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Double-hairpin molecular-beacon-based amplification detection for gene diagnosis linked to cancer

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Abstract

A powerful double-hairpin molecular beacon (DHMB) was developed for cancer-related KRAS gene detection based on the one-to-two stoichiometry. During target DNA detection, DHMB can execute signal transduction even if no any exogenous element is involved. Unlike the conventional molecular beacon based on the one-to-one interaction, one target DNA not only hybridizes with one DHMB and opens its hairpin but also promotes the interaction between two DHMBs, causing the separation of two fluorophores from quenchers. This leads to an enhanced fluorescence signal. As a result, the target KRAS gene is able to be detected within a wide dynamic range from 0.05 to 200 nM with the detection limit of 50 pM, indicating a dramatic improvement compared with traditional molecular beacons. Moreover, the point mutations existing in target DNAs can be easily screened. The potential application for target species in real samples was indicated by the analysis of PCR amplicons of DNAs from the DNA extracted from SW620 cell. Besides becoming a promising candidate probe for molecular biology research and clinical diagnosis of genetic diseases, the DHMB is expected to provide a significant insight into the design of DNA probe-based homogenous sensing systems.

A powerful double-hairpin molecular beacon (DHMB) was developed for cancer-related gene KRAS detection based on the one-to-two stoichiometry. Without the help of any exogenous probe, the point mutation is easily screened, and the target DNA can be quantified down to 50 pM, indicating a dramatic improvement compared with traditional molecular beacons

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Acknowledgments

This work was supported by National Natural Science Foundation of China (NSFC) (grant no: 21275002), Zhejiang Province Natural Science Foundation of China (LY16C07002), and Independent Research Project of State Key Laboratory of Photocatalysis on Energy and Environment (No. 2014CO1).

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Authors and Affiliations

  1. Key laboratory of watershed science and health of Zhejiang Province, Institute of Functional Nucleic Acids and Personalized Cancer Theranostics, Key Laboratory of Laboratory Medicine, Ministry of Education of China, and Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Huo Xu, Rongbo Zhang, Feng Li, Yingying Zhou, Ting Peng, Xuedong Wang & Zhifa Shen

  2. Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis of State Key Laboratory of Photocatalysis on Energy and Environment, and Fujian Provincial Key Laboratory of Cancer Metastasis Chemoprevention and Chemotherapy, College of Chemistry, Fuzhou University, Fuzhou, 350002, China

    Huo Xu

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  1. Huo Xu
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  2. Rongbo Zhang
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Correspondence to Xuedong Wang or Zhifa Shen.

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Xu, H., Zhang, R., Li, F. et al. Double-hairpin molecular-beacon-based amplification detection for gene diagnosis linked to cancer. Anal Bioanal Chem 408, 6181–6188 (2016). https://doi.org/10.1007/s00216-016-9729-z

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  • DOI: https://doi.org/10.1007/s00216-016-9729-z

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