Establishment of a Chip Digital PCR Detection Method for Canine Circovirus

Abstract

Canine circovirus (CanineCV), which is a new mammalian circovirus first reported in the United States in 2012, mainly causes diarrhea and vomiting in dogs. As CanineCV itself evolves and new subtypes emerge, there is an urgent need for new detection technologies to improve the sensitivity and detection rates of viruses in complex scenarios. A cdPCR assay was established for the detection of canine CV in this study. The results showed good reproducibility, specificity and a linear relationship; the minimum detection limit of Canine CV by cdPCR was 6.62 copies/μL, which is 10 times more sensitive than q-PCR. The q-PCR-positive detection rate was 1%, while CanineCV ddPCR (2.1%) exhibited a greater positive detection rate. Fifteen complete genomes were sequenced and subdivided into CanineCV-1 and CanineCV-3. In conclusion, we developed a rapid, reliable, and specific cdPCR method for the screening and monitoring of canine CV.

Keywords: Canine circovirus (CanineCV), chip digital PCR (cdPCR), real-time quantitative PCR (qRT-PCR).