Development and Application of Highly Sensitive Anti-immune Complex ELISAs for Microcystins in Tap Water
Development and Application of Highly Sensitive Anti-immune Complex ELISAs for Microcystins in Tap Water
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针对自来水中微囊藻毒素的高灵敏抗免疫复合物 ELISA 的开发和应用
DOI：
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发表时间：
2000
期刊：
影响因子：
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通讯作者：
K. Harada
中科院分区：
文献类型：
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作者：
T. Tsutsumi;S. Nagata;F. Yoshida;Y. Ueno;K. Harada
我们开发了一种抗免疫复合物 (IC) ELISA，适用于直接测定自来水中的痕量微囊藻毒素 (MC)。两种检测形式的比较表明，在包被步骤中使用抗免疫复合物单克隆抗体 (MAB) 来捕获抗 MC MABMC 复合物可提高灵敏度和精确度。 IC ELISA 的检测限和定量范围分别为 2 pg ml−1 和 2100 pg ml−1 微囊藻毒素LR (MCLR)，表明是迄今为止报道的所有检测 MC 的方法中最灵敏的。此外，IC ELISA 在其定量范围内保持了良好的可靠性，低变异系数（测定内和测定间分别为 5.010.8 和 4.910.2%）证明了这一点。 IC ELISA 显示与微囊藻毒素-RR 和微囊藻毒素-YR 具有良好的交叉反应性，表明该方法可以检测环境中发现的主要 MC。将定量范围的 MCLR 添加到自来水中的回收率测试结果平均回收率为 99%，平均标准偏差为 5.7%；因此，IC ELISA 在自来水样品分析中表现良好。对中国和日本采集的自来水样本进行 ELISA 分析显示，在检测的 17 个样本中，中国采集的两个样本的 MCs 呈阳性，浓度为 4.914 pg ml−1。这些结果表明新开发的 IC ELISA 可用于监测自来水中的微量 MC。
We developed an anti-immune complex (IC) ELISA applicable to direct determination of trace amounts of microcystins (MCs) in tap water. Comparison of two assay formats revealed that the use of anti-immune complex monoclonal antibody (MAB) in the coating step to trap anti-MC MABMC complexes improved the sensitivity as well as precision. The detection limit and quantitative range of the IC ELISA was 2 pg ml−1 and 2100 pg ml−1 of microcystinLR (MCLR), respectively, indicating the most sensitive of all the methods for detecting MCs reported to date. Additionally, the IC ELISA maintained good reliability through its quantitative range, as evidenced by low coefficients of variation (5.010.8 and 4.910.2% for intra- and interassay, respectively). The IC ELISA showed good cross-reactivity to microcystin-RR and microcystin-YR, suggesting major MCs found in the environment can be detected by this method. Recovery tests in which quantitative range of MCLR were added to tap water resulted in a mean recovery of 99%, with a mean standard deviation of 5.7%; therefore the IC ELISA performed well in the analysis of tap water samples. ELISA analysis of tap water samples collected in China and Japan revealed that, among 17 samples tested, two samples collected in China were positive for MCs at 4.914 pg ml−1. These results suggest that the newly developed IC ELISA can be used to monitor trace amounts of MCs in tap water.
DOI：
10.1016/s0021-9258(17)45384-1
发表时间：
1990-11
期刊：
The Journal of biological chemistry
影响因子：
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作者：
R. Honkanen;J. Zwiller;R. Moore;S. L. Daily;B. Khatra;M. Dukelow;A. Boynton
通讯作者：
R. Honkanen;J. Zwiller;R. Moore;S. L. Daily;B. Khatra;M. Dukelow;A. Boynton