Concentrating rotaviruses from water samples using monolithic chromatographic supports.
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 1M NaCl. Using this approach, rotavirus particles spiked in 1l 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.
Gutiérrez-Aguirre
Ion
I
Department of Biotechnology and Systems Biology, National Institute of Biology, Vecna pot 111, SI-1000 Ljubljana, Slovenia. ion.gutierrez@nib.si
Banjac
Marko
M
Steyer
Andrej
A
Poljsak-Prijatelj
Mateja
M
Peterka
Matjaz
M
Strancar
Ales
A
Ravnikar
Maja
M
D002852:Chromatography, Ion Exchange / Q000379:methods*; D004784:Environmental Monitoring / Q000379:methods*; D005243:Feces / Q000821:virology; D005618:Fresh Water / Q000032:analysis / Q000821:virology*; D006801:Humans; D008689:Methacrylates / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D012401:Rotavirus / Q000302:isolation & purification* / Q000648:ultrastructure
D008689:Methacrylates