC, YK and KY. Project administration: KF, WNE, YK and KY. Funding acquisition: KF, WNE, YK and KY. All authors have read and agreed to the published version of the manuscript.
Funding This research was funded by the Foundation for Australia-Japan Studies (FAJS).
Competing interests HH, KF and WNE are inventors of a patent application covering the immuno-flowFISH technique. The remaining authors declare no competing interests.
Provenance and peer review Not commissioned; externally peer reviewed.