Preliminary evaluation of the ELISA as a tool for the detection of rotaviruses in activated sewage sludge.

F Agbalika, M Wullenweber, J Prévot
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Abstract

Samples of activated sludge were taken in three different treatment plants near Nancy (France) and in West Berlin (F.R.G.), and were investigated for their content of rotaviruses. After elution with beef extract (0.25%, pH 7.3) and ultrasonication, the samples were concentrated by protamine flocculation and, in a few cases, in parallel by ultracentrifugation. A commercially available ELISA test kit (Rotazyme, Abbott Diagnostics) was routinely used for rotavirus detection. Although both the Rotazyme and a laboratory-designed ELISA gave positive results for the suldge concentrates, confirmation with a blocking test and electron microscope (EM) was negative in all cases so far investigated. Therefore, the exclusive use of ELISA cannot be recommended for similar investigations as it may lead to false positive results. In this light, an additional verification step with neutralizing antibodies should be included by the ELISA kits' manufacturers to enable a quick confirmation of the user's results.

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ELISA检测活性污泥中轮状病毒的初步评价。
在南希(法国)和西柏林(F.R.G.)附近的三个不同的处理厂采集了活性污泥样本,并对其轮状病毒含量进行了调查。样品经牛肉提取物(0.25%,pH 7.3)和超声波洗脱后,用鱼精蛋白絮凝浓缩,少数情况下用超离心平行浓缩。市售的ELISA检测试剂盒(Rotazyme,雅培诊断公司)常规用于轮状病毒检测。虽然Rotazyme和实验室设计的ELISA对污泥浓缩物的检测结果都是阳性的,但到目前为止,通过阻断试验和电子显微镜(EM)确认的所有病例都是阴性的。因此,不建议在类似的调查中单独使用ELISA,因为它可能导致假阳性结果。在这种情况下,ELISA试剂盒的制造商应该包括一个额外的中和抗体验证步骤,以便能够快速确认用户的结果。
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