Blais Burton W, Leggate Johanna, Bosley Jessica, Martinez-Perez Amalia
Canadian Food Inspection Agency, Ottawa Laboratory (Carling), Building 22, Central Experimental Farm, Ottawa, Ontario, Canada.
J Food Prot. 2005 Feb;68(2):233-8. doi: 10.4315/0362-028x-68.2.233.
An enzyme-linked immunosorbent assay (ELISA) system was developed using polymyxin immobilized in the wells of a microtiter plate as a high-affinity adsorbent for Escherichia coli O157 lipopolysaccharide (LPS) antigens. Extracts from cell suspensions were reacted with polymyxin-coated microwells followed by immunoenzymatic detection of captured LPS antigens using a commercially available anti-E. coli O157 antibody-peroxidase conjugate and a 3,3',5,5'-tetramethylbenzidine substrate. The polymyxin ELISA was highly sensitive and specific for E. coli strains bearing the O157 antigen. When this ELISA was combined with enrichment, results were in complete agreement with those of standard culture techniques for the detection of this pathogen in a variety of artificially inoculated and naturally contaminated foods. The polymyxin ELISA is a simple and inexpensive assay for E. coli O157 with a 96-well microtiter plate format, making this system ideally suited for processing large numbers of samples.
开发了一种酶联免疫吸附测定（ELISA）系统，该系统使用固定在微量滴定板孔中的多粘菌素作为大肠杆菌O157脂多糖（LPS）抗原的高亲和力吸附剂。细胞悬液提取物与包被有多粘菌素的微孔板反应，然后使用市售的抗大肠杆菌O157抗体-过氧化物酶偶联物和3,3',5,5'-四甲基联苯胺底物对捕获的LPS抗原进行免疫酶检测。多粘菌素ELISA对携带O157抗原的大肠杆菌菌株具有高度敏感性和特异性。当该ELISA与富集相结合时，结果与标准培养技术在各种人工接种和自然污染食品中检测该病原体的结果完全一致。多粘菌素ELISA是一种使用96孔微量滴定板形式对大肠杆菌O157进行的简单且廉价的检测方法，使该系统非常适合处理大量样品。
