Quantitative and Sensitive Detection of Cancer Genome Amplifications from Formalin Fixed Paraffin Embedded Tumors with Droplet Digital PCR

摘要: For the analysis of cancer, there is great interest in rapid and accurate detection cancer genome amplifications containing oncogenes that are potential therapeutic targets. The vast majority tissue samples formalin fixed paraffin embedded (FFPE) which enables histopathological examination long term archiving. However, FFPE genomic DNA oftentimes degraded generally a poor substrate for many molecular biology assays. To overcome issues quality from detect oncogenic copy number with high accuracy sensitivity, we developed novel approach. Our assay requires nanogram amounts DNA, thus facilitating study small clinical samples. Using droplet digital PCR (ddPCR), can determine relative specific loci even presence intermingled normal tissue. We used control dilution series to limits ddPCR report its improved sensitivity on minimal compared standard real-time PCR. develop this approach, designed an fibroblast growth factor receptor 2 gene (FGFR2) amplified gastric breast cancers as well others. successfully utilized ascertain FGFR2 FFPE-preserved gastrointestinal adenocarcinomas.