Key Laboratory of Food Quality and Safety of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou, 510642, China.
Key Laboratory of Food Quality and Safety of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou, 510642, China.
Toxicon. 2020 Feb;175:44-48. doi: 10.1016/j.toxicon.2019.12.003. Epub 2019 Dec 6.
A highly sensitive and broadly specific competitive indirect enzyme-linked immunosorbent assay (ciELISA) method was developed for the simultaneous detection of nine microcystins (MCs) and nodularin (NOD) using MC-LR-keyhole limpet hemocyanin (KLH) for New Zealand white rabbit immunization to produce antibodies. The MC-LR-bovine serum albumin (BSA) and NOD-BSA coating antigens were compared and heterogeneous coating strategy was found to significantly improve the sensitivity of detection, as evident from the appropriate structure. Comparison of the half-maximum inhibitory concentration (IC50) with MC-LR and MC-LR-BSA coating techniques (0.29 ng/mL) revealed the superior performance of 0.054 ng/mL for NOD-BSA coating. NOD-BSA was selected as the coating antigen, because it showed ultrahigh sensitivity for the detection of MC-LR with a limit of detection (LOD) of 0.0016 ng/mL, which was below the maximum residue level (MRL) of 1 ng/mL. In addition, high reproducibility, good stability, and acceptable spiked sample detection, as validated by liquid chromatography tandem mass spectrometry (LC-MS/MS), indicated the possible application of this method for the analysis of MCs and NOD in water sample.
建立了一种高灵敏度、广特异性的竞争间接酶联免疫吸附测定（ciELISA）方法，用于同时检测九种微囊藻毒素（MCs）和节球藻毒素（NOD）。采用 MC-LR-钥孔戚血蓝蛋白（KLH）免疫新西兰白兔制备抗体。比较了 MC-LR-牛血清白蛋白（BSA）和 NOD-BSA 包被抗原，发现异质包被策略可显著提高检测灵敏度，这从合适的结构中可以明显看出。与 MC-LR 和 MC-LR-BSA 包被技术（0.29ng/mL）的半抑制浓度（IC50）比较表明，NOD-BSA 包被的性能更优，IC50 为 0.054ng/mL。选择 NOD-BSA 作为包被抗原，因为它对 MC-LR 的检测具有超高灵敏度，检测限（LOD）为 0.0016ng/mL，低于 1ng/mL 的最大残留限量（MRL）。此外，通过液相色谱串联质谱（LC-MS/MS）验证，该方法具有良好的重现性、稳定性和可接受的加标样品检测能力，表明该方法可能适用于水样中 MCs 和 NOD 的分析。
