Ultraviolet Light Inactivation of Murine Norovirus and Human Norovirus GII: PCR May Overestimate the Persistence of Noroviruses Even When Combined with Pre-PCR Treatments

Abstract

Transmission of gastroenteritis-causing noroviruses may be significant via contaminated surfaces. Measures for control, e.g. disinfection with ultraviolet irradiation (UV), are therefore necessary for interrupting this transmission. Human norovirus (HuNoV) GII.4 and Murine norovirus (MuNoV) were used to study the efficacy of UV for virus inactivation on dry glass surfaces. MuNoV inactivation was measured using viability assay and the reduction in viral RNA levels for both viruses using reverse transcription quantitative PCR (RT-QPCR). For each UV dose, two parallel sample groups were detected using RT-QPCR: one group was enzymatically pre-PCR treated with Pronase and RNAse enzymes, while the other was not treated enzymatically. In the viability assay, loss of infectivity and a 4-log reduction of MuNoV were observed when the viruses on glass slides were treated with a UV dose of 60 mJ/cm(2) or higher. In the RT-QPCR assay, a steady 2-log decline of MuNoV and HuNoV RNA levels was observed when UV doses were raised from 0 to 150 mJ/cm(2). A distinct difference in RNA levels of pretreated and non-pretreated samples was observed with UV doses of 450-1.8 × 10(3) mJ/cm(2): the RNA levels of untreated samples remained over 1.0 × 10(3) PCR units (pcr-u), while the RNA levels of enzyme-treated samples declined below 100 pcr-u. However, the data show a prominent difference between the persistence of MuNoV observed with the infectivity assay and that of viral RNA detected using RT-QPCR. Methods based on genome detection may overestimate norovirus persistence even when samples are pretreated before genome detection.

Keywords: Environmental surfaces; Murine norovirus; Norovirus; UV irradiation.