COMPARATIVE-ANALYSIS OF DETECTION SYSTEMS FOR EVALUATION OF PCR AMPLIFIED IMMUNOGLOBULIN HEAVY-CHAIN GENE REARRANGEMENTS

Four different detection systems were compared for evaluation of polym erase chain reaction (PCR) amplified immunoglobulin heavy-chain gene r earrangements in acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphoma (NHL) of B-cell lineage. In 63.0% of the fragments detected b y ethidium bromide stained agarose gel electrophoresis (Agarose-EtBr) the sensitivity was insufficient to separate the specific clonal popul ation from the background of normal B cells. Using polyacrylamide gel electrophoresis (PAGE), PAGE combined with single-strand conformation polymorphism (PAGE-SSCP) and PhastGel-SSCP (Phast-SSCP) analysis with silver staining, the resolution was improved and the majority of the i nconclusive amplicons were elucidated. However, Phast-SSCP displayed a slightly higher detection level compared to PAGE and PAGE-SSCP. Accor ding to our findings PAGE-SSCP and Phast-SSCP were superior to agarose -EtBr and PAGE in detecting new emerging clones and clonal evolution.