Ultrasensitive detection of microRNAs based on hairpin fluorescence probe assisted isothermal amplification.
A hairpin fluorescence probe assisted isothermal amplification strategy was used for microRNAs (miRNAs) detection. The fluorescence hairpin probe was rationally designed by software NUPACK to reduce background signal. This isothermal amplification method consisted of two circuits. The amplification strategy not only could detect miRNA, but also amplified and reversely transcribed miRNA into DNA to enhance the stability of the target. The approach was ultrasensitive and as low as 8.5×10(-15)mol/L miR-Let-7a, corresponding to 8.5×10(-20)mol miR-Let-7a in 10µL, was able to be detected within 20min at 37°C. Moreover, successful detection of miR-Let-7a in a total RNA sample was also achieved. Thus, the rapid, simple, isothermal, and highly sensitive approach should be a promising tool for on-the-spot detection.
['Ma|Cuiping|C|', 'Liu|Sen|S|', 'Shi|Chao|C|']
D001483:Base Sequence D015374:Biosensing Techniques D004867:Equipment Design D019544:Equipment Failure Analysis D005456:Fluorescent Dyes D035683:MicroRNAs D008832:Microchemistry D008969:Molecular Sequence Data D021141:Nucleic Acid Amplification Techniques D013050:Spectrometry, Fluorescence D013696:Temperature
MicroRNAs Isothermal amplification Hairpin probe Cascade amplification Real-time fluorescence