水产品中孔雀石绿AlphaLISA检测方法的建立

Quantitative determination of malachite green in aquatic products using amplified luminescent proximity homogeneous assay

建立孔雀石绿(malachite green,MG)的光激化学发光纳米均相时间分辨荧光免疫法(AlphaLISA)分析方法。将待测物,生物素化-MG-BSA,抗体加入到孔板中,再加入供体微珠和受体微珠。生物素化-MG-BSA和游离的待测物对抗体的竞争使得信号值减小,从而优化检测条件并进行方法学验证。结果表明:该方法特异性良好,与无色孔雀石绿,无色结晶紫无交叉反应,灵敏度为0.42 ng/m L,MG在2种鱼样品中的回收率在81.0%~105.0%和84.0%~90.8%之间,批内精密度RSD<9.5%,批间精密度RSD<16.0%。MG-AlphaLISA方法可以简单、快速的对鱼类进行MG测定,具有特异性强、灵敏性高、稳定等特点。

A new method was established for analyzing malachite green （MG） by amplified luminescent proximi- ty homogeneous assay （AlphaLISA）. Samples, botinylated MG-BSA and MG antibody were added to a 384-well mi- croplate, then donor beads and acceptor beads were added. Antibody competition between Biotinylated -MG-BSA and disassociate sample substances caused signal value decrease, thus optimized the test condition and analytical performance. The cross-reactivity of the MG-AlphaLISA with Leucomalachite green and Leucomalachite crystal violet were 11.74% and 7.24% , respectively; the sensitivity was 0.42 ng/mL; the recoveries of the determination for MG in two kinds of fish samples were 81.0% -105.0% and 84.0% -90.8% , respectively; the RSD of intra-day and inter-assay were 〈9.5% and 〈 16.0% , respectively; the MG-AlphaLISA method is excellent in specificity, sensitivity, and stability. It can be widely used in rapid and simple screening for MG contamination in fish.