title: Ultrasensitive cDNA detection of dengue virus RNA using electrochemical nanoporous membrane-based biosensor.
abstract: A nanoporous alumina membrane-based ultrasensitive DNA biosensor is constructed using 5'-aminated DNA probes immobilized onto the alumina channel walls. Alumina nanoporous membrane-like structure is carved over platinum wire electrode of 76 µm diameter dimension by electrochemical anodization. The hybridization of complementary target DNA with probe DNA molecules attached inside the pores influences the pore size and ionic conductivity. The biosensor demonstrates linear range over 6 order of magnitude with ultrasensitive detection limit of 9.55×10(-12) M for the quantification of ss-31 mer DNA sequence. Its applicability is challenged against real time cDNA PCR sample of dengue virus serotype1 derived from asymmetric PCR. Excellent specificity down to one nucleotide mismatch in target DNA sample of DENV3 is also demonstrated.
authors.affiliation: Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University, Singapore, Singapore.
authors.forename: Varun
authors.initials: V
authors.lastname: Rai
authors.forename: Hapuarachchige C
authors.initials: HC
authors.lastname: Hapuarachchi
authors.forename: Lee Ching
authors.initials: LC
authors.lastname: Ng
authors.forename: Siew Hwa
authors.initials: SH
authors.lastname: Soh
authors.forename: Yee Sin
authors.initials: YS
authors.lastname: Leo
authors.forename: Chee-Seng
authors.initials: CS
authors.lastname: Toh
mesh_terms: D000537:Aluminum Oxide / Q000737:chemistry; D020137:Base Pair Mismatch; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D003716:Dengue Virus / Q000145:classification / Q000235:genetics*; D004563:Electrochemistry; D016679:Genome, Viral / Q000235:genetics; D006358:Hot Temperature; D008567:Membranes, Artificial; D058608:Nanopores; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D004279:DNA, Viral; D008567:Membranes, Artificial; D012367:RNA, Viral; D000537:Aluminum Oxide