RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection
Issued Date
2024-12-01
Resource Type
eISSN
20452322
Scopus ID
2-s2.0-85189791965
Journal Title
Scientific Reports
Volume
14
Issue
1
Rights Holder(s)
SCOPUS
Bibliographic Citation
Scientific Reports Vol.14 No.1 (2024)
Suggested Citation
Sutipatanasomboon A., Wongsantichon J., Sakdee S., Naksith P., Watthanadirek A., Anuracpreeda P., Blacksell S.D., Saisawang C. RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection. Scientific Reports Vol.14 No.1 (2024). doi:10.1038/s41598-024-58169-6 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/97969
Title
RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection
Corresponding Author(s)
Other Contributor(s)
Abstract
Anaplasma marginale infection is one of the most common tick-borne diseases, causing a substantial loss in the beef and dairy production industries. Once infected, the pathogen remains in the cattle for life, allowing the parasites to spread to healthy animals. Since clinical manifestations of anaplasmosis occur late in the disease, a sensitive, accurate, and affordable pathogen identification is crucial in preventing and controlling the infection. To this end, we developed an RPA-CRISPR/Cas12a assay specific to A. marginale infection in bovines targeting the msp4 gene. Our assay is performed at one moderately high temperature, producing fluorescent signals or positive readout of a lateral flow dipstick, which is as sensitive as conventional PCR-based DNA amplification. This RPA-CRISPR/Cas12a assay can detect as few as 4 copies/μl of Anaplasma using msp4 marker without cross-reactivity to other common bovine pathogens. Lyophilized components of the assay can be stored at room temperature for an extended period, indicating its potential for field diagnosis and low-resource settings of anaplasmosis in bovines.