Notermans S, Wernars K
Laboratory for Water- and Food Microbiology, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
Int J Food Microbiol. 1991 Jan;12(1):91-102. doi: 10.1016/0168-1605(91)90050-y.
Improved methods to detect microorganisms and their toxins introduced during the last decade involve among others recombinant DNA techniques and various immuno-assays such as the enzyme-linked immunosorbent assay and the latex agglutination. Immuno-assays are based on a quantitative reaction of an antigen (bacterial metabolite, e.g., toxin) with its antibody. Therefore, they are suited for detection of microorganisms based on their production of specific antigens and for quantitative detection of bacterial toxins. Sensitivity and specificity of immuno-assays are mainly determined by the antiserum used. In this respect the use of well selected monoclonal antibodies can be of advantage. With the enzyme-linked immunosorbent assay and latex agglutination test quantities of 0.1-1 ng of antigen/ml can be detected. Of both techniques the latex agglutination method has several advantages; the method is simple, inexpensive and rapid. Since each immuno-assay is sensitive to non-specific reactions, recognition of false positive results is necessary. The most appropriate method for this is to add an inhibitor to the test sample which blocks specifically the paratope of the immunoglobulin. Another general disadvantage of immuno-assays is that only the antigenicity is determined and this may differ from the actual toxicity. Therefore, antibodies should be used that react with the toxic centre(s) of the molecule, which can be accomplished by using well selected monoclonal antibodies.
过去十年间出现的用于检测微生物及其毒素的改进方法包括重组DNA技术以及各种免疫测定法,如酶联免疫吸附测定法和乳胶凝集试验等。免疫测定法基于抗原(细菌代谢产物,如毒素)与其抗体的定量反应。因此,它们适用于基于微生物产生特定抗原的检测以及细菌毒素的定量检测。免疫测定法的灵敏度和特异性主要取决于所使用的抗血清。在这方面,使用精心挑选的单克隆抗体可能具有优势。通过酶联免疫吸附测定法和乳胶凝集试验,可以检测到0.1 - 1纳克/毫升的抗原量。乳胶凝集法在这两种技术中具有几个优点;该方法简单、廉价且快速。由于每种免疫测定法都对非特异性反应敏感,因此识别假阳性结果很有必要。最合适的方法是向测试样品中添加一种抑制剂,该抑制剂能特异性地阻断免疫球蛋白的互补位。免疫测定法的另一个普遍缺点是仅测定了抗原性,而这可能与实际毒性不同。因此,应使用与分子毒性中心反应良好的抗体,这可以通过使用精心挑选的单克隆抗体来实现。