Concentrating rotaviruses from water samples using monolithic chromatographic supports

被引：38

作者：

Gutierrez-Aguirre, Ion ^{[1
]}

Banjac, Marko ^{[2
]}

Steyer, Andrej ^{[3
]}

Poljsak-Prijatelj, Mateja ^{[3
]}

Peterka, Matjaz ^{[2
]}

Strancar, Ales ^{[2
]}

Ravnikar, Maja ^{[1
]}

机构：

[1] Natl Inst Biol, Dept Biotechnol & Syst Biol, SI-1000 Ljubljana, Slovenia

[2] BIA Separat doo, SI-1000 Ljubljana, Slovenia

[3] Univ Ljubljana, Fac Med, Inst Microbiol & Immunol, SI-1000 Ljubljana, Slovenia

来源：

JOURNAL OF CHROMATOGRAPHY A | 2009年 / 1216卷 / 13期

关键词：

Rotavirus; Methacrylate monoliths; Concentrating viruses; Ion-exchange chromatography; Water contamination; RT q-PCR; ENTERIC VIRUSES; RT-PCR; PURIFICATION; SEPARATION; SPECIMENS; COLUMNS; ASSAY;

D O I：

10.1016/j.chroma.2008.10.106

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Rotaviruses are the leading cause of diarrhoea in infants around the globe and, under certain conditions they can be present in drinking water sources and systems. ingestion of 10-100 viral particles is enough to cause disease, emphasizing the need for sensitive diagnostic methods. In this study we have optimized the concentration of rotavirus particles using methacrylate monolithic chromatographic supports. Different surface chemistries and mobile phases were tested. A strong anion exchanger and phosphate buffer (pH 7) resulted in the highest recoveries after elution of the bound virus with I M NaCl. Using this approach, rotavirus particles spiked in 11 volumes of tap or river water were efficiently concentrated. The developed concentration method in combination with a real time quantitative polymerase chain reaction assay detected rotavirus concentrations as low as 100 rotavirus particles/ml. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：2700 / 2704

页数：5

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