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dc.contributor.authorDeane, M
dc.contributor.authorMcCarthy, K P
dc.contributor.authorWiedemann, L M
dc.contributor.authorNorton, John D
dc.date.accessioned2010-06-11T15:58:30Z
dc.date.available2010-06-11T15:58:30Z
dc.date.issued1991-08
dc.identifier.citationAn improved method for detection of B-lymphoid clonality by polymerase chain reaction. 1991, 5 (8):726-30 Leukemiaen
dc.identifier.issn0887-6924
dc.identifier.pmid1909411
dc.identifier.urihttp://hdl.handle.net/10541/104752
dc.description.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.
dc.language.isoenen
dc.subjectB Cell Leukemiaen
dc.subject.meshBase Sequence
dc.subject.meshClone Cells
dc.subject.meshGene Rearrangement, B-Lymphocyte, Heavy Chain
dc.subject.meshGenes, Immunoglobulin
dc.subject.meshHumans
dc.subject.meshImmunoglobulin Heavy Chains
dc.subject.meshImmunoglobulin Variable Region
dc.subject.meshLeukemia, B-Cell
dc.subject.meshLymphoma, B-Cell
dc.subject.meshMolecular Sequence Data
dc.subject.meshOligonucleotides
dc.subject.meshPolymerase Chain Reaction
dc.titleAn improved method for detection of B-lymphoid clonality by polymerase chain reaction.en
dc.typeArticleen
dc.contributor.departmentDepartment of Haematology, Royal Free Hospital, London, UK.en
dc.identifier.journalLeukemiaen
html.description.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.


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