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Volume 24, Number 8—August 2018
Research

Human Norovirus Replication in Human Intestinal Enteroids as Model to Evaluate Virus Inactivation

Veronica CostantiniComments to Author , Esther K. Morantz, Hannah Browne, Khalil Ettayebi, Xi-Lei Zeng, Robert L. Atmar, Mary K. Estes, and Jan Vinjé
Author affiliations: Centers for Disease Control and Prevention, Atlanta, Georgia, USA (V. Costantini, J. Vinjé); Synergy America, Inc., Atlanta (E.K. Morantz); Oak Ridge Institute for Science and Education, Oak Ridge, Tennessee, USA (H. Browne); Baylor College of Medicine, Houston, Texas, USA (K. Ettayebi, X.-L. Zeng, R.L. Atmar, M.K. Estes)

Main Article

Figure 8

Inactivation of human norovirus by chlorine compared with 70% alcohol solutions. A) Ten percent fecal filtrates (G3868 [GII.4 Den Haag], 2.04 × 106 RNA copies; G3829 [GII.4 New Orleans], 4.14 × 106 RNA copies; A5413 [GII.4 Sydney], 1.58 × 107 RNA copies) were either treated or not treated with 70% ethanol for 1 min or 5 min at room temperature. We added complete media without growth factors supplemented with 10% fetal bovine serum to neutralize remaining ethanol. B) Ten percent fecal filtrates f

Figure 8. Inactivation of human norovirus with 70% alcohol or chlorine solution in suspension. A) Ten percent fecal filtrates (G3868 [GII.4 Den Haag], 2.04 × 106 RNA copies; G3829 [GII.4 New Orleans], 4.14 × 106 RNA copies; A5413 [GII.4 Sydney], 1.58 × 107 RNA copies) were either treated or not treated with 70% ethanol for 1 min or 5 min at room temperature. We added complete media without growth factors supplemented with 10% fetal bovine serum to neutralize remaining ethanol. B) Ten percent fecal filtrates from 3 GII.4 Sydney strains (CDC830, 2.89 × 107 RNA copies; R3702, 7.73 × 107 RNA copies; A5413, 1.58 × 107 RNA copies) were either treated or not treated with 70% ethanol or 70% isopropanol for 5 min at room temperature and neutralized with complete media without growth factors supplemented with 10% fetal bovine serum. C, D) Ten percent fecal filtrates (G3868, 2.04 × 106 RNA copies [C]; A5413, 1.58 × 107 RNA copies [D]) were either treated or not treated with freshly prepared chlorine solutions of increasing concentrations (5, 50, 100, 200, 400, 600, 800, 1,000, and 5,000 ppm) for 1 min at room temperature. Sodium thiosulfate (final concentration 50 mg/L) was added to neutralize the remaining free chlorine. For all experiments, HIEs were then infected with 100 μL of treated or not treated fecal filtrate. After 1 h at 37°C in 5% CO2, we washed the monolayers, added differentiation media, and incubated for 3 d. Data represent mean ± SD of 2 experiments with 3 wells for each treatment and time point. For each fecal filtrate, we performed 1-way analysis of variance followed by Dunnett’s test. Dotted lines represent RT-qPCR limit of detection. p values are compared with the nontreated fecal filtrate: *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. dpi, days postinfection; hpi, hours postinfection.

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Page created: July 18, 2018
Page updated: July 18, 2018
Page reviewed: July 18, 2018
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
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