Neusticomys peruviensis is a poorly known sigmodontine rodent of the tribe Ichthyomyini, represented in collections by only five specimens collected in five localities from lowland forests of central and southern Peru. Recent expeditions in Llanchama, in northern Peru, north of the Río Amazonas, and near Allpahuayo Mishana Natural Reserve (Loreto, Peru), were successful in obtaining three specimens of Neusticomys. Based on morphological and meristic data, we found the population at Llanchama is distinct from the allopatric populations of N. peruviensis, and other species of Neusticomys. A species distribution model also shows the population at Llanchama is not highly predicted by the set of variables of the known localities of N. peruviensis. However, sequence data from the mitochondrial cytochrome-b gene indicate that genetic distinctiveness is low. Because intraspecific variability is important to understand evolutionary and biogeographic processes, and in concordance with the polytypic species concept, we interpret the population at Llanchama to represent a new subspecies of N. peruviensis that we describe in this paper.