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Polystyrene Microspheres Coupled with Hybridization Chain Reaction for Dual-Amplified Chemiluminescence Detection of Specific DNA Sequences

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Abstract

We report on a highly sensitive chemiluminescence (CL) assay for the detection of specific DNA sequence, where streptavidin-modified polystyrene microspheres (PS–SA) coupled with hybridization chain reaction (HCR) were employed as dual amplification platform. Briefly, a “sandwich-type” detection strategy was proposed in our design, which involved capture probe DNA immobilized on the surface of carboxyl-terminated magnetic beads (MB) and the reporter DNA combined with the HCR trigger immobilized on the surface of PS–SA. After that, two hairpin-type monomers were added to initiate the HCR. In the detection system, CL signal was obtained via the instantaneous derivatization reaction between 3,4,5-trimethoxylphenylglyoxal (TMPG) and guanine bases in the target and the HCR complex binding on the magnetic beads. As compared to traditional sandwich type (capture/target/reporter) assays, this dual-amplified strategy for sequence-specific DNA detection showed better specificity, lower detection limit and wider linear response range (linear range of 0.01 fmol–10 pmol and a low detection limit of 5 amol). Moreover, this approach could be easily extended to detect a wide range of specific DNA sequences by modification of the hybridization region. We believe this simple technique will present a significant step towards early diagnoses of diseases.

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Acknowledgements

This work was supported by the Natural Science Foundation of China (Nos. 21375025 and 21675030).

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Authors and Affiliations

  1. School of Pharmacy, Fudan University, 826 Zhangheng Road, Shanghai, 201203, China

    Ying Zhou, Yinan Wang, Xin Wang & Jianzhong Lu

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  1. Ying Zhou
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  2. Yinan Wang
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  3. Xin Wang
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Correspondence to Jianzhong Lu.

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Zhou, Y., Wang, Y., Wang, X. et al. Polystyrene Microspheres Coupled with Hybridization Chain Reaction for Dual-Amplified Chemiluminescence Detection of Specific DNA Sequences. J. Anal. Test. 1, 306–314 (2017). https://doi.org/10.1007/s41664-017-0042-4

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  • DOI: https://doi.org/10.1007/s41664-017-0042-4

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