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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aluminum Oxide / chemistry
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Base Pair Mismatch
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Base Sequence
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Biosensing Techniques / methods*
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DNA, Complementary / genetics*
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DNA, Viral / genetics
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Dengue Virus / classification
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Dengue Virus / genetics*
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Electrochemistry
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Genome, Viral / genetics
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Hot Temperature
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Membranes, Artificial*
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Nanopores*
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RNA, Viral / analysis*
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RNA, Viral / genetics
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Real-Time Polymerase Chain Reaction
Substances
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DNA, Complementary
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DNA, Viral
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Membranes, Artificial
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RNA, Viral
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Aluminum Oxide
Grants and funding
We thankfully acknowledge financial support from Nanyang Technological University grant, the Singapore Immunology Network-Agency for Science (SIgN-A*STAR) for research grant (SIgN 09-023), National Environment Agency and the Ministry of Education - NUS scholarship for VR. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.