ID07 - Comparison of qPCR and Centrifugal Flotation for the Detection of Ancylostoma in experimentally Infected Beagles

Abstract: Background: Ova and parasite (O&P) by centrifugal flotation is a common method for the detection of hookworm eggs in fecal samples. Molecular diagnostics using real-time quantitative PCR (qPCR) is a highly sensitive method to detect nucleic acids of infectious agents, including parasites.

Objective: This study compared O&P with qPCR for the detection of Ancylostoma caninum in experimentally and Ancylostoma spp. in naturally infected dogs and cats.

Animals: Five beagles treated with an anthelmintic drug at day minus 10 were experimentally infected on day 0 with 300 Ancylostoma caninum infective larvae. Fecal samples were collected at regular intervals.

Field Samples: A total of 790 fecal samples were included in the comparison of O&P with qPCR to detect Ancylostoma spp.

Methods: Fecal samples from experimentally infected beagles were analyzed within 48 hours using O&P zinc sulfate centrifugation and flotation and qPCR for Ancylostoma. The O&P Ancylostoma positive field samples were tested for Ancylostoma spp. by qPCR.

Results: In the experimentally infected beagles, all five dogs tested qPCR positive by day 17 whereas all five dogs tested positive by O&P by day 28. In the field samples, overall agreement was 97.5% between O&P and qPCR.

Conclusions and clinical importance: The high correlation of O&P Ancylostoma positive samples with qPCR confirms the high sensitivity of this molecular diagnostic assay. In experimentally infected dogs, qPCR indicates earlier and more consistent detection of Ancylostoma caninum eggs in feces than O&P.