Improved real-time quantitative reverse transcription PCR detection of norovirus following removal of inhibitors

Human norovirus (HuNoV) is an important enteric virus that can cause large gastroenteritis outbreaks via the fecal-oral route from contaminated water and produce. Real-time quantitative reverse transcription PCR (RT-qPCR) is the only method to apply the routine detection of HuNoV in various samples, however, inhibitors present in the samples can affect the accuracy and sensitivity of RT-qPCR results. Here, we suggest an inhibitor-removal treatment for two types of noroviruses using two commercial kits. Two types of water sample (surface and seawater) and four types of produce (green onions, lettuces, radishes, and strawberries) were evaluated. The recovery efficiencies of noroviruses in water samples clearly increased in surface and seawater samples with the inhibitor-removal treatment compared to untreated samples. Moreover, murine norovirus-1 was well recovered from the four types of produce with the inhibitor-removal treatment. The mean recovery efficiencies of HuNoV genogroup II genotype 4 in lettuces and strawberries were also increased in the treated samples. Therefore, we suggest that the inhibitor-removal treatment could be useful for improving the accuracy and sensitivity of RT-qPCR methods for noroviruses in water and produce.