Flow cytometric immunophenotyping test for staging/monitoring neuroblastoma patients.
Michael J Warzynski1, David M Graham, Richard A Axtell
1Department of Pathology and Laboratory Medicine, Spectrum Health, Grand Rapids, Michigan, USA. Michael.Warzynski@Spectrum-Health.org
Cytometry
|December 24, 2002
Summary
A novel flow cytometry assay using CD56 and GD2 markers accurately detects neuroblastoma cells in patient samples. This rare event analysis aids in staging and monitoring neuroblastoma, achieving 100% sensitivity.
Area of Science:
- Pediatric Oncology
- Immunophenotyping
- Flow Cytometry
Background:
- Incidental observation of neoplastic CD45(-) cells with bright CD56 expression in neuroblastoma samples.
- Distinguishing these cells from natural killer lymphocytes and other tumors is crucial.
Purpose of the Study:
- To develop a sensitive and specific flow cytometry assay for detecting neuroblastoma cells.
- To establish a clinically applicable method for rare event analysis in neuroblastoma diagnosis and monitoring.
Main Methods:
- Development of a flow cytometry "rare event" assay using CD45, CD56, and neuron-specific enolase (NSE).
- Modification to a lysed whole blood assay using CD45, CD56, and ganglioside GD2 for improved clinical application.
- Further refinement by adding CD81 as a fourth marker for enhanced staging and monitoring.
Main Results:
- The assay demonstrated 83% specificity and 100% sensitivity in clearly interpretable samples.
- The ganglioside GD2 assay detected neuroblastoma cells at a level of 0.002% (1 in 100,000 cells).
- The four-color assay is now used clinically for staging and monitoring all neuroblastoma patients.
Conclusions:
- A highly sensitive and specific flow cytometry assay for neuroblastoma detection has been developed.
- The assay, particularly the three-color GD2 panel, is effective for identifying rare neuroblastoma cells.
- This rare event analysis method significantly aids in the clinical management of neuroblastoma.
