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Detection of Immunoglobulin κ Light Chain Rearrangements by Polymerase Chain Reaction : An Improved Method for Detecting Clonal B-Cell Lymphoproliferative Disorders

Authors
  • Jerry Z. Gong
  • Sherman Zheng
  • Roberto Chiarle
  • Christine De Wolf-Peeters
  • Giorgio Palestro
  • Glauco Frizzera
  • Giorgio Inghirami
Publisher
American Society for Investigative Pathology
Publication Date
Aug 01, 1999
Source
PMC
Keywords
Disciplines
  • Biology
  • Design
License
Unknown

Abstract

The clonal determination of B-cell lymphoproliferative disorders by immunoglobulin heavy chain (IgH) rearrangement by polymerase chain reaction (PCR) is widely used. However, few attempts have been made to detect immunoglobulin κ light chain (Igκ) gene rearrangement using PCR. We studied 145 cases of B-cell neoplasms, along with 58 atypical and 18 reactive lymphoproliferative disorders, using newly designed degenerate oligoprimers recognizing the framework 3 (FR3κ) and the joint (Jκ) regions of the Igκ gene. PCR products were analyzed on nondenaturing polyacrylamide gel (ndPAGE). Clonal B-cell determination was further investigated using IgH rearrangement and t(11:14) or t(14:18). By combining these methods, we detected either clonality or translocation in 117 of 137 cases (85%) in mature B-cell neoplasms. The additional analysis of Igκ rearrangement improved sensitivity from 66% to 85%. To investigate whether the Ig gene configuration could be characterized using Igκ PCR in B-cell neoplasms showing severe breakdown of genomic DNA, 18 selected cases were analyzed. Successful amplification was detected in 72% of the cases using either FR3/2-JH and/or FR3Jκ oligoprimers. Finally, clonality was detected in 21 of 58 atypical B-cell proliferations, and among them, the atypical marginal cell (54%) and atypical large cell (50%) proliferations showed the highest frequency of clonal immunoglobulin gene products. We concluded that PCR/ndPAGE analysis of Igκ is a sensitive, rapid, and efficient method for assessing clonality in conjunction with IgH and specific translocation analysis. This approach is particularly useful in the characterization of B-cell lymphoproliferative disorders in archival material with poor preservation of the genomic DNA.

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