MOLECULAR ANALYSIS OF T-CELL RECEPTOR V-BETA CHAIN TO DETECT LEUKEMIA-CELL CLONALITY IN PATIENTS BY ADAPTER LIGATION-MEDIATED POLYMERASE CHAIN-REACTION

We have developed a simple and rapid method to analyze the clonality o f leukemia cells. After three rounds of amplification by adaptor-ligat ion polymerase chain reaction (PCR), the cDNA is cut with AluI, HaeIII , RsaI, and Sau3AI, and analyzed by polyacrylamide gel electrophoresis . The size of the restriction fragments is compared to that of the pub lished restriction fragments size each TCR-beta subfamily V region. Th e sensitivity of adaptor-ligation PCR restriction enzyme analysis (AL- PCR-REA) was 10(-4) MOLT-4 T-ALL cell population in the normal periphe ral blood lymphocytes (PBL). Application of AL-PCR-REA to PBL and bone marrow (BM) cells from eight clinical leukemia samples indicated that a detection sensitivity was rather low, but revealed the clonality of all eight clinical samples. This AL-PCR-REA method can detect clonali ty without the need for either radioisotopes or sequencing procedures.