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A PCR-free electrochemical method for messenger RNA detection in cancer tissue samples

Journal Article


Abstract


  • Despite having reliable and excellent diagnostic performances, the currently available messenger RNA (mRNA)

    detection methods mostly use enzymatic amplification steps of the target mRNA which is generally affected by

    the sample manipulations, amplification bias and longer assay time. This paper reports an amplification-free

    electrochemical approach for the sensitive and selective detection of mRNA using a screen-printed gold

    electrode (SPE-Au). The target mRNA is selectively isolated by magnetic separation and adsorbed directly onto

    an unmodified SPE-Au. The surface-attached mRNA is then measured by differential pulse voltammetry (DPV)

    in the presence of [Fe(CN)6]

    4-/3- redox system. This method circumvents the PCR amplification steps as well as

    simplifies the assay construction by avoiding multiple steps involved in conventional biosensing approaches of

    using recognition and transduction layers. Our method has demonstrated good sensitivity (LOD = 1.0 pM) and

    reproducibility (% RSD = < 5%, for n = 3) for detecting FAM134B mRNA in two cancer cell lines and a small

    cohort of clinical samples (number of samples = 26) collected from patients with oesophageal cancer. The

    analytical performance of our method is validated with a standard qRT-PCR analysis. We believe that our PCRfree

    approach holds a great promise for the analysis of tumor-specific mRNA in clinical samples.

Authors


  •   Islam, Md Nazmul (external author)
  •   Gopalan, Vinod (external author)
  •   Haque, Md Hakimul. (external author)
  •   Masud, Mostafa (external author)
  •   Hossain, Md Shahriar
  •   Yamauchi, Yusuke (external author)
  •   Nguyen, Nam-Trung (external author)
  •   Lam, Alfred (external author)
  •   Shiddiky, Muhammad J. A. (external author)

Publication Date


  • 2017

Citation


  • Islam, M., Gopalan, V., Haque, M. Hakimul., Masud, M. Kamal., Hossain, M. Al., Yamauchi, Y., Nguyen, N., Lam, A. K. & Shiddiky, M. J. A. (2017). A PCR-free electrochemical method for messenger RNA detection in cancer tissue samples. Biosensors and Bioelectronics, 98 227-233.

Scopus Eid


  • 2-s2.0-85021751203

Ro Metadata Url


  • http://ro.uow.edu.au/aiimpapers/2628

Number Of Pages


  • 6

Start Page


  • 227

End Page


  • 233

Volume


  • 98

Place Of Publication


  • United States

Abstract


  • Despite having reliable and excellent diagnostic performances, the currently available messenger RNA (mRNA)

    detection methods mostly use enzymatic amplification steps of the target mRNA which is generally affected by

    the sample manipulations, amplification bias and longer assay time. This paper reports an amplification-free

    electrochemical approach for the sensitive and selective detection of mRNA using a screen-printed gold

    electrode (SPE-Au). The target mRNA is selectively isolated by magnetic separation and adsorbed directly onto

    an unmodified SPE-Au. The surface-attached mRNA is then measured by differential pulse voltammetry (DPV)

    in the presence of [Fe(CN)6]

    4-/3- redox system. This method circumvents the PCR amplification steps as well as

    simplifies the assay construction by avoiding multiple steps involved in conventional biosensing approaches of

    using recognition and transduction layers. Our method has demonstrated good sensitivity (LOD = 1.0 pM) and

    reproducibility (% RSD = < 5%, for n = 3) for detecting FAM134B mRNA in two cancer cell lines and a small

    cohort of clinical samples (number of samples = 26) collected from patients with oesophageal cancer. The

    analytical performance of our method is validated with a standard qRT-PCR analysis. We believe that our PCRfree

    approach holds a great promise for the analysis of tumor-specific mRNA in clinical samples.

Authors


  •   Islam, Md Nazmul (external author)
  •   Gopalan, Vinod (external author)
  •   Haque, Md Hakimul. (external author)
  •   Masud, Mostafa (external author)
  •   Hossain, Md Shahriar
  •   Yamauchi, Yusuke (external author)
  •   Nguyen, Nam-Trung (external author)
  •   Lam, Alfred (external author)
  •   Shiddiky, Muhammad J. A. (external author)

Publication Date


  • 2017

Citation


  • Islam, M., Gopalan, V., Haque, M. Hakimul., Masud, M. Kamal., Hossain, M. Al., Yamauchi, Y., Nguyen, N., Lam, A. K. & Shiddiky, M. J. A. (2017). A PCR-free electrochemical method for messenger RNA detection in cancer tissue samples. Biosensors and Bioelectronics, 98 227-233.

Scopus Eid


  • 2-s2.0-85021751203

Ro Metadata Url


  • http://ro.uow.edu.au/aiimpapers/2628

Number Of Pages


  • 6

Start Page


  • 227

End Page


  • 233

Volume


  • 98

Place Of Publication


  • United States