dPCR assay for monitoring donor-derived cfDNA
SOtrack® is an HLA-informed, digital PCR (dPCR)-based assay currently under development for monitoring donor-derived cell-free DNA (cfDNA) in solid organ transplantation research. Using HLA-specific markers and the Absolute Q dPCR platform, SOtrack aims to provide sensitive and rapid quantification of cfDNA without the need for pre-transplant donor DNA.
Donor-derived cfDNA is widely being evaluated as a biomarker for monitoring graft injury in solid organ transplantation. Monitoring cfDNA levels in peripheral blood could be used to confirm and even substitute invasive, expensive and subjective biopsies. SOtrack™, uses informative markers determined based on HLA typing in combination with dPCR. This eliminates the need for (additional) donor pre-transplant DNA typing.
SOtrack utilizes allele group-specific HLA-DRB1 and HLA-DQB1 markers selected based on donor/recipient typing. The assay involves a pre-amplification step followed by a dual-channel (FAM/HEX) dPCR reaction on the QuantStudio Absolute Q platform. Each channel represents either donor or recipient alleles, allowing precise quantification of donor-derived cfDNA in peripheral blood.
Performance
Accurate detection of low-level chimerism (validated down to 0.2%)
R² > 0.99 for heterozygous samples, R² > 0.98 for homozygous samples
Turnaround time < 3.5 hours
Hands-on time ~30 minutes
Successfully adapted from QX200 protocol to Absolute Q with assay-specific optimizations
Currently includes 12 DRB1 and 4 DQB1 markers; additional loci under evaluation
SOtrack is currently in development.
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