Development and Single Laboratory Evaluation of a Refined and specific Real-time PCR Detection Method, Using Mitochondrial Primers (Mit1C), for the Detection of Cyclospora cayetanensisin Produce

•A specific real-time PCR assay (Mit1C) was developed for the detection of C. cayetanensis.•The Mit1C assay targets a conserved region of the C. cayetanensismitochondrial genome.•A single laboratory validation was performed in spiked produce samples.•Evaluation against closely related organisms revealed specificity to C. cayetanensis.•As few as 5 oocysts were detected in spiked produce samples, demonstrating sensitivity.