Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent...

Area of Science:

• Environmental Virology
• Water Quality Monitoring
• Public Health

Background:

• Wastewater treatment plants (WWTPs) primarily monitor bacterial indicators, often overlooking human pathogenic viruses in treated effluent.
• The presence of viruses in reused wastewater poses a significant public health risk due to incomplete removal during treatment.
• Existing virus concentration methods often require specialized equipment like ultracentrifuges, limiting their accessibility.

Purpose of the Study:

• To evaluate an aluminum adsorption-precipitation protocol for concentrating human enteric viruses (norovirus genogroup I and II, hepatitis A virus, astrovirus, rotavirus).
• To assess the method's applicability in different laboratories for monitoring virus presence in wastewater.
• To determine the potential infectivity of viruses in influent and effluent waters using viability PMAxx-RT-qPCR.

Main Methods:

• Concentration of enteric viruses using aluminum adsorption-precipitation.
• Detection and quantification via RT-qPCR for norovirus GI, norovirus GII, and hepatitis A virus.
• Viability assessment using PMAxx-RT-qPCR to differentiate between infectious and non-infectious viral particles.

Main Results:

• The aluminum adsorption-precipitation method demonstrated effective concentration of target viruses with low limits of detection.
• The protocol was successfully validated across two independent laboratories for analyzing influent and effluent samples from five WWTPs.
• Viability RT-qPCR indicated potential viral infectivity in both influent and effluent samples, with statistically significant differences compared to standard RT-qPCR.

Conclusions:

• The aluminum adsorption-precipitation method offers a practical and accessible approach for concentrating and detecting enteric viruses in wastewater.
• This method is suitable for resource-limited laboratories lacking advanced equipment, facilitating improved water quality surveillance.
• The findings highlight the continued presence and potential infectivity of viruses in WWTP effluents, underscoring the need for robust viral monitoring.