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A Membrane-Based Electro-Separation Method (MBES) for Sample Clean-Up and Norovirus Concentration.

Kang W, Cannon JL - PLoS ONE (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Center for Food Safety, The University of Georgia, Griffin, Georgia, United States of America.

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.

No MeSH data available.


Related in: MedlinePlus

Illustration of viral particle movement in the MBES device.
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pone.0141484.g002: Illustration of viral particle movement in the MBES device.

Mentions: The union (3.5 ml) was connected to two dialyzer chambers (0.5 ml and 1.5 ml) on each side to create a linked chamber. A pore-size selective polycarbonate membrane (1 μm; SterliTech Corp, Kent, WA), was fitted into the linked chamber and two 300 kDa MWCO cellular acetate restricting membranes (Harvard Apparatus) were flanked on both sides of the linked chambers to create two membrane-separated chambers; one for sample addition (4.0 ml sample chamber) and one for sample collection (1.5 ml collection chamber) (Fig 2). Both chambers were filled with sodium phosphate buffer solutions (pH 7.5; concentration range from 20 mM to 100 mM) with or without the addition of 0.01% SDS or other norovirus elution buffers (PBS, tris-glycine, or TGBE), all with 0.01% SDS addition.


A Membrane-Based Electro-Separation Method (MBES) for Sample Clean-Up and Norovirus Concentration.

Kang W, Cannon JL - PLoS ONE (2015)

Illustration of viral particle movement in the MBES device.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4625962&req=5

pone.0141484.g002: Illustration of viral particle movement in the MBES device.
Mentions: The union (3.5 ml) was connected to two dialyzer chambers (0.5 ml and 1.5 ml) on each side to create a linked chamber. A pore-size selective polycarbonate membrane (1 μm; SterliTech Corp, Kent, WA), was fitted into the linked chamber and two 300 kDa MWCO cellular acetate restricting membranes (Harvard Apparatus) were flanked on both sides of the linked chambers to create two membrane-separated chambers; one for sample addition (4.0 ml sample chamber) and one for sample collection (1.5 ml collection chamber) (Fig 2). Both chambers were filled with sodium phosphate buffer solutions (pH 7.5; concentration range from 20 mM to 100 mM) with or without the addition of 0.01% SDS or other norovirus elution buffers (PBS, tris-glycine, or TGBE), all with 0.01% SDS addition.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Center for Food Safety, The University of Georgia, Griffin, Georgia, United States of America.

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.

No MeSH data available.


Related in: MedlinePlus