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Updated: Jun 18, 2026

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay
Published on: September 11, 2016
Capsid Integrity Detection of Enteric Viruses in Reclaimed Waters
Pablo Puchades-Colera1, Azahara Díaz-Reolid1, Inés Girón-Guzmán1
1VISAFELab, Department of Preservation and Food Safety Technologies, Institute of Agrochemistry and Food Technology, IATA-CSIC, Av. Agustín Escardino 7, 46980 Valencia, Spain.
Wastewater reuse for irrigation often fails to remove enteric viruses, posing risks to public health. Most reclaimed water samples analyzed did not meet EU standards, indicating a need for improved treatment to ensure water safety.
Area of Science:
- Environmental microbiology
- Water quality engineering
- Virology
Background:
- Climate change and drought necessitate wastewater reuse for agriculture.
- EU regulations mandate reduced enteric viruses in reclaimed water for irrigation.
- Current wastewater treatment may not sufficiently inactivate all viral pathogens.
Purpose of the Study:
- To assess the prevalence of human enteric viruses (norovirus, rotavirus) and crAssphage in wastewater.
- To evaluate somatic coliphages as a culturable indicator of viral inactivation.
- To optimize a method for detecting infectious viral particles in reclaimed water.
Main Methods:
- (RT)-qPCR for detecting human norovirus genogroup I and II, rotavirus, and crAssphage.
- Somatic coliphage analysis for culturable viral indicators.
- Optimization of PMAxx-RT-qPCR to assess viral capsid integrity and infectivity.
Main Results:
- Somatic coliphages were found in 60% of reclaimed water, suggesting incomplete inactivation.
- 66% of samples tested positive for enteric viruses using PMAxx-RT-qPCR.
- Detected viral concentrations ranged from 2.79 to 7.30 Log10 genome copies/L.
Conclusions:
- Most analyzed reclaimed water samples did not meet EU water reuse standards.
- Infectious viral particles were detected in a significant portion of reclaimed water.
- Current wastewater treatment processes require enhancement for effective viral pathogen removal.
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