Sobsey M D, Wallis C, Melnick J L
Appl Microbiol. 1975 Jan;29(1):21-6. doi: 10.1128/am.29.1.21-26.1975.
The development of a simple method for concentrating enteroviruses from oysters is described. In this method viruses in homogenized oyster tissues are efficiently absorbed to oyster solids at pH 5.5 and low salt concentration. After low-speed centrifugation, the supernatant is discarded and viruses are eluted from the sedimented oyster solids by resuspending them in pH 3.5 glycine-buffered saline. The solids are then removed by low-speed centrifugation, and the virus-containing supernatant is filtered through a 0.2-micronm porosity filter to remove bacteria and other small particulates without removing viruses. The virus-containing filtrate is then concentrated to a volume of a few milliliters by ultrafiltration, and the concentrate obtained is inoculated directly into cell cultures for virus assay. When tested with pools of oysters experimentally contaminated with small amounts of different enteroviruses, virus recovery efficiency averaged 63%.
本文描述了一种从牡蛎中浓缩肠道病毒的简单方法。在此方法中,匀浆后的牡蛎组织中的病毒在pH 5.5和低盐浓度下能有效地吸附到牡蛎固体上。低速离心后,弃去上清液,将沉淀的牡蛎固体重新悬浮于pH 3.5的甘氨酸缓冲盐溶液中,从中洗脱病毒。然后通过低速离心去除固体,含病毒的上清液通过0.2微米孔径的过滤器过滤以去除细菌和其他小颗粒,而不除去病毒。然后通过超滤将含病毒的滤液浓缩至几毫升体积,所得浓缩物直接接种到细胞培养物中进行病毒检测。当用实验性感染少量不同肠道病毒的牡蛎池进行测试时,病毒回收效率平均为63%。