Obert G, Gloeckler R, Burckard J, van Regenmortel M H
J Virol Methods. 1981 Sep;3(2):99-107. doi: 10.1016/0166-0934(81)90006-9.
The detection of human rotaviruses by routine electron microscopy examination of stool specimens has been compared with the sensitivity of detection obtainable by three different immunoassays. These assays are: 1) immunosorbent electron microscopy (ISEM), which consists of the serological trapping of viruses on electron microscopy grids coated with protein A and specific viral antiserum; 2) an enzyme-linked immunosorbent assay (ELISA), in which the primary antibody is rabbit anti-rotavirus immunoglobulin, the secondary antibody is chicken anti-rotavirus immunoglobulin extracted from egg yolk of immunized hens, and the indicator antibody is alkaline phosphatase-conjugated rabbit anti-chicken immunoglobulin; 3) counterimmunoelectrophoresis (CIE). A total of 63 stool specimens from infants with gastroenteritis were examined. Of these, 23 and 24 specimens were found to contain rotavirus by electron microscopy and CIE, respectively. When scored by ELISA and ISEM, 37 and 39 were found to be positive, respectively. Confirmatory inhibition assays were necessary to eliminate some false positive reactions in ELISA. Detection of human rotaviruses in stools by ISEM is as sensitive as by ELISA, but in weakly positive specimens, ISEM offers the additional advantage of a direct visual demonstration of the presence of the aetiological agent.
通过对粪便标本进行常规电子显微镜检查来检测人类轮状病毒，并与三种不同免疫测定法的检测灵敏度进行了比较。这些测定法分别是：1）免疫吸附电子显微镜法（ISEM），该方法是在涂有蛋白A和特异性病毒抗血清的电子显微镜网格上通过血清学方法捕获病毒；2）酶联免疫吸附测定法（ELISA），其中一抗是兔抗轮状病毒免疫球蛋白，二抗是从免疫母鸡的蛋黄中提取的鸡抗轮状病毒免疫球蛋白，指示抗体是碱性磷酸酶偶联的兔抗鸡免疫球蛋白；3）对流免疫电泳法（CIE）。总共检查了63份来自患肠胃炎婴儿的粪便标本。其中，通过电子显微镜检查和CIE分别发现23份和24份标本含有轮状病毒。用ELISA和ISEM评分时，分别发现37份和39份为阳性。需要进行确证抑制试验以消除ELISA中的一些假阳性反应。通过ISEM检测粪便中的人类轮状病毒与ELISA一样灵敏，但在弱阳性标本中，ISEM具有直接直观显示病原体存在的额外优势。
