Quantitative and rapid detection of microcystin-LR using time-resolved fluorescence immunochromatographic assay based on europium nanospheres,Analytical Methods

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Quantitative and rapid detection of microcystin-LR using time-resolved fluorescence immunochromatographic assay based on europium nanospheres
Analytical Methods ( IF 2.7 ) Pub Date : 2017-10-25 00:00:00 , DOI: 10.1039/c7ay02036h
Yi Zhang _{1,

2,

3,

4,

5} , Xin-Liang Ding _{5,

6,

7,

8} , Ming-Ming Guo _{1,

2,

3,

4,

5} , Ting-Ting Han _{1,

2,

3,

4,

5} , Zi-Jian Huang _{8,

9,

10,

11} , Hong-Tao Shang _{8,

12,

13,

14} , Biao Huang _{1,

2,

3,

4,

5}

Affiliation

In the present study, a novel time-resolved fluorescence immunochromatographic assay was established for the rapid quantitative detection of microcystin-leucine arginine (MC-LR). In this method, the europium nanoshpere labelled with anti-MC-LR antibodies was used as the luminescent tracer, dissolved in the running buffer and then added with the sample solution on the pad. MC-LR-BSA and goat anti-mouse antibody were dispensed on the nitrocellulose membrane for the test and the control line, respectively. The optimal parameters were 0.05 g L⁻¹ MC-LR-BSA, 1 : 100 colloidal europium–antibody conjugate, and 10 min reaction time. The linear working range for MC-LR was 0.1–5 μg L⁻¹ with an IC50 of 0.78 μg L⁻¹ and a sensitivity of 0.035 μg L⁻¹. The low cross-reactivity was observed with MC-YR and MC-LF. The assay accuracy was confirmed by the HPLC method with a correlation coefficient of 0.99. When the variable coefficients were 4.4% and 5.4%, the average recoveries of tap and lake water were 94.6% and 102.8%, respectively. The time-resolved fluorescence immunochromatographic assay provides a sensitive, simple, and speedy performance for MC-LR quantitative determination and has a potential use for water sample screening.

中文翻译：

基于time纳米球的时间分辨荧光免疫色谱法定量和快速检测微囊藻毒素-LR

在本研究中，建立了一种新型的时间分辨荧光免疫色谱分析方法，用于快速定量检测微囊藻毒素-亮氨酸精氨酸（MC-LR）。在这种方法中，将标记有抗MC-LR抗体的nano纳米晶用作发光示踪剂，将其溶解在运行缓冲液中，然后将样品溶液添加到垫上。将MC-LR-BSA和山羊抗小鼠抗体分配在硝酸纤维素膜上，分别用于测试和对照品系。最佳参数为0.05 g L ^-1 MC-LR-BSA，1：100胶体euro抗体结合物和10分钟的反应时间。MC-LR的线性工作范围为0.1–5μgL ^-1，IC50为0.78μgL ^-1，灵敏度为0.035μgL ^-1。使用MC-YR和MC-LF观察到较低的交叉反应性。通过HPLC方法确认测定准确性，相关系数为0.99。当变量系数为4.4％和5.4％时，自来水和湖水的平均回收率分别为94.6％和102.8％。时间分辨荧光免疫色谱分析法为MC-LR定量测定提供了灵敏，简单且快速的性能，并有可能用于水样筛查。

更新日期：2017-11-09

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