Long F, Shi H C, He M, Sheng J W, Wang J F
Environmental Simulate and Pollution Control State Key Joint Laboratory, Department of Environment Science and Engineering, Tsinghua University, Beijing 100084, China.
Anal Chim Acta. 2009 Sep 1;649(1):123-7. doi: 10.1016/j.aca.2009.07.026. Epub 2009 Jul 14.
A highly sensitive, specific, simple, and rapid chemiluminescence enzyme immunoassay (CLEIA) was developed for the determination of microcystin-LR (MC-LR). Several physicochemical parameters such as the chemiluminescent assay mediums, the dilution ratio of MC-LR-OVA conjugate, monoclonal antibody concentration, and peroxidase labeled antibody concentration were studied and optimized. Under optimum conditions, calibration curve obtained for MC-LR had detection limits of 0.032+/-0.003 microg L(-1), the 50% inhibition concentration (IC50) was 0.20+/-0.02 microg L(-1) and the quantitative detection range was 0.062-0.65 microg L(-1). The proposed methods was successfully applied to the monitoring of MC-LR in spiked water samples without significant effect of the matrix, and the recovery of MC-LR added to water samples at different concentrations ranged from 80% to 115% with the coefficients of variation (CVs) less than 9%. The LOD attained from the calibration curves and the results obtained for the real samples demonstrate the potential use of CLEIA as a screening tool for the analysis of MC-LR in environmental samples.
开发了一种高灵敏度、高特异性、简单且快速的化学发光酶免疫分析法（CLEIA）用于测定微囊藻毒素-LR（MC-LR）。研究并优化了几个物理化学参数，如化学发光检测介质、MC-LR-OVA偶联物的稀释比例、单克隆抗体浓度和过氧化物酶标记抗体浓度。在最佳条件下，MC-LR的校准曲线检测限为0.032±0.003 μg L(-1)，50%抑制浓度（IC50）为0.20±0.02 μg L(-1)，定量检测范围为0.062 - 0.65 μg L(-1)。所提出的方法成功应用于加标水样中MC-LR的监测，不受基质显著影响，不同浓度添加到水样中的MC-LR回收率在80%至115%之间，变异系数（CVs）小于9%。校准曲线得到的检测限以及实际样品的结果表明，CLEIA有潜力作为环境样品中MC-LR分析的筛选工具。
