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

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 (JH) 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.

• Authors: I Ramasamy, M Brisco, A Morley
• Journal: Journal of clinical pathology (J Clin Pathol) Vol. 45 Issue 9 Pg. 770-5 (Sep 1992) ISSN: 0021-9746 [Print] England
• PMID: 1401205 (Publication Type: Case Reports, Journal Article)

Chemical References

• Immunoglobulin Heavy Chains
• DNA

Topics

• Aged
• B-Lymphocytes (physiology)
• Base Sequence
• DNA (analysis)
• Gene Rearrangement, B-Lymphocyte, Heavy Chain (genetics)
• Humans
• Immunoglobulin Heavy Chains (genetics)
• Lymphoproliferative Disorders (genetics)
• Male
• Molecular Sequence Data
• Polymerase Chain Reaction (methods)