Development of an RT-RPA assay for La Crosse virus detection provides insights into age-dependent neuroinvasion in mice
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La Crosse virus (LACV) is a mosquito-borne arbovirus that causes pediatric encephalitis in North America, primarily affecting children under the age of 16 years of age. Early and accurate diagnosis is crucial to reducing morbidity in this vulnerable population; however, existing molecular and serological methods face limitations in sensitivity, specificity, and accessibility. Here, we present the development of a reverse transcription recombinase polymerase amplification (RT-RPA) assay for LACV detection. Our assay detects LACV within 20 minutes with a limit of detection of 100-1000 viral copies, demonstrating high specificity without cross-reactivity against closely related or geographically relevant arboviruses. We further integrate the RT-RPA assay into a lateral flow format, potentially enabling simple and inexpensive point-of-care diagnosis. To complement our assay development, we investigated viral pathogenesis in a mouse model that recapitulates the age-dependent susceptibility observed in human populations. Using this RT-RPA assay, we reveal that LACV invades the brains of weanling mice as early as 4 days post-infection (dpi) and adult mice by 5 dpi. Surviving adult mice had no detectable virus within their brains by 20 dpi. These results underscore the utility of our RT-RPA-based platform for LACV diagnosis and offer novel insights into the age-dependent temporal dynamics of LACV neuroinvasion and viral clearance.
