Establishment of PCR based methods for detection of ctDNA in blood

TL;DRAbstract

Cell-free DNA appears in the circulation when cells undergo apoptosis and necrosis. This is a natural process and small amounts of cell-free DNA can be found in the blood of healthy individuals. When tumors are present, a variable part of the cell-free DNA will derive from tumor. Monitoring ctDNA is attractive in solid cancers because a blood sample is less invasive than a biopsy and it might represent all lesions including metastasis. “Liquid biopsies” also give the opportunity for serial monitoring during therapy and follow up when tumor biopsies are difficult to achieve. Little is still known about the stability of ctDNA in vitro, so one of the subprojects in this thesis was to elucidate this. Cell-free DNA is extracted from plasma and ctDNA is often a minor fraction of the total plasma DNA. Our results showed that blood can be left for 24 hours in room temperature without significantly affecting the total plasma-DNA concentration. It was however not clear if ctDNA showed a slight d

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