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Glutaraldehyde-modified electrode for nonlabeling voltammetric detection of p16 INK4A gene

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Abstract

A nonlabeling electrochemical detection method for analyzing the polymerase-chain-reaction-amplified sequence-specific p16 INK4A gene, in which the basis for the covalent immobilization of deoxyribonucleic acid (DNA) probe is described, has been developed. The self-assembly process was based on the covalent coupling of glutaraldehyde (GA) as an arm molecule onto an amino-functional surface. The p16 INK4A gene was used as the model target for the methylation detection of early cancer diagnosis. An amino-modified DNA probe was successfully assembled on the GA-coupling surface through the formation of Schiff base under potential control. The hybridization of amino-modified DNA probes with the target was investigated by means of electrochemical measurements, including cyclic voltammetry and square wave voltammetry. Furthermore, the functions of GA coupling for sequence-specific detection were compared with those obtained based on mercaptopropionic acid. Hybridization experiments indicated that the covalent coupling of GA was suitable for the immobilization of DNA probe and was sensitive to the electrochemical detection of single-base mismatches of label-free DNA targets in hybridization. Moreover, reported probe-modified surfaces exhibited excellent stability, and the hybridization reactions were found to be completely reversible and highly specific for recognition in subsequent hybridization processes. The strategy provided the potential for taking full advantage of existing modified electrode technologies and was verified in microarray technology, which could be applied as a useful and powerful tool in electrochemical biosensor and microarray technology.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant nos. 90406023, 60371027, 60121101) and the Natural Science Foundation of Jiangsu (China; grant nos. 03KJD310177 and JHB04-005).

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Authors and Affiliations

  1. School of Chemistry and Chemical Engineering, Nantong University, Nantong, 226007, People's Republic of China

    Cunwang Ge

  2. Chien-Shiung Wu Laboratory, Department of Biological Science and Medical Engineering, Research Center for Nano-scale Science and Technology, Southeast University, Nanjing, 210096, People's Republic of China

    Cunwang Ge, Meiju Ji & Ning Gu

  3. Department of Chemistry and Biochemistry, The University of Southern Mississippi, Hattiesburg, MS, 39406-5043, USA

    Wujian Miao

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  1. Cunwang Ge
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  2. Wujian Miao
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Correspondence to Cunwang Ge.

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Ge, C., Miao, W., Ji, M. et al. Glutaraldehyde-modified electrode for nonlabeling voltammetric detection of p16 INK4A gene. Anal Bioanal Chem 383, 651–659 (2005). https://doi.org/10.1007/s00216-005-0032-7

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  • DOI: https://doi.org/10.1007/s00216-005-0032-7

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