Human norovirus (HuNoV), which causes gastroenteritis, can be transmitted to food and food contact surfaces via viruscontaminated hands. To investigate this transmission in food processing environments, we developed a swabbing protocol for environmental samples, evaluated the stability of HuNoV in the swabs, and applied the method in the food industry. Swabs made of polyester, flocked nylon, cotton wool, and microfiber were moistened in either phosphate-buffered saline (PBS) or glycine buffer (pH 9.5) and used to swab four surfaces (latex, plastic, stainless steel, and cucumber) inoculated with HuNoV. HuNoV was eluted with either PBS or glycine buffer and detected with quantitative reverse transcription PCR. HuNoV recoveries were generally higher with an inoculation dose of 100 PCR units than 1,000 PCR units. The highest recoveries were obtained when surfaces were swabbed with microfiber cloth moistened in and eluted with glycine buffer after a HuNoV inoculation dose of 100 PCR units: 66% ± 18% on latex, 89% ±2% on plastic, and 79% ±10% on stainless steel. The highest recovery for cucumber, 45% ±5%, was obtained when swabbing the surface with microfiber cloth and PBS. The stability of HuNoV was tested in microfiber cloths moistened in PBS or glycine buffer. HuNoV RNA was detected from swabs after 3 days at 4 and 22°C, although the RNA levels decreased more rapidly in swabs moistened with glycine buffer than in those moistened with PBS at 22°C. In the field study, 172 microfiber and 45 cotton wool swab samples were taken from environmental surfaces at three food processing companies. Five (5.6%) of 90 swabs collected in 2010 and 7 (8.5%) of 82 swabs collected in 2012 were positive for HuNoV genogroup II; all positive samples were collected with microfiber swabs. Three positive results were obtained from the production line and nine were obtained from the food workers' break room and restroom areas. Swabbing is a powerful tool for HuNoV RNA detection from environmental surfaces and enables investigation of virus transmission during food processing.

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.

No abstract

cHuman noroviruses (HuNoVs), a leading cause of food-borne gastroenteritis worldwide, are easily transferred via ready-to-eat (RTE) foods, often prepared by infected food handlers. In this study, the transmission of HuNoV and murine norovirus (MuNoV) from virus-contaminated hands to latex gloves during gloving, as well as from virus-contaminated donor surfaces to recipient surfaces after simulated preparation of cucumber sandwiches, was inspected. Virus transfer was investigated by swabbing with polyester swabs, followed by nucleic acid extraction from the swabs with a commercial kit and quantitative reverse transcription-PCR. During gloving, transfer of MuNoV dried on the hand was observed 10/12 times. HuNoV, dried on latex gloves, was disseminated to clean pairs of gloves 10/12 times, whereas HuNoV without drying was disseminated 11/12 times. In the sandwich-preparing simulation, both viruses were transferred repeatedly to the first recipient surface (left hand, cucumber, and knife) during the preparation. Both MuNoV and HuNoV were transferred more efficiently from latex gloves to cucumbers (1.2% ؎ 0.6% and 1.5% ؎ 1.9%) than vice versa (0.7% ؎ 0.5% and 0.5% ؎ 0.4%). We estimated that transfer of at least one infective HuNoV from contaminated hands to the sandwich prepared was likely to occur if the hands of the food handler contained 3 log 10 or more HuNoVs before gloving. Virus-contaminated gloves were estimated to transfer HuNoV to the food servings more efficiently than a single contaminated cucumber during handling. Our results indicate that virus-free food ingredients and good hand hygiene are needed to prevent HuNoV contamination of RTE foods.

No abstract