Applying rigor and reproducibility standards to assay donor-derived cell-free DNA as a non-invasive method for detection of acute rejection and graft injury after heart transplantation.

Published

Journal Article

BACKGROUND: Use of new genomic techniques in clinical settings requires that such methods are rigorous and reproducible. Previous studies have shown that quantitation of donor-derived cell-free DNA (%ddcfDNA) by unbiased shotgun sequencing is a sensitive, non-invasive marker of acute rejection after heart transplantation. The primary goal of this study was to assess the reproducibility of %ddcfDNA measurements across technical replicates, manual vs automated platforms, and rejection phenotypes in distinct patient cohorts. METHODS: After developing and validating the %ddcfDNA assay, we subjected the method to a rigorous test of its reproducibility. We measured %ddcfDNA in technical replicates performed by 2 independent laboratories and verified the reproducibility of %ddcfDNA patterns of 2 rejection phenotypes: acute cellular rejection and antibody-mediated rejection in distinct patient cohorts. RESULTS: We observed strong concordance of technical-replicate %ddcfDNA measurements across 2 independent laboratories (slope = 1.02, R2 > 0.99, p < 10-6), as well as across manual and automated platforms (slope = 0.80, R2 = 0.92, p < 0.001). The %ddcfDNA measurements in distinct heart transplant cohorts had similar baselines and error rates. The %ddcfDNA temporal patterns associated with rejection phenotypes were similar in both patient cohorts; however, the quantity of ddcfDNA was significantly higher in samples with severe vs mild histologic rejection grade (2.73% vs 0.14%, respectively; p < 0.001). CONCLUSIONS: The %ddcfDNA assay is precise and reproducible across laboratories and in samples from 2 distinct types of heart transplant rejection. These findings pave the way for larger studies to assess the clinical utility of %ddcfDNA as a marker of acute rejection after heart transplantation.

Full Text

Duke Authors

Cited Authors

  • Agbor-Enoh, S; Tunc, I; De Vlaminck, I; Fideli, U; Davis, A; Cuttin, K; Bhatti, K; Marishta, A; Solomon, MA; Jackson, A; Graninger, G; Harper, B; Luikart, H; Wylie, J; Wang, X; Berry, G; Marboe, C; Khush, K; Zhu, J; Valantine, H

Published Date

  • September 2017

Published In

Volume / Issue

  • 36 / 9

Start / End Page

  • 1004 - 1012

PubMed ID

  • 28624139

Pubmed Central ID

  • 28624139

Electronic International Standard Serial Number (EISSN)

  • 1557-3117

Digital Object Identifier (DOI)

  • 10.1016/j.healun.2017.05.026

Language

  • eng

Conference Location

  • United States