title: Double-Stranded Ligation Assay for the Rapid Multiplex Quantification of MicroRNAs.
abstract: MicroRNAs are auspicious candidates for a new generation of biomarkers. The detection of microRNA panels in body fluids promises early diagnosis of many diseases, including cancer or acute coronary syndrome. For a fast, sensitive, and specific detection of microRNA panels on the bedside, medical point-of-care systems that measure those biomarkers are required. As microchips are promising technical tools for a robust signal measurement at biochemical interfaces, we developed an assay for the electrochemical multiplex quantification of microRNAs on a CMOS chip with interdigitated gold electrode sensor positions. The method is based on the formation of a tripartite hybridization complex and subsequent both-sided ligation of the target nucleic acid to a reporter and capture strand. With a time to results of 30 min, the reported assay achieves a limit of detection below 1 pM, at a specificity down to single mismatch discrimination. It also offers very good signal dynamics between 1 pM and 1 nM, thus, allowing reliable quantification of the detected microRNAs and easy implementation into automated devices due to a simple workflow.
authors.affiliation: Department of Bioinformatics, University of Würzburg , Würzburg, 97074, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hofmann
authors.affiliation: Technology Center, Siemens Healthcare, Erlangen, 91058, Germany.
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Technology Center, Siemens Healthcare, Erlangen, 91058, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Paulicka
authors.affiliation: Technology Center, Siemens Healthcare, Erlangen, 91058, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kappel
authors.affiliation: Internal Medicine III, University Hospital Heidelberg , Heidelberg, 69120, Germany.
authors.forename: Hugo A
authors.initials: HA
authors.lastname: Katus
authors.affiliation: Clinical Bioinformatics, Medical Faculty, Saarland University , Saarbrücken, 66123, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Keller
authors.affiliation: Internal Medicine III, University Hospital Heidelberg , Heidelberg, 69120, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Meder
authors.affiliation: Strategy, Siemens Healthcare, Erlangen, 91052, Germany.
authors.forename: Cord Friedrich
authors.initials: CF
authors.lastname: Stähler
authors.affiliation: Technology Center, Siemens Healthcare, Erlangen, 91058, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Gumbrecht
mesh_terms: D005821:Genetic Techniques; D057230:Limit of Detection; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D013997:Time Factors
chemical_list: D035683:MicroRNAs