Australian Chronic Allograft Dysfunction I

The Australian Chronic Allograft Dysfunction (AUSCAD) study is an ongoing single centre cohort study at Westmead hospital in Australia. In this section of the study, we aimed to identify biomarkers for rejection phenotypes in kidney transplant recipients, 3-months after their transplant. Our study recruited 70 patients, each having whole blood taken at the time of their 3-month protocol biopsy. 70 patients had paired renal allograft biopsies and blood draws collected 3-months post-transplant at Westmead hospital. Biopsies collected from patients were scored by a single pathologist using the Banff19 scoring schema. A composite definition of rejection (rej) was used for this study. Specially, patients with T-cell mediated rejection (TCMR), antibody mediated rejection (ABMR) and mixed rejection phenotypes were classified as a rejection blood sample. Peripheral blood was collected into PAXgene Blood RNA tubes (Qiagen, Germany), left at room temperature for 5 hours, and stored at -80°C until RNA extraction. RNA was extracted by using PAXgene Blood miRNA kit (Qiagen, Germany). RNA samples were frozen and stored at -80°C.Frozen RNA samples were then sent to Australian Genome Research Facility (AGRF), Melbourne, Australia. Sample QC and library preparation were performed in-house by AGRF, and the resultant libraries sequenced using the NovaSeq 6000 platform (Illumina).