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斑点酶联免疫吸附测定法在免疫寄生虫学中的最新应用。

Recent applications of the Dot-ELISA in immunoparasitology.

作者信息

Pappas M G

机构信息

Covalent Technology Corporation, Ann Arbor, MI 48106.

出版信息

Vet Parasitol. 1988 Sep;29(2-3):105-29. doi: 10.1016/0304-4017(88)90120-3.

Abstract

The dot enzyme-linked immunosorbent assay (Dot-ELISA) is a highly versatile solid-phase immunoassay for antibody or antigen detection. The assay uses minute amounts of reagent dotted onto solid surfaces such as nitrocellulose and other paper membranes which avidly bind proteins. After incubation with antigen-specific antibody and enzyme-conjugated anti-antibody, the addition of a precipitable, chromogenic substrate causes the formation of a colored dot on the solid phase which is visually read. The Dot-ELISA has been used extensively in the detection of human and veterinary protozoan and metazoan parasitic diseases, including amebiasis, babesiosis, fascioliasis, cutaneous and visceral leishmaniasis, cysticercosis, echinococcosis, malaria, schistosomiasis, toxocariasis, toxoplasmosis, trichinosis, trypanosomiasis and even ixodid tick infestation. The technique is rapid, easy to perform and interpret, reagent conservative, cost effective and field portable. In addition, the Dot-ELISA may be configured to detect antibodies or parasite antigen in either microtiter plates for large-batch testing or with dipsticks for small numbers of determinations. A slight modification of the Dot-ELISA procedure allows the determination of infection rates of vectors such as ticks and sandflies with parasites.

摘要

斑点酶联免疫吸附测定(Dot-ELISA)是一种用于检测抗体或抗原的高度通用的固相免疫测定法。该测定法使用微量试剂点样于固相表面,如硝酸纤维素膜和其他能强烈结合蛋白质的纸质膜上。在用抗原特异性抗体和酶标记的抗抗体孵育后,加入可沉淀的显色底物会在固相上形成一个有色斑点,通过目视读取。Dot-ELISA已广泛用于检测人和兽医的原生动物和后生动物寄生虫病,包括阿米巴病、巴贝斯虫病、肝片吸虫病、皮肤和内脏利什曼病、囊尾蚴病、棘球蚴病、疟疾、血吸虫病、弓蛔虫病、弓形虫病、旋毛虫病、锥虫病,甚至硬蜱感染。该技术快速、易于操作和解读,试剂消耗少,成本效益高且便于携带到现场。此外,Dot-ELISA可配置用于在微量滴定板中检测抗体或寄生虫抗原以进行大批量检测,或使用试纸条进行少量测定。对Dot-ELISA程序稍作修改即可测定蜱和白蛉等病媒被寄生虫的感染率。

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