Assessment of a Virus Sampling and Detection Method for Water Testing Using Pepino mosaic virus

Abstract

The aim of this research is to study water sampling as a critical step for detection of unwanted waterborne plant pathogenic viruses that can seriously damage agriculture and the environment. Water sources, such as reservoirs, lakes, farm ponds, tanks, and irrigation systems are vulnerable to inadvertent (most frequent) and intentional (rare), but equally relevant, contamination by microbial pathogens. Pathogen detection in large bodies of water is complex because volume, dilution, and water dynamics. This project assesses the development and adaption of a method for water sampling and detection of waterborne plant viruses of concern for water-security. Rapid detection and monitoring of high consequence plant pathogens in water systems present a significant challenge where viruses occur at low titers. This project seeks statistical validation of a water sampling model that uses q-PCR as the method to detected Potexvirus, a group of plant viruses reported to have water borne species such as Pepino mosaic virus selected as model virus for this study. To determine the minimum amount of virus detectable in water, sensitivity tests were conducted using ELISA and qPCR. A range of pH were tested during virus filtration using self-made glass-wool filters. Virus capturing based on the isoelectric point affinity between the virus capsid proteins with glass wool was hypothesized. This research supports water-biosecurity decision-making, and will be useful for survey analysis, prevention, detection, and mitigation of unwanted waterborne plant viruses.