Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B-lymphocyte clones in a polyclonal population.
A method has been developed to use the polymerase chain reaction to amplify and sequence the chain determining region 3 (CDR 3) of the human immunoglobulin heavy-chain gene, and to use the sequence as a marker for rare neoplastic B lymphocytes. Consensus primers for the Variable and Joining regions of the gene were constructed and shown to enable efficient amplification, directed cloning, and sequencing of CDR 3. Using leukaemic cell line PFMC as a test system, CDR 3 was sequenced, specific primers synthesized, and PFMC DNA was detected down to a dilution of 1:1300 in DNA from normal lymphocytes. This strategy should be useful for monitoring therapy and detecting early disease relapse in B lymphoproliferative disease.
Brisco
M J
MJ
Department of Haematology, Flinders Medical Centre, Bedford Park, South Australia.
Tan
L W
LW
Orsborn
A M
AM
Morley
A A
AA
D001402:B-Lymphocytes / Q000502:physiology*; D001483:Base Sequence; D002460:Cell Line; D004273:DNA, Neoplasm / Q000032:analysis*; D005784:Gene Amplification; D005803:Genes, Immunoglobulin / Q000235:genetics; D006801:Humans; D007938:Leukemia / Q000235:genetics; D008232:Lymphoproliferative Disorders / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D004273:DNA, Neoplasm