Analysis of Rearranged T-cell Receptor /?-Chain Genes by Polymerase Chain Reaction (PCR) DNA Sequencing and Automated High Resolution PCR Fragment Analysis

Polymerase chain reaction (PCR)-directed amplification and sequencing of rearranged immune genes for identification of clone-specific markers are increasingly being used in acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphoma INHL) patients instead of the time consuming and labor intensive Southern analysis. In previous reports, no single common VP and JP sequence had been identified that allowed reliable amplification of the majority of rearranged Tcell antigen receptor (TCR)-p V-D-J junctions at the DNA level because of the relatively large number of possible TCRP variable (VPl and joining [JP) gene segments involved in the rearrangement processes. In the present study we designed highly degenerate PCR primers directed against conserved sequences of the JP genes. In combination with a previously published consensus Vp primer, these Jp primers specifically amplify TCR-/3 V-NID)N-J junctions from genomic DNA. Using this approach we studied DNA extracted from biopsy material of nine patients with T-cell lymphoproliferative disorders, one c-ALL patient, and five patients with