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全小鼠器官系统中DNA和RNA病毒基因组的检测:多瘤病毒对小鼠器官的感染模式

Detection of DNA and RNA virus genomes in organ systems of whole mice: patterns of mouse organ infection by polyomavirus.

作者信息

Dubensky T W, Murphy F A, Villarreal L P

出版信息

J Virol. 1984 Jun;50(3):779-83. doi: 10.1128/JVI.50.3.779-783.1984.

Abstract

A technique which detects viral DNA or RNA in situ in the organ systems of whole mice is described. Frozen thin sections from whole mice were transferred directly to nitrocellulose and hybridized to labeled viral DNA, allowing the detection of viral DNA or RNA. By this procedure, polyomavirus infection of newborn mice inoculated intranasally was followed. We found that the initial inoculum could be detected in the nasal cavity, lungs, and stomach lining after a 5-h absorption period. Primary replication of virus was observed in the nasal cavity, submaxillary gland, and lungs, followed by a systemic phase of infection in which the liver, spleen, kidney, and large colon also became infected. Viral RNA as well as DNA could also be detected as shown by infecting mice intracerebrally with vesicular stomatitis virus. Vesicular stomatitis virus-specific RNA was observed only in the brains of these mice. It is most likely that this technique can be applied to general molecular studies of mice. With this method we should be able to detect all viruses, bacteria, plasmids, and organ-specific transcripts to which a cloned probe exists.

摘要

本文描述了一种在完整小鼠的器官系统中原位检测病毒DNA或RNA的技术。将完整小鼠的冷冻薄切片直接转移至硝酸纤维素膜上,并与标记的病毒DNA杂交,从而实现病毒DNA或RNA的检测。通过该程序,对经鼻接种的新生小鼠的多瘤病毒感染情况进行了跟踪。我们发现,在5小时的吸附期后,可在鼻腔、肺部和胃黏膜中检测到初始接种物。在鼻腔、下颌下腺和肺部观察到病毒的初次复制,随后进入感染的全身阶段,在此阶段肝脏、脾脏、肾脏和大肠也被感染。通过脑内接种水疱性口炎病毒感染小鼠的实验表明,病毒RNA以及DNA均可被检测到。仅在这些小鼠的大脑中观察到水疱性口炎病毒特异性RNA。极有可能该技术可应用于小鼠的一般分子研究。通过这种方法,我们应该能够检测到所有存在克隆探针的病毒、细菌、质粒和器官特异性转录本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beda/255737/aa7844ead93c/jvirol00135-0118-a.jpg

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