CRISPR-powered BOVDS: a next-generation diagnostic platform for brucellosis with abortifacient pathogens screening and vaccine/field strain differentiation

BOVDS specifically identified ten clinically abortifacient pathogens with minimal equipment and simple isothermal incubation, enabling point-of-care diagnostics. 2. Unprecedented species-level resolution: enables precise epidemiological tracing and outbreak source attribution by differentiating B. melitensis , B. abortus , and B. suis . 3. Vaccine/wild-type differentiation: this capability is essential for monitoring vaccine efficacy and distinguishing between natural infections and vaccine-induced immune responses in animals. Brucellosis, caused primarily by Brucella abortus , Brucella melitensis , and Brucella suis , remains a critical global public health challenge, particularly in regions where these pathogens persist in livestock and wildlife reservoirs. Despite decades of control measures-including vaccination, test-and-removal programs, and biosecurity protocols-persistent human and animal cases highlight the limitations of existing diagnostic and intervention strategies. CRISPR-based diagnostics have emerged as a transformative tool, offering rapid, ultrasensitive, and field-deployable pathogen detection. Here, we present BOVDS ( Brucella melitensis/abortus/suis -other pathogens-vaccine detection and differentiation system), an innovative CRISPR/Cas13a-based platform that integrates ultrahigh sensitivity (10 copies/μL), screening for 10 major abortifacient pathogens, and precise strain differentiation-overcoming key challenges in Brucella diagnostics. By incorporating mismatched spacer designs, BOVDS achieves robust discrimination between B. melitensis , B. abortus , and B. suis despite their high genomic conservation. Additionally, the platform enables differentiation between vaccine and wild-type strains, addressing critical gaps in vaccination monitoring and epidemiological surveillance. Uniting laboratory-level accuracy with on-farm practicality, BOVDS facilitates real-time outbreak management, targeted culling, and environmental decontamination, advancing One Health initiatives toward sustainable brucellosis prevention and control. This system sets a new benchmark for next-generation zoonotic disease diagnostics, with broad applicability in global public and veterinary health.