SIMPLE, RELIABLE DETECTION OF T-CELL CLONES BY PCR-LIS-SSCP ANALYSIS OF TCR-GAMMA REARRANGEMENT

Clonal populations of T cells can be identified by polymerase chain re action (PCR) amplification of the rearranged T cell receptor gamma (TC RG) chain gene. However, because of the limited combinatorial diversit y of this locus it is necessary to separate the PCR product on the bas is of sequence as well as size to distinguish clonal and polyclonal T cell populations. A simple method is described which achieves this by analysing the PCR product on a single-stranded conformation polymorphi sm (SSCP) gel. Sensitivity has been improved by denaturing the DNA usi ng a low ionic strength (LIS) method rather than the more conventional alkali or formamide. Results from the PCR-LIS-SSCP method on a wide r ange of disorders and types of tissue samples show that clonality coul d be demonstrated in 40/44 cases. (C) 1998 Academic Press Limited.