title: Quantum dots and duplex-specific nuclease enabled ultrasensitive detection and serotyping of Dengue viruses in one step in a single tube.
abstract: Leveraging on the enzymatic processing of Dengue virus (DV) RNA hybridized quantum dot-capped DNA capture probes (QD-CPs), an ultrasensitive assay for the detection and serotyping of DVs is described in the report. Briefly, DV-specific DNA CPs are first capped by QDs and then conjugated to magnetic beads. In a sample solution, strands of DV RNA form heteroduplexes with the QD-CPs on the magnetic beads. The CPs together with the QDs in the heteroduplexes are subsequently cleaved off the magnetic beads by a duplex-specific nuclease (DSN), releasing the QDs to the solution, freeing the target RNA strands, and availing them for another around of hybridization with the remaining QD-CPs. After removing the magnetic beads along with unreacted (uncleaved) QD-CPs by using a permanent magnet, ultrasensitive fluorescent detection of DV is realized through the cleaved QDs. Serotyping of DV is accomplished by a judicious design of the QD-CPs. The assay combines excellent signal generation by the highly fluorescent QDs and the effortlessness of utilizing magnetic beads in the removal of the unreacted QD-CPs. The highly efficient DSN cleavage in conjunction with its excellent mismatch discrimination ability permits serotyping of DVs in one tube with excellent sensitivity and selectivity.
authors.affiliation: Department of Chemistry, National University of Singapore, Singapore 117543, Singapore.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.affiliation: Department of Chemistry, National University of Singapore, Singapore 117543, Singapore. Electronic address: chmgaoz@nus.edu.sg.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Gao
mesh_terms: D000818:Animals; D033381:Anomura / Q000201:enzymology; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods; D015342:DNA Probes / Q000737:chemistry*; D003715:Dengue / Q000175:diagnosis / Q000821:virology*; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D004720:Endonucleases / Q000737:chemistry; D006801:Humans; D059346:Magnets / Q000737:chemistry; D045663:Quantum Dots / Q000737:chemistry*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012703:Serotyping / Q000191:economics / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000191:economics / Q000379:methods
chemical_list: D015342:DNA Probes; D012367:RNA, Viral; D004720:Endonucleases
keywords: Biosensor.; Dengue virus; Duplex-specific nuclease; Fluorescence; Quantum dot