Human airway epithelial cell culture to identify new respiratory viruses: coronavirus NL63 as a model.
Human airway epithelial cell culture to identify new respiratory viruses: coronavirus NL63 as a model.
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DOI：
10.1016/j.jviromet.2008.10.022
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发表时间：
2009-03
影响因子：
3.1
通讯作者：
Baker, Susan C.
中科院分区：
文献类型：
--
作者：
Banach, Bridget S.;Orenstein, Jan M.;Fox, Linda M.;Randell, Scott H.;Rowley, Anne H.;Baker, Susan C.
新的人类呼吸道病毒病原体在已建立的细胞系中的传播受到细胞系许可性缺乏可预测性以及缺乏合适的抗体试剂来检测不表现出显着细胞病变效应的细胞系感染的阻碍。最近，分子方法已被用来直接从临床样本中扩增和鉴定新的核酸序列，但这些方法可能会受到感染发作后不同时间点呼吸道分泌物中存在的病毒数量的阻碍。人气道上皮 (HAE) 培养物可有效模拟人支气管环境，可培养多种人呼吸道病毒病原体。这里描述的实验的目的是确定是否可以通过接种 HAE 培养物，然后进行全转录组扩增 (WTA) 和序列分析来实现人类呼吸道病毒的传播和识别。为了建立原理验证，对人类冠状病毒 NL63 (HCoV-NL63) 进行了评估，并首次报道了通过透射电子显微镜 (TEM) 对 HCoV-NL63 病毒进行可视化。将人类呼吸道分泌物初始繁殖到 HAE 培养物上，然后对培养物上清液进行 TEM 和 WTA，这可能是可视化和检测传统方法无法识别的新人类呼吸道病原体的有用方法。
Propagation of new human respiratory virus pathogens in established cell lines is hampered by a lack of predictability regarding cell line permissivity and by availability of suitable antibody reagents to detect infection in cell lines that do not exhibit significant cytopathic effect. Recently, molecular methods have been used to amplify and identify novel nucleic acid sequences directly from clinical samples, but these methods may be hampered by the quantity of virus present in respiratory secretions at different time points following the onset of infection. Human airway epithelial (HAE) cultures, which effectively mimic the human bronchial environment, allow for cultivation of a wide variety of human respiratory viral pathogens. The goal of the experiments described here was to determine if propagation and identification of a human respiratory virus may be achieved through inoculation of HAE cultures followed by whole transcriptome amplification (WTA) and sequence analysis. To establish proof-of-principle human coronavirus NL63 (HCoV-NL63) was evaluated, and the first visualization of HCoV-NL63 virus by transmission electron microscopy (TEM) is reported. Initial propagation of human respiratory secretions onto HAE cultures followed by TEM and WTA of culture supernatant may be a useful approach for visualization and detection of new human respiratory pathogens that have eluded identification by traditional approaches.
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影响因子：
82.9
作者：
van den Hoogen BG;de Jong JC;Groen J;Kuiken T;de Groot R;Fouchier RA;Osterhaus AD
通讯作者：
Osterhaus AD
影响因子：
3.6
作者：
Suzuki, A;Okamoto, M;Nishimura, H
通讯作者：
Nishimura, H
影响因子：
5.4
作者：
Thompson, Catherine I.;Barclay, Wendy S.;Pickles, Raymond J.
通讯作者：
Pickles, Raymond J.
DOI：
10.1073/pnas.0504666102
发表时间：
2005-09-06
影响因子：
11.1
作者：
Allander, T;Tammi, MT;Andersson, B
通讯作者：
Andersson, B
影响因子：
5.4
作者：
Sims, AC;Baric, RS;Pickles, RJ
通讯作者：
Pickles, RJ