INL, International Iberian Nanotechnology Laboratory, Av. Mestre José Veiga s/n, 4715-330, Braga, Portugal.
CIIMAR, Interdisciplinary Centre of Marine and Environmental Research of the University of Porto, 4450-208, Matosinhos, Portugal.
Biosens Bioelectron. 2019 Oct 1;142:111550. doi: 10.1016/j.bios.2019.111550. Epub 2019 Jul 30.
Microcystins are the most worldwide extended and common toxins produced by cyanobacteria in freshwater. Microcystin-leucine arginine (MC-LR), associated with the most toxic incidents involving microcystins, are within the cyanobacteria (intracellular) until released into the surrounding waters (extracellular) during cell lysis. Therefore, the relationship between intracellular and extracellular cyanotoxins will allow a comprehensive risk of cyanobacteria-containing waters, preventing disease and improving human safety. In this work, we present the development of a novel portable microfluidic sensing platform for the simultaneous detection of free (extracellular) and total MC-LR (intracellular and extracellular). The integrated system contains the sample processing and detection modules capable of performing the chemical lysis, filtration, sample mixing with antibodies, and electrochemical detection of MC-LR based on an indirect strategy. The performance of the immunosensors was evaluated by electrochemical impedance spectroscopy, showing a linear dynamic range between 3.3 × 10 and 10 g L and a limit of detection of 5.7 × 10 g L. The results demonstrate the potential of the developed portable biosensor platform and its suitable application for the analysis of MC-LR at regulated levels for drinking water. Finally, the integrated system was able to simultaneously detect the free and total MC-LR on a Microcystis aeruginosa culture. To the best of our knowledge this is the first described system that can differentiate between intracellular and extracellular concentration of MC-LR. This novel electrochemical sensing platform avoids the multiple processing steps typically needed for standard MC-LR analysis in the laboratory and provides an early warning system for MC-LR remote monitoring in water.
微囊藻毒素是蓝藻在淡水中产生的分布最广、最常见的毒素。与涉及微囊藻毒素的最有毒事件相关的亮氨酸精氨酸微囊藻毒素(MC-LR),在细胞溶解之前一直存在于蓝藻内(细胞内),然后释放到周围水中(细胞外)。因此,细胞内和细胞外蓝藻毒素之间的关系将允许对含有蓝藻的水进行全面的风险评估,从而预防疾病并提高人类安全性。在这项工作中,我们提出了一种用于同时检测游离(细胞外)和总 MC-LR(细胞内和细胞外)的新型便携式微流控传感平台的开发。集成系统包含能够执行化学裂解、过滤、与抗体混合样品以及基于间接策略的 MC-LR 电化学检测的样品处理和检测模块。通过电化学阻抗谱评估了免疫传感器的性能,显示出 3.3×10 和 10 g L 之间的线性动态范围和 5.7×10 g L 的检测限。结果表明,所开发的便携式生物传感器平台具有潜力,并且非常适合用于分析饮用水中规定水平的 MC-LR。最后,该集成系统能够同时检测微囊藻培养物中的游离和总 MC-LR。据我们所知,这是第一个能够区分 MC-LR 细胞内和细胞外浓度的系统。这种新型电化学传感平台避免了实验室中通常进行的标准 MC-LR 分析所需的多个处理步骤,并为水中 MC-LR 的远程监测提供了早期预警系统。