Introduction: Pancreatic ductal adenocarcinoma (PDAC) accounts for approximately 90-95% exocrine malignant tumors of the pancreas.The high prevalence of metastasis and the difficulty of early diagnosis lead to a dismal prognosis.MicroRNAs (miRNAs) play a critical role in extensive biological processes.The purpose of this study was to evaluate the feasibility of stool miRNAs as novel biomarker for PDAC screening.Materials and Methods: MiRNAs were extracted from clinical specimens which included cancer and matched adjacent benign pancreatic tissues of 30 PDAC patients, pancreatic juice of 20 from the 30 PDAC patients and 10 chronic pancreatitis (CP) patients,stool samples of the 30 PDAC patients,the 10 CP patients and 15 healthy volunteers.Relative expression of a panel of 5 dysregulated miRNAs (miR-21 ,miR-155 ,miR-196a,miR-216 and miR-217) was analyzed with qRT-PCR.Receiver operating characteristic curve (ROC) analysis was performed to assess the diagnosing value of stool miRNAs in PDAC patients.Results:The study showed that our methods of extracting and detecting miRNAs from pancreatic juice and stool specimens had high reproducibility.Compared to matched adjacent benign pancreatic tissues and pancreatic juice of CP patients,the expression of miR-21 (P =0.0021 and P =0.0027) as well as miR-155 (P =0.0087 and P =0.0067) was significantly higher and the expression of miR-216 (P <0.0001 and P =0.0044) was significantly lower in primary tumor tissues and pancreatic juice of PDAC patients.PDAC patients had a significantly higher stool miR-21 and miR-155 (P =0.0049 and P =0.0112) and lower miR-216 level (P =0.0002) compared to normal controls.The same results were obtained in the expression levels of stool miR-21, miR-155 and miR-216 between PDAC and CP patients (P =0.0337 ,P =0.0388 and P =0.0117 ,respectively).Receiver operating characteristic (ROC) analysis by using stool miRNAs expression indicated that combination of miR-21 and miR-155 had best sensitivity of 93.33% while the combination of miR-21 ,miR-155 and miR-216 would be best for detecting and screening PDAC with area under the curve (AUC) of 0.8667 (95% CI:0.7722-0.9612) and a better balance of sensitivity and specificity (83.33 % vs.83.33%).Conclusion:Our data indicate that miRNAs could be extracted and detected from pancreatic juice and stool efficiently and reproducibly.MiR-21 ,miR-155 and miR-216 in stool have the potential of becoming biomarkers for screening PDAC.