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A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications. / Farkas, Kata; Pang, Liping; Lin, Susan; Williamson, Wendy; Easingwood, Richard ; Fredericks, Rayleen ; Jaffer, Mohamed A ; Varsani, Arvind.

In: Food and Environmental Virology, Vol. 5, No. 4, 01.12.2013, p. 231-235.

Research output: Contribution to journalArticle

HarvardHarvard

Farkas, K, Pang, L, Lin, S, Williamson, W, Easingwood, R, Fredericks, R, Jaffer, MA & Varsani, A 2013, 'A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications', Food and Environmental Virology, vol. 5, no. 4, pp. 231-235. https://doi.org/10.1007/s12560-013-9122-4

APA

Farkas, K., Pang, L., Lin, S., Williamson, W., Easingwood, R., Fredericks, R., Jaffer, M. A., & Varsani, A. (2013). A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications. Food and Environmental Virology, 5(4), 231-235. https://doi.org/10.1007/s12560-013-9122-4

CBE

Farkas K, Pang L, Lin S, Williamson W, Easingwood R, Fredericks R, Jaffer MA, Varsani A. 2013. A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications. Food and Environmental Virology. 5(4):231-235. https://doi.org/10.1007/s12560-013-9122-4

MLA

VancouverVancouver

Farkas K, Pang L, Lin S, Williamson W, Easingwood R, Fredericks R et al. A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications. Food and Environmental Virology. 2013 Dec 1;5(4):231-235. https://doi.org/10.1007/s12560-013-9122-4

Author

Farkas, Kata ; Pang, Liping ; Lin, Susan ; Williamson, Wendy ; Easingwood, Richard ; Fredericks, Rayleen ; Jaffer, Mohamed A ; Varsani, Arvind. / A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications. In: Food and Environmental Virology. 2013 ; Vol. 5, No. 4. pp. 231-235.

RIS

TY - JOUR

T1 - A Gel Filtration-Based Method for the Purification of Infectious Rotavirus Particles for Environmental Research Applications

AU - Farkas, Kata

AU - Pang, Liping

AU - Lin, Susan

AU - Williamson, Wendy

AU - Easingwood, Richard

AU - Fredericks, Rayleen

AU - Jaffer, Mohamed A

AU - Varsani, Arvind

PY - 2013/12/1

Y1 - 2013/12/1

N2 - This article describes a rapid method for purifying infectious rotavirus particles from cell culture for environmental research. The method is based on size-exclusion chromatography using TOSOH TSKgel® G5000PWXL-CP with a TSKgel® Size Exclusion G2500PWxl guard column, set up on an AKTA Explorer10. Four peaks were identified from the chromatogram and the corresponding fractions were collected and analysed by electron microscopy, 1-step quantitative reverse transcription polymerase chain reaction (RT-PCR) and qNano measurement. Infectivity potential of the recovered virus particles was determined using cell culture. Our analysis reveals that the first fraction contains majority of the intact triple-layered infectious virions while the other three fractions contain mixtures of empty capsids and intact infectious virions. Our results also indicate that there is a gross overestimation of rotaviruses in crude extracts due to encapsidated RNA in the order of 2.3 × 1011 particles and we note that estimates by qNano are similarly skewed (1.36 × 1013 particle) possibly due to empty capsids and cellular debris. In summary we present a method for purification (~12 h) of rotaviruses for a more robust and accurate quantification of virus size, surface charge and particle concentration in environmental contexts.

AB - This article describes a rapid method for purifying infectious rotavirus particles from cell culture for environmental research. The method is based on size-exclusion chromatography using TOSOH TSKgel® G5000PWXL-CP with a TSKgel® Size Exclusion G2500PWxl guard column, set up on an AKTA Explorer10. Four peaks were identified from the chromatogram and the corresponding fractions were collected and analysed by electron microscopy, 1-step quantitative reverse transcription polymerase chain reaction (RT-PCR) and qNano measurement. Infectivity potential of the recovered virus particles was determined using cell culture. Our analysis reveals that the first fraction contains majority of the intact triple-layered infectious virions while the other three fractions contain mixtures of empty capsids and intact infectious virions. Our results also indicate that there is a gross overestimation of rotaviruses in crude extracts due to encapsidated RNA in the order of 2.3 × 1011 particles and we note that estimates by qNano are similarly skewed (1.36 × 1013 particle) possibly due to empty capsids and cellular debris. In summary we present a method for purification (~12 h) of rotaviruses for a more robust and accurate quantification of virus size, surface charge and particle concentration in environmental contexts.

U2 - 10.1007/s12560-013-9122-4

DO - 10.1007/s12560-013-9122-4

M3 - Article

VL - 5

SP - 231

EP - 235

JO - Food and Environmental Virology

JF - Food and Environmental Virology

SN - 1867-0334

IS - 4

ER -