Detection and Quantitation of Norovirus Genome Using Real-Time RT-PCR

Abstract

We have studied the two assay methods of the 1Step Real-Time RT-PCR and conventional Real-Time RT-PCR (2Step) using ABI PRISM 7000 Sequence Detection System (ABI PRISM 7000) and Thermal Cycler Dice Real Time System (Thermal Cycler Dice) for the detection of Norovirus (NV) genogroups I (G1) and II (GII). Thermal Cycler Dice showed the same performance with the sensitivity and the quantitation compared with ABI PRISM 7000 that the official assay presented for the detection of NV. Both of the 1Step and 2Step Real-Time RT-PCR enabled to get the rapid results about 90 minutes. Excepting the probes with MGB, the primers and probes used in this study were identical with those used in the official assay. Results of field samples using both of the instruments and both of the assays were the same. Hence, these assays are very useful laboratory techniques for the detection of NV from field samples.