In Vitro Cell Culture Infectivity Assay for Human Noroviruses

Journal Article · Tue Jan 30 00:00:00 EST 2007 · Emerging Infectious Diseases, 13(3):396-403

DOI:https://doi.org/10.3201/eid1303.060549· OSTI ID:907937

Straub, Tim M; Honer Zu Bentrup, Kerstin A; Orosz Coghlan, Patricia A; Dohnalkova, Alice; Mayer, Brooke K; Bartholomew, Rachel A; Valdez, Catherine O; Bruckner-Lea, Cindy J; Gerba, Charles P; Abbaszadegan, Morteza; Nickerson, Cheryl A

Human noroviruses (NoV) cause severe, self-limiting gastroenteritis that typically lasts 24 - 48 hours. The true nature of NoV pathogenesis remains unknown due to the lack of suitable tissue culture or animal models. Here we show, for the first time, that NoV can infect and replicate in an organoid, three-dimensional (3-D) model of human small intestinal epithelium (INT-407). Cellular differentiation for this model was achieved by growing the cells in 3-D on porous collagen I-coated microcarrier beads under conditions of physiological fluid shear in rotating wall vessel bioreactors. Microscopy, PCR, and fluorescent in-situ hybridization were employed to provide evidence of NoV infection. CPE and norovirus RNA was detected at each of the five cell passages for both genogroup I and II viruses. Our results demonstrate that the highly differentiated 3-D cell culture model can support the natural growth of human noroviruses, whereas previous attempts using differentiated monolayer cultures failed.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 907937
• Report Number(s): PNNL-SA-53115; 16297; 16297a; TRN: US200721%%507
• Journal Information: Emerging Infectious Diseases, 13(3):396-403, Vol. 13, Issue 3
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
ANIMALS
BIOREACTORS
CELL CULTURES
COLLAGEN
EPITHELIUM
IN VITRO
INFECTIVITY
IN-SITU HYBRIDIZATION
MICROSCOPY
PATHOGENESIS
RNA
SHEAR
TISSUE CULTURES
VIRUSES
Norovirus
Cytopathogenic Effect
Viral (CPE)
Polymerase Chain Reaction
In Situ Hybridization
Fluorescence
Microscopy
Electron
three-dimensional cell culture
Organoids
RWV bioreactor
Environmental Molecular Sciences Laboratory