A rapid isothermal RPA–CRISPR/Cas12a assay for detection of Rickettsia rickettsii

Introduction: infections is a potentially fatal tick-borne disease affecting humans and dogs in the Americas. It is difficult to confirm the diagnosis early in the laboratory on account of low-level and inconsistent rickettsemia in whole blood. Methods: . Two independent crRNA primer sets (104 bp and 92 bp) were developed to independently amplify the gene target to enhance the reliability and specificity of the assay. Results: = 6), the performance was intermittent for known limitations of whole blood testing in RMSF and limited to specific post-infection days. Discussion: detection, while indicating that broader validation in additional clinically relevant and removed tick sample types is still needed.