Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, IN 46556, United States.
National Calicivirus Laboratory, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, United States.
J Appl Microbiol. 2024 Feb 1;135(2). doi: 10.1093/jambio/lxae033.
This study aimed to compare the heat inactivation kinetics of viable human norovirus with the surrogate, MS2 bacteriophage as well as assess the decay of the RNA signal.
Human intestinal enteroids were used to analyze the heat inactivation kinetics of viable human norovirus compared to the surrogate MS2 bacteriophage, which was cultured using a plaque assay. Norovirus decay rates were 0.22 min-1, 0.68 min-1, and 1.11 min-1 for 50°C, 60°C, and 70°C, respectively, and MS2 bacteriophage decay rates were 0.0065 min-1, 0.045 min-1, and 0.16 min-1 for 50°C, 60°C, and 70°C, respectively. Norovirus had significantly higher decay rates than MS2 bacteriophage at all tested temperatures (P = .002-.007). No decrease of RNA titers as measured by reverse transcription-PCR for both human norovirus and MS2 bacteriophage over time was observed, indicating molecular methods do not accurately depict viable human norovirus after heat inactivation and treatment efficiency is underestimated.
Overall, our data demonstrate that MS2 bacteriophage is a conservative surrogate to measure heat inactivation and potentially overestimates the infectious risk of norovirus. Furthermore, this study corroborates that measuring viral RNA titers, as evaluated by PCR methods, does not correlate with the persistence of viable norovirus under heat inactivation.
本研究旨在比较人类诺如病毒与替代物 MS2 噬菌体的热失活动力学,并评估 RNA 信号的衰减。
使用人肠类器官分析与替代物 MS2 噬菌体相比,活的人类诺如病毒的热失活动力学,MS2 噬菌体使用噬菌斑测定法培养。诺如病毒的衰减率分别为 50°C、60°C 和 70°C 时为 0.22 min-1、0.68 min-1 和 1.11 min-1,而 MS2 噬菌体的衰减率分别为 50°C、60°C 和 70°C 时为 0.0065 min-1、0.045 min-1 和 0.16 min-1。在所有测试温度下,诺如病毒的衰减率均显著高于 MS2 噬菌体(P 均=0.002-0.007)。未观察到时间推移时逆转录-PCR 测量的人类诺如病毒和 MS2 噬菌体 RNA 滴度下降,这表明分子方法不能准确描述热失活后的活的人类诺如病毒,并且低估了处理效率。
总体而言,我们的数据表明 MS2 噬菌体是衡量热失活和潜在高估诺如病毒感染风险的保守替代物。此外,本研究证实,如 PCR 方法评估的病毒 RNA 滴度与热失活下活的诺如病毒的持续存在无关。