Abstract
Noroviruses are the leading cause of acute gastroenteritis and foodborne illnesses in the United States. Enhanced methods for detecting noroviruses in food matrices are needed as current methods are complex, labor intensive and insensitive, often resulting in inhibition of downstream molecular detection and inefficient recovery. Membrane-based electro-separation (MBES) is a technique to exchange charged particles through a size-specific dialysis membrane from one solution to another using electric current as the driving force. Norovirus has a net negative surface charge in a neutrally buffered environment, so when placed in an electric field, it moves towards the anode. It can then be separated from the cathodic compartment where the sample is placed and then collected in the anodic compartment for downstream detection. In this study, a MBES-based system was designed, developed and evaluated for concentrating and recovering murine norovirus (MNV-1) from phosphate buffer. As high as 30.8% MNV-1 migrated from the 3.5 ml sample chamber to the 1.5 ml collection chamber across a 1 μm separation membrane when 20 V was applied for 30 min using 20 mM sodium phosphate with 0.01% SDS (pH 7.5) as the electrolyte. In optimization of the method, weak applied voltage (20 V), moderate duration (30 min), and low ionic strength electrolytes with SDS addition were needed to increase virus movement efficacy. The electric field strength of the system was the key factor to enhance virus movement, which could only be improved by shortening the electrodes distance, instead of increasing system applied voltage because of virus stability. This study successfully demonstrated the norovirus mobility in an electric field and migration across a size-specific membrane barrier in sodium phosphate electrolyte. With further modification and validation in food matrixes, a novel, quick, and cost-effective sample clean-up technique might be developed to separate norovirus particles from food matrices by electric force.
Highlights
Norovirus (NoV) is recognized as the major cause of foodborne illnesses in the U.S each year, responsible for 58% of foodborne gastroenteritis and 95% of nonbacterial gastroenteritis [1, 2].PLOS ONE | DOI:10.1371/journal.pone.0141484 October 29, 2015Membrane-Based Electro-Separation for NorovirusThe economic impact of norovirus related outbreaks is estimated to be $5.8 billion annually in the U.S [3]
When another electrolyte was used, the virus recovery in the collection chamber ranged from 5.6% to 19.3% with 20V applied voltage for 30 min
Because virus inactivation can occur when too high of a concentration of sodium dodecyl sulfate (SDS) (0.1%) is used, SDS addition was decreased to 0.01%, which resulted in a 14.2% virus recovery in the collection chamber when 20V was applied for 30 min using a 20 mM sodium phosphate electrolyte buffer
Summary
Norovirus (NoV) is recognized as the major cause of foodborne illnesses in the U.S each year, responsible for 58% of foodborne gastroenteritis and 95% of nonbacterial gastroenteritis [1, 2].PLOS ONE | DOI:10.1371/journal.pone.0141484 October 29, 2015Membrane-Based Electro-Separation for NorovirusThe economic impact of norovirus related outbreaks is estimated to be $5.8 billion annually in the U.S [3]. Indirect norovirus transmission occurs by consumption of fecally contaminated foods or water, or contact with contaminated environmental surfaces [5]. Ready-to-eat deli meats, leafy-green salad mixes, and shellfish are susceptible to norovirus contamination [6]. Oysters can bio-concentrate virus in their tissue when grown in contaminated water. High-level post-handling processing of ready-to-eat deli meats and salad mixes is the primary contamination route of food handlers. Vegetables, and raw oysters have higher chances of causing norovirus infection because they are often consumed without cooking. The National Outbreak Reporting System (NORS) reported 1,008 norovirus foodborne outbreaks in the U.S from 2008–2012, wherein 30% were caused by consumption of norovirus contaminated leafy vegetables, 21% by fruits, and 19% by shellfish [7]
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