Sevars A H, Schasfoort R B, Salden M H
Netherlands Organization for Applied Scientific Research (TNO).
Biosens Bioelectron. 1993;8(3-4):185-9. doi: 10.1016/0956-5663(93)85031-i.
In this paper the development of a surface plasmon resonance (SPR) immunosensor for syphilis screening is described. This immunosensor is based on the detection of antibodies in serum against the causative organism Treponema pallidum. In order to achieve selectivity a recombinant Treponema pallidum membrane protein A (TmpA) was used. This antigen can react with antibodies to T. pallidum, present in serum of syphilitic patients. Reproducible results have been obtained, using a 'sandwich SPR' method: binding of a sandwich antibody to the treponemal antibody after serum incubation was measured in real time while the binding was taking place. The SPR results obtained from ten blind-coded sera corresponded well with classical syphilis tests (Treponema pallidum haemagglutination assay (TPHA), fluorescent treponemal antibody-absorbed test (FTA-ABS), venereal diseases research laboratory flocculation test (VDRL) and TmpA-based enzyme-linked immunosorbent assay (TmpA-ELISA)). Preliminary experiments showed that direct measurement of serum (in the 'one step SPR') is not yet possible, probably as a result of non-uniformity of serum samples. The application of latex beads is considered to solve this problem.
本文描述了一种用于梅毒筛查的表面等离子体共振(SPR)免疫传感器的研制。这种免疫传感器基于检测血清中针对致病微生物梅毒螺旋体的抗体。为了实现选择性,使用了重组梅毒螺旋体膜蛋白A(TmpA)。这种抗原能与梅毒患者血清中存在的抗梅毒螺旋体抗体发生反应。采用“夹心SPR”方法获得了可重复的结果:在血清孵育后,实时测量夹心抗体与梅毒螺旋体抗体的结合情况。从10份盲编码血清获得的SPR结果与经典梅毒检测(梅毒螺旋体血凝试验(TPHA)、荧光梅毒螺旋体抗体吸收试验(FTA-ABS)、性病研究实验室絮凝试验(VDRL)和基于TmpA的酶联免疫吸附试验(TmpA-ELISA))结果吻合良好。初步实验表明,直接测量血清(“一步SPR”)目前还不可能,这可能是血清样本不均匀所致。考虑应用乳胶珠来解决这个问题。