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Analytical and Bioanalytical Chemistry

, Volume 411, Issue 10, pp 1935–1941 | Cite as

A rapid bioanalytical tool for detection of sequence-specific circular DNA and mitochondrial DNA point mutations

  • Yuqian Zhang
  • Ahmet Kaynak
  • Taosheng Huang
  • Leyla EsfandiariEmail author
Communication

Abstract

Mutations in mitochondrial DNA (mtDNA) have been an essential cause of numerous diseases, making their identification critically important. The majority of mtDNA screening techniques require polymerase chain reaction (PCR) amplification, enzymatic digestion, and denaturation procedures, which are laborious and costly. Herein, we developed a sensitive PCR-free electrokinetic-based sensor combined with a customized bis-peptide nucleic acid (bis-PNA) and gamma-PNA (γ-PNA) probes immobilized on beads, for the detection of mtDNA point mutations and sequence-specific supercoiled plasmid DNA at the picomolar range. The probes are capable of invading the double-stranded circular DNA and forming a stable triplex structure. Thus, this method can significantly reduce the sample preparation and omit the PCR amplification steps prior to sensing. Further, this bioanalytical tool can open up a new paradigm in clinical settings for the screening of double-stranded circular nucleic acids with a single-base mismatch specificity in a rapid and sensitive manner.

Keywords

Bis-PNA Gamma-PNA Mitochondrial DNA Circular double-stranded DNA Electrokinetic sensor 

Notes

Acknowledgements

The authors thank Shiyu Luo from Dr. Huang’s group for the generous help on mitochondrial DNA preparation.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

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

Authors and Affiliations

  • Yuqian Zhang
    • 1
  • Ahmet Kaynak
    • 2
  • Taosheng Huang
    • 3
  • Leyla Esfandiari
    • 1
    • 2
    Email author
  1. 1.Department of Electrical Engineering and Computer ScienceUniversity of CincinnatiCincinnatiUSA
  2. 2.Department of Biomedical EngineeringUniversity of CincinnatiCincinnatiUSA
  3. 3.Department of Human GeneticsCincinnati Children’s HospitalCincinnatiUSA

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