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Two-stage Nicking Enzyme Signal Amplification Combined with DNAzyme Amplification for the Detection of Bone Morphogenetic Protein 6 mRNA

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Abstract

Highly sensitive detection of bone morphogenetic protein 6 (BMP6) mRNA is essential to monitor bone regeneration in the regenerating defects. In this work, we proposed a quantitative approach based on two-stage nicking enzyme signal amplification (NESA) and DNAzyme amplification for highly sensitive detection of BMP6 mRNA. The two-stage NESA involves two templates and two-stage amplification reactions under isothermal conditions. The first template contains two repeat sequences that could hybridize to the target RNA, triggering an exponential amplification. The amplified product was a short single-stranded DNA with the same sequence as the target RNA. The single-stranded DNA can trigger another linear NESA and produces a large amount of horseradish peroxidase (HRP)-mimicking G-quadruplex DNAzyme. This proposed assay showed a quantitative analysis of BMP6 mRNA in a wide range from 1 fM to 100 nM with a detection limit of 0.01 fM.

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Authors and Affiliations

  1. Department of Orthopaedics, Liuzhou People’s Hospital, 8 Wenchang Road, 545006, Liuzhou, Guangxi Province, P. R. China

    Zhaohui Hu, Yanhong Zhou, Xiangtao Xie, Renjie Jiang & Ningning Li

Authors
  1. Zhaohui Hu
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  2. Yanhong Zhou
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  3. Xiangtao Xie
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  4. Renjie Jiang
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  5. Ningning Li
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Correspondence to Ningning Li.

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Hu, Z., Zhou, Y., Xie, X. et al. Two-stage Nicking Enzyme Signal Amplification Combined with DNAzyme Amplification for the Detection of Bone Morphogenetic Protein 6 mRNA. ANAL. SCI. 30, 1039–1044 (2014). https://doi.org/10.2116/analsci.30.1039

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  • DOI: https://doi.org/10.2116/analsci.30.1039

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