Anusz K Z, Mason P H, Riggs M W, Perryman L E
Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040.
J Clin Microbiol. 1990 Dec;28(12):2770-4. doi: 10.1128/jcm.28.12.2770-2774.1990.
A monoclonal antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect Cryptosporidium parvum oocysts in bovine feces. Fecal oocysts were concentrated by centrifugation through Formalin-ethyl acetate solution and captured with monoclonal antibody 18.280.2 reactive with C. parvum oocysts. Captured oocysts were detected with goat anti-oocyst serum, following the addition of a peroxidase conjugate of rabbit anti-goat immunoglobulin and O-phenylenediamine substrate. The assay was specific for Cryptosporidium sp. oocysts and did not detect oocysts of Eimeria auburnensis, Eimeria bovis, Eimeria ellipsoidalis, or Eimeria zuernii. Assay sensitivity allowed detection of 3 x 10(5) oocysts per ml of feces, compared with 1 x 10(6) oocysts per ml detected by examination of acid-fast-stained fecal smears and 1 x 10(3) oocysts per ml detected by indirect immunofluorescence. The capture ELISA was suitable for diagnostic analysis of bovine fecal samples and for assessment of oocyst shedding in experimentally infected calves. This assay may also prove useful for diagnostic assessment of humans in which cryptosporidiosis is suspected.
已开发出一种单克隆抗体酶联免疫吸附测定法（ELISA），用于检测牛粪中的微小隐孢子虫卵囊。粪便中的卵囊通过福尔马林 - 乙酸乙酯溶液离心浓缩，并用与微小隐孢子虫卵囊反应的单克隆抗体18.280.2捕获。在加入兔抗山羊免疫球蛋白的过氧化物酶结合物和邻苯二胺底物后，用山羊抗卵囊血清检测捕获的卵囊。该测定法对隐孢子虫属卵囊具有特异性，未检测到奥氏艾美耳球虫、牛艾美耳球虫、椭圆艾美耳球虫或邱氏艾美耳球虫的卵囊。该测定法的灵敏度可检测到每毫升粪便中3×10⁵个卵囊，相比之下，通过抗酸染色粪便涂片检查每毫升可检测到1×10⁶个卵囊，通过间接免疫荧光法每毫升可检测到1×10³个卵囊。捕获ELISA适用于牛粪样本的诊断分析以及对实验感染小牛的卵囊排出情况进行评估。该测定法可能对疑似隐孢子虫病的人类诊断评估也有用。
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