There is a lack of study of Extracellular vesicle-derived microRNAs (EV miRNAs) as biomarkers in nonalcoholic fatty liver disease (NAFLD) and NAFLD-associated hepatocellular carcinoma (NAFLD-HCC). In the present study, we investigated the EV miRNAs as diagnostic biomarkers for NAFLD and NAFLD-HCC by miRNA profiling using the NanoString miRNA Expression Assay in a discovery set and validating in an independent set of plasma samples by quantitative RT-PCR. Pre-treatment plasma samples of NAFLD and NAFLD-HCC along with healthy controls were collected to isolate EVs and characterized by nanoparticle tracking analysis, western blot, and transmission electron microscopy. The expression profiles of EV miRNAs in the plasma of the patients with NAFLD and NAFLD-HCC to healthy controls were compared (n=9 per group). Differentially expressed miRNAs were observed from the discovery set, and the most significant upregulated miRNA in the NAFLD-HCC group compared with the other groups, miR-19b-3p, was identified. To examine the expression level of miR-19b-3p in the validation cohort, a total of 194 participants were preliminary enrolled. NAFLD patients were sub-group analyzed via Fibroscan for grading of steatosis using controlled attenuation parameter (CAP). S1-S2 NAFLD and S3 NAFLD were patients with CAP of 249-280 dB/m and 249-280 dB/m, respectively. miR-19b-3p from the plasma EVs showed significantly higher in NAFLD-HCC group (n=74) than S1-S2 NAFLD (n=37) (p=0.001), S3 NAFLD (n=39) (p=0.010), and healthy control group (n=44) (p<0.001). Our data demonstrated that the expression profiles of EV miRNAs of patients with NAFLD-HCC and NAFLD were different from the healthy control group. Indeed, EV miRNA miR-19b-3p might act as a biomarker for the diagnosis of NAFLD-HCC.