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Updated: Aug 28, 2025
Interphase Fluorescence in situ Hybridization of Bone Marrow Smears of Multiple Myeloma
Published on: April 15, 2022
IGH cytogenetic abnormalities can be detected in multiple myeloma by imaging flow cytometry
Henry Hui1, Kathy A Fuller1, Luna Eresta Jaya2
1School of Biomedical Sciences, The University of Western Australia, WA Australia.
Insights
An automated imaging flow cytometry method accurately detects IGH abnormalities in multiple myeloma patients. This high-throughput technique identifies translocations and trisomy without cell isolation, improving precision in genetic lesion detection.
Area of Science:
- Hematology
- Cytogenetics
- Flow Cytometry
Background:
- Cytogenetic abnormalities of the IGH gene occur in up to 55% of multiple myeloma patients.
- Current IGH abnormality testing relies on manual fluorescence in situ hybridization (FISH) on isolated plasma cells.
Purpose of the Study:
- To evaluate an automated imaging flow cytometric method for identifying IGH abnormalities.
- To assess if this method can detect IGH abnormalities without prior plasma cell isolation.
Main Methods:
- Bone marrow aspirates from 10 multiple myeloma patients were analyzed.
- Plasma cells were identified by CD38 and CD138 coexpression.
- Imaging flow cytometry acquired thousands of cells, analyzed with IGH FISH probes for numerical/structural abnormalities.
Main Results:
- IGH chromosomal abnormalities were detected in 5 of 10 samples.
- Identified translocations (t(4;14), t(11;14)) and trisomy 14.
- The lowest detection limit for an IGH abnormality was 0.05% of all cells.
Conclusions:
- Automated high-throughput immuno-flowFISH successfully identified IGH translocations and trisomy in multiple myeloma plasma cells.
- The method analyzes thousands of cells without isolation, achieving a low detection limit of 0.05%.
- This approach offers enhanced precision for detecting critical genetic lesions involving IGH and other chromosomal defects in multiple myeloma.
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.
