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Original Articles

Ultrasensitive immunoassay of microcystins-LR using G-quadruplex DNAzyme as an electrocatalyst

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Pages 988-1000 | Received 02 Aug 2013, Accepted 09 Dec 2013, Published online: 27 Feb 2014
 

Abstract

An electrochemical immunoassay for microcystin-LR (MC-LR) detection was developed using multi-labeled horseradish peroxidase-mimicking DNAzyme on carbon nanotubes (CNTs) as electrocatalyst for signal amplification. CNTs were covalently conjugated to multiple DNAzyme along with MC-LR for a competitive immunoassay. The as-prepared DNAzyme/CNTs/MC-LR biolabel was specifically captured on the electrode surface, and current responses were obtained upon the electro-catalytic reduction of hydrogen peroxide by the captured biolabels. Under optimal conditions, the electro-catalytic current decreased linearly with the increase amount of MC-LR in the range from 0.01 to 7.0 µg L−1. The linear regression equation was I (µA) = 12.96 − 1.48 X [MC–LR] (µg L−1), with a correlation coefficient of 0.989. The limit of detection of MC-LR was 2.31 ng L−1. Application of the immunoassay method and LC/MS/MS method for MC-LR determination on spiked reservoir water gave recovery range of 91.7–105.2% and 94.0–105.0%, respectively. The resulting versatile immunoassay exhibited high sensitivity, good precision and satisfactory reproducibility, which could have vast potential in routine water quality monitoring for various environmental toxins.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (21277016) and the China Environmental Protection Foundation (CEPF2009-123-1-9).

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