Development of a Novel Molecular Method for Detection of La Crosse Virus from Mosquito Vectors

La Crosse virus (LACv) is transmitted via the bite of primary vector, Aedes triseriatus, and secondary vectors Ae.albopictus and Ae.japonicus. The virus is the leading cause of arboviral encephalitis in children. The standard molecular method for LACv screening is reverse transcriptase polymerase chain reaction (RT-PCR), an effective yet time consuming and expensive procedure. No vaccines or antivirals exist, making accurate surveillance and prevention methods crucial. We hypothesize that a more economical, isothermal, sensitive, specific, and less time consuming molecular method for the detection of LACv from vectors, without the need for an expensive thermalcycler, can be developed. Positive controls were tested using the known/standard RT-PCR protocol and compared with a newly designed procedure. Varying concentrations of reagents, including primers, buffer, and enzymes, in the novel reaction mix were tested against RT-PCR for efficacy and sensitivity. Once virus was routinely detected from positive controls the novel method was compared to RT-PCR for detection of LACv from vectors and reservoirs collected from sites around Knox County, Tennessee. The development of a new method for LACv detection provides a rapid and cost-effective monitoring tool for mosquito abatement districts and county health departments, which may not have the resources for expensive RT-PCR equipment.