In vitro cell culture infectivity assay for human noroviruses

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

Research output: Contribution to journalArticle

205 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)396-403
Number of pages8
JournalEmerging Infectious Diseases
Volume13
Issue number3
StatePublished - Mar 2007

Fingerprint

Norovirus
Cell Culture Techniques
Organoids
Somatostatin-Secreting Cells
Gastroenteritis
Bioreactors
Intestinal Mucosa
Fluorescence In Situ Hybridization
In Vitro Techniques
Microscopy
Collagen
Animal Models
Genotype
RNA
Viruses
Polymerase Chain Reaction
Growth
Infection

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Straub, T. M., Zu Bentrup, K. H., Orosz-Coghlan, P., Dohnalkova, A., Mayer, B. K., Bartholomew, R. A., ... Nickerson, C. (2007). In vitro cell culture infectivity assay for human noroviruses. Emerging Infectious Diseases, 13(3), 396-403.

In vitro cell culture infectivity assay for human noroviruses. / Straub, Timothy M.; Zu Bentrup, Kerstin Höner; Orosz-Coghlan, Patricia; Dohnalkova, Alice; Mayer, Brooke K.; Bartholomew, Rachel A.; Valdez, Catherine O.; Bruckner-Lea, Cynthia J.; Gerba, Charles P.; Abbaszadegan, Morteza; Nickerson, Cheryl.

In: Emerging Infectious Diseases, Vol. 13, No. 3, 03.2007, p. 396-403.

Research output: Contribution to journalArticle

Straub, TM, Zu Bentrup, KH, Orosz-Coghlan, P, Dohnalkova, A, Mayer, BK, Bartholomew, RA, Valdez, CO, Bruckner-Lea, CJ, Gerba, CP, Abbaszadegan, M & Nickerson, C 2007, 'In vitro cell culture infectivity assay for human noroviruses', Emerging Infectious Diseases, vol. 13, no. 3, pp. 396-403.
Straub TM, Zu Bentrup KH, Orosz-Coghlan P, Dohnalkova A, Mayer BK, Bartholomew RA et al. In vitro cell culture infectivity assay for human noroviruses. Emerging Infectious Diseases. 2007 Mar;13(3):396-403.
Straub, Timothy M. ; Zu Bentrup, Kerstin Höner ; Orosz-Coghlan, Patricia ; Dohnalkova, Alice ; Mayer, Brooke K. ; Bartholomew, Rachel A. ; Valdez, Catherine O. ; Bruckner-Lea, Cynthia J. ; Gerba, Charles P. ; Abbaszadegan, Morteza ; Nickerson, Cheryl. / In vitro cell culture infectivity assay for human noroviruses. In: Emerging Infectious Diseases. 2007 ; Vol. 13, No. 3. pp. 396-403.
@article{032b549dacce4dad9a97f464843bf93c,
title = "In vitro cell culture infectivity assay for human noroviruses",
abstract = "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.",
author = "Straub, {Timothy M.} and {Zu Bentrup}, {Kerstin H{\"o}ner} and Patricia Orosz-Coghlan and Alice Dohnalkova and Mayer, {Brooke K.} and Bartholomew, {Rachel A.} and Valdez, {Catherine O.} and Bruckner-Lea, {Cynthia J.} and Gerba, {Charles P.} and Morteza Abbaszadegan and Cheryl Nickerson",
year = "2007",
month = "3",
language = "English (US)",
volume = "13",
pages = "396--403",
journal = "Emerging Infectious Diseases",
issn = "1080-6040",
publisher = "Centers for Disease Control and Prevention (CDC)",
number = "3",

}

TY - JOUR

T1 - In vitro cell culture infectivity assay for human noroviruses

AU - Straub, Timothy M.

AU - Zu Bentrup, Kerstin Höner

AU - Orosz-Coghlan, Patricia

AU - Dohnalkova, Alice

AU - Mayer, Brooke K.

AU - Bartholomew, Rachel A.

AU - Valdez, Catherine O.

AU - Bruckner-Lea, Cynthia J.

AU - Gerba, Charles P.

AU - Abbaszadegan, Morteza

AU - Nickerson, Cheryl

PY - 2007/3

Y1 - 2007/3

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=33847339029&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33847339029&partnerID=8YFLogxK

M3 - Article

VL - 13

SP - 396

EP - 403

JO - Emerging Infectious Diseases

JF - Emerging Infectious Diseases

SN - 1080-6040

IS - 3

ER -