Ramia S, Sattar S A
Appl Environ Microbiol. 1980 Mar;39(3):493-9. doi: 10.1128/aem.39.3.493-499.1980.
There is mounting evidence for the waterborne transmission of diarrhea caused by rotaviruses. As a result, proper techniques are required for their recovery from samples of incriminated water. The combined efficiency of the talc-Celite technique and polyethylene glycol 6000 hydroextraction was, therefore, tested for this purpose, using Simian rotavirus SA-11 and MA-104 cells. Conditioning of the dechlorinated tap water samples was carried out by pH adjustment to 6.0 and the addition of Earle balanced salt solution to a final concentration of 1:100. Passage of a 1-liter volume of such a conditioned sample through a layer containing a mixture of talc (300 mg) and Celite 503 (100 mg) led to the adsorption of nearly 93% of the added SA-11 plaque-forming units. For the recovery of the layer-adsorbed virus, 3% beef extract and 1x tryptose phosphate broth were found to be superior to a variety of other eluents tested. When we tested 100-liter sample volumes, layers containing 1.2 g of talc and 0.4 g of Celite were employed. Virus elution was carried out with 100 ml of tryptose phosphate broth. The eluate was concentrated 10-fold by overnight (4 degrees C) hydroextraction with polyethylene glycol. With a total input virus of 7.0 x 10(5) and 1.4 x 10(2) plaque-forming units, the recoveries were about 71 and 59%, respectively.
有越来越多的证据表明轮状病毒引起的腹泻可通过水传播。因此,需要适当的技术从受污染的水样中分离出病毒。为此,使用猿猴轮状病毒SA - 11和MA - 104细胞,测试了滑石粉 - 硅藻土技术和聚乙二醇6000水相萃取法的联合效率。对脱氯自来水样品进行预处理,将pH值调至6.0,并加入Earle平衡盐溶液,使其终浓度达到1:100。使1升这样预处理过的样品通过一层含有滑石粉(300毫克)和硅藻土503(100毫克)混合物的滤层,导致近93%添加的SA - 11空斑形成单位被吸附。对于从滤层吸附的病毒的回收,发现3%牛肉浸出液和1倍胰蛋白胨磷酸盐肉汤优于所测试的其他多种洗脱剂。当我们测试100升样品体积时,使用含有1.2克滑石粉和0.4克硅藻土的滤层。用100毫升胰蛋白胨磷酸盐肉汤进行病毒洗脱。洗脱液通过在4℃下用聚乙二醇过夜水相萃取浓缩10倍。对于总输入病毒量分别为7.0×10⁵和1.4×10²空斑形成单位的情况,回收率分别约为71%和59%。