Andrews W D, Al-Chalabi A, Garson J A
Department of Virology, University College London Medical School, UK.
J Neurol Sci. 1997 Dec 9;153(1):86-90. doi: 10.1016/s0022-510x(97)00199-8.
It has recently been claimed (Ferrante et al., 1995. HTLV tax-rex DNA and antibodies in idiopathic amyotrophic lateral sclerosis. J. Neurol. Sci. 129 (Suppl.) 140-144) that human T-lymphotropic virus (HTLV) tax-rex sequences are detectable in the peripheral blood mononuclear cells (PBMCs) of 40% of patients with motor neurone disease (MND). In an attempt to confirm this we employed a highly sensitive 'nested' polymerase chain reaction (PCR) assay, capable of detecting single molecules of HTLV proviral DNA, to look for tax-rex sequences in the PBMCs of 43 patients with MND. We were unable to detect the presence of HTLV tax-rex in any of 43 MND patients tested, using three different PCR primer sets under both high and low stringency conditions. Using the same DNA samples we were able to detect the presence of the single-copy pyruvate dehydrogenase gene, thus demonstrating that the extracted DNA was indeed amplifiable by PCR. To further exclude the possibility that the extracted DNA samples contained unrecognised inhibitory factors we conducted spiking experiments with trace amounts (approximately 10 copies) of HTLV proviral DNA. Spiked samples yielded PCR products of the expected size. We are therefore unable to confirm the presence of HTLV tax-rex sequences in this disease.
最近有人声称（费兰特等人，1995年。《特发性肌萎缩侧索硬化症中的人类嗜T淋巴细胞病毒tax-rex DNA和抗体》。《神经科学杂志》129（增刊）140 - 144页），在40%的运动神经元病（MND）患者的外周血单个核细胞（PBMC）中可检测到人类嗜T淋巴细胞病毒（HTLV）的tax-rex序列。为了证实这一点，我们采用了一种高度灵敏的“巢式”聚合酶链反应（PCR）检测方法，该方法能够检测HTLV前病毒DNA的单分子，以在43例MND患者的PBMC中寻找tax-rex序列。在高严谨度和低严谨度条件下，使用三种不同的PCR引物对，我们在检测的43例MND患者中均未能检测到HTLV tax-rex的存在。使用相同的DNA样本，我们能够检测到单拷贝的丙酮酸脱氢酶基因的存在，从而证明提取的DNA确实可通过PCR扩增。为了进一步排除提取的DNA样本中含有未识别的抑制因子的可能性，我们用痕量（约10个拷贝）的HTLV前病毒DNA进行了加标实验。加标样本产生了预期大小的PCR产物。因此，我们无法证实在这种疾病中存在HTLV tax-rex序列。
