Secondary Logo

Institutional members access full text with Ovid®

Sensitive Detection of Clonal Immunoglobulin Rearrangements in Frozen and Paraffin Embedded Tissues by Polymerase Chain Reaction Heteroduplex Analysis

Ranheim, Erik A. M.D., Ph.D.; Jones, Carol D. B.S.; Zehnder, James L. M.D.

Diagnostic Molecular Pathology: December 2000 - Volume 9 - Issue 4 - pp 177-183

Molecular detection of a clonal population of B or T cells through analysis of rearranged antigen receptor genes is an essential adjunct to the morphologic, flow cytometric, and immunohistochemical evaluation of tissue specimens for the presence of leukemia or lymphoma. Combining polymerase chain reaction (PCR) with heteroduplex annealing and polyacrylamide gel electrophoresis (PAGE) has been used to detect clonal T-cell receptor rearrangements, particularly in skin biopsy specimens. The authors have developed a similar PCR heteroduplex assay for detection of clonal VDJ immunoglobulin gene rearrangements using two sets of primers based on relatively conserved consensus regions in the JH and framework 1 and 2 regions of the immunoglobulin heavy chain V region gene. This method is able to detect a clonal rearrangement when the clone comprises as little as 1% of the population in a polyclonal B-cell background. It may be used on fresh, frozen, or paraffin-embedded tissue and detects a clonal population in a majority of lymphoma subtypes. Compared with conventional PCR analysis, this method requires only a short additional cycle of denaturation and slow renaturation before PAGE. Interpretation is simplified as the clonal PCR product migrates away from the polyclonal background products.

From Department of Pathology, Stanford University Medical Center, Stanford, California, U.S.A.

Address correspondence and reprint requests to Dr. Erik A. Ranheim, Department of Pathology, Stanford University Medical Center, L-235, Stanford, CA 94305.

© 2000 Lippincott Williams & Wilkins, Inc.