Gow J, Cash P, Behan W, McGarry F, Simpson K, Behan P
Department of Neurology, University of Glasgow.
Electrophoresis. 1995 Mar;16(3):338-40. doi: 10.1002/elps.1150160156.
Attempts to evaluate the relative levels of enteroviral genomic and template RNA strands in small biopsy tissue samples from patients have yielded ambiguous data, largely due to the limited amount of RNA available. A novel semi-nested polymerase chain reaction (PCR) technique was developed to enable RNA levels to be examined more accurately. PCR products were visualised by horizontal agarose gel electrophoresis. This technique was demonstrated on linear single-stranded plasmid DNA; viral RNA isolated from a human rhabdomyosarcoma (RD) cell line persistently infected with a mutated coxsackie B5 virus (piRD) and two cell lines, RD and HEp2 cells, acutely infected with a wild-type clinical isolate of coxsackie B5 virus.
评估患者小活检组织样本中肠道病毒基因组和模板RNA链的相对水平的尝试产生了不明确的数据，这主要是由于可用RNA的量有限。开发了一种新型半巢式聚合酶链反应（PCR）技术，以使RNA水平能够得到更准确的检测。PCR产物通过水平琼脂糖凝胶电泳进行可视化。该技术在线性单链质粒DNA上得到了验证；从持续感染突变柯萨奇B5病毒的人横纹肌肉瘤（RD）细胞系（piRD）以及急性感染柯萨奇B5病毒野生型临床分离株的两个细胞系RD和HEp2细胞中分离出的病毒RNA也验证了该技术。
