To illustrate these goals, a literature study was performed in chapter 1.
Finally, a screening study on a selection of fruit produce products towards NoV presence was the third main goal of this PhD. The first goal consisted of the development and evaluation of a methodology for detection of noroviruses (NoV) in ready-to-eat (RTE) foods and soft red fruits while the second main goal included the evaluation of the murine norovirus 1 (MNV-1) as control reagent for different steps throughout the NoV detection protocols. In this PhD, three main goals were defined. Once developed, such technologies provide a way forward to minimize public health risks associated with the shellfish consumption. However, more collaborative efforts on research and development will be required to commercialize products. Many of these approaches have the potential to be developed as user-friendly onsite detection kits with minimal costs. Other emerging technologies include microfluidic, aptamer and biosensor-based detection methods developed to detect norovirus with high sensitivity from a simple matrix. Furthermore, some omics techniques have the potential to simultaneously detect multiple enteric viruses that cause human disease. Recent advances in omics-based detection approaches have the potential to identify novel biomarkers that can be incorporated into rapid detection kits for onsite use. The focus of this review is to evaluate current detection methods and discuss emerging approaches. farm-to-plate) to ensure shellfish quality. Due to the lack of rapid, user-friendly and onsite/infield methods, it has been difficult to establish an effective virus monitoring regime that is able to identify contamination points across the production line (i.e.
Current virus recovery and PCR detection methods can be expensive and time consuming. Reports of norovirus infections associated with the consumption of contaminated bivalve molluscan shellfish negatively impact both consumers and commercial shellfish operators. The overall procedure for detecting HAV could be then simplify avoiding virus losses during manipulation. Real-time RT-PCR detection could allow to eliminate some purification and concentration steps that are required for conventional RT-nested PCR detection. In contrast, the HAV recovery percentage was higher after the virus elution step while the RNA purity was lower. virus concentration by ultracentrifugation, the RNA purity was high but the estimated HAV recovery efficiency was however low, probably due to virus losses and the presence of RT-PCR inhibitors in sample concentrates. Besides the improvements in detection sensitivity, the real-time RT-PCR, by quantifying HAV RNA, allowed to check the overall extraction procedure and the recovery efficiency after each processing step. Real-time RT-PCR yielded higher detection sensitivity than the obtained by conventional RT-nested PCR. The efficiency of this method to recover Hepatitis A virus (HAV) from oysters seeded with this virus, was assessed by real-time RT-PCR and conventional RT-nested PCR after extracting viral RNA by a commercial isolation kit. This procedure consists of an alkaline elution with a glycine buffer, solids removal by slow speed centrifugation, purification by chloroform extraction and virus concentration by ultracentrifugation. In the present study, we evaluated a rapid and simple extraction method to concentrate and purify enteric viruses from shellfish tissues for their detection by real-time RT-PCR. Consumption of virus-contaminated shellfish has caused numerous outbreaks of gastroenteritis and hepatitis worldwide.
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