Main content area

A Simple Modification to the Mosquito Homogenization Protocol Safely Inactivates West Nile Virus and Allows Virus Detection by the Rapid Analyte Measurement Platform (RAMP®) ASSAY

Burkhalter, Kristen L., Biggerstaff, Brad J., Horiuchi, Kalanthe, Savage, Harry M.
Journal of the American Mosquito Control Association 2016 v.32 no.2 pp. 77-82
Culicidae, West Nile virus, albumins, biocontainment, cattle, cell culture, culture media, detergents, homogenization, octoxynol, viruses
We evaluated the ability of the Rapid Analyte Measurement Platform (RAMP®) mosquito-grinding buffer to inactivate West Nile virus (WNV) by subjecting WNV-positive samples ground in RAMP buffer to incubation intervals ranging from 5 min to 60 min. At each time point an aliquot was removed and serially diluted in bovine albumin (BA)-1 cell culture media to stop the inactivation process by RAMP buffer. Each BA-1 sample was tested for viable virus using Vero 6-well cell culture plaque assay and observed for plaques. We observed very limited inactivation of WNV (1–2 log₁₀ plaque-forming units/ml) by RAMP buffer. Concerned for RAMP operators who may be using this assay in low-level biocontainment facilities, we developed an alternate sample homogenization protocol using Triton X-100 detergent that ensures complete WNV inactivation without compromising the performance of the RAMP assay.