Rossi C L, Tsang V C, Pilcher J B
Department of Clinical Pathology, Faculty of Medical Sciences, State University of Campinas, São Paulo, Brazil.
J Clin Microbiol. 1991 Sep;29(9):1836-41. doi: 10.1128/jcm.29.9.1836-1841.1991.
Simple and rapid polystyrene- and nitrocellulose-based enzyme-linked immunosorbent assays were developed for detecting antibodies against adult Schistosoma mansoni microsomal antigens. The polystyrene test uses the Nunc Immuno Stick System. A single dilution of the antibody source being tested, the conjugate, and the substrate (3,3',5,5'-tetramethylbenzidine) are placed in tubes. Dried, antigen-coated polystyrene sticks are then exposed to the reagents by immersion. Once the sticks are sensitized, an entire assay can be completed in 8 min. Positive reactions result in a rich blue color in the substrate tube and can be distinguished with the naked eye. In the nitrocellulose-based test, a nitrocellulose sheet with antigen drawn in a line by pen is cut to produce identical strips. The ligand-binding steps and washings are performed in the troughs of incubation trays. The exposure times required for a single dilution of the antibody source being tested, the conjugate, and the substrate (3,3'-diaminobenzidine) are 5 min, 5 min, and 7.5 min, respectively. Once sensitized strips are available, an entire assay can be run in 50 min. Both techniques can assay serum or whole blood. The characteristics of polystyrene- and nitrocellulose-based techniques allow them to be used successfully in field studies and in minimally equipped laboratories.
已开发出基于聚苯乙烯和硝酸纤维素的简单快速酶联免疫吸附测定法，用于检测抗曼氏血吸虫成虫微粒体抗原的抗体。聚苯乙烯检测使用Nunc免疫棒系统。将单一稀释度的待测抗体源、结合物和底物（3,3',5,5'-四甲基联苯胺）置于试管中。然后将干燥的、包被有抗原的聚苯乙烯棒浸入试剂中。一旦棒被致敏，整个检测可在8分钟内完成。阳性反应会使底物管呈现浓郁的蓝色，肉眼即可辨别。在基于硝酸纤维素的检测中，用钢笔在硝酸纤维素膜上画一条线作为抗原，然后将其切割成相同的条带。配体结合步骤和洗涤在孵育托盘的槽中进行。单一稀释度的待测抗体源、结合物和底物（3,3'-二氨基联苯胺）所需的暴露时间分别为5分钟、5分钟和7.5分钟。一旦有了致敏条带，整个检测可在50分钟内完成。这两种技术均可检测血清或全血。基于聚苯乙烯和硝酸纤维素的技术特点使其能够成功应用于现场研究和设备简陋的实验室。
