This review addresses recent progress made in the use of nanofibers for analyte detection and sample preparation within analytical devices. The unique characteristics of nanofibers make them ideal for incorporation within sensors designed to allow for sensitive detection of clinical, environmental, and food safety analytes. In particular, the extremely large surface area provided by nanofiber mats and arrays drastically increases the availability of immobilization sites within biosensors. Additionally, nanofibers can be made from a variety of biocompatible materials and can be functionalized through the incorporation of nanoscale materials within spinning dopes or polymerization solutions. Finally, methods of nanofiber formation are largely well understood, allowing for controlled synthesis of nanofiber mats with specific sizes, shapes, pore sizes, and tensile strengths. In this paper, we present a survey of the different materials that are currently being used to produce nanofibers for use within sensing devices. In addition, we compare the limits of detection and linear ranges of nanofiber-based sensors and conventional sensors to determine if detection is improved by the inclusion of nanoscale materials (review).