Consensus primers for detecting monoclonal immunoglobulin heavy chain rearrangement in B cell lymphomas
- Department of Pathology, and Division of Hematology, Third Department of Internal Medicine, Iwate Medical University School of Medicine, Morioka 020–8505, Japan
- Correspondence to: Dr C Maesawa, Department of Pathology, Iwate Medical University School of Medicine, Uchimaru 19–1, Morioka 020–8505, Japan; chihaya{at}iwate-med.ac.jp
- Accepted 27 March 2003
Abstract
Aims: To demonstrate the usefulness of polymerase chain reaction (PCR) methodology with both the FR2A/LJH/VLJH and the FR1c/LJH/VLJH primer sets for detecting monoclonal immunoglobulin heavy chain (IgH) gene rearrangement in B cell non-Hodgkin lymphomas (B-NHLs).
Methods: Eighty three patients with B-NHL were enrolled in this study. DNA was isolated from paraffin wax embedded sections and amplified by PCR to determine the sequences of the rearranged IgH gene. Each PCR product was subcloned. Cycle sequences and sequence analyses were done to determine the clone specific IgH variable region (VH) sequences.
Results: Clonal IgH gene rearrangements were detected in 45 cases with FR2a/JH/VLJH and in 14 of the remaining cases with FR1c/JH/VLJH. Most of the cases detectable with FR2a/JH/VLJH were derived from VH3 and VH4 families. Five of six cases in the VH1 family and one in the VH7 family were amplified with the FR1c/JH/VLJH primer set only.
Conclusion: The detection rate of IgH rearrangement in B-NHLs can be increased by using both FR2A/LJH/VLJH and FR1c/LJH/VLJH, and these two primer sets are suitable for routine PCR methodology. Moreover, each primer set appears to be closely related to VH family specificity.
- B-NHL, B cell non-Hodgkin lymphoma
- FR, framework
- GC, germinal centre
- IgH, immunoglobulin heavy chain
- PCR, polymerase chain reaction
- VH, immunoglobulin heavy chain variable region
