分析化学2012，Vol.40Issue(9)：1347-1352,6.DOI:10.3724/SP.J.1096.2012.11253
细交链孢菌酮酸酶联免疫吸附分析方法研究
Development of an Enzyme-linked Immunosorbent Assay Method for Detection of Tenuazonic Acid
摘要
Abstract
To develop an immunoassay method for tenuazonic acid (TeA), two haptens, 2-(2-(l-(5-(sec-butyl)-2-oxo-2,5-dihydro-lH-pyrrol-3-yl)ethylidene)-hydrazinyl)acetic acid (TeAHGA) and 5-(sec-butyl)-3-(l-hydrazonoethyl-4-hydroxy-lH-pyrrol-2 (5H)-one (TeAH) derived from TeA by hydrazine hydrate and glyoxylic acid were synthesized. Then TeAHGA-BSA conjugate was used as immunogen and the specific polyclonaL antibody against TeAH was prepared. Furthermore, an indirect competitive ELISA (icELISA) method for TeA with TeAH as target analyte was established. The optimized assay conditions were 0. 156 μg/L of heterologous coating antigen (TeAH-OVA, ovalbumin), with PBS as assay buffer, the incubation times for the primary antibody and the secondary antibody were 40 and 20 min, respectively. The icELISA results showed IC50 value, limit of detection (LOD) and linear range as 1. 61 ng/mL, 0. 08 μg/L and 0. 19 - 12. 89 μg/L, respectively. The average recovery rates for standard addition of TeA from tomato and flour samples were 67. 2% -89.8% and 74. 8%-93. 7%, respectively.关键词
细交链孢菌酮酸/异源包被/半抗原/多克隆抗体/免疫分析Key words
Tenuazonic acid/ Heterologous coating/ Hapten/ Polyclonal antibody/ Immunoassay引用本文复制引用
杨星星,刘细霞,王弘,徐振林,沈玉栋,孙远明..细交链孢菌酮酸酶联免疫吸附分析方法研究[J].分析化学,2012,40(9):1347-1352,6.基金项目
本文系国家自然科学基金(Nos.30871755,30901005)资助项目 （Nos.30871755,30901005）
