Agroathelia rolfsii (anamorph: Sclerotium rolfsii) is a soilborne fungal pathogen that can cause disease on over 500 documented host species, including economically important field and vegetable crops. The pathogen commonly infects the stem or crown of most hosts, but it is also capable of damaging fruit and root structures that are near the soil line, resulting in wilting, stunting, and plant death. Two diseases caused by this pathogen are sclerotial blight and circular spot, both of which are detrimental for sweetpotato production. A. rolfsii is a necrotrophic pathogen and can be cultured from susceptible hosts and on artificial media. The purpose of this diagnostic guide is to provide characteristic traits for identifying A. rolfsii in sweetpotato as well as outline methods for pathogen isolation, morphological and molecular characterization, culture maintenance and long-term storage, and pathogenicity testing.