Australian Chronic Allograft Dysfunction II

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 chronic allograft dysfunction in kidney transplant recipients. Our study recruited 136 patients, each having protocol renal allograft biopsies taken pre transplantation. 136 patients had renal allograft back table biopsies collected pre-transplant at Westmead hospital. A combined criteria was used to classify the early function of the graft as either immediate or delayed. Kidney biopsy specimens were left in RNAlater (Sigma-Aldrich) overnight at 4°C before removal and storage at -80°C until RNA extraction. Specimens were chemically and physically lysed by using 2-Mercaptoethanol (Sigma-Aldrich) and TissueLyser II (Qiagen, Germany), follow by RNA extraction using AllPrep DNA/RNA/microRNA and MiniElute clean- up kits (Qiagen, Germany). All RNA samples were frozen and stored at -80°C and then sent in bulk to the Australian Genome Research Facility (AGRF, Melbourne, Australia). Sample QC and library preparation were performed in-house, and the resultant libraries sequenced using the NovaSeq 6000 platform (Illumina)