A highly sensitive electrochemical assay for microRNA expression profiling

The Analyst
Zhiqiang Gao

Abstract

A simple and highly sensitive electrochemical assay for ligation-free and polymerase chain reaction (PCR)-free microRNA (miRNA) expression profiling is described in this work. The electrode used in the assay was made of a monolayer of stem-looped capture probes (CPs) comprising of a miRNA complementing region at one end and detection probes (DPs) receiving region at the other. It engaged an electrocatalytic reaction between electrochemically activated glucose oxidase (GOx) and glucose to enhance its sensitivity. Briefly, upon hybridizing to its target miRNA, the stem loop is unlocked exposing the DP receiving region. A subsequent hybridization with the DPs brought them together with an amplifier, the activated GOx, onto the electrode. The activated GOx exhibited excellent catalytic activity towards electrooxidation of glucose. MicroRNA detection could therefore be conducted in 60 mM glucose in phosphate-buffered saline. A detection limit of 4.0 fM and a linear calibration curve up to 10 pM were obtained under optimal conditions. The assay was applied to profile human let-7 miRNA expressions in cultured cancer cells.

References

Dec 9, 1998·Analytical Chemistry·A B SteelM J Tarlov
Oct 27, 2001·Science·M Lagos-QuintanaT Tuschl
Apr 4, 2003·Genes & Development·Lee P LimDavid P Bartel
Sep 26, 2003·Nature·Yoontae LeeV Narry Kim
Feb 27, 2004·Nucleic Acids Research·Thomas D SchmittgenLiuqing Yang
Apr 3, 2004·Genome Génome / Conseil National De Recherches Canada·Ke-Jing WangYoshihito Takahata
Jun 23, 2004·RNA·Hatim T AllawiVictor I Lyamichev
Jun 24, 2004·Proceedings of the National Academy of Sciences of the United States of America·Chang-Gong LiuCarlo M Croce
Jul 31, 2004·Proceedings of the National Academy of Sciences of the United States of America·George Adrian CalinCarlo M Croce
Mar 23, 2005·Nature Methods·Peter T NelsonZissimos Mourelatos
Jun 23, 2005·Frontiers in Bioscience : a Journal and Virtual Library·Hong Xie, Zhiqiang Gao
Oct 21, 2005·BioTechniques·Rui Shi, Vincent L Chiang
Nov 30, 2005·Nucleic Acids Research·Caifu ChenKarl J Guegler
Mar 1, 2006·Analytical Chemistry·Zhiqiang Gao, Zichao Yang
Apr 7, 2007·Journal of the American Chemical Society·Yi FanZhiqiang Gao
Nov 13, 2007·Angewandte Chemie·Alastair W WarkRobert M Corn
Dec 26, 2007·Methods : a Companion to Methods in Enzymology·Thomas D Schmittgen
Dec 26, 2007·Methods : a Companion to Methods in Enzymology·Thomas D SchmittgenCaifu Chen
Jan 15, 2008·Trends in Biotechnology·James Q YinKevin V Morris
Aug 4, 2010·Analytical and Bioanalytical Chemistry·Abraham J QaviRyan C Bailey
Mar 23, 2011·Biosensors & Bioelectronics·Zhiqiang Gao, Yanfen Peng

❮ Previous
Next ❯

Citations

May 24, 2013·Chemical Reviews·Haifeng DongXueji Zhang
Jan 7, 2015·Biosensors & Bioelectronics·Mahmoud Labib, Maxim V Berezovski
Dec 24, 2014·Bioanalysis·Ilaria Palchetti
Oct 31, 2018·The Analyst·Philip GillespieDanny O'Hare

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.