Sharer L R, Saito Y, Da Cunha A, Ung P C, Gelbard H A, Epstein L G, Blumberg B M
Department of Pathology, New Jersey Medical School, Newark 07103, USA.
Hum Pathol. 1996 Jun;27(6):614-7. doi: 10.1016/s0046-8177(96)90172-0.
To examine whether latent infection by HIV-1 occurs in the central nervous system, we optimized a procedure for amplification and detection of HIV-1 DNA in situ, in formalin-fixed brain tissue from a child with severe HIV-1-associated progressive encephalopathy and severe HIV-1 encephalitis. By the use of a two-step technique, which involved polymerase chain reaction with incorporation of digoxigenin-labeled nucleotides followed by in situ hybridization with biotinylated probes, we found infection of numerous mononuclear cells and astrocytes in the cerebral white matter as well as of perineuronal satellite cells in basal ganglia, but not of neurons. Following PCR amplification, nuclear signal was found in 10 to 20 times as many cells as in parallel, control experiments using conventional, unamplified in situ hybridization.
为了检测HIV-1潜伏感染是否发生在中枢神经系统，我们优化了一种程序，用于在一名患有严重HIV-1相关进行性脑病和严重HIV-1脑炎儿童的福尔马林固定脑组织中对HIV-1 DNA进行原位扩增和检测。通过使用两步技术，该技术包括掺入地高辛标记核苷酸的聚合酶链反应，随后用生物素化探针进行原位杂交，我们发现在脑白质中有大量单核细胞和星形胶质细胞受到感染，在基底神经节中有神经元周围卫星细胞受到感染，但神经元未受感染。PCR扩增后，与使用传统未扩增原位杂交的平行对照实验相比，发现有核信号的细胞数量多10到20倍。
