The developed procedure combining elution, ultrafiltration, and real-time RT-PCR detected average limits of 1 TCID50 for HAV, 54 RT-PCR units for NV, and 0.02 TCID50 for RV per 15 g of food within 6 hours.
The developed procedure allows for rapid and sensitive detection and quantification of enteric viruses in fresh and frozen produce within 6 hours.
Several hepatitis A virus (HAV) and norovirus (NV) outbreaks due to consumption of berries and vegetables have been reported during recent years. To facilitate the detection of enteric viruses that may be present on different fresh and frozen products, we developed a rapid and sensitive detection method for HAV, NV, and rotavirus (RV). Initial experiments focused on optimizing the composition of the elution buffer, improving the viral concentration method, and evaluating the performance of various extraction kits. Viruses were extracted from the food surface by a direct elution method in a glycine-Tris (pH 9.5) buffer containing 1% beef extract and concentrated by ultrafiltration. Occasionally, PCR inhibitors were present in the processed berry samples, which gave relatively poor detection limits. However, this problem was overcome by adding a pectinase treatment in the protocol, which markedly improved the sensitivity of the method. After optimization, this concentration method was applied in combination with real-time reverse transcription-PCR (RT-PCR) using specific primers in various types of berries and vegetables. The average detection limits were 1 50% tissue culture infective dose (TCID(50)), 54 RT-PCR units, and 0.02 TCID(50) per 15 g of food for HAV, NV, and RV, respectively. Based on our results, it is concluded that this procedure is suitable to detect and quantify enteric viruses within 6 h and can be applied for surveillance of enteric viruses in fresh and frozen products.
Butot et al. (Tue,) conducted a other in Enteric virus contamination in food. Elution, ultrafiltration, and real-time RT-PCR vs. PEG precipitation was evaluated on Detection limits of HAV, NV, and RV per 15 g of food. The developed procedure combining elution, ultrafiltration, and real-time RT-PCR detected average limits of 1 TCID50 for HAV, 54 RT-PCR units for NV, and 0.02 TCID50 for RV per 15 g of food within 6 hours.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: