APPLICATION OF LONG-DISTANCE POLYMERASE CHAIN-REACTION TO DETECTION OF JUNCTIONAL SEQUENCES CREATED BY CHROMOSOMAL TRANSLOCATION IN MATURE B-CELL NEOPLASMS

Junctional sequences created by chromosomal translocations in mature B -cell neoplasms, which involve immunoglobulin gene loci (IG) and putat ive proto-oncogenes on reciprocal partner chromosomes, are unique to n eoplastic cells characterized by particular histological and immunolog ical phenotypes. To establish a rapid and sensitive method to detect n eoplastic cells carrying a specific chromosomal translocation, we have developed a novel strategy based on long-distance polymerase chain re action (LD-PCR) amplification. Genomic DNA was extracted from tumor ce lls carrying t(14;19)(q32;q13), t(8;14)(q24;q32), t(3;22)(q27;q11), t( 2;3)(p12;q27), or t(3;14)(q27;q32). Thirty-two to 35-mer oligonucleoti de primer pairs were designed to be complementary to exons or flanking sequences of the BCL3, c-MYC and BCL6 oncogenes, and to IG constant r egion genes. LD-PCR with a newly available Tag polymerase for longer p roduct synthesis successfully amplified fragments representing BCL3/C alpha junctional sequences for t(14;19); c-MYC/C mu, c-MYC/C gamma, an d c-MYC/C alpha for t(8;14); BCL6/C lambda for t(3;22); BCL6/C kappa f or t(2;3); 5'-BCL6/C mu, and 5'-BCL6/C gamma for t(3;14), In Burkitt's lymphoma/leukemia, all materials in which c-MYC rearrangements were d etectable by conventional Southern blot hybridization showed positive LD-PCR amplification, The sizes of the amplified fragments varied from 1.8 kb to 12 kb, and these were specific to each material. Serial dil ution of tumor cells or DNA in negative materials demonstrated a singl e band on agarose gel electrophoresis stained with ethidium bromide at a level of sensitivity of 10(-3), and hybridization with radioactive probe improved the level by one order of magnitude (1 cell in 10(4)), indicating that this LD-PCR approach is a sensitive technique capable of detecting minimal residual disease. Thus, the present study provide d a useful tool for diagnosis and subsequent management of B-cell neop lasms characterized by specific chromosomal translocations. (C) 1996 b y The American Society of Hematology.