RPA/CRISPR-cas12a as a specific, sensitive and rapid method for diagnosing Ehrlichia canis and Anaplasma platys in dogs in Thailand

In this study, a recombinase polymerase amplification (RPA) coupled with CRISPR-Cas12a methods was established to detectE. canis andA. platys infection in dogs based on the 16S rRNA. The optimal condition for DNA amplification by RPA was 37  °C for 20 min, followed by CRISPR-Cas12a digestion at 37 °C for one hour. A combination of RPA and the cas12a detection method did not react with other pathogens and demonstrated strong sensitivity, detecting as low as 100 copies of bothE. canis andA. platys. This simultaneous detection method was significantly more sensitive than conventional PCR. The RPA-assisted cas12a assay provides specific, sensitive, rapid, simple and appropriate detection of rickettsial agents in canine blood at the point-of-care for diagnostics, disease prevention and surveillance.
Source: Veterinary Research Communications - Category: Veterinary Research Source Type: research