Major advances in molecular detection are being driven by goals associated with the development of methods that are amenable to miniaturization and automation, and that have high sensitivity and low interference. The new detection methods are confronted by many interfacial issues, which when properly addressed can lead to improved performance. One interfacial property, special wettability, can facilitate precise delivery and local enrichment of molecules to sensing elements. This review summarizes applications of unique features of special wettability in molecular detection including (1) chemical and electrochemical reactions in anchored microdroplets on superwetting surfaces, (2) enrichment of analytes and active materials at low contact areas between droplets and superwetting surfaces, (3) complete opposite affinities of superwetting surfaces toward nonpolar/polar solutes and oil/water phases, and (4) directional droplet transportation on asymmetric superwetting surfaces. The challenges and opportunities that exist in design and applications of special wettability in interfacial delivery and enrichment for detection of a few molecules are also discussed. This review highlights interfacial design to reach major objectives in the field of molecular detection, including high sensitivity, low interference, miniaturization and automation.