Bloch P, Simonsen P E
Danish Bilharziasis Laboratory, Charlottenlund.
Acta Trop. 1998 Jun 15;70(1):73-86. doi: 10.1016/s0001-706x(98)00019-9.
To explore the possibility for development of an immunodiagnostic test for Dracunculus medinensis infections, the antibody responses (IgG1, IgG2, IgG3, IgG4 and IgE) to antigen preparations made from adult female worms (ADGW) and first stage larvae (LVGW) of D. medinensis were analysed. By sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting, sera from individuals with patent D. medinensis infection reacted extensively and similarly with ADGW and LVGW over a broad molecular weight range (5-200 kDa). Sera from individuals infected with Onchocerca volvulus (the major cross-reacting infection) had a more pronounced reaction with ADGW than LVGW, and only with antigens of molecular weights above 23 kDa. These findings were used to design an enzyme-linked immunosorbent assay (ELISA) with high sensitivity and specificity for D. medinensis infection. The most promising results were obtained when detecting for specific IgG4. Thus, when using LVGW, the assay had a sensitivity of 83% and a specificity of 97%. Refining the antigen into a low molecular weight filtrate improved the sensitivity to 92%, and the sensitivity was further improved (to 96%) when combining the results from two or more antibody types measured simultaneously, without affecting the specificity. The results were encouraging for the prospects for developing an applicable immunodiagnostic test for patent D. medinensis infections based on detection of specific serum antibodies.
为探索开发麦地那龙线虫感染免疫诊断检测方法的可能性，分析了针对麦地那龙线虫成年雌虫（ADGW）和第一期幼虫（LVGW）制备的抗原制剂的抗体反应（IgG1、IgG2、IgG3、IgG4和IgE）。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）/蛋白质印迹法，麦地那龙线虫显性感染个体的血清在较宽分子量范围（5-200 kDa）内与ADGW和LVGW发生广泛且相似的反应。感染盘尾丝虫（主要交叉反应感染）个体的血清与ADGW的反应比与LVGW更明显，且仅与分子量高于23 kDa的抗原发生反应。这些发现被用于设计一种对麦地那龙线虫感染具有高灵敏度和特异性的酶联免疫吸附测定（ELISA）。检测特异性IgG4时获得了最有前景的结果。因此，使用LVGW时，该检测方法的灵敏度为83%，特异性为97%。将抗原提纯为低分子量滤液可将灵敏度提高到92%，同时检测两种或更多种抗体类型的结果时，灵敏度进一步提高（至96%），且不影响特异性。这些结果对于基于检测特异性血清抗体开发适用于麦地那龙线虫显性感染的免疫诊断检测方法的前景而言令人鼓舞。
