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Assessment of clonal evolution at Ig/TCR loci in acute lymphoblastic leukaemia by single‐strand conformation polymorphism studies and highly resolutive PCR derived methods: implication for a general strategy of minimal residual disease detection

Junctional sequences of immunoglobulin (Ig)/T‐cell receptor (TCR) gene rearrangements are used as patient‐specific PCR targets for the detection of minimal residual disease (MRD) in acute lymphoblastic leukaemias (ALLs). Clonal evolution of gene rearrangements is a major pitfall of this strategy. Using high‐resolution PCR‐based analyses (including denaturing gel electrophoresis and single‐stranded conformation polymorphism (SSCP)) we have compared Ig/TCR gene rearrangements at presentation and relapse in a series of ALLs. These methods allow an unambigous comparison of rearrangements taking into account junctional size and nucleotide sequence information and allow a precise assessment of the clonal evolution. V7‐J7 and V6j‐J6i rearrangements were analysed in 12 T‐ALLs. VH‐JH, V7‐J7, V62‐D63 and, in selected cases, DH‐JH rearrangements were studied in 14 B‐lineage ALLs. Clonal evolution, regarding major rearrangements, occurs for at least one of these loci in 2/12 T‐ALLs and in 5/14 B‐lineage ALLs. Clonal evolution is more marked for minor rearrangements than for major ones. As shown using SSCP analysis, rearrangements observed at relapse are sometimes found in minor clones at presentation which are therefore selected in vivo by a proliferative advantage. These data, as well as those from the available literature, suggest the use of at least two patient‐specific probes to detect MRD in ALLs. A general strategy including selected Ig/TCR rearrangements and chromosomal abnormalities as PCR targets is proposed.