title: Multiplexed microRNA detection by capillary electrophoresis with laser-induced fluorescence.
abstract: In this study, we developed a novel assay that simultaneously detects multiple miRNAs (microRNAs) within a single capillary by combining a tandem adenosine-tailed DNA bridge-assisted splinted ligation with denaturing capillary gel electrophoresis with laser-induced fluorescence. This proposed method not only represents a significant improvement in resolution but also allows for the detection of multiple miRNAs within a single capillary based on the length differences of specified target bridge DNA. The assay's linear range covers three orders of magnitude (1.0 nM to 1.0 pM) with a limit of detection (S/N=3) as low as 190 fM (2.5 zmol). Five miRNAs of Epstein-Barr virus (EBV) were also detected in EBV-infected nasopharyngeal carcinoma cells, while they did not appear in non-virus infected cells. Moreover, the electropherogram indicated that the screening of isomiRs (isomer of miRNA) of BART2 by CE-LIF is feasible by our proposed method. The developed electrophoresis-based method for miRNA detection is fast, amplification-free, multiplexed and cost-effective, making it potentially applicable to large-scale screening of isomiRs.
authors.affiliation: Department of Chemistry, Tunghai University, Taitung, Taiwan.
authors.forename: Ruei-Min
authors.initials: RM
authors.lastname: Jiang
authors.forename: Yu-Sun
authors.initials: YS
authors.lastname: Chang
authors.forename: Shu-Jen
authors.initials: SJ
authors.lastname: Chen
authors.forename: Jian-Hung
authors.initials: JH
authors.lastname: Chen
authors.forename: Hua-Chien
authors.initials: HC
authors.lastname: Chen
authors.forename: Po-Ling
authors.initials: PL
authors.lastname: Chang
mesh_terms: D000241:Adenosine / Q000378:metabolism; D002277:Carcinoma; D019075:Electrophoresis, Capillary / Q000379:methods*; D000431:Ethanol; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016014:Linear Models; D035683:MicroRNAs / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000821:virology; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured
chemical_list: D035683:MicroRNAs; D000431:Ethanol; D000241:Adenosine