组成图示
点击图片查看大图 · 依据论文自动绘制
传感器类型
其他（光盘光学免疫微传感器）
检测对象
微囊藻毒素LR（microcystin-LR, MC-LR）；样品基质：河水（Júcar、Turia河水及加标河水）
检测原理
该传感器采用间接竞争微免疫分析。BSA-MC偶联物预先固定在DVD聚碳酸酯表面，样品中游离MC-LR与固定BSA-MC竞争结合mAb-MC。MC-LR浓度越高，mAb-MC与固定BSA-MC结合越少，后续GAM-Au金标记二抗结合越少，银增强液催化沉积的银量越少，光盘表面光学密度越低。DVD驱动器以650 nm激光扫描，免疫复合物改变反射特性，光电二极管获得与光密度相关的模拟信号，信号强度随MC-LR浓度升高而降低。OVA-BT/PAb-BT内标和MIgG/RIgG阳性对照用于校正免疫反应与检测步骤，银增强提供信号放大。
检测灵敏度
IC50: 1.04 µg/L；线性范围: 0.12-2.00 µg/L；r^2 = 0.991；PBST 回归斜率: -6345；河水回归斜率: -6036
效应效果
体系对MC-LY、MC-LA、MC-LF、MC-LW、MC-YR和节球藻毒素交叉反应为144%、125%、119%、102%、83%和94%。盘内RSD 1.8%–2.6%，盘间RSD 3.3%–5.2%。预包被光盘4 ℃保存至少7周；免疫复合物5天保留90%，7天保留73%。12份河水加标0.50、1.00、1.50 µg/L，回收率78%–113%，无需预浓缩。河水IC50 0.96 µg/L，动态范围0.12–2.00 µg/L。单盘分析42样品，37 min，较LC/MS和ELISA更便携、快速，适合现场高通量监测。
传感器的构成
- 基底/换能器：DVD-R光盘聚碳酸酯（polycarbonate）表面，作为微阵列载体和光学反射面，供DVD驱动器读取
- 捕获/识别层：BSA-MC偶联物（30 mg/L）固定于聚碳酸酯表面，作为竞争免疫分析中捕获微囊藻毒素的抗原
- 内标/质控层：OVA-BT（卵白素-生物素偶联物，0.2 mg/L）用于内标校准和免疫反应阳性对照
- 识别抗体：mAb-MC（抗MC-LR单克隆抗体，1/5000）与PAb-BT（兔抗生物素多克隆抗体，1/1000）混合，识别MC-LR和OVA-BT
- 阳性对照层：MIgG（非免疫小鼠血清，1/1000）和RIgG（兔血清，1/4000）固定于盘上，用于检测步骤阳性对照
- 信号标记物：GAM-Au（金标记山羊抗小鼠IgG，1/50）和GAR-Au（金标记山羊抗兔IgG，1/250）二抗，结合一抗形成免疫复合物
- 信号放大/显色：银增强液A/B（silver enhancer solutions A and B）催化银沉积，放大光学信号
- 缓冲/洗涤：PBS-T（磷酸盐缓冲液含Tween 20）用于抗体孵育和洗涤；打印缓冲液（0.1 M碳酸钠，pH 9.6，1%甘油）用于点样固定
- 读取系统：DVD驱动器650 nm激光和光电二极管，配合Diskpick软件采集模拟信号并转换为光密度图像
中文摘要
针对水体中关键目标物的简单、准确、快速多样品分析方法需求迫切。本文在光盘聚碳酸酯侧制备光学微传感器阵列，用于检测河水中微囊藻毒素。传感器基于间接竞争微免疫分析：样品中游离微囊藻毒素LR（MC-LR）与固定偶联物竞争特异性单克隆抗体。免疫反应结果由DVD驱动器读取，数分钟内获得读数。该方法对MC-LR的灵敏度（IC50）为1.04 µg/L，线性范围为0.12–2.00 µg/L，可低于世界卫生组织饮用水限值进行测定。方法简便、灵敏、高通量，现场使用仅需37 min。优化体系对MC-LY、MC-LA、MC-LF、MC-LW、MC-YR和节球藻毒素的交叉反应分别为144%、125%、119%、102%、83%和94%。河水样品分析回收率为78%–113%。预包被光盘至少稳定7周。系统便携，可现场定量，节省时间和资源。据作者所知，这是首个用于现场微囊藻毒素定量的便携式阵列系统，可同时分析42个样品及校准曲线，达到微克/升级别灵敏度。
英文摘要
The development of simple, accurate, and rapid multisample analytical methodologies to find out critical targets in waters is highly demanded. Optical microsensor arrays to determine microcystins in river waters are developed on the polycarbonate side of compact discs. The working principle of the sensors relied on an indirect competitive microimmunoassay, where free microcystin LR (MC-LR) competes with immobilized conjugate for specific monoclonal antibody. The results of the immunoreaction are detected with a DVD drive, showing the readouts in minutes. The method reached a sensitivity (IC(50)) for MC-LR of 1.04 μg/L and a linear response in the range 0.12-2.00 μg/L, allowing its determination below the upper limit proposed by the World Health Organization in drinking water. The developed analytical approach shows simplicity, good sensitivity, high throughput capability, and rapidity (37 min) in field use. The optimized assay showed also high congener reactivity to MC-LY (144%), MC-LA (125%), MC-LF (119%), MC-LW (102%), MC-YR (83%), and nodularin (94%). Furthermore, the suitability of the disk biosensor to quantify MC-LR was successfully evaluated analyzing river water samples, obtaining excellent recoveries (78-113%). Precoated discs are stable for at least seven weeks without loosing their analytical performances. Also, the portability of the analytical system permits on-site analysis and quantification, saving time and other resources. To our knowledge, this is the only work where a portable, easy-to-use, array based system has been developed for on-site microcystin quantification and applied to simultaneously analyze 42 samples plus the calibration curve, reaching microgram per liter sensitivity.