Improved PCR method for detecting monoclonal immunoglobulins heavy chain rearrangement in B cell neoplasms

I. Ramasamy, M. Brisco, A. Morley

Research output: Contribution to journalArticlepeer-review

248 Citations (Scopus)

Abstract

Aims: To develop a simple, optimised, polymerase chain reaction (PCR) based method for detecting the rearranged immunoglobulin heavy chain (IgH). Methods: Using as primers oligonucleotides (Fr2A, Fr2B) homologous to the conserved sequences to the framework II region and the joining (J(H)) region, 25 patients with B cell lymphoproliferative disorders, previously characterised by Southern blotting, and three patients with light chain myeloma were studied. Results: The PCR product from a polyclonal B cell population showed a broad band when analysed on a 3% agarose gel; DNA from B cell lines and B lymphoproliferative disorders showed a discrete band. Specificity of the amplification was confirmed by cloning and sequencing the amplified product as well as by Southern blotting with an internal probe homologous to the framework 3 region. Primers Fr2A and Fr2B detected monoclonality in three patients with light chain myeloma, while primers directed against the FrIII region showed a polyclonal response. Conclusions: Deletions and extensive somatic mutations within the FrIII region may give false negative results with primers homologous to the region. A PCR using the method described, with a repertoire of primers homologous to the FrII and FrIII regions, will therefore increase the frequency of detection of monoclonality.

Original languageEnglish
Pages (from-to)770-775
Number of pages6
JournalJournal of Clinical Pathology
Volume45
Issue number9
DOIs
Publication statusPublished - 1992
Externally publishedYes

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