Alterations in microRNA (miRNA) expression in postmortem brain tissue or peripheral blood have been linked to schizophrenia. Cerebrospinal fluid might provide an in vivo biomarker more directly reflecting functional changes in the brain. The goals of this study were to determine the feasibility of detecting miRNAs in cerebrospinal fluid and to compare miRNA levels in cerebrospinal fluid versus blood. Four healthy volunteers and four patients with psychotic disorders underwent a lumbar puncture and a blood draw. Expression of 378 validated miRNAs was assessed from each biofluid type for each subject using microarray technology. Five miRNAs were chosen for validation with quantitative polymerase chain reaction. A substantial number of miRNAs (n = 95) were exclusively or predominately detected in cerebrospinal fluid (CSF). Levels of 35 miRNAs detected in both CSF and blood samples in all subjects were poorly correlated. The investigation of miRNAs in CSF can help advance the understanding of psychiatric diseases and particularly schizophrenia.