+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Amplification-free detection of microRNAs via a rapid microarray-based sandwich assay



Amplification-free detection of microRNAs via a rapid microarray-based sandwich assay



Analytical and Bioanalytical Chemistry 409(14): 3497-3505



The detection and profiling of microRNAs are of great interest in disease diagnosis and prognosis. In this paper, we present a method for the rapid amplification-free detection of microRNAs from total RNA samples. In a two-step sandwich assay approach, fluorescently labeled reporter probes were first hybridized with their corresponding target microRNAs. The reaction mix was then added to a microarray to enable their specific capture and detection. Reporter probes were Tm equalized, enabling specificity by adjusting the length of the capture probe while maintaining the stabilizing effect brought about by coaxial base stacking. The optimized assay can specifically detect microRNAs in spiked samples at concentrations as low as 1 pM and from as little as 100 ng of total RNA in 2 h. The detection signal was linear between 1 and 100 pM (R2 = 0.99). Our assay data correlated well with results generated by qPCR when we profiled a select number of breast cancer related microRNAs in a total RNA sample.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 059349931

Download citation: RISBibTeXText

PMID: 28349168

DOI: 10.1007/s00216-017-0298-6


Related references

Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay. Bmc Biotechnology 10: 74, 2010

An enzyme-free flow cytometric bead assay for the sensitive detection of microRNAs based on click nucleic acid ligation-mediated signal amplification. Analyst 142(16): 2967-2973, 2017

Rapid and equipment-free detection of Phytophthora capsici using lateral flow strip-based recombinase polymerase amplification assay. Letters in Applied Microbiology 69(1): 64-70, 2019

Multiplexed detection of microRNAs by a competitive DNA microarray-based resonance light scattering assay. Analyst 142(23): 4529-4535, 2017

Label-free quantification of microRNAs using ligase-assisted sandwich hybridization on a DNA microarray. Plos one 9(3): E90920, 2014

Detection and Quantification of MicroRNAs by Ligase-Assisted Sandwich Hybridization on a Microarray. Methods in Molecular Biology 1368: 53-65, 2016

Sensitive detection of microRNAs with hairpin probe-based circular exponential amplification assay. Analytical Chemistry 84(16): 7037-7042, 2012

Ultrasensitive Electrochemical Detection of Glycoprotein Based on Boronate Affinity Sandwich Assay and Signal Amplification with Functionalized SiO 2 @Au Nanocomposites. Acs Applied Materials and Interfaces 9(16): 13855-13864, 2017

Dumbbell probe-mediated cascade isothermal amplification: a novel strategy for label-free detection of microRNAs and its application to real sample assay. Analytica Chimica Acta 760: 69-74, 2013

Microsphere-based rolling circle amplification microarray for the detection of DNA and proteins in a single assay. Analytical Chemistry 81(14): 5777-5782, 2009

Rapid and sensitive detection of norovirus genomes in oysters by a two-step isothermal amplification assay system combining nucleic acid sequence-based amplification and reverse transcription-loop-mediated isothermal amplification assays. Applied and Environmental Microbiology 74(12): 3912-3914, 2008

Rapid and label-free amplification and detection assay for genotyping of cancer biomarker. Biosensors and Bioelectronics 68: 107-114, 2015

Enzyme-free detection of sequence-specific microRNAs based on nanoparticle-assisted signal amplification strategy. Biosensors and Bioelectronics 77: 995, 2016

Non-instrumented DNA isolation, amplification and microarray-based hybridization for a rapid on-site detection of devastating Phytophthora kernoviae. Analyst 140(19): 6610-6618, 2015

A simple and label-free electrochemical biosensor for DNA detection based on the super-sandwich assay. Analyst 137(21): 5071-5075, 2012