Concentrating rotaviruses from water samples using monolithic chromatographic supports

Area of Science:

• Environmental Microbiology
• Virology
• Analytical Chemistry

Background:

• Rotaviruses are a primary cause of infant diarrhea globally.
• Low viral loads (10-100 particles) in water can cause infection, necessitating sensitive detection methods.

Purpose of the Study:

• To optimize a concentration method for rotavirus particles in water samples.
• To develop a sensitive diagnostic approach for detecting rotavirus in drinking water.

Main Methods:

• Methacrylate monolithic chromatographic supports with varying surface chemistries were employed.
• Optimization involved testing different mobile phases, with a strong anion exchanger and phosphate buffer (pH 7) yielding optimal results.
• Elution was achieved using 1M NaCl, and the concentrated virus was analyzed using real-time quantitative polymerase chain reaction (qPCR).

Main Results:

• The optimized method efficiently concentrated rotavirus particles spiked in 1-liter volumes of tap and river water.
• The combined concentration and real-time qPCR assay detected rotavirus at concentrations as low as 100 particles/ml.
• High viral recoveries were achieved using a strong anion exchanger and phosphate buffer.

Conclusions:

• The developed chromatographic concentration method is effective for detecting low levels of rotavirus in water.
• This approach significantly enhances the sensitivity of rotavirus detection in environmental water samples.
• The method offers a promising tool for ensuring the safety of drinking water sources.