Yang Yu,* Xiao Feng,* Shundong Cang Department of Oncology, Henan Province People’s Hospital, Henan University, Zhengzhou, Henan, China *These authors contributed equally to this work Background and aim: Some cancer-specific miRNAs are dysregulated in pancreatic adenocarcinoma (PAAD) and involved in cell autophagy, differentiation, proliferation, migration, invasion, and malignant transformation. The aim of our study was to determine a panel of new diagnostic and prognostic biomarkers for PAAD. Methods: We conducted a comprehensive analysis of global miRNA-expression profiles and corresponding prognosis information of 168 PAAD patients from the Cancer Genome Atlas data set. A total of 16 differentially expressed miRNAs were identified as aberrantly expressed in PAAD, and six of these were evaluated for use as diagnostic markers for PAAD. Next, we confirmed a two-miRNA signature significantly associated with PAAD patient diagnosis and outcome prediction. Results: The panel of two miRNAs showed outstanding diagnostic performance, with sensitivity of 100% and specificity of 87.5%. Finally, we divided the PAAD patients into high-risk and low-risk groups based on the expression profile of the two miRNAs. Kaplan–Meier analysis demonstrated that patients in the high-risk group had significantly worse prognosis than patients in the low-risk group. Univariate and multivariate Cox regression analysis showed that the two-miRNA signature was an independent prognostic factor for the overall survival of PAAD patients. Conclusion: Taken together, the two-miRNA signature may serve as an accurate and sensitive biomarker for diagnosis and PAAD-outcome prediction, facilitating the diagnosis and potentially improving treatment outcome of PAAD. Keywords: pancreatic adenocarcinoma, microRNA signature, TCGA, prognosis, diagnosis