A device comprising a filter attached to a vacuum cleaner was purpose-built to sample rust spores from three potentially high-risk pathways in Australia: passengers, fresh flowers, and sea cargo. The proportion of spores recovered from eight surfaces comparable with those on each pathway (cotton, denim, roses, carnations, chrysanthemums, wood, plastic, and metal) was estimated in the laboratory. Spore recovery percentages were highest for denim clothing (61% Puccinia triticina Erikss. and 62% Uromycladium tepperianum) and lowest for carnations (4% P. triticina Erikss. and 5% U. tepperianum). Subsequently, the device was tested at several locations on the Central Coast of New South Wales, Australia, recently affected by a “myrtle rust” outbreak. Symptomatic and asymptomatic myrtle rust hosts, myrtle rust nonhosts, and inanimate objects (e.g., clothing and vehicles) were sampled in conjunction with the emergency response to the outbreak. A polymerase chain reaction (PCR) assay developed for P. psidii established the presence of myrtle rust, and visual inspections provided spore count estimations. All samples from symptomatic myrtle rust hosts produced positive PCR results and spore count estimations were generally much greater. Several samples from asymptomatic myrtle rust hosts, myrtle rust nonhosts, and inanimate objects also produced positive PCR results; however, there were discrepancies between PCR results and spore count estimations in some of these samples, all of which had <100 spores. This study highlights the utility of the device and analytical methodology, especially during the early stages of a disease outbreak when infection symptoms on plants and contamination on objects is not visible upon gross examination.