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Factors determining whether diffuse large B-cell lymphoma samples are detected by flow cytometry
David Peng1, Aruna Kodituwakku2, Steven Le2
1St Vincent's Healthcare Clinical Campus, University of NSW, Sydney, Australia.
Insights
Flow cytometry (FCM) often misses diffuse large B-cell lymphoma (DLBCL) diagnoses. Factors like lower lymphocyte percentages and higher T-cell percentages in samples increase the missed diagnosis rate for DLBCL.
Area of Science:
- Hematology
- Immunophenotyping
- Oncology
Background:
- Flow cytometry (FCM) is a standard diagnostic tool for mature B-cell neoplasms (MBN).
- Diffuse large B-cell lymphoma (DLBCL), a prevalent MBN, is sometimes misdiagnosed or missed by FCM.
- Understanding factors contributing to FCM detection failure in DLBCL is crucial for improving diagnostic accuracy.
Purpose of the Study:
- To investigate factors associated with the failure of flow cytometry (FCM) to detect diffuse large B-cell lymphoma (DLBCL).
- To identify specific sample characteristics and protein expression levels that correlate with missed DLBCL diagnoses by FCM.
Main Methods:
- Retrospective analysis of 135 DLBCL cases diagnosed using eight-colour FCM.
- Comparison of clinical, FCM, histopathological, and genetic data between DLBCL cases detected and not detected by FCM.
- Statistical analysis to identify significant differences and correlations.
Main Results:
- 16% (22 out of 135) of DLBCL cases were not detected by FCM.
- Samples with a lower percentage of total events as lymphocytes and a higher percentage of T cells among lymphocytes were less likely to be detected.
- High MYC protein expression on immunohistochemistry was associated with a lower likelihood of DLBCL being missed by FCM.
Conclusions:
- Several factors influence the rate at which DLBCL is missed by FCM, even with advanced eight-colour analysis.
- The study highlights the limitations of FCM in detecting all DLBCL cases, emphasizing the need for complementary diagnostic methods.
- Diagnostic yield of FCM for DLBCL can be affected by sample composition and specific protein expression profiles.
Introduction:
Flow cytometry (FCM) is widely used in the diagnosis of mature B-cell neoplasms (MBN), and FCM data are usually consistent with morphological findings. However, diffuse large B-cell lymphoma (DLBCL), a common MBN, is sometimes not detected by FCM. This study aimed to explore factors that increase the likelihood of failure to detect DLBCL by FCM.
Methods:
Cases with a final diagnosis of DLBCL that were analysed by eight-colour FCM were retrospectively collated. Clinical, FCM, histopathological and genetic data were compared between cases detected and cases not detected by FCM.
Results:
DLBCL cases from 135 different patients were analysed, of which 22 (16%) were not detected by FCM. In samples not detected by flow cytometry, lymphocytes were a lower percentage of total events (p = 0.02), and T cells were a higher percentage of total lymphocytes (p = 0.01). Cases with high MYC protein expression on immunohistochemistry were less likely to be missed by FCM (p = 0.011). Detection of DLBCL was not different between germinal centre B-cell (GCB) and non-GCB subtypes, not significantly affected by the presence of necrosis or fibrosis, and not significantly different between biopsy specimens compared to fine-needle aspirates, or between samples from nodal compared to extranodal tissue.
Conclusion:
The study identifies several factors which affect the likelihood of DLBCL being missed by FCM. Even with eight-colour analysis, FCM fails to detect numerous cases of DLBCL.
