title: Ultrasensitive detection of microRNA with isothermal amplification and a time-resolved fluorescence sensor.
abstract: MicroRNAs (miRNAs) play important roles in a plethora of biological and cellular processes. The levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis, thus the method for sensitive and selective detection of miRNAs is imperative to miRNA discovery, study, and clinical diagnosis. Here we develop a novel method to quantify miRNA expression levels as low as attomolar sensitivity by two-stage exponential amplification reaction (EXPAR) and a time-resolved fluorescence sensor in real samples. The method reveals superior sensitivity with a detection limit of miRNA of 0.1 aM under pure conditions. The method also shows the high selectivity for discriminating differences between miRNA family members, thus providing a promising alternative to standard approaches for quantitative detection of miRNA.
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China. Electronic address: wangke@jsinm.org.
authors.forename: Ke
authors.initials: K
authors.lastname: Wang
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China. Electronic address: zhangkai@jsinm.org.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Wuxi People׳s Hospital Affiliated to Nanjing Medical University, Wuxi, Jiangsu 214023, China.
authors.forename: Zhuwu
authors.initials: Z
authors.lastname: Lv
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Save Sight Institute, University of Sydney, NSW 2000, Australia.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Faculty of Pharmacy, University of Sydney, NSW 2006, Australia.
authors.forename: Fanfan
authors.initials: F
authors.lastname: Zhou
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D057230:Limit of Detection; D035683:MicroRNAs / Q000032:analysis / Q000097:blood* / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D035683:MicroRNAs
keywords: Human serum; Isothermal Amplification; MiRNA; Time-resolved fluorescence