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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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Jun 2016
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Source: J Am Mosq Control Assoc. 32(2):77-82
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Alternative Title:J Am Mosq Control Assoc
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Description: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.
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Pubmed ID:27280345
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Pubmed Central ID:PMC7111560
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