Abstract

Rapid molecular detection of Babesia gibsoni is needed in veterinary settings where conventional polymerase chain reaction (PCR) is limited by equipment requirements, technical complexity and turnaround time. We developed a closed-tube, colourimetric loop-mediated isothermal amplification (LAMP) assay targeting the 18 S rRNA gene of B. gibsoni and integrated the assay with a palm-sized portable incubator. Using quantified B. gibsoni DNA, the assay achieved the limit of detection (LoD) of 100 copies/reaction, matching that of an ITS-based TaqMan qPCR comparator, with visual results after 20 min of amplification. Diagnostic performance was assessed using DNA extracted from 200 archived canine blood samples submitted for suspected tick-borne infection. Compared with qPCR, the portable LAMP assay detected all 41 qPCR-positive and all 159 qPCR-negative samples, yielding 100% sensitivity (95% CI: 91.4–100%), 100% specificity (95% CI: 97.7–100%) and 100% overall accuracy/agreement. Inter-reader agreement for visual interpretation was also complete. This portable colourimetric LAMP platform provides a technically simple molecular assay for targeted detection of B. gibsoni using extracted canine blood DNA and warrants further evaluation in prospective near-patient workflows.