使用生物素化单克隆抗体通过酶免疫测定法检测血吸虫循环阴极抗原。

Detection of the schistosome circulating cathodic antigen by enzyme immunoassay using biotinylated monoclonal antibodies.

作者信息

de Jonge N, Kremsner P G, Krijger F W, Schommer G, Fillié Y E, Kornelis D, van Zeyl R J, van Dam G J, Feldmeier H, Deelder A M

机构信息

Laboratory of Parasitology, Medical Faculty, University of Leiden, The Netherlands.

出版信息

Trans R Soc Trop Med Hyg. 1990 Nov-Dec;84(6):815-8. doi: 10.1016/0035-9203(90)90094-u.

DOI:10.1016/0035-9203(90)90094-u

PMID:2128984

Abstract

We have developed an enzyme immunoassay (ELISA) for the quantification of the schistosome circulating cathodic antigen (CCA), a glycoprotein associated with the syncitium lining the gut of the parasite. A mouse monoclonal antibody of IgG3 isotype was used as coating (antigen-capture) antibody, while a biotinylated mouse monoclonal IgM was used as second (antigen-detecting) antibody. Streptavidin-alkaline phosphatase was used as enzyme label. The lower detection limit of the assay was 1.0 ng of the trichloroacetic acid soluble fraction of adult worm antigen (AWA-TCA) per ml, which corresponds to approximately 0.2 ng CCA per ml. The ELISA showed a linear range from 1.0 to 62.5 ng AWA-TCA per ml. Serum and urine samples of 16 individuals infected with Schistosoma mansoni (egg counts ranging from 5 to 4820 eggs per gram of faeces) were tested in the assay. Antigen titres ranged from less than 4-8192. This assay represents a considerable advantage in diagnosis of Schistosoma infections as it allows the detection and quantification of CCA in serum and urine in even lightly infected individuals.

摘要

我们开发了一种酶免疫测定法（ELISA），用于定量检测血吸虫循环阴极抗原（CCA），这是一种与寄生虫肠道内衬合胞体相关的糖蛋白。使用IgG3同种型的小鼠单克隆抗体作为包被（抗原捕获）抗体，而生物素化的小鼠单克隆IgM用作第二（抗原检测）抗体。链霉亲和素-碱性磷酸酶用作酶标记。该测定法的最低检测限为每毫升1.0纳克成虫抗原的三氯乙酸可溶部分（AWA-TCA），这相当于每毫升约0.2纳克CCA。ELISA显示每毫升AWA-TCA的线性范围为1.0至62.5纳克。对16名感染曼氏血吸虫的个体（每克粪便中的虫卵计数范围为5至4820个虫卵）的血清和尿液样本进行了该测定法检测。抗原滴度范围为小于4至8192。该测定法在血吸虫感染诊断中具有相当大的优势，因为它能够检测和定量即使是轻度感染个体血清和尿液中的CCA。