Comprehensive Biomarker Approach By Utilizing qPCR, NGS, in situ PCR: A Tool Box To Decipher Cancer

Robert P. Loewe
GeneWake GmbH, Germany

In many indications like oncology there is an increasing demand to stratify patient populations with reliable biomarkers to ensure effective therapies. Taking colorectal cancer as an example, multiple methods are demonstrated in a combinational approach to analyze biomarkers from different angles to provide a larger picture. After a short overview of routine methods like pathological examination, micro-dissection and purification of DNA/RNA, mutation analyses of known markers via next generation sequencing (NGS) and high resolution melting (HRM) are highlighted. Subsequently, gene expression analysis (mRNA and microRNA) and epigenetic analysis with its impact on cancer therapy are shown. Finally, the use of in situ PCR will be demonstrated, including the development of a highly innovative method to visualize mutations directly in formalin fixed tissue sections. Taken together, this combinational approach can help to understand complex patterns of biomarkers and the interaction of genes and also to evaluate and find those biomarkers that are useful as companion diagnostics or those that are therapeutically relevant for an improved and personalized treatment of cancer.

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