title: Label-free detection of microRNA based on coupling multiple isothermal amplification techniques.
abstract: MicroRNA (miRNA) was a promising class of cancer biomarkers. Here we developed a label-free method for sensitive measurement of let-7d miRNA based on multiple amplification techniques. The primer will bind to the duplex strand DNA that was formed by stem-loop template and target let-7d to initiate strand displacement amplification (SDA) in tandem. The released single strand DNA will be a primer to bind the circular template to initiate rolling circle amplification (RCA). The products based on multiple amplifications will be detected by a standard fluorimeter with N-methyl mesoporphyrin IX (NMM) as the fluorescent indicator. The proposed method exhibited excellent selectivity and high sensitivity with a detection limit of as low as 1.5 × 10-13M. Moreover, this methodology was used for the determination of biomolecules in real serum samples with satisfying results.
authors: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, P. R. China.|Shandong Provincial Key Laboratory of Detection Technology for Tumor Makers, Research Institute of Biochemical Analysis, Linyi University, Linyi 276000, P. R. China.
authors: Xiangjiang
authors: X
authors: Zheng
authors: Shandong Provincial Key Laboratory of Detection Technology for Tumor Makers, Research Institute of Biochemical Analysis, Linyi University, Linyi 276000, P. R. China.
authors: Li
authors: L
authors: Niu
authors: Shandong Provincial Key Laboratory of Detection Technology for Tumor Makers, Research Institute of Biochemical Analysis, Linyi University, Linyi 276000, P. R. China.
authors: Di
authors: D
authors: Wei
authors: Shandong Provincial Key Laboratory of Detection Technology for Tumor Makers, Research Institute of Biochemical Analysis, Linyi University, Linyi 276000, P. R. China.
authors: Xuemei
authors: X
authors: Li
authors: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, P. R. China.
authors: Shusheng
authors: S
authors: Zhang
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D009369:Neoplasms / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs; C487931:mirnlet7 microRNA, human