McCrae M A
Ciba Found Symp. 1987;128:24-48. doi: 10.1002/9780470513460.ch3.
Simple genome profile studies on polyacrylamide gels allow all non-group A rotaviruses isolated so far to be recognized by the absence of the tight triplet (7-9) of RNA segments seen in all group A viruses. However, reliance solely on genome profile studies for rotavirus grouping can be misleading and, for virus group definition, additional corroborating nucleic acid and serological studies are essential. Terminal fingerprint analysis was the first generation of nucleic acid-based assays that allowed discrimination between the various rotavirus groups. By means of this technique the clear definition of five rotavirus groups (A-E), correlating exactly with those found by a serological assay, has been possible, with preliminary evidence for at least two additional groups. The technical sophistication of fingerprinting techniques prevents their widespread use in epidemiological studies; the development of a second generation of nucleic acid-based assays is therefore under way. These employ molecularly cloned cDNA probes to the genomes of non-group A viruses which can be widely distributed for use in 'dot-blot' screening of faecal samples and, if expressed as protein in Escherichia coli, should provide a ready source of viral antigen for use in surveying viral prevalence through the screening of serum antibody levels.
在聚丙烯酰胺凝胶上进行的简单基因组图谱研究显示,所有迄今分离出的非A组轮状病毒,都可通过缺乏A组病毒中所见的紧密RNA片段三联体(7 - 9)来识别。然而,仅依靠基因组图谱研究对轮状病毒进行分组可能会产生误导,对于病毒组的定义,额外的核酸和血清学确证研究必不可少。末端指纹分析是第一代基于核酸的检测方法,可用于区分不同的轮状病毒组。借助这项技术,已经能够明确界定五个轮状病毒组(A - E),这与血清学检测结果完全相符,并且初步证据表明至少还有另外两个组。指纹分析技术的技术复杂性限制了其在流行病学研究中的广泛应用;因此,第二代基于核酸的检测方法正在研发中。这些方法使用针对非A组病毒基因组的分子克隆cDNA探针,这些探针可以广泛分发用于粪便样本的“斑点印迹”筛查,如果在大肠杆菌中表达为蛋白质,应该能为通过血清抗体水平筛查来调查病毒流行情况提供现成的病毒抗原来源。