CRISPR/Cas-based detection of Spiroplasma citri for citrus stubborn disease management

Spiroplasma citri, the causal agent of citrus stubborn disease, remains a challenging plant pathogen to control, causing substantial economic loss in global citrus production. Rapid and efficient detection assays are essential for timely and effective mitigation interventions. This research focused on developing a CRISPR/Cas-based detection system using the DETECTR method to create a point-of-care assay for S. citri. Preliminary results demonstrated the successful design of RPA primers, crRNAs, and a protocol for detecting S. citri at a concentration as low as 1 fM in two hours. Further optimization will be performed to enhance field applicability, including developing lateral flow strips and enabling direct, equipment-free, detection in the field.