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在肌萎缩侧索硬化症患者及对照患者的神经组织中检测微小核糖核酸病毒序列。

Detection of picornavirus sequences in nervous tissue of amyotrophic lateral sclerosis and control patients.

作者信息

Brahic M, Smith R A, Gibbs C J, Garruto R M, Tourtellotte W W, Cash E

出版信息

Ann Neurol. 1985 Sep;18(3):337-43. doi: 10.1002/ana.410180311.

Abstract

We used in situ hybridization to look for picornavirus ribonucleic acid (RNA) sequences in frozen sections of central nervous system (CNS) tissue of amyotrophic lateral sclerosis (ALS) and control patients. Using reconstruction experiments, we concluded that 30 copies of viral RNA per cell could be detected with the assay. RNA which hybridized to DNAs complementary (cDNAs) to both poliovirus and Theiler's virus was found at several levels in the CNS of 2 patients, 1 ALS patient, and 1 control. In transverse sections of the spinal cord, these sequences predominated in cells of the anterior horns. We assessed the specificity of hybridization by several criteria: no hybridization was observed with heterologous visna virus cDNA probes; hybridization was abolished by pretreatment of the sections with ribonuclease; chemography artifacts were ruled out; and the results were reproduced in three independent experiments. We concluded that RNA molecules, possibly belonging to a picornavirus having sequences in common with poliovirus and Theiler's virus, were present in the CNS of these 2 patients. On the other hand, 14 cases of classic ALS, 2 cases of Guamanian parkinsonian dementia, and 5 controls had negative results. However, the presence of picornavirus sequences in our series could be underestimated because in many cases autolysis times were 10 hours or longer.

摘要

我们采用原位杂交技术,在肌萎缩侧索硬化症(ALS)患者及对照患者的中枢神经系统(CNS)组织冰冻切片中寻找微小核糖核酸病毒核糖核酸(RNA)序列。通过重建实验,我们得出该检测方法可检测到每个细胞中有30个病毒RNA拷贝。在2例患者(1例ALS患者和1例对照)的中枢神经系统多个层面发现了与脊髓灰质炎病毒和泰勒氏病毒的互补DNA(cDNA)杂交的RNA。在脊髓横切片中,这些序列在前角细胞中占主导地位。我们通过几个标准评估了杂交的特异性:用异源维斯纳病毒cDNA探针未观察到杂交;用核糖核酸酶预处理切片后杂交被消除;排除了化学成像假象;并且结果在三个独立实验中得到重现。我们得出结论,这2例患者的中枢神经系统中存在RNA分子,可能属于一种与脊髓灰质炎病毒和泰勒氏病毒有共同序列的微小核糖核酸病毒。另一方面,14例经典ALS患者、2例关岛帕金森痴呆患者及5例对照结果均为阴性。然而,我们研究系列中微小核糖核酸病毒序列的存在情况可能被低估了,因为在许多情况下自溶时间为10小时或更长。

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