Gallimore C I, Appleton H, Lewis D, Green J, Brown D W
Enteric and Respiratory Virus Laboratory, Virus Reference Division, London, United Kingdom.
J Med Virol. 1995 Feb;45(2):135-40. doi: 10.1002/jmv.1890450204.
The prevalence of picobirnaviruses (PBVs) in human stools was investigated by polyacrylamide gel electrophoresis (PAGE) analysis of 832 faecal specimens collected between 1982 and 1993 from patients in various clinical groups. Similar prevalences (9-13%) were detected in patients with or without gastroenteritis and throughout the age range of 3 to > 65 years. Two methods for the extraction of nucleic acid, a phenol/chloroform method and a guanidinium thiocynate (GTC)/silica method, were compared. Detection of PBVs by PAGE was three times more sensitive following RNA extraction by the GTC/silica method. Characterisation of three strains was carried out. Segment sizes ranged from 1.625 to 1.95 kilo base pairs (Kbp) and 2.2 to 2.5 Kbp for the fast and slow migrating bands, respectively. The nuclic acid was shown to be double-stranded RNA (dsRNA) by nuclease digestion. PBV-like particles were detected by electron microscopy in two PAGE-positive stools. Virion diameters ranged from 35 to 41 nm and a buoyant density of 1.38-1.4 g/ml in caesium chloride (CsCl) was demonstrated. These findings suggest that PBVs are widespread in humans in the United Kingdom. However, no disease association could be demonstrated.
采用聚丙烯酰胺凝胶电泳（PAGE）分析法，对1982年至1993年间从不同临床组患者中收集的832份粪便标本进行检测，以调查人粪便中微小双股RNA病毒（PBV）的流行情况。在患有或未患肠胃炎的患者以及3岁至65岁以上的所有年龄段患者中，均检测到相似的流行率（9%-13%）。比较了两种核酸提取方法，即酚/氯仿法和异硫氰酸胍（GTC）/硅胶法。采用GTC/硅胶法提取RNA后，通过PAGE检测PBV的灵敏度提高了三倍。对三株病毒进行了特性鉴定。快速迁移带和慢速迁移带的片段大小分别为1.625至1.95千碱基对（Kbp）和2.2至2.5 Kbp。通过核酸酶消化证明核酸为双链RNA（dsRNA）。在两份PAGE阳性粪便中通过电子显微镜检测到PBV样颗粒。病毒体直径范围为35至41 nm，在氯化铯（CsCl）中的浮力密度为1.38-1.4 g/ml。这些发现表明，PBV在英国人群中广泛存在。然而，未发现其与疾病的关联。
