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Microchimica Acta

, 186:286 | Cite as

Fluorometric determination of HIV DNA using molybdenum disulfide nanosheets and exonuclease III-assisted amplification

  • Lele Wang
  • Lianhua Dong
  • Gang Liu
  • Xizhong Shen
  • Jing Wang
  • Changfeng ZhuEmail author
  • Min Ding
  • Yanli WenEmail author
Original Paper
  • 48 Downloads

Abstract

A convenient and ultrasensitive fluorometric method is described for the determination of HIV DNA. It exploits the strong difference in the affinities of MoS2 nanosheets for long ssDNA versus short oligonucleotide fragments. In addition, efficient signal amplification is accomplished by exonuclease III-assisted target recycling. When absorbed on the MoS2 nanosheets, the fluorescence of the FAM-labeled ssDNA probe (FP) is quenched. However, in the presence of HIV DNA, the FP hybridizes with target to form a duplex. As a result, the FP in the duplex will be stepwise hydrolyzed into short fragments by Exo III, and the fluorescence signal thus is retained because short fragments have low affinity for the MoS2 nanosheets. By using the Exo III-assisted target recycling amplification, the detection sensitivity is strongly improved. The sensor can detect DNA in a concentration as low as 5.3 pM (at an S/N ratio of 3), and the analytical range extends from 0.01 nM to 10 nM. The assay is simple, sensitive and specific, and conceivably represents a valuable tool in clinical studies related to the HIV.

Graphical abstract

Schematic presentation of fluorometric determination of HIV DNA based on molybdenum disulfide nanosheets and Exo III. When the fluorescence-tagged ssDNA probe hybridized with target to form a duplex, the Exo III-assisted target recycling amplification is generated. The method can detect as low as 5.3 pM HIV DNA.

Keywords

Layered transition metal dichalcogenide nanosheets Enzyme amplification Fluorometric assay DNA biosensor 

Notes

Acknowledgments

This work was financially supported by National Natural Science Foundation of China (No. 21775104, 21605026, 81672720), the National Quality Infrastructure Program of China (2017YFF0204605), and Shanghai Rising-Star Program (16QB1403100).

Author contributions

L.W., Y.W., G.L., and C.Z. designed the experiments and wrote the main manuscript ;L. W.,L.D., M. D. and Y.W. performed experiments; X.S. and J.W. contributed to datacuration and formal analysis. All authors reviewed the manuscript.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3368_MOESM1_ESM.doc (9.7 mb)
ESM 1 (DOC 9.73 mb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Lele Wang
    • 1
  • Lianhua Dong
    • 2
  • Gang Liu
    • 1
  • Xizhong Shen
    • 3
    • 4
  • Jing Wang
    • 2
  • Changfeng Zhu
    • 3
    Email author
  • Min Ding
    • 1
  • Yanli Wen
    • 1
    Email author
  1. 1.Laboratory of BiometrologyShanghai Institute of Measurement and Testing TechnologyShanghaiPeople’s Republic of China
  2. 2.Division of Medical and Biological MeasurementNational Institute of MetrologyBeijingPeople’s Republic of China
  3. 3.Department of Gastroenterology and Hepatology, Zhongshan HospitalFudan UniversityShanghaiPeople’s Republic of China
  4. 4.Shanghai Institute of Liver Diseases, Zhongshan HospitalFudan UniversityShanghaiPeople’s Republic of China

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