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J Clin Pathol. 1992 Sep;45(9):770-5.

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

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  • 1Department of Biochemistry, Repatriation General Hospital, Daw Park, SA.

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.

PMID:
1401205
[PubMed - indexed for MEDLINE]
PMCID:
PMC495101
Free PMC Article
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