High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms

Abstract

There are four species of blood-sucking hookworms that infect dogs, namely Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala. Each hookworm species differs in its geographical distribution, impact on canine health and more importantly its ability to infect and produce disease in humans. For example, dogs are the primary reservoir for A. ceylanicum, the second most common hookworm species infecting humans in the Asia Pacific. Ancylostoma braziliense is more geographically confined and responsible for chronic skin eruptions in humans. Accurate diagnosis of hookworm species in dogs is therefore essential for understanding the relative risk each species pose to human populations living in close association with dogs. As microscopic methods are unable to distinguish between species of hookworms in stool, highly efficient real time PCR assays capable of detecting and differentiating all four canine hookworm species in canine faeces were developed. These new assays are superior to previously published gel-based PCR assay displaying higher throughput, and ability to correctly identify single and mixed hookworm species infections with higher accuracy.