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Label-free supersandwich electrogenerated chemiluminescence biosensor for the determination of the HIV gene

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Abstract

We describe a highly sensitive electrochemiluminescence (ECL) based method for the determination of the human immunodeficiency virus-1 (HIV-1) gene. A long-range self-assembled double strand DNA (ds-DNA) is used as a carrier, and the ruthenium complex Ru(phen)3 2+ as an ECL indicator for signal amplification. The thiolated ss-DNA serving as a capture probe is firstly self-assembled on the surface of a gold electrode. After the target HIV-1 gene is completely hybridized with the capture probe, two previously hybridized auxiliary probes are hybridized with the target HIV-1 gene to form long-range supersandwich ds-DNA polymers on the surface of the electrode. Finally, the ECL indicator is intercalated into the supersandwich ds-DNA grooves. This results in a strongly increased ECL in tripropylamine solution because a large fraction of the intercalator is intercalated into supersandwich ds-DNA. The results showed that the increased ECL intensity is directly related to the logarithm of the concentration of the HIV-1 gene in the range from 0.1 pM to 0.1 nM, with a detection limit of 0.022 pM and using only 10 μL of analyte samples. The method can effectively discriminate target HIV-1 gene (a perfectly matched ss-DNA) from a 2-base mismatched ss-DNA. This work demonstrates that the high sensitivity and selectivity of an ECL DNA biosensor can be largely improved by using supersandwich ds-DNA along with ECL indicators.

The method can effectively discriminate target HIV-1 gene (a perfectly matched ss-DNA) from a 2-base mismatched ss-DNA. The high sensitivity and selectivity of electrochemiluminescence DNA biosensor can be largely improved by using supersandwich ds-DNA along with ECL indicators.

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Acknowledgments

Financial supports from the National Science Foundations of China (No. 21027007, 21275095) are gratefully acknowledged.

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Authors and Affiliations

  1. Key Lab of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi’an, 710062, China

    Sanpeng Ruan, Zhejian Li, Honglan Qi, Qiang Gao & Chengxiao Zhang

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  1. Sanpeng Ruan
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  2. Zhejian Li
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  3. Honglan Qi
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  4. Qiang Gao
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  5. Chengxiao Zhang
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Correspondence to Chengxiao Zhang.

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Ruan, S., Li, Z., Qi, H. et al. Label-free supersandwich electrogenerated chemiluminescence biosensor for the determination of the HIV gene. Microchim Acta 181, 1293–1300 (2014). https://doi.org/10.1007/s00604-014-1252-3

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  • DOI: https://doi.org/10.1007/s00604-014-1252-3

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