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Sustained poor survival rate in pancreatic ductal adenocarcinoma (PDAC) calls for an earlier diagnosis to assure curative treatment. New powerful biomarkers are necessary because the currently used CA19-9 is not sensitive enough to distinguish PDAC, especially from chronic pancreatitis (CP). Expressions of miRNA-21, -30 -192, -196, -200, and -423 were measured in 77 patients with PDAC, 26 patients with CP and 64 non-cancer/non-CP subjects (39 patients with type 2 diabetes mellitus and 25 control healthy persons). Eleven patients with PDAC had CP at the background. The expressions of all microRNAs were significantly 1.4-3.7 times higher in the PDAC group compared to non-cancer/non-CP subjects and 2.2-6.1 times higher compared to CP patients. No difference in miRNA expressions was found between diabetic and non-diabetic patients. CA19-9 did not distinguish CP from PDAC patients with the history of CP, whereas all six miRNAs were able to do it. Adding miR-196, -200 and -423 to current marker CA19-9 improved sensitivity by 7 % (to 93 %) and specificity by 8 % (to 89 %). MicroRNA-423 could significantly distinguish PDAC from CP with both sensitivity and specificity 96 %. Panel of six miRNAs could be used as reliable marker in differentiating PDAC from chronic pancreatitis with the most impressive difference in miR-196 and miR-423. Key words microRNA " Pancreatic ductal adenocarcinoma " Chronic pancreatitis " Biomarker " CA19-9.