抄録
Junctional sequences created by chromosomal translocation in mature B-cell neoplasms, which involve immunoglobulin gene (IG) loci and oncogenes on reciprocal partner chromosomes, are unique to neoplastic cells characterized by particular histological and immunological phenotypes. We present here a novel strategy for detection of the junctional sequences based on long-distance polymerase chain reaction (LD-PCR) amplification. LD-PCR is a general method using primer pairs designed for distinctive regions of the oncogene and IG involved in translocations, and amplifying sequences encompassing the oncogene/IG junction of 2kb up to 30kb in size. LD-PCR is capable of detecting virtually all the important translocations in B-cell neoplasms, including t(8;14)(q24;q32), t(14;18)(q32;q21) and t(3;14)(q27;q32) and its variants. In Burkitt's lymphoma, all materials with the sporadic type c-MYC/IGH fusion showed positive LD-PCR amplification. Restriction analysis and sequencing of the LD-PCR products from BCL2/IGH fusion revealed that breakpoints on BCL2 were distributed over a large region downstream of the BCL2 coding region. 3q27 translocations involving BCL6 and three IGs showed heterogeneous structure at the molecular level. The present ent study suggested that LD-PCR can substitute for time-consuming Southern blot hybridization and cytogenetic analysis in the rapid and sensitive detection of these translocations. Furthermore, as amplified fragments obtained by the LD-PCR contained distinctive regions of oncogenes and IGs, restriction analysis and nucleotide sequencing of the products refined the characteristics of translocations.
