Determination of microRNA-122 in hepatocytes by two-step amplification of duplex-specific nuclease with laser-induced fluorescence detection

Abstract

MicroRNAs (miRNAs) play important roles in physiological and pathological processes of cells. To develop a fast, simple and sensitive method to determine miRNAs is significant for miRNA studies. In this work, determination of microRNA-122 (miR-122) was achieved by laser-induced fluorescence (LIF) detection. A vial–LIF interface was first applied for sample analysis. A two-step amplification of the fluorescence signal for miR-122 was designed and realized by applying duplex-specific nuclease in the cleaving of two sensing probes. Under optimized conditions, the analysis of a miR-122 sample could be completed in less than 50 min. Only 10 μL sample was required for each test and the detection limit for the method was 0.60 pM equal to 1.2 amol of miR-122 in 10 μL solution. Lastly, the developed method was successfully applied to determine miR-122 in chicken and duck liver. The developed method was fast, selective, sensitive and sample-saving for the determination of miRNAs.