Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, P.O. Box 66, University of Helsinki, 00014 Helsinki, Finland.
J Food Prot. 2013 Aug;76(8):1421-8. doi: 10.4315/0362-028X.JFP-12-371.
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.
人诺如病毒(HuNoV)可引起肠胃炎,可通过污染病毒的手传播至食物和食物接触面。为了研究食品加工环境中的这种传播,我们开发了一种环境样本拭子采集方案,评估了 HuNoV 在拭子中的稳定性,并将该方法应用于食品工业。聚酯、植绒尼龙、棉花和微纤维制成的拭子分别用磷酸盐缓冲盐水(PBS)或甘氨酸缓冲液(pH9.5)润湿,用于擦拭涂有 HuNoV 的四个表面(乳胶、塑料、不锈钢和黄瓜)。用 PBS 或甘氨酸缓冲液洗脱 HuNoV,并用定量逆转录 PCR 检测。与接种剂量为 1000PCR 单位相比,接种剂量为 100PCR 单位时 HuNoV 的回收率普遍更高。用 100PCR 单位接种剂量的微纤维布擦拭并在甘氨酸缓冲液中洗脱后,HuNoV 的回收率最高:乳胶表面为 66%±18%,塑料表面为 89%±2%,不锈钢表面为 79%±10%。用微纤维布和 PBS 擦拭黄瓜表面时,回收率最高,为 45%±5%。在 PBS 或甘氨酸缓冲液润湿的微纤维布中检测到 HuNoV 的稳定性。HuNoV RNA 在 4°C 和 22°C 下 3 天从拭子中检出,尽管在 22°C 下,甘氨酸缓冲液润湿的拭子中 RNA 水平下降速度快于 PBS 润湿的拭子。在现场研究中,从三家食品加工公司的环境表面采集了 172 个微纤维和 45 个棉拭子样本。2010 年采集的 90 个拭子中有 5 个(5.6%)和 2012 年采集的 82 个拭子中有 7 个(8.5%)检测到 II 型 HuNoV 基因组;所有阳性样本均用微纤维拭子采集。从生产线中获得了 3 个阳性结果,从食品加工人员的休息室和卫生间区域获得了 9 个阳性结果。拭子采样是从环境表面检测 HuNoV RNA 的有力工具,可用于研究食品加工过程中的病毒传播。