Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
J Agric Food Chem. 2013 May 29;61(21):5031-6. doi: 10.1021/jf400803q. Epub 2013 May 15.
A lateral flow dual immunoassay (LFDIA) was developed for rapid quantitative detection of zearalenone (ZEN) and fumonisin B1 (FB1) in corn and wheat samples on a single test strip. Two test lines and the control line on the nitrocellulose membrane were coated with ZEN and FB1 conjugates and goat anti-mouse IgG, respectively. Colloidal gold nanoparticles were conjugated with monoclonal antibodies against ZEN or FB1. The intensity of the test lines was analyzed by a photometric strip reader to determine the concentrations of ZEN and FB1 based on the calibration curves of known concentrations versus intensity readings. Test parameters such as types of buffers, ratio of the two gold-labeled antibodies, and dilution of the sample extracts and the gold-labeled antibodies were optimized. The detection limit was 0.35 and 5.23 ng/mL for ZEN and FB1, respectively, and the corresponding detection ranges were 0.94-7.52 and 9.34-100.45 ng/mL, respectively. Spiked and natural samples were analyzed using both LFDIA and liquid chromatography-tandem mass spectrometry. The two methods had a good correlation (R(2) = 0.96). The dual quantitative LFDIA is sensitive, rapid, and easy-to-use for on-site testing of a large number of samples.
开发了一种侧向流双免疫测定法（LFDIA），用于在单个测试条上快速定量检测玉米和小麦样品中的玉米赤霉烯酮（ZEN）和伏马菌素 B1（FB1）。硝酸纤维素膜上的两条测试线和控制线分别包被 ZEN 和 FB1 缀合物以及山羊抗小鼠 IgG。胶体金纳米颗粒与针对 ZEN 或 FB1 的单克隆抗体缀合。通过光度计条读数器分析测试线的强度，根据已知浓度与强度读数的校准曲线确定 ZEN 和 FB1 的浓度。优化了测试参数，如缓冲液类型、两种金标记抗体的比例以及样品提取物和金标记抗体的稀释度。ZEN 和 FB1 的检测限分别为 0.35 和 5.23 ng/mL，相应的检测范围分别为 0.94-7.52 和 9.34-100.45 ng/mL。使用 LFDIA 和液相色谱-串联质谱法分析了加标和天然样品。两种方法相关性良好（R(2) = 0.96）。双定量 LFDIA 灵敏、快速，易于现场测试大量样品。
