Digital PCR for absolute quantification of cell-free DNA introduces new
aspects in the surveillance of patients after heart transplantation:
clinical scenarios
Abstract
Background: Heart transplantation (HTx) relies on an endomyocardial
biopsy for rejection surveillance. Cell-free DNA (cfDNA) is released
into the circulatory system from the cells of the recipient
(recipient-derived rd-cfDNA) as well as from the graft (donor-derived
dd-cfDNA) and is determined by the analysis of a blood test. There is
growing evidence that the donor fraction (DF, the ratio of dd-cfDNA to
all cfDNA) corresponds to the integrity of the graft. However, it is
important that rd-cfDNA levels vary greatly in different, sometimes
clinically silent circumstances, which in turn makes DF the sole measure
of graft status vulnerable to bias. Methods: Using a digital PCR-based
method that allowed us to quantify separate values of rd-cfDNA,
dd-cfDNA, and DF, we investigated a prospective cohort of 52 patients
during their first year after HTx. Results: We found median DF levels of
approximately 0.1%, which is in accordance with the literature. We
present patients with various clinical scenarios after HTx, including an
uncomplicated clinical course, several infections (viral and bacterial),
acute and chronic rejection, and false-negative and false-positive
rejections. Conclusions: Quantification of circulating cfDNA reflects a
live-view of cell turnaround in the recipient and graft. Measuring the
DF alone may not be sufficient to fully understand the complex biology
of cfDNA after HTx.