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使用不同纯化方法对包虫囊肿抗原的血清学识别

Serologic recognition of hydatid cyst antigens using different purification methods.

作者信息

Sbihi Y, Janssen D, Osuna A

机构信息

Instituto de Biotecnología, Universidad de Granada, Spain.

出版信息

Diagn Microbiol Infect Dis. 1996 Apr;24(4):205-11. doi: 10.1016/0732-8893(96)00061-2.

Abstract

The specificity and sensitivity of enzyme-linked immunosorbent assays (ELISA) and Western immunoblot assays in detecting antibodies in serum from patients suffering cystic hydatid disease (Echinococcus granulosus) are compared using either crude antigen preparations (total sheep hydatid fluid and homogenates of protoscoleces), purified fractions enriched in Antigens 5 and B, and glycoproteins from hydatid fluid. Polyprotein bands of 12-14, 20, and 34 kDa, when purified from hydatid fluid by applying changes in the ionic strength, yielded a sensitive (95%) immunodiagnostic test that was also extremely specific (100%) when assayed with sera from noninfected humans and from patients suffering from other parasitic diseases. However, subjecting hydatid fluid to chromatography through a concanavilin A column rendered a 42 kDa band that was sensitive (95%) as well as highly specific (100%) for hydatidosis. Therefore purification procedures can strongly affect the diagnostic value of antigens with identical electrophoretic behavior in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

摘要

使用粗抗原制剂(全羊包虫液和原头蚴匀浆)、富含抗原5和B的纯化组分以及包虫液糖蛋白,比较酶联免疫吸附测定(ELISA)和Western免疫印迹测定检测囊性包虫病(细粒棘球绦虫)患者血清中抗体的特异性和敏感性。通过改变离子强度从包虫液中纯化出的12 - 14 kDa、20 kDa和34 kDa的多蛋白条带,产生了一种敏感的(95%)免疫诊断试验,当用未感染人类和其他寄生虫病患者的血清进行检测时,其特异性也极高(100%)。然而,将包虫液通过伴刀豆球蛋白A柱进行层析,得到了一条42 kDa的条带,该条带对包虫病敏感(95%)且特异性高(100%)。因此,纯化程序会强烈影响在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)中具有相同电泳行为的抗原的诊断价值。

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