使用原位聚合酶链反应和DNA杂交技术在石蜡包埋组织中鉴定登革病毒感染细胞。

Identification of dengue virus-infected cells in paraffin-embedded tissue using in situ polymerase chain reaction and DNA hybridization.

作者信息

Lucia H L, Kangwanpong D

机构信息

Department of Pathology, University of Mississippi Medical Center, Jackson 39216.

出版信息

J Virol Methods. 1994 Jun;48(1):1-8. doi: 10.1016/0166-0934(94)90083-3.

DOI:10.1016/0166-0934(94)90083-3

PMID:7525622

Abstract

Using tissue from suckling mice infected with Dengue virus, the following improved method for detecting Dengue viral RNA in tissue sections was devised. Reverse transcription of the viral RNA to DNA was followed by the in situ polymerase chain reaction to amplify the viral nucleic acid. This was followed by DNA hybridization with an alkaline phosphatase-labelled probe. The enzyme was then reacted with Fast red and counterstained with hematoxylin. Viral nucleic acid was readily demonstrated within glial cells and macrophages in brains of animals infected with two different serotypes of Dengue.

摘要

利用感染登革病毒的乳鼠组织，设计了以下用于检测组织切片中登革病毒RNA的改进方法。病毒RNA逆转录为DNA后，进行原位聚合酶链反应以扩增病毒核酸。接着用碱性磷酸酶标记的探针进行DNA杂交。然后该酶与固红反应，并用苏木精复染。在感染两种不同血清型登革病毒的动物大脑的神经胶质细胞和巨噬细胞内很容易检测到病毒核酸。