In Vitro Cell Culture Infectivity Assay for Human Noroviruses

T. M. Straub; Kerstin Hner zu Bentrup; Patricia Orosz-Coghlan; Alice Dohnalkova; Brooke K Mayer; Rachel A Bartholomew; Catherine OValdez; Cynthia J Bruckner-Lea; Charles P Gerba; Morteza Abbaszadegan; Cheryl A Nickerson

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In Vitro Cell Culture Infectivity Assay for Human Noroviruses

机译：人诺如病毒的体外细胞培养感染性测定

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Human noroviruses cause severe, self-limiting gas-troenteritis that typically lasts 24–48 hours. Because of thelack of suitable tissue culture or animal models, the truenature of norovirus pathogenesis remains unknown. Weshow, for the first time, that noroviruses can infect and repli-cate in a physiologically relevant 3-dimensional (3-D),organoid model of human small intestinal epithelium. Thislevel of cellular differentiation was achieved by growing thecells on porous collagen-I coated microcarrier beads underconditions of physiological fluid shear in rotating wall ves-sel bioreactors. Microscopy, PCR, and fluorescent in situhybridization provided evidence of norovirus infection.Cytopathic effect and norovirus RNA were detected at eachof the 5 cell passages for genogroup I and II viruses. Ourresults demonstrate that the highly differentiated 3-D cellculture model can support the natural growth of humannoroviruses, whereas previous attempts that used differen-tiated monolayer cultures failed

机译：人类诺如病毒会引起严重的自限性胃肠炎，通常持续24-48小时。由于缺乏合适的组织培养物或动物模型，诺如病毒发病机理的真实性仍然未知。我们首次表明，诺如病毒可以感染并复制人类小肠上皮的生理相关的3维（3-D）有机样模型。这种水平的细胞分化是通过在旋转壁容器生物反应器中在生理流体剪切的条件下使细胞在胶原蛋白I包覆的多孔微载体珠上生长来实现的。显微镜检查，PCR和荧光原位杂交提供了诺如病毒感染的证据。基因组I和II的5个细胞传代中均检测到细胞致病作用和诺如病毒RNA。我们的结果表明，高度分化的3-D细胞培养模型可以支持人类诺如病毒的自然生长，而先前使用差异化单层培养的尝试却失败了

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《Emerging Infectious Diseases》 |2007年第3期|共页
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T. M. Straub; Kerstin Hner zu Bentrup; Patricia Orosz-Coghlan; Alice Dohnalkova; Brooke K Mayer; Rachel A Bartholomew; Catherine OValdez; Cynthia J Bruckner-Lea; Charles P Gerba; Morteza Abbaszadegan; Cheryl A Nickerson;
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1. Cell Culture Assay for Human Noroviruses [J] . M. Chan, YP Wong, Wai K Leung Emerging Infectious Diseases . 2007,第7期

机译：人类诺如病毒的细胞培养测定
2. Development of an Infectivity Assay for Noroviruses in Cells [Project #3027] [J] . Journal of the New England Water Works Association . 2011,第2期

机译：诺如病毒在细胞中的感染性测定方法的开发[项目＃3027]
3. Temperature inactivation of Feline calicivirus vaccine strain FCV F-9 in comparison with human noroviruses using an RNA exposure assay and reverse transcribed quantitative real-time polymerase chain reaction-A novel method for predicting virus infect [J] . Topping JR, Schnerr H, Haines J, Journal of Virological Methods . 2009,第1a2期

机译：与人诺如病毒相比，猫杯状病毒疫苗株FCV F-9的温度失活使用RNA暴露测定法和逆转录定量实时聚合酶链反应-一种预测病毒感染的新方法
4. Titration of infectivity in buffy coat of sheep experimentally-infected with 127S scrapie using an in vitro cell culture assay [C] . Segarra C, You B, Bougard D, International PRION Congress . 2010

机译：使用体外细胞培养测定试图用127S瘙痒病进行实验感染羊毛中的感染性的滴定
5. Prevention of Respiratory Syncytial Virus Attachment Protein Cleavage in Vero Cells Rescues Infectivity of Progeny Virions for Primary Human Airway Cultures. [D] . Corry, Jacqueline D. 2015

机译：预防Vero细胞中呼吸道合胞病毒附着蛋白的切割可拯救子代病毒对原代人气道培养物的感染性。
6. In Vitro Cell Culture Infectivity Assay for Human Noroviruses [O] . Timothy M. Straub, Kerstin Höner zu Bentrup, Patricia Orosz Coghlan, 2007

机译：人诺如病毒的体外细胞培养感染性测定
7. In Vitro Cell Culture Infectivity Assay for Human Noroviruses [O] . Straub, Timothy M., Höner zu Bentrup, Kerstin, Coghlan, Patricia Orosz, 2007

机译：人诺如病毒的体外细胞培养感染性测定
8. Results of an in Vitro Corrosion Assay in Cultured Human Epidermal Cells. [R] . 2012

机译：培养的人表皮细胞的体外腐蚀测定结果。

In Vitro Cell Culture Infectivity Assay for Human Noroviruses

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