MULTIPLEX PCR FOR SIMULTANEOUS DETECTION OF THE MOST FREQUENT T-CELL RECEPTOR DELTA-GENE REARRANGEMENTS IN CHILDHOOD ALL

A rapid and simple multiplex polymerase chain reaction (PCR) is descri bed that is capable of identifying the six most frequent rearrangement s of the T cell receptor (TCR)-delta gene segments in childhood acute lymphoblastic leukemia (ALL). The PCR products amplified in a single r eaction are of different size for each TCR-delta gene rearrangement. T herefore, they are readily and unambiguously distinguished after agaro se gel electrophoresis and assigned to a specific V-D-J gene rearrange ment. There is no need for labor-intensive and time-consuming Southern blot hybridization or nested PCR. To evaluate the multiplex assay we chose 45 DNA samples of childhood ALL analyzed beforehand for TCR-delt a gene rearrangements by Southern blot and single PCR of which 30 show ed TCR-delta gene rearrangements. The multiplex PCR results correspond ed to the Southern blot and single PCR analyses. The described multipl ex PCR enables the detection of clonal markers in about 50% of patient s in order to monitor minimal residual disease (MRD) in prospective st udies with a high turnover of samples.