» Articles » PMID: 22788545

Direct Quantification of MicroRNA at Low Picomolar Level in Sera of Glioma Patients Using a Competitive Hybridization Followed by Amplified Voltammetric Detection

Overview
Journal Anal Chem
Specialty Chemistry
Date 2012 Jul 14
PMID 22788545
Citations 14
Authors
Affiliations
Soon will be listed here.
Abstract

MicroRNAs (miRNAs), acting as oncogenes or tumor suppressors in humans, play a key role in regulating gene expression and are believed to be important for developing novel therapeutic treatments and clinical prognoses. Due to their short lengths (17-25 nucleotides) and extremely low concentrations (typically < picomolar) in biological samples, quantification of miRNAs has been challenging to conventional biochemical methods, such as Northern blotting, microarray, and quantitative polymerase chain reaction (qPCR). In this work, a biotinylated miRNA (biotin-miRNA) whose sequence is the same as that of a miRNA target is introduced into samples of interest and allowed to compete with the miRNA target for the oligonucleotide (ODN) probe preimmobilized onto an electrode. Voltammetric quantification of the miRNA target was accomplished after complexation of the biotin-miRNA with ferrocene (Fc)-capped gold nanoparticle/streptavidin conjugates. The Fc oxidation current was found to be inversely proportional to the concentration of target miRNA between 10 fM and 2.0 pM. The method is highly reproducible (relative standard deviation (RSD) < 5%), regenerable (at least 8 regeneration/assay cycles without discernible signal decrease), and selective (with sequence specificity down to a single nucleotide mismatch). The low detection levels (10 fM or 0.1 attomoles of miRNA in a 10 μL solution) allow the direct quantification of miRNA-182, a marker correlated to the progression of glioma in patients, to be performed in serum samples without sample pretreatment and RNA extraction and enrichment. The concentration of miRNA-182 in glioma patients was found to be 3.1 times as high as that in healthy persons, a conclusion in excellent agreement with a separate qPCR measurement of the expression level. The obviations of the requirement of an internal reference in qPCR, simplicity, and cost-effectiveness are other additional advantages of this method for detection of nucleic acids in clinical samples.

Citing Articles

Multiplexed smFRET Nucleic Acid Sensing Using DNA Nanotweezers.

Kaur A, Mahmoud R, Megalathan A, Pettit S, Dhakal S Biosensors (Basel). 2023; 13(1).

PMID: 36671954 PMC: 9856376. DOI: 10.3390/bios13010119.


DNA walking system integrated with enzymatic cleavage reaction for sensitive surface plasmon resonance detection of miRNA.

Chen S, He Y, Liu L, Wang J, Yi X Sci Rep. 2022; 12(1):16093.

PMID: 36167754 PMC: 9515148. DOI: 10.1038/s41598-022-20453-8.


Blood-Based Biomarkers for Glioma in the Context of Gliomagenesis: A Systematic Review.

Ali H, Harting R, de Vries R, Ali M, Wurdinger T, Best M Front Oncol. 2021; 11:665235.

PMID: 34150629 PMC: 8211985. DOI: 10.3389/fonc.2021.665235.


Hybridizing clinical translatability with enzyme-free DNA signal amplifiers: recent advances in nucleic acid detection and imaging.

Borum R, Jokerst J Biomater Sci. 2020; 9(2):347-366.

PMID: 32734995 PMC: 7855509. DOI: 10.1039/d0bm00931h.


Voltammetric determination of the Alzheimer's disease-related ApoE 4 gene from unamplified genomic DNA extracts by ferrocene-capped gold nanoparticles.

Lu H, Wu L, Wang J, Wang Z, Yi X, Wang J Mikrochim Acta. 2018; 185(12):549.

PMID: 30426239 DOI: 10.1007/s00604-018-3087-9.


References
1.
Asadi-Moghaddam K, Chiocca E, Lawler S . Potential role of miRNAs and their inhibitors in glioma treatment. Expert Rev Anticancer Ther. 2010; 10(11):1753-62. PMC: 7330712. DOI: 10.1586/era.10.168. View

2.
Zhou W, Chen Y, Corn R . Ultrasensitive microarray detection of short RNA sequences with enzymatically modified nanoparticles and surface plasmon resonance imaging measurements. Anal Chem. 2011; 83(10):3897-902. PMC: 3097511. DOI: 10.1021/ac200422u. View

3.
Zhang J, Fu Y, Mei Y, Jiang F, Lakowicz J . Fluorescent metal nanoshell probe to detect single miRNA in lung cancer cell. Anal Chem. 2010; 82(11):4464-71. PMC: 2878973. DOI: 10.1021/ac100241f. View

4.
Zhang G, Chua J, Chee R, Agarwal A, Wong S . Label-free direct detection of MiRNAs with silicon nanowire biosensors. Biosens Bioelectron. 2009; 24(8):2504-8. DOI: 10.1016/j.bios.2008.12.035. View

5.
Yu C, Wan Y, Yowanto H, Li J, Tao C, James M . Electronic detection of single-base mismatches in DNA with ferrocene-modified probes. J Am Chem Soc. 2001; 123(45):11155-61. DOI: 10.1021/ja010045f. View