Chimeric external control to quantify cell free DNA in plasma samples by real time PCR

ABSTRACT

Background: DNA isolation procedure can significantly influence the quantification of DNA by real time PCR specially when cell free DNA [cfDNA] is the subject. To assess the extraction efficiency, linearity of the extraction yield, presence of co-purified inhibitors and to avoid problems with fragment size relevant to cfDNA, development of appropriate External DNA Control [EDC] is challenging. Using non-human chimeric nucleotide sequences, an EDC was developed for standardization of qPCR for monitoring stability of cfDNA concentration in blood samples over time