In Vitro Cell Culture Infectivity Assay for Human Noroviruses

Straub, Timothy M.; Höner zu Bentrup, Kerstin; Coghlan, Patricia Orosz; Dohnalkova, Alice; Mayer, Brooke K.; Bartholomew, Rachel A.; Valdez, Catherine O.; Bruckner-Lea, Cynthia J.; Gerba, Charles P.; Abbaszadegan, Morteza A.; Nickerson, Cheryl A.

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In Vitro Cell Culture Infectivity Assay for Human Noroviruses

Supporting Files Public Domain

Mar 2007
By Straub, Timothy M. ; Höner zu Bentrup, Kerstin ; Coghlan, Patricia Orosz ; ...

Details

Alternative Title:

Emerg Infect Dis
Personal Author:

Straub, Timothy M. ; Höner zu Bentrup, Kerstin ; Coghlan, Patricia Orosz ; Dohnalkova, Alice ; Mayer, Brooke K. ; Bartholomew, Rachel A. ; Valdez, Catherine O. ; Bruckner-Lea, Cynthia J. ; Gerba, Charles P. ; Abbaszadegan, Morteza A. ; Nickerson, Cheryl A.
Description:

Human noroviruses cause severe, self-limiting gastroenteritis that typically lasts 24-48 hours. Because of the lack of suitable tissue culture or animal models, the true nature of norovirus pathogenesis remains unknown. We show, for the first time, that noroviruses can infect and replicate in a physiologically relevant 3-dimensional (3-D), organoid model of human small intestinal epithelium. This level of cellular differentiation was achieved by growing the cells 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 provided evidence of norovirus infection. Cytopathic effect and norovirus RNA were detected at each of the 5 cell passages for 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 that used differentiated monolayer cultures failed.
Subjects:

3-dimensional Cell Culture Caliciviridae Infections Cell Culture Techniques Cell Line Collagen Type I Cytopathic Effect Cytopathogenic Effect, Viral Electron Microscopy Fluorescence Humans Intestinal Mucosa Microspheres Norovirus Organoids Research RNA, Viral RWV Bioreactor
Source:

Emerg Infect Dis. 13(3):396-403.
Document Type:

Journal Article
Volume:

13
Issue:

3
Collection(s):

Emerging Infectious Diseases
Main Document Checksum:

urn:sha256:3620560172544459f1b57199e9d68a93a8cd77f980e8ca046228217842c90c81
Download URL:

https://stacks.cdc.gov/view/cdc/16362/cdc_16362_DS1.pdf
File Type:

[PDF - 388.74 KB ]

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