Visible and rapid detection of feline chaphamaparvovirus using multienzyme isothermal rapid amplification and lateral flow dipstick assay.

Abstract

Feline chaphamaparvovirus (FeChPV) is a novel parvovirus previously reported in Canadian cats and Chinese dogs with diarrhea in 2019 and 2020, respectively. Herein, we aimed to establish a simple detection method for FeChPV in field clinics. The primers and probes for the multienzyme isothermal rapid amplification and lateral flow dipstick (MIRA-LFD) assay were designed to target the conserved regions of the FeChPV genome and determine the optimal reaction temperature and time. Without relying on precision instruments, FeChPV detection using the MIRA-LFD assay was completed within 20 min at 37°C, without any cross-reaction with other reference viruses. The newly established MIRA-LFD assay had a detection limit of 32.3 copies/μL, which was 10-fold lower than that of the nested polymerase chain reaction (PCR) assay. Furthermore, the MIRA-LFD assay detected 29 FeChPV-positive samples among 417 cats with diarrhea, providing a slightly higher positivity rate than the nested PCR assay. These results indicate that the newly developed MIRA-LFD assay for FeChPV detection is an efficient, economical, reliable, and simple method that can help in the early prevention and control of FeChPV infection.