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12.1 What are the key challenges to setting up molecular tests in histopathology?
Answer: To have a good understanding of the clinical relevance and of the limitations of the real life samples (size; quality of DNA; amount of tumour).
12.2 What would be the challenges to face in practice to replace formalin by an alternative fixative
Answer: Replacement fixative should preserve DNA, RNA and proteins better than formalin. It should not induce major differences to the morphology of tissues and be similar to formalin in this regard thus making it possible for the pathologists to be able to make an accurate diagnosis. It would need to be universally validated by the pathologist community. It would allow pathologists to use commercialised kits for ICC, ISH and FISH (which are currently only validated on FFPE samples).
12.3 What kind of molecular tests could be used to look for EGFR mutation in lung cancer and for RAS mutation in colorectal cancer?
Answer: Non-targeted tests (Sanger sequencing) or targeted tests (Pyrosequencing, real time PCR, NGS with panel including the genes of interest).
12.4 What are the ideal strategies for multiplex tests in molecular pathology?
Answer: It is yet to be established. Short tumour-specific panels using multiplex real time PCR-based assays or short NGS panels or extensive genome testing with targeted interpretation for what is clinically relevant are most likely developments.