A Novel Next-Generation Sequencing Approach without Donor-Derived Material for Acute Rejection and Infection Monitoring in Solid Organ Transplantation

Background: Donor-derived cell free DNA (ddcfDNA) has been reported as a universal noninvasive biomarker for rejection monitoringin heart, kidney, liver, and lung transplantation. Current approaches based on next-generationsequencing for quantification of ddcf DNA, although promising, may be restrictedby the requirement for donor material, as donor samples may not be available. Methods:We proposed a novel next-generation sequencing approach without donor-derived material and compared the non-donor-derived approach and the donor-derived approach using simulation testing and 69 clinicalspecimens. We also evaluated the performance for acute rejection and infection monitoring in lung transplantation. Results: The non-donor-derived approach reached similar efficacy as the donor-derivedapproach with a significant linear correlation of R2 = 0.98. Subsequent validation in clinical specimensdemonstrated significant difference between the acute rejection group (4.83% ± 2.11%, mean± SD) and the non-rejection group (1.61% ± 0.63%, mean ± SD) (P ’s t test). With the cut-offvalue of 2.999, our approach had 90.48% sensitivity (95% CI, 69.62% - 98.83%), 100%specificity (95% CI, 91.59% - 100%), and AUC 0.9266 (95% CI, 0.8277 - 1.026). Thetest also had the ability to simultaneously detect infectious agents, especiallycytomegalovirus, as compared with the clinical test. Conclusion: The proposedapproach without donor-derived material could potentially be used to monitor acuterejection and infection in lung transplantation and may be applied to other typesof solid organ transplantation.