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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: 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.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.

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

The impact of applied voltage strength on MNV—1 recovery using the MBES system.Applying a constant voltage of 20V, 40V, or 60V and no voltage (control) to the system for 30 min, 7 log genomic copies of MNV-1 were added to the sample chamber prior to voltage application using 20 mM sodium phosphate with 0.01% as the electrolyte buffer. Error bars represent standard deviations, n = 2.
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pone.0141484.g009: The impact of applied voltage strength on MNV—1 recovery using the MBES system.Applying a constant voltage of 20V, 40V, or 60V and no voltage (control) to the system for 30 min, 7 log genomic copies of MNV-1 were added to the sample chamber prior to voltage application using 20 mM sodium phosphate with 0.01% as the electrolyte buffer. Error bars represent standard deviations, n = 2.

Mentions: MNV-1 recovery rates in the collection chambers were 30.8% and 31.3% when the applied voltages were 20V and 40V, respectively; while MNV-1 recovery in sample chambers were also similar, 60.7% and 65.5%, respectively. However, when the applied voltage increased to 60V, no MNV-1 was detected in the collection chamber and only 31.6% was recovered in the sample chamber. Almost 70% of MNV-1 was lost after 60V was applied for 30 min (Fig 9). Fig 9 results indicated that when a high voltage is applied for prolonged durations, this could cause viral destabilization and RNA degradation (see also Table F in S1 Text).


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

Kang W, Cannon JL - PLoS ONE (2015)

The impact of applied voltage strength on MNV—1 recovery using the MBES system.Applying a constant voltage of 20V, 40V, or 60V and no voltage (control) to the system for 30 min, 7 log genomic copies of MNV-1 were added to the sample chamber prior to voltage application using 20 mM sodium phosphate with 0.01% as the electrolyte buffer. Error bars represent standard deviations, n = 2.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0141484.g009: The impact of applied voltage strength on MNV—1 recovery using the MBES system.Applying a constant voltage of 20V, 40V, or 60V and no voltage (control) to the system for 30 min, 7 log genomic copies of MNV-1 were added to the sample chamber prior to voltage application using 20 mM sodium phosphate with 0.01% as the electrolyte buffer. Error bars represent standard deviations, n = 2.
Mentions: MNV-1 recovery rates in the collection chambers were 30.8% and 31.3% when the applied voltages were 20V and 40V, respectively; while MNV-1 recovery in sample chambers were also similar, 60.7% and 65.5%, respectively. However, when the applied voltage increased to 60V, no MNV-1 was detected in the collection chamber and only 31.6% was recovered in the sample chamber. Almost 70% of MNV-1 was lost after 60V was applied for 30 min (Fig 9). Fig 9 results indicated that when a high voltage is applied for prolonged durations, this could cause viral destabilization and RNA degradation (see also Table F in S1 Text).

Bottom Line: 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.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.

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