This project aimed to determine the robustness and uses of in situ mutation detection using specific probes to RNAs carrying cancer driver mutations in human tumour samples. Possible ultimate uses included identifying residual disease in apparently excellent responders to radio-/chemotherapy and for identifying mutations (for example, in transgenic mouse models) in scenarios in which antibody-based detection was infeasible and DNA-based detection provided insufficint cellular resolution. The potential of offering the analyses using the underpinning BaseScope technology as a commercial venture was to be explored. We found that the reliability of the probes was limited by the quality and age of the tissue specimen, such that recent specimens could be analysed reliably. Despite these problems, we have extended mutation detection to driver mutations including “atypical” BRAF mutations at codons 594 and 466. We have also recently designed probes for driver mutations in the IDH1 and IDH2 genes (R132H, R132C, R132G, R172K, R140H) and will analyse these intially in mouse models of cancer. We are also analusing competing technologies, such as single cell sequencing, alongide the in situ methods. Commerical diagnostic use of the probes and method does not seem to have a market currently, although this will be re-assessed over the coming 12 months.
