Surface Plasmon Resonance Imaging Analysis Of MicroRNA Based On Nanomaterials And Isothermal Amplification Technology

Non-coding RNA(ncRNA),a class of RNA,is capable of transcription but not encoding proteins,playing an important role in various diseases.It has been received particular attention and become the focus of the functional genomics.MicroRNA,a class of ncRNA,are known to be closely associated with the pathogenesis of human diseases,including acute kidney injury(AKI),polycystic kidney disease(PKD)and diabetic nephropathy(DN).The development of human diseases is often accompanied by abnormal expression of multiple microRNAs.Therefore,simultaneous and highly sensitive detection of multiple microRNAs facilitates clinicians? diagnosis and treatment of diseases.Surface plasmon resonance imaging(SPRi),a biosensing technology based on the principle of surface plasmon resonance(SPR),which integrates array sensor chip and charge-coupled device(CCD).It has been proven to be an attractivetechnology for simultaneous monitor of molecular interaction and quantitative detection of biomolecule owing to its advantages of label-free,real-time,and high-throughput detection.However,the sensitivity of SPRi biosensing method is not satisfying for direct detection of low abundance biomolecule in complex matrix.So in recent years,many researches have been reported to combine isothermal amplification technology with nucleic acid detection to achieve rapid,highly sensitive nucleic acid detection.In this study,a simple surface plasmon resonance imaging biosensing method was designed by integrating strand displacement amplification(SDA)and DNA functionalized gold nanoparticles.We chose the nephropathy related microRNA(microRNA-21,microRNA-192)as the target microRNA,the strand displacement amplification could be triggered by microRNA to generate a lot of triggers,and further amplified by DNA functionalized gold nanoparticles,providing an alternative tool for fast and sensitive multiple microRNAs detection.The established biosensor was capable of simultaneously detecting multiplex miRNAs with a limit of detection down to 0.15 pM.Moreover,the method exhibited good specificity,low matrix effect and acceptable reproducibility.Thus,this study proposed a simple SPRi biosensing method for simultaneous and rapid detection of multiple microRNAs,providing a potential alternative platform toward multiple microRNAs detection for early diagnosis,treatment,or prevention of clinical disease.