Establecimiento de un método rápido de detección visual de un solo paso para el circovirus de la paloma basado en el...

Pigeon circovirus (PiCV) is an important pathogen that infects pigeons, which can induce multiple disorders such as immunosuppression and respiratory symptoms, posing a serious threat to the pigeon industry. In this study, we combined the RAA and CRISPR/Cas12a assay to establish a highly sensitive and accurate detection method for PiCV. This detection method amplifies the target nucleic acids through RAA; and the resultant dsDNA is specifically recognized by crRNA, the trans-cleavage activity of Cas12a is activated, which further cleaves the fluorescent reporter group to generate a fluorescent signal that can be visually observed under blue light. The method established in this study exhibited high sensitivity, with a minimum detection limit of 6.08 copies/µL. It showed no cross-reactivity with non-PiCV samples, demonstrating high specificity. When 40 clinical samples were tested by this method and quantitative polymerase chain reaction (qPCR) respectively, the coincidence rate was 92.5%, and the method developed herein achieved a higher positive detection rate. In conclusion, we successfully developed a rapid, on-site operable, one-step visual detection method for PiCV, which holds promising application prospects.