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Sensitivity of PCR in detecting monoclonal B cell proliferations
F C Ling1, C E Clarke, W E Corbett
1Department of Pathology, Queen's University, Kingston, Ontario, Canada.
Insights
Polymerase chain reaction (PCR) rapidly detects monoclonal B cell proliferations by identifying immunoglobulin gene rearrangements. This method is useful for difficult diagnoses and limited samples, complementing Southern blotting.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Monoclonal B cell proliferations require accurate diagnostic methods.
- Current diagnostic techniques may have limitations in sensitivity and sample requirements.
Purpose of the Study:
- To evaluate the polymerase chain reaction (PCR) for rapid detection of monoclonal B cell proliferations.
- To assess PCR's ability to identify clonal immunoglobulin heavy chain genomic rearrangements.
Main Methods:
- Examined 34 B cell lymphomas and 22 other lymphoproliferative/non-lymphoproliferative disorders.
- Utilized PCR to detect immunoglobulin heavy chain gene rearrangements.
- Compared PCR results with Southern blot analysis.
Main Results:
- PCR identified monoclonal rearrangements in 56% of B cell lymphomas.
- PCR showed higher sensitivity for non-follicular center cell lesions (67%) compared to follicular center cell lymphomas (54%).
- PCR detected 3 cases missed by Southern blotting, highlighting complementary roles.
Conclusions:
- PCR analysis detects approximately 55% of clonal lymphoproliferative proliferations.
- PCR is valuable for diagnostically challenging cases and limited sample material.
- PCR serves as a complementary diagnostic tool alongside standard methods.
Aims:
To evaluate the rapid detection of various forms of monoclonal B cell proliferations by using the polymerase chain reaction (PCR) to identify clonal immunoglobulin heavy chain genomic rearrangements.
Methods:
Thirty four B cell lymphomas defined by morphology, immunophenotyping, and positive immunoglobulin heavy chain gene rearrangements detected by Southern blot analysis were examined. An additional 22 cases representing miscellaneous lymphoproliferative and non-lymphoproliferative disorders were also studied.
Results:
Monoclonal rearrangements were identified in 19 (56%) cases of B cell lymphoma. The method was less sensitive in the detection of follicular centre cell lymphomas (15 of 28, or 54%) than non-follicular centre cell lesions (four of six, or 67%). Monoclonal rearrangement was not identified in 19 control cases, including T cell lymphomas, Hodgkin's disease, reactive lymphadenopathy and metastatic carcinoma. Three cases showed positive immunoglobulin gene rearrangement by PCR but were negative on Southern blotting. Two of these cases had definite clinical, morphological, and immunophenotypic evidence of monoclonal B cell proliferation suggesting that PCR could, on occasion, pick up cases missed by Southern blotting and that the two methods are complementary in clonal lymphoproliferative disease diagnosis. The third case represented a "false positive" PCR reaction involving a colonic adenocarcinoma.
Conclusions:
PCR analysis, using the primer sequences outlined in this study, will detect about 55% of clonal lymphoproliferative proliferations with increased sensitivity for non-follicular centre cell lesions. With these levels of detection in mind, this testing strategy can still be especially useful in cases which prove diagnostically problematic with standard morphological and immunophenotypic analysis, and in instances where the quantity and type of diagnostic material is limiting (needle aspirates and cellular fluids).
