Sensitive detection of clonal immunoglobulin rearrangements in frozen and paraffin embedded tissues by polymerase chain reaction heteroduplex analysis

Molecular detection of a clonal population of B or T cells through analysis of rearranged antigen receptor genes is an essential adjunct to the morpho logic, flow cytometric, and immunohistochemical evaluation of tissue specim ens for the presence of leukemia or lymphoma. Combining polymerase chain re action (PCR) with heteroduplex annealing and polyacrylamide Sd electrophore sis (PAGE) has been used to detect clonal T-cell receptor rearrangements, p articularly in skin biopsy specimens. The authors have developed a similar PCR heteroduplex assay fur detection of clonal VDJ immunoglobulin gene rear rangements using two sets of primers based on relatively conserved consensu s regions in the J(H) and framework 1 and 2 regions of the immunoglobulin h eavy chain V region gene. This method is able to detect a clonal rearrangem ent when the clone comprises as little as 1% of the population in a polyclo nal B-cell background. It may be used on fresh, frozen, or paraffin-embedde d tissue and detects a clonal population in a majority of lymphoma subtypes . Compared with conventional PCR analysis, this method requires only a shor t additional cycle of denaturation and slow renaturation before PAGE. inter pretation is simplified as the clonal PCR product migrates away from the po lyclonal background products.