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1 June 2016 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
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Abstract

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.

Copyright © 2016 by The American Mosquito Control Association, Inc.
Kristen L. Burkhalter, Brad J. Biggerstaff, Kalanthe Horiuchi, and Harry M. Savage "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," Journal of the American Mosquito Control Association 32(2), 77-82, (1 June 2016). https://doi.org/10.2987/moco-32-02-77-82.1
Published: 1 June 2016
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