一种用于检测抗溶组织内阿米巴抗体的斑点酶联免疫吸附测定法。
A dot enzyme-linked immunosorbent assay for detection of antibodies against Entamoeba histolytica.
作者信息
Kumar S, Band A H, Samantaray J C, Dang N, Talwar G P
出版信息
J Immunol Methods. 1985 Oct 24;83(1):125-33. doi: 10.1016/0022-1759(85)90065-1.
A visual micro-dot enzyme-linked immunosorbent assay (ELISA) based on detection of antibodies against soluble antigens of axenically grown cultures of Entamoeba histolytica is described. The antigen was spotted on a nitrocellulose sheet, the unsaturated sites blocked with bovine serum albumin (BSA) and incubated with 3-fold dilutions of patient sera followed by incubation with protein A conjugated to peroxidase. Enzymic activity was evidenced using the substrate 4-chloro-1-naphthol. A positive reaction produced a blue spot. The sensitivity of the assay was better and comparable to the indirect haemagglutination assay (IHA) and plate ELISA. The entire assay could be completed within 3 h. Antigen-loaded and pre-blocked nitrocellulose strips could be stored up to 3 months at room temperature and 37 degrees C.
描述了一种基于检测针对无菌培养的溶组织内阿米巴可溶性抗原抗体的视觉微点酶联免疫吸附测定（ELISA）。将抗原点样在硝酸纤维素膜上，用牛血清白蛋白（BSA）封闭未饱和位点，与患者血清的3倍稀释液孵育，随后与缀合过氧化物酶的蛋白A孵育。使用底物4-氯-1-萘酚证明酶活性。阳性反应产生蓝色斑点。该测定的灵敏度较好，与间接血凝试验（IHA）和平板ELISA相当。整个测定可在3小时内完成。加载抗原并预先封闭的硝酸纤维素条在室温及37℃下可保存长达3个月。
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