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Published on: July 6, 2014
Evaluation of lymphoid cell populations in cytology specimens using flow cytometry and polymerase chain reaction
B Davidson1, B Risberg, A Berner
1Department of Pathology, The Norwegian Radium Hospital, University of Oslo.
Insights
Flow cytometry (FCM) and polymerase chain reaction (PCR) are valuable for diagnosing lymphomas in cytology specimens. Combining FCM and PCR improves diagnostic sensitivity for B-cell malignancies, aiding in differentiating lymphomas from reactive proliferations.
Area of Science:
- Hematology
- Oncology
- Cytopathology
Background:
- Distinguishing lymphomas from reactive lymphoid proliferations in cytology is challenging.
- Flow cytometry (FCM) and polymerase chain reaction (PCR) detect monoclonal B-cell populations, aiding diagnosis.
- Ancillary techniques are crucial for accurate morphologic evaluation in lymphoid lesions.
Purpose of the Study:
- To compare the sensitivity and specificity of FCM and PCR in evaluating cytology specimens for B-cell monoclonality.
- To assess the combined diagnostic utility of FCM and PCR in identifying lymphomas.
- To evaluate the impact of these techniques on fine needle aspiration sensitivity.
Main Methods:
- Eighty-six cytology specimens from 81 patients were analyzed.
- Specimens were tested using FCM for kappa:lambda ratio and PCR for immunoglobulin heavy chain (IgH) gene rearrangements.
- Histologic diagnosis was used as the gold standard for 51 samples.
Main Results:
- FCM demonstrated higher sensitivity (77%) than PCR (64%) for detecting B-cell malignancy.
- The combined use of FCM and PCR achieved 93% sensitivity in diagnosing B-cell lymphoma.
- Combined methods increased fine needle aspiration sensitivity by 48%.
Conclusions:
- Both FCM and PCR are effective in diagnosing lymphoid lesions in cytology specimens.
- Their combined application is recommended due to complementary detection of monoclonal B-cell populations.
- These techniques help identify subtle B-cell monoclonality that might be missed by morphology alone.
Abstract:
Differential diagnosis between lymphomas and reactive lymphoid proliferations often requires ancillary techniques and morphologic evaluation. Flow cytometry (FCM) and polymerase chain reaction (PCR) can aid the detection of monoclonal B-cell populations. In the present study, the sensitivity and specificity of these two methods in the study of cytology specimens were compared. Eighty-six cytologic specimens from 81 patients (lymph nodes, solid organs, and body cavities) were evaluated. These specimens were taken from three groups of patients: those who underwent an initial evaluation for suspected lymphoma; those who were previously diagnosed with B-cell lymphoma and were now evaluated for possible disease recurrence; and those who were diagnosed with a nonhematologic malignancy. Histologic diagnosis was available for 51 samples. All samples were tested by FCM for the detection of monoclonality using kappa:lambda ratio and for clonal immunoglobulin heavy chain (IgH) gene rearrangements using a single-round PCR after cytologic evaluation. Tissue morphology, FCM and PCR results, and clinical findings in specimens without histologic diagnosis were correlated. Histologic evaluation (N = 51) revealed 44 specimens with B-cell malignancy. Twenty of the 44 lymphoma specimens (45%) were accurately diagnosed in cytologic smears, 18 (41%) were classified as suspicious of lymphoma, and 6 (14%) were diagnosed as reactive. FCM had superior sensitivity compared with PCR (77% vs. 64%). Fifty-six percent of specimens with B-cell malignancy were FCM+/PCR+, 23% were FCM+/PCR-, 14% were FCM-/PCR+, and 7% were FCM-/PCR-. The combined use of FCM and PCR resulted in a diagnosis of B-cell lymphoma in 41 (93%) of 44 B-cell lymphoma specimens and increased the sensitivity of fine needle aspiration by 48%. Both FCM and PCR aid in the diagnosis of lymphoid lesions in cytology specimens, and both can detect monoclonal B-cell populations that may be interpreted in cytology smears as reactive, even by experienced cytologists. Although FCM had higher sensitivity than PCR test in the present study, their combined use should be considered because of a relatively large number of specimens that were detected as monoclonal only with PCR.
