An improved method for detection of B-lymphoid clonality by polymerase chain reaction.
AffiliationDepartment of Haematology, Royal Free Hospital, London, UK.
MetadataShow full item record
AbstractSeveral groups have recently described methods for the detection of clonal immunoglobulin heavy chain (IgH) gene rearrangements in B-cell malignancies by polymerase chain reaction (PCR) gene amplification using variable region-(VH) and joining (JH) region-specific primers. The simplest methods utilize a single VH primer specific for sequences present in most VH regions corresponding to the third framework region (FR3). An alternative approach is to use a panel of VH family-specific primers specific for the first framework regions (FR1). In the course of nucleotide sequence analysis of IgH gene rearrangements amplified using a VH FR1 primer panel, these authors previously observed 3' VH region deletion and/or base mis-matches sufficient to prevent efficient priming from the VH FR3 primer target sequence in a significant minority of cases of B-lineage malignancy. An improved PCR method has therefore been developed by using a panel of seven VH FR1 family-specific primers incorporated in a single reaction. By using this method clonal IgH gene rearrangement is detected in 15 of 16 cases of B-lineage malignancy. Significantly, this series included four cases of B-lymphoma in which previous attempts to detect PCR clonal IgH gene rearrangements using a VH FR3 primer were unsuccessful. In two of these cases, nucleotide sequence analysis of the amplified DNA showed that failure to prime with the VH FR3 primer was likely to be attributable to insufficient homology with the target sequence. The use of the approach described in this paper should significantly improve the reliability of detection of B-lymphoid clonality by PCR.
CitationAn improved method for detection of B-lymphoid clonality by polymerase chain reaction. 1991, 5 (8):726-30 Leukemia
- Description of a novel FR1 IgH PCR strategy and its comparison with three other strategies for the detection of clonality in B cell malignancies.
- Authors: Aubin J, Davi F, Nguyen-Salomon F, Leboeuf D, Debert C, Taher M, Valensi F, Canioni D, Brousse N, Varet B
- Issue date: 1995 Mar
- Use of UITma DNA polymerase improves the PCR detection of rearranged immunoglobulin heavy chain CDR3 junctions.
- Authors: Linke B, Bolz I, Pott C, Hiddemann W, Kneba M
- Issue date: 1995 Dec
- Analysis of clonal rearrangements of the Ig heavy chain locus in acute leukemia.
- Authors: Height SE, Swansbury GJ, Matutes E, Treleaven JG, Catovsky D, Dyer MJ
- Issue date: 1996 Jun 15
- Detection of immunoglobulin gene rearrangement of B cell non-Hodgkin's lymphomas and leukemias in fresh, unfixed and formalin-fixed, paraffin-embedded tissue by polymerase chain reaction.
- Authors: Inghirami G, Szabolcs MJ, Yee HT, Corradini P, Cesarman E, Knowles DM
- Issue date: 1993 Jun
- Clonal evolution in B-lineage acute lymphoblastic leukemia by contemporaneous VH-VH gene replacements and VH-DJH gene rearrangements.
- Authors: Choi Y, Greenberg SJ, Du TL, Ward PM, Overturf PM, Brecher ML, Ballow M
- Issue date: 1996 Mar 15