Periodic Reporting for period 1 - CHAMPION (Catalytic Hairpin Assembly-based Point of Need Detection Device for Noroviruses)
Période du rapport: 2022-12-16 au 2025-02-15
Foodborne diseases remain a major global public health challenge, with viruses accounting for a significant proportion of outbreaks. Among these, noroviruses (NoV) are the leading cause of non-bacterial gastroenteritis in Europe and worldwide. NoV is highly contagious and frequently leads to significant outbreaks, particularly through the consumption of contaminated food items, such as fresh produce, shellfish, and ready-to-eat products. It poses a considerable burden on healthcare systems and has a serious impact on the agri-food industry, particularly in sectors dealing with perishable goods. The European Food Safety Authority (EFSA) has highlighted the urgent need for improved monitoring and detection of foodborne viruses to protect public health and enhance food safety in the European Union (EU).
Despite its prevalence, the detection of norovirus in food remains a significant scientific and technical challenge. Current testing methods are often time-consuming, costly, and require complex laboratory procedures that are not easily scalable or suitable for routine food safety monitoring. In addition, norovirus is often present in very low concentrations in food samples and may be unevenly distributed, making its reliable detection difficult. There is also a lack of harmonized sample preparation methods across food matrices, which further limits consistent detection efforts.
In this context, the CHAMPION project was launched to develop improved, rapid, and robust methods for the detection of norovirus in food, contributing directly to EU food safety goals and supporting the implementation of more effective risk management strategies. It focuses on integrating novel sample enrichment techniques with cutting-edge detection technologies to deliver more sensitive, accurate, and efficient diagnostic tools that can be applied across various food types and testing environments. It tested and validated a range of sample enrichment methods, including immunomagnetic separation, ultrafiltration, and enzymatic treatments, aiming to increase viral recovery rates from complex food matrices.
CHAMPION also investigated the feasibility of portable, field-deployable diagnostic platforms, which can provide rapid on-site results, reducing the time between sample collection and decision-making. This supports more agile responses in food production and distribution chains, potentially preventing contaminated products from reaching consumers.
Despite its prevalence, the detection of norovirus in food remains a significant scientific and technical challenge. Current testing methods are often time-consuming, costly, and require complex laboratory procedures that are not easily scalable or suitable for routine food safety monitoring. In addition, norovirus is often present in very low concentrations in food samples and may be unevenly distributed, making its reliable detection difficult. There is also a lack of harmonized sample preparation methods across food matrices, which further limits consistent detection efforts.
In this context, the CHAMPION project was launched to develop improved, rapid, and robust methods for the detection of norovirus in food, contributing directly to EU food safety goals and supporting the implementation of more effective risk management strategies. It focuses on integrating novel sample enrichment techniques with cutting-edge detection technologies to deliver more sensitive, accurate, and efficient diagnostic tools that can be applied across various food types and testing environments. It tested and validated a range of sample enrichment methods, including immunomagnetic separation, ultrafiltration, and enzymatic treatments, aiming to increase viral recovery rates from complex food matrices.
CHAMPION also investigated the feasibility of portable, field-deployable diagnostic platforms, which can provide rapid on-site results, reducing the time between sample collection and decision-making. This supports more agile responses in food production and distribution chains, potentially preventing contaminated products from reaching consumers.
In the context of CHAMPION, the overall activities are described below.
1. The sample enrichment methods for the detection of NoV was optimized using Magnetic nanostructures and aptamer based recognition molecules. For this different concentrations of the magnetic nanostructures, aptamers are optimized. The conjugation methods are also optimized to get suitable conditions for enrichment.~
2. The sample pre concentrations methods are also validated using different samples such as buffer spiked with NoV VLP. The pH and the buffer conditions are also optimized
3. The amplification methods are also optimized using the synthetic NoV RNA. Different conditions of amplification are performed and validated.
4. The colorimetric assay for the detection of NoV was also performed and validated
1. The sample enrichment methods for the detection of NoV was optimized using Magnetic nanostructures and aptamer based recognition molecules. For this different concentrations of the magnetic nanostructures, aptamers are optimized. The conjugation methods are also optimized to get suitable conditions for enrichment.~
2. The sample pre concentrations methods are also validated using different samples such as buffer spiked with NoV VLP. The pH and the buffer conditions are also optimized
3. The amplification methods are also optimized using the synthetic NoV RNA. Different conditions of amplification are performed and validated.
4. The colorimetric assay for the detection of NoV was also performed and validated
The CHAMPION project has advanced the development of rapid, sensitive, and field-deployable diagnostic tools for norovirus detection, with a strong focus on improving detection limits, sample processing, and real-time monitoring capabilities.
Optimized Sample Pre-concentration Protocols: The affinity-based concentration techniques have been successfully validated for different food samples. These methods have significantly increased the sensitivity of downstream assays. The colorimetric assay also developed which is highly sensitive and specific towards the detection of NoV RNA. The future research direction is to integrate the both the sample enrichment and amplification methods to the portable and user friendly detection devices. which can offer minimal user input. Field trials in food processing environments and clinical labs have demonstrated robustness, reliability, and reduced false positives compared to conventional qPCR methods.
Optimized Sample Pre-concentration Protocols: The affinity-based concentration techniques have been successfully validated for different food samples. These methods have significantly increased the sensitivity of downstream assays. The colorimetric assay also developed which is highly sensitive and specific towards the detection of NoV RNA. The future research direction is to integrate the both the sample enrichment and amplification methods to the portable and user friendly detection devices. which can offer minimal user input. Field trials in food processing environments and clinical labs have demonstrated robustness, reliability, and reduced false positives compared to conventional qPCR methods.