Evaluation of a norovirus detection methodology for soft red fruits

Abstract

In the present study, a proposed methodology for detection of GI and GII noroviruses (NoV) in soft red fruits was evaluated. The murine norovirus-1 (MNV-1), a recently described cultivable NoV surrogate was integrated in the detection methodology as full process control, reverse transcription control and real-time PCR internal amplification control. Both the performance and robustness of the proposed methodology were analyzed. Firstly, the performance of the method was examined by analysis of the recovery of MNV-1, GI and/or GII NoV inoculated on frozen raspberry crum samples. Results showed that the recovery of MNV-1 was not significantly influenced by the inoculum incubation time (30 min or overnight incubation) or the inoculum level (10(6) or 10(8) genomic MNV-1 copies/10 g of frozen raspberry crum sample). In contrast, a significant influence of the GI and GII NoV inoculum level (10(4) or 10(6) genomic MNV-1 copies/10 g of frozen raspberry crum sample) was noticed on the recovery of respectively GI and GII NoV from frozen raspberry crum samples. Secondly, the robustness of the methodology was evaluated by subjecting three types of artificially MNV-1, GI and/or GII NoV contaminated soft red fruit products (deepfrozen forest fruit mix, fresh raspberries and fresh strawberry puree) to the method. Results showed a significant influence of the soft red fruit product type on the recovery efficiency of GI NoV and MNV-1, while no significant differences could be shown for GII NoV. In general, the recovery of GI and GII NoV in strawberry puree was more efficient from the strawberry puree compared to the two other soft red fruit types. In conclusion, results show that this methodology can be used for detection of NoV in different soft red fruits, although NoV recovery efficiencies can be influenced by (1) the NoV concentration on the soft red fruit type and (2) the tested soft red fruit type.