Abstract The estimated incidence of genitourinary (GU) cancers (prostate, bladder, and kidney) will be over 2 million worldwide and responsible for ∼800 000 deaths by 2020. Current diagnosis and monitoring methods of GU cancer patients are often invasive and/or lack sensitivity and specificity. The recent tremendous success in cell-free DNA (cfDNA) blood test has laid the groundwork for using body fluids such as blood and urine as minimally invasive “liquid biopsies” to identify RNA- and DNA-based molecular biomarkers in urologic malignancies. Here we report the development and clinical application of PrediSeq panel, a urine- and blood-based next generation sequencing-based diagnostics assay that offers integrated genomic cfDNA and cell-free RNA (cfRNA) profiling of genitourinary cancers to detect RNA-based splicing variants (e.g. AR-V7) and rearrangement (e.g. TMPRSS2-ERG), DNA/RNA-based point mutation (e.g. p53, PIK3CA, FGFRs), DNA-based copy number change (e.g. AR) as well as tumor mutation burden (TMB) and microsatellite instability (MSI) using a single tube of blood and/or urine sample in prostate, bladder, and kidney cancers, respectively. In addition to NGS-based multiplex test, we also developed a Bio-Rad digital PCR assay measuring drug response and resistance biomarkers including AR-V7 and PD-L1. Collectively, PrediSeq NGS and ddPCR assays offer comprehensive genomic profiling of both cfDNA and cfRNA using blood and/or urine samples in patients with genitourinary cancers. The successful development and clinical validation of these tests has potential to enable early diagnosis, cancer screening, and precision medicine in genitourinary cancers. Citation Format: Amy Wang, Zhixin Zhao, Cheng Xie, Tak Cheung, Carlos Montesinos, Phoebe Zhang, Pan Du, Jianjun Yu, Shidong Jia. Urine- and blood-based DNA+RNA liquid biopsy tests in prostate, bladder and kidney cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3670.