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Abstract
Background
Immunoglobulin (Ig) gene rearrangement analysis is a useful additional tool to detect clonality of B-lymphoproliferative disease and the method to detect immunoglobulin gene rearrangement is required the high sensitivity and specificity. BIOMED-2 multiplex PCR was designed for the evaluation of molecular clonality of lymphoid lesions. We evaluated the usefulness of the BIOMED-2 multiplex PCR by comparing it with conventional nested PCR.
Methods
Sixteen patients with malignant lymphoma and 5 with reactive lymph nodes were examined to assess the sensitivity, specificity, and accuracy between conventional nested PCR and BIOMED-2. All 3 tests performed using the BIOMED-2 kit for immunoglobulin (Ig) heavy chain (IGH) gene, Igκ light chain (IGK) gene, and Igλ light chain (IGL) gene, were used to analyze clonality.
Results
Both the methods showed 100% specificity (95% confidence interval, 56.6-100.0). The combination of IGH and IGK BIOMED-2 tests with or without IGL revealed the highest sensitivity (87.5%; range, 64.0-96.5%) and accuracy (90%; range, 0.70-0.97). Compared to the conventional method, the BIOMED-2 test for IGH showed a higher sensitivity (62.5%; range, 38.6-81.5%) and accuracy (71%, 0.50-0.86).
Figures and Tables
Fig. 1
Results of both the methods in a single case. The case (case no. 9) with nodal marginal zone lymphoma (A) showed an uncertain band in the conventional method (B), but a definite peak in BIOMED-2 (C). (A, hematoxylin and eosin [H&E] stain ×40; Inset of A, H&E stain ×400; Arrow indicates uncertain band).
Table 1
Results of clonality analysis in 16 cases of malignant lymphoma
*B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and classic Hodgkin lymphoma.
Abbreviations: IGH, immunoglobulin heavy chain; IGK, immunoglobulin kappa light chain; IGL, immunoglobulin lambda light chain; M, monoclonal; P, polyclonal; ND, not determined; LN, lymph node; BM, bone marrow; MZL, marginal zone B-cell lymphoma; DLBCL, diffuse large B-cell lymphoma; CI, chronic inflammation; CLL/SLL, chronic lymphocytic leukemia/small lymphocytic lymphoma.
Table 2
Sensitivity, specificity, accuracy, and effect of BIOMED-2 to detect monoclonality
*The interpretation of 4 cases was unavailable; †The P values were calculated by Fisher's exact test to show the effect of BIOMED-2 and to compare it with that of conventional nested PCR.
Abbreviations: CI, confidence interval; IGH, immunoglobulin heavy chain; IGK, immunoglobulin kappa light chain; IGL, immunoglobulin lambda light chain; M, monoclonal; P, polyclonal.
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