New sensitive and specific method for microRNA analysis

Abstract
MicroRNAs (miRNAs) are small non-coding RNAs that function as important biological regulators in viruses, plants, and animals (1). The single-stranded miRNAs regulate gene expression at the post-transcription level and the dysregulation of miRNAs is associated with various human diseases (2-4). Recent reports show microRNAs are abundant not only in tissues but also in body fluids and they show great potential as minimum-invasive biomarkers in the diagnosis and prognosis of cancers and other diseases (5, 6). However, there are two main challenges when analyzing miRNAs by molecular methods: 1) miRNAs are short, usually not more than 22 nucleotides, which is the length of conventional PCR primers; 2) closely related miRNAs may differ in only a single nucleotide position. Current methods approach these challenges by extending the length of the miRNA using either miRNA specific RT primers or by non-specific RT primers, which compromises specificity and sensitivity (7). Here we present a novel method for the detection and quantification of short nucleic acids that has higher sensitivity than current approaches with concomitant enhanced specificity.
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