Enzyme enhanced quantitative determination of multiple DNA targets based on capillary electrophoresis |
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Authors: | Xuemei Li Zhiming Zhan Shusheng Zhang Hongyuan Chen |
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Institution: | 1. Department of Chemistry, Nanjing University, Nanjing 210093, China;2. Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China |
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Abstract: | In the current paper, enzyme enhanced simultaneous quantitative determination of multiple DNA targets based on capillary electrophoresis (CE) was described. We used three biotin-modified DNA probes, which reacted with avidin-conjugated horseradish peroxidase (avidin-HRP) conjugate to obtain the HRP labeled probes, to hybridize with three corresponding targets. The resulting mixture containing double-strand DNA (dsDNA)-HRP, excess single-strand DNA (ssDNA)-HRP and remaining avidin-HRP was separated by capillary electrophoresis, and then the system of HRP catalyzing H2O2/o-aminophenol (OAP) reaction was adopted. The catalytic product was detected with electrochemical detection. With this protocol, the limits of quantification for the hybridization assay of 21-, 39- and 80-mer DNA fragments were of 1.2 × 10−11, 2.4 × 10−11 and 3.0 × 10−11 M, respectively. The multiplex assay also provided good specificity without any cross-reaction. |
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Keywords: | Capillary electrophoresis DNA Multianalyte determination Enzyme enhancement Electrochemical detection |
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