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一种用于定量念珠菌对人血管内皮细胞黏附的酶联免疫吸附测定法。

An enzyme-linked immunosorbent assay for quantifying adherence of Candida to human vascular endothelium.

作者信息

Filler S G, Der L C, Mayer C L, Christenson P D, Edwards J E

出版信息

J Infect Dis. 1987 Oct;156(4):561-6. doi: 10.1093/infdis/156.4.561.

Abstract

Success in elucidating the pathogenesis of certain bacterial infections through studies of bacterial adherence to host cells has stimulated interest in parallel investigations of fungal adherence. Fungal adherence differs from bacterial adherence, especially when fungal coadherence (adherence of fungal cells to each other) is a factor. Using human umbilical vein endothelial cells cultured in a living monolayer in microtiter plates, we developed an ELISA to study adherence of Candida albicans to endothelial cells in the absence of yeast coadherence. A rabbit antibody to Candida detected the adherent Candida, and an alkaline phosphatase-conjugated antibody to rabbit IgG was the developing antibody. A linear relationship between the log of the optical density and the log of the number of adherent organisms was seen for wells containing 3 X 10(4)-1 X 10(6) organisms (r = .923- .965). In addition to measuring adherence of living Candida to living target cells and avoiding Candida coadherence, this assay makes it possible to investigate adherence limited to lumenal surfaces, conserves reagents, and facilitates the testing of large numbers of potential adherence modifiers.

摘要

通过研究细菌对宿主细胞的黏附作用来阐明某些细菌感染发病机制的成功,激发了人们对真菌黏附平行研究的兴趣。真菌黏附不同于细菌黏附,尤其是当真菌共黏附(真菌细胞彼此之间的黏附)是一个因素时。利用在微量滴定板中活单层培养的人脐静脉内皮细胞,我们开发了一种酶联免疫吸附测定法(ELISA),以研究白色念珠菌在无酵母共黏附情况下对内皮细胞的黏附。一种抗念珠菌的兔抗体检测黏附的念珠菌,一种碱性磷酸酶偶联的抗兔IgG抗体作为显色抗体。对于含有3×10⁴ - 1×10⁶个生物体的孔,光密度的对数与黏附生物体数量的对数之间呈线性关系(r = 0.923 - 0.965)。除了测量活念珠菌对活靶细胞的黏附并避免念珠菌共黏附外,该测定法还能够研究仅限于腔表面的黏附,节省试剂,并便于对大量潜在的黏附调节剂进行测试。

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