The rapid advances of Next-generation sequencing (NGS) technologies have greatly facilitated clinical genetic diagnosis of sporadic disease genome-widely. However, the vast amount of mutations generated by NGS poses multiple challenges for identification of functional mutations and candidate genes. We devolopped a web server named mirTrios to accurately detect de novo mutations (DNMs) based on Expectation-maximization (EM) model. In addition, to facilitate the interpretation of diverse mutations, mirTrios also surports identification of rare inherited mutations, known diagnostic variants, as well as the prioritization of novel and promising candidate genes. mirTrios provides an intuitive interface for the general geneticist and clinicians, which is freely available at http://centre.bioinformatics.zj.cn/mirTrios/. (By Dr. Jinyu Wu, http://jmg.bmj.com/content/early/2015/01/16/jmedgenet-2014-102656 )