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Volume 24, Number 8—August 2018

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)

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Figure 7

Determination of ID50 in study of norovirus replication in human intestinal enteroids (HIEs). We inoculated HIE monolayers in triplicate with 10-fold serial diluted of the indicated A) A5413_GII.4 Sydney, B) G3868_GII.4 Den Haag, or C) N741656 _GII.3 RNA copies and incubated them for 1 h at 37°C. We washed the monolayers 3 times and cultured them in differentiation media for 3 d. We extracted RNA and quantified it by quantitative reverse transcription PCR from frozen lysates (cells and supernata

Figure 7. Determination of ID50 required for human norovirus replication in human intestinal enteroids (HIEs). We inoculated HIE monolayers in triplicate with 10-fold serial diluted fecal filtrates A) A5413_GII.4 Sydney, B) G3868_GII.4 Den Haag, or C) N741656 _GII.3 RNA copies and incubated them for 1 h at 37°C. We washed the monolayers 3 times and cultured them in differentiation media for 3 d. We extracted RNA and quantified it by quantitative reverse transcription PCR from frozen lysates (cells and supernatant) at 1 hour postinfection and 3 days postinfection. We calculated ID50 using the Reed-Muench method (28). ID50, 50% infectious dose.

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