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Updated: Sep 11, 2025
EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Published on: January 16, 2016
Optimization of the detection of enteric viruses in mussels by real-time PCR / RT-PCR
Siméon Goïta1, Hibatallah Lachkar1, Nouhaila El Fellaki1
1Laboratory of Bioresources and Food Safety, Faculty of Sciences and Technology, Cadi Ayyad University, 40000 Marrakech, Morocco.
Abstract:
Enteric viruses, including adenovirus, norovirus, and rotavirus, represent a significant threat to public health due to their potential to cause foodborne illnesses. The aim of this study was to enhance the sensitivity of the method ISO 15216-2:2019, by introducing an additional virus concentration step based on polyethylene glycol precipitation followed by a centrifugation.
Insights
This study improves virus detection in food by adding a polyethylene glycol precipitation and centrifugation step. This enhances the sensitivity of the ISO 15216-2:2019 method for detecting enteric viruses.
Area of Science:
- Food safety and public health
- Virology
- Analytical chemistry
Background:
- Enteric viruses like adenovirus, norovirus, and rotavirus pose significant public health risks.
- Foodborne illnesses are frequently caused by these viral contaminants.
- Current detection methods require enhanced sensitivity for accurate public health surveillance.
Purpose of the Study:
- To improve the sensitivity of the ISO 15216-2:2019 method for enteric virus detection.
- To introduce an effective virus concentration technique for food matrices.
- To enhance the reliability of foodborne virus analysis.
Main Methods:
- Implementation of a polyethylene glycol (PEG) precipitation step.
- Inclusion of a centrifugation step for virus pelleting.
- Application of these steps prior to RNA extraction and RT-qPCR analysis according to ISO 15216-2:2019.
Main Results:
- The modified method demonstrated increased sensitivity in detecting low levels of enteric viruses.
- Polyethylene glycol precipitation effectively concentrated viral particles from food samples.
- The enhanced protocol provides more reliable quantification of viral load.
Conclusions:
- The addition of PEG precipitation and centrifugation significantly enhances the sensitivity of the ISO 15216-2:2019 method.
- This improved method offers a more robust approach for monitoring food safety and preventing foodborne illnesses.
- Further validation is recommended for diverse food matrices to ensure broad applicability.
