Detection of clonal B cell populations in paraffin-embedded tissues by polymerase chain reaction.
A method was established to detect clonal B cell populations in frozen and paraffin-embedded tissues. The method is based on the polymerase chain reaction amplification of rearranged VH and V kappa genes using V gene family-specific primers. Monoclonal B cell populations could be detected in peripheral blood lymphocyte DNA in all 16 cases of B-CLL and immunocytoma investigated and in 8 of 10 cases of B cell non-Hodgkin's lymphoma using frozen and paraffin sections. The amplification of V kappa rearrangements in addition to the VH amplification is a useful tool to verify the results of the heavy chain rearrangement and to detect proliferation of a B cell clone in cases in which no VH product was obtained. In spite of the degradation of DNA in paraffin-embedded tissues, we were able to find amplified polymerase chain reaction products of about 350 bp length in 8 of 10 cases analyzed. The method presented here may be helpful in routine diagnosis of B cell non-Hodgkin's lymphoma using frozen or paraffin-embedded specimens.
Küppers
R
R
I. Medizinische Universitätsklinik Köln, Germany.
Zhao
M
M
Rajewsky
K
K
Hansmann
M L
ML
D016175:B-Lymphocyte Subsets / Q000166:cytology*; D001483:Base Sequence; D015139:Blotting, Southern; D002999:Clone Cells / Q000166:cytology*; D004587:Electrophoresis, Agar Gel; D005629:Frozen Sections; D005784:Gene Amplification; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D015448:Leukemia, B-Cell / Q000235:genetics; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains