A novel DNA binding protein-based platform for electrochemical detection of miRNA

Muhammad Umer; Nahian B. Aziz; Rabbee G. Mahmudunnabi; Yoon Bo Shim; Carlos Salomon; Muhammad J.A. Shiddiky

doi:10.1039/d1an00935d

A novel DNA binding protein-based platform for electrochemical detection of miRNA

Muhammad Umer
, Nahian B. Aziz
, Rabbee G. Mahmudunnabi
, Yoon Bo Shim
, Carlos Salomon
, Muhammad J.A. Shiddiky

Griffith University
Pusan National University
University of Queensland
University of Queensland Centre for Clinical Research

Research output: Contribution to journal › Article › peer-review

10 Citations (Scopus)

Abstract

We present a novel amplification-free sandwich type platform assay for electrochemical detection of miRNA. The assay is based on T4 DNA polymerase mediated synthesis of the p53 binding DNA sequence at the 3′ end of target miRNA. The resulting miRNA-DNA chimera is detected via an electrochemical sandwich hybridization assay where HRP-labelled p53 binds to its recognition sequence and an amperometric signal is generated by hydroquinone-mediated enzymatic reduction of H2O2. The limit of detection of our assay was estimated to be 22 fM with a linear dynamic range of 100 fM-1 nM. This new platform method of detecting miRNA shows superior performance to conventional electrochemical miRNA biosensors and has the potential for amplification-free analysis of miRNA with high specificity and sensitivity.

Original language	English
Pages (from-to)	5496-5501
Number of pages	6
Journal	Analyst
Volume	146
Issue number	18
Early online date	08 Jul 2021
DOIs	https://doi.org/10.1039/d1an00935d
Publication status	Published - 21 Sept 2021

Access to Document

10.1039/d1an00935d

Cite this

@article{4e9e45e7321b451d9a29a7855ca2171d,

title = "A novel DNA binding protein-based platform for electrochemical detection of miRNA",

abstract = "We present a novel amplification-free sandwich type platform assay for electrochemical detection of miRNA. The assay is based on T4 DNA polymerase mediated synthesis of the p53 binding DNA sequence at the 3′ end of target miRNA. The resulting miRNA-DNA chimera is detected via an electrochemical sandwich hybridization assay where HRP-labelled p53 binds to its recognition sequence and an amperometric signal is generated by hydroquinone-mediated enzymatic reduction of H2O2. The limit of detection of our assay was estimated to be 22 fM with a linear dynamic range of 100 fM-1 nM. This new platform method of detecting miRNA shows superior performance to conventional electrochemical miRNA biosensors and has the potential for amplification-free analysis of miRNA with high specificity and sensitivity.",

author = "Muhammad Umer and Aziz, \{Nahian B.\} and Mahmudunnabi, \{Rabbee G.\} and Shim, \{Yoon Bo\} and Carlos Salomon and Shiddiky, \{Muhammad J.A.\}",

note = "Publisher Copyright: {\textcopyright} The Royal Society of Chemistry.",

year = "2021",

month = sep,

day = "21",

doi = "10.1039/d1an00935d",

language = "English",

volume = "146",

pages = "5496--5501",

journal = "Analyst",

issn = "0003-2654",

publisher = "Royal Society of Chemistry",

number = "18",

}

TY - JOUR

T1 - A novel DNA binding protein-based platform for electrochemical detection of miRNA

AU - Umer, Muhammad

AU - Aziz, Nahian B.

AU - Mahmudunnabi, Rabbee G.

AU - Shim, Yoon Bo

AU - Salomon, Carlos

AU - Shiddiky, Muhammad J.A.

N1 - Publisher Copyright: © The Royal Society of Chemistry.

PY - 2021/9/21

Y1 - 2021/9/21

N2 - We present a novel amplification-free sandwich type platform assay for electrochemical detection of miRNA. The assay is based on T4 DNA polymerase mediated synthesis of the p53 binding DNA sequence at the 3′ end of target miRNA. The resulting miRNA-DNA chimera is detected via an electrochemical sandwich hybridization assay where HRP-labelled p53 binds to its recognition sequence and an amperometric signal is generated by hydroquinone-mediated enzymatic reduction of H2O2. The limit of detection of our assay was estimated to be 22 fM with a linear dynamic range of 100 fM-1 nM. This new platform method of detecting miRNA shows superior performance to conventional electrochemical miRNA biosensors and has the potential for amplification-free analysis of miRNA with high specificity and sensitivity.

AB - We present a novel amplification-free sandwich type platform assay for electrochemical detection of miRNA. The assay is based on T4 DNA polymerase mediated synthesis of the p53 binding DNA sequence at the 3′ end of target miRNA. The resulting miRNA-DNA chimera is detected via an electrochemical sandwich hybridization assay where HRP-labelled p53 binds to its recognition sequence and an amperometric signal is generated by hydroquinone-mediated enzymatic reduction of H2O2. The limit of detection of our assay was estimated to be 22 fM with a linear dynamic range of 100 fM-1 nM. This new platform method of detecting miRNA shows superior performance to conventional electrochemical miRNA biosensors and has the potential for amplification-free analysis of miRNA with high specificity and sensitivity.

UR - https://www.scopus.com/pages/publications/85115175252

UR - https://www.scopus.com/pages/publications/85115175252#tab=citedBy

U2 - 10.1039/d1an00935d

DO - 10.1039/d1an00935d

M3 - Article

C2 - 34346412

AN - SCOPUS:85115175252

SN - 0003-2654

VL - 146

SP - 5496

EP - 5501

JO - Analyst

JF - Analyst

IS - 18

ER -

A novel DNA binding protein-based platform for electrochemical detection of miRNA

Abstract

Access to Document

Other files and links

Fingerprint

Cite this