Swanson N R, Fox S A, Mastaglia F L
University Department of Medicine, Queen Elizabeth II Medical Centre, Nedlands, Australia.
Neuromuscul Disord. 1995 Nov;5(6):457-65. doi: 10.1016/0960-8966(95)00002-5.
A longstanding hypothesis proposes that amyotrophic lateral sclerosis-motor neurone disease (ALS-MND) is a late consequence of subclinical poliovirus (PV) infection. In this study, RNA extracts of CNS tissue from 28 patients with ALS-MND and 7 controls were assayed by nested polymerase chain reaction (PCR) using primers to the 5'-untranslated region (UTR) of the enterovirus (EV) genome which is highly conserved between EVs including PV, echovirus and coxsackie viruses. The integrity of RNA extracted from either archival paraffin-embedded or frozen CNS tissue was assessed by detection of constitutive Ableson tyrosine kinase (ABL) mRNA by PCR. Of 63 tissue samples assayed, 81% (51/63) were ABL-positive corresponding to 78% (22/28) of the ALS-MND cases and all controls. None of the 27 ALS-MND cases (i.e. 21 ABL+ and 6 ABL-) in which paraffin-embedded tissue was used nor any of the age and sex matched controls were positive for specific PV/EV RNA. Moreover, CNS tissue from 14 different locations obtained from one patient < 2 hrs after death and immediately frozen, showed no evidence of PV/EV at any site by PCR. Disease duration, degree of tissue autolysis and duration of tissue storage were all excluded as factors which may predispose to negative results. The sensitivity of the PV PCR was determined to be 40-400 copies (12.5 - 125 ag) of synthetic EV RNA transcripts in 1 microgram of cellular RNA and the assay was shown to detect all types of PV and and other EVs tested. Thus it seems unlikely that a persistent PV or related EV infection is implicated in ALS-MND unless there has been alteration in the 5'-UTR of the virus genome.
一个长期存在的假说是，肌萎缩侧索硬化症-运动神经元病（ALS-MND）是亚临床脊髓灰质炎病毒（PV）感染的晚期后果。在本研究中，使用针对肠道病毒（EV）基因组5'-非翻译区（UTR）的引物，通过巢式聚合酶链反应（PCR）对28例ALS-MND患者和7例对照的中枢神经系统（CNS）组织RNA提取物进行检测，该区域在包括PV、埃可病毒和柯萨奇病毒在内的肠道病毒之间高度保守。通过PCR检测组成型阿贝尔森酪氨酸激酶（ABL）mRNA来评估从存档石蜡包埋或冷冻的中枢神经系统组织中提取的RNA的完整性。在所检测的63个组织样本中，81%（51/63）为ABL阳性，对应78%（22/28）的ALS-MND病例和所有对照。使用石蜡包埋组织的27例ALS-MND病例（即21例ABL阳性和6例ABL阴性）以及任何年龄和性别匹配的对照均未检测到特异性PV/EV RNA阳性。此外，在一名患者死亡后<2小时立即获取并立即冷冻的来自14个不同部位的中枢神经系统组织，通过PCR在任何部位均未显示PV/EV的迹象。疾病持续时间、组织自溶程度和组织储存时间均被排除为可能导致阴性结果的因素。PV PCR的灵敏度被确定为在1微克细胞RNA中可检测到40-400个合成EV RNA转录本拷贝（12.5-125阿克），并且该检测方法显示可检测所有类型的PV和所测试的其他EV。因此，除非病毒基因组的5'-UTR发生改变，否则持续性PV或相关EV感染似乎不太可能与ALS-MND有关。
