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利用原位杂交技术在体外对鼻病毒复制进行定位

Localization of rhinovirus replication in vitro with in situ hybridization.

作者信息

de Arruda E, Mifflin T E, Gwaltney J M, Winther B, Hayden F G

机构信息

Department of Internal Medicine, University of Virginia Health Sciences Center, Charlottesville 22908.

出版信息

J Med Virol. 1991 May;34(1):38-44. doi: 10.1002/jmv.1890340107.

Abstract

To facilitate understanding of human rhinovirus (HRV) pathogenesis, methods were developed for detection of HRV infection in vitro using in situ hybridization (ISH). HRV-14 RNA probes and oligonucleotide probes representing conserved sequences in the 5'-non-translated region were labeled with 35S and used to detect infected HeLa or WI-38 strain human embryonic lung cells in cytological preparations. ISH was shown to be specific for detection of HRV on a single-cell basis. Subsequently, in human nasal polyps infected in vitro, both oligonucleotide- and ribo-probes produced a strong signal in association with ciliated epithelial cells. In human adenoids infected in vitro, a signal was observed in non-ciliated epithelial cells. This study shows that HRV replicates in ciliated cells in the epithelium of human nasal polyps infected in vitro, and the presence of viral RNA in non-ciliated cells of the human adenoid infected in vitro suggests that other cell types may also support rhinovirus replication.

摘要

为了便于理解人鼻病毒(HRV)的发病机制,人们开发了利用原位杂交(ISH)在体外检测HRV感染的方法。用35S标记HRV - 14 RNA探针和代表5'-非翻译区保守序列的寡核苷酸探针,用于在细胞学制剂中检测感染的HeLa或WI - 38株人胚肺细胞。ISH被证明在单细胞水平上对HRV检测具有特异性。随后,在体外感染的人鼻息肉中,寡核苷酸探针和核糖探针均在纤毛上皮细胞中产生强烈信号。在体外感染的人腺样体中,在非纤毛上皮细胞中观察到信号。这项研究表明,HRV在体外感染的人鼻息肉上皮的纤毛细胞中复制,并且在体外感染的人腺样体的非纤毛细胞中存在病毒RNA表明其他细胞类型也可能支持鼻病毒复制。

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