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通过生物素化探针原位杂交在常规石蜡包埋组织中进行人类和病毒基因检测：疱疹性脑炎中的病毒定位

Human and viral gene detection in routine paraffin embedded tissue by in situ hybridisation with biotinylated probes: viral localisation in herpes encephalitis.

作者信息

Burns J, Redfern D R, Esiri M M, McGee J O

出版信息

J Clin Pathol. 1986 Oct;39(10):1066-73. doi: 10.1136/jcp.39.10.1066.

DOI:10.1136/jcp.39.10.1066
PMID:3023452
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC500224/
Abstract

A simple reproducible protocol for detecting multiple copy human genes and viral DNA in routine formalin fixed paraffin embedded tonsil and brain, by in situ hybridisation with biotinylated probes, is described. The protocol consists of digestion of formalin fixed paraffin sections, with 0.4% pepsin in 0.01 M hydrochloric acid for one hour at 37 degrees C, followed by hybridisation with biotinylated probes. The biotinylated probes used for establishing the conditions for in situ localisation of DNA were total placental DNA (TG1), pHY 2.1 (a Y chromosome probe), and herpes simplex virus I and II. In human male tonsil TG1 labelled all nuclei and pHY 2.1 reacted only with nuclear Y bodies. In herpes encephalitis the virus was detected in some glial cells and neurones.

摘要

本文描述了一种简单且可重复的方案，用于通过与生物素化探针进行原位杂交，在常规福尔马林固定石蜡包埋的扁桃体和脑组织中检测多拷贝人类基因和病毒DNA。该方案包括用0.4%胃蛋白酶于0.01 M盐酸中在37℃消化福尔马林固定的石蜡切片1小时，随后与生物素化探针杂交。用于确定DNA原位定位条件的生物素化探针有胎盘总DNA（TG1）、pHY 2.1（Y染色体探针）以及单纯疱疹病毒I和II。在人类男性扁桃体中，TG1标记所有细胞核，而pHY 2.1仅与核Y小体反应。在疱疹性脑炎中，在一些神经胶质细胞和神经元中检测到了病毒。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/0f4047941734/jclinpath00205-0023-b.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/c62a6cfb60a2/jclinpath00205-0021-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/2107bcc03c49/jclinpath00205-0021-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/a2d64eed72d5/jclinpath00205-0022-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/66cc5082af76/jclinpath00205-0022-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/230107521b3b/jclinpath00205-0022-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/9e1c86e9229b/jclinpath00205-0023-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6718/500224/0f4047941734/jclinpath00205-0023-b.jpg

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本文引用的文献

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Chromosoma. 1982;87(5):491-502. doi: 10.1007/BF00333470.
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Biotinated probe containing a long-terminal repeat hybridized to a mouse colon tumor and normal tissue.
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In situ hybridization: alkaline phosphatase visualization of biotinylated probes in cryostat and paraffin sections.原位杂交：在冷冻切片机切片和石蜡切片中对生物素化探针进行碱性磷酸酶显色。
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Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.生物素标记多核苷酸的酶促合成：新型核酸亲和探针。
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