Lei La-Mei, Wu Ying-Song, Gan Nan-Qin, Song Li-Rong
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, CAS, Donghu Nanlu No. 7, Wuchang, Wuhan 430072, PR China.
Clin Chim Acta. 2004 Oct;348(1-2):177-80. doi: 10.1016/j.cccn.2004.05.019.
A time-resolved fluorescence immunoassay (TRFIA), based on anti-microcystin-LR (MCLR) monoclonal antibodies (MAbs) and europium-labeled antimouse IgG conjugate, was first developed for microcystin detection.
Anti-MCLR MAbs were prepared by a standard method, and the attained MAbs showed a good cross reactivity with MCLR, MCRR and MCYR. The TRFIA was performed in an indirect competitive mode. The detection method of TRFIA was compared with indirect competitive enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC).
The TRFIA exhibited a typical sigmoidal response for MCLR at concentrations of 0.005-50 ng/ml, with a wide quantitative range between 0.01 and 10 ng/ml, indicating the broadest detective range and the most sensitive of all the methods for microcystins (MCs) detection. Additionally, the TRFIA maintained good reliability through its quantitative range, as evidenced by low coefficients of variation (1.6-12.2%). The toxin data of algal samples assayed from TRFIA were in the same range as those with ELISA and HPLC, implying that the method was reliable and practical for the detection of MCs.
The TRFIA may offer a valuable alternative or a substitute for conventional ELISA for microcystin detection.
基于抗微囊藻毒素-LR（MCLR）单克隆抗体（MAbs）和铕标记的抗小鼠IgG缀合物，首次开发了一种时间分辨荧光免疫分析（TRFIA）用于微囊藻毒素检测。
采用标准方法制备抗MCLR单克隆抗体，所获得的单克隆抗体与MCLR、MCRR和MCYR具有良好的交叉反应性。TRFIA采用间接竞争模式进行。将TRFIA检测方法与间接竞争酶联免疫吸附测定（ELISA）和高效液相色谱（HPLC）进行比较。
TRFIA在0.005-五十纳克/毫升浓度下对MCLR呈现典型的S形响应，定量范围在0.01至10纳克/毫升之间，表明其在所有微囊藻毒素（MCs）检测方法中检测范围最广且最灵敏。此外，TRFIA在其定量范围内保持良好的可靠性，变异系数较低（1.6-12.2%）证明了这一点。TRFIA测定的藻类样品毒素数据与ELISA和HPLC测定的数据在同一范围内，这意味着该方法对于MCs的检测是可靠且实用的。
TRFIA可能为微囊藻毒素检测提供一种有价值的替代方法或替代传统ELISA的方法。
