A novel and integrated membrane sensing platform for DNA detection is developed based on an anodic aluminum oxide (AAO) membrane. Platinum electrodes (~50–100 nm thick) are coated directly on both sides of the alumina membrane to eliminate the solution resistance outside the nanopores. The electrochemical impedance technique is employed to monitor the impedance changes within the nanopores upon DNA binding. Pore resistance ( R p) linearly increases in response towards the increasing concentration of the target DNA in the range of 1 × 10 ？12 to 1 × 10 ？6 M. Moreover, the biosensor selectively differentiates the complementary sequence from single base mismatched (MM-1) strands and non-complementary strands. This study reveals a simple, selective and sensitive method to fabricate a label-free DNA biosensor.
References
[1]
Bian, G.; Xu, Y.; Lu, P.; Xie, Y.; Xi, Z. The endosymbiotic bacterium Wolbachia induces resistance to dengue virus in Aedes aegypti. PLoS Pathog. 2010, 6, doi:10.1371/journal.ppat.1000833.
[2]
Wilson, M.E.; Chen, L.H. Dengue in the Americas. Dengue Bull. 2002, 26, 44–61.
[3]
Vaughn, D.W.; Nisalak, A.; Solomon, T.; Kalayanarooj, S.; Nguyen, M.D.; Kneen, R.; Cuzzubbo, A.; Devine, P.L. Rapid serologic diagnosis of dengue virus infection using a commercial capyure ELISA that distinguishes primary and secondary infections. Am. J. Trop. Med. Hyg. 1999, 60, 693–698.
[4]
Conceicao, T.M.; da Poian, A.T.; Sorgine, M.H.F. A real-time PCR procedure for detection of dengue virus serotypes 1, 2, and 3, and their quantitation in clinical and laboratory samples. J. Virol. Methods 2010, 163, 1–9.
[5]
De la Escosura-Mu？iz, A.; Merko？i, A. Nanochannels preparation and application in biosensing. ACS Nano 2012, 6, 7556–7583.
[6]
Santos, A.; Kumeria, T.; Losic, D. Nanoporous anodic aluminum oxide for chemical sensing and biosensors. Trends Anal. Chem. 2013, 44, 25–38.
[7]
Wang, L.; Liu, Q.; Hu, Z.; Zhang, Y.; Wu, C.; Yang, M.; Wang, P. A novel electrochemical biosensor based on dynamic polymerase-extending hybridization for E. coli O157:H7 DNA detection. Talanta 2009, 78, 647–652.
[8]
Takmakov, P.; Vlassiouk, I.; Smirnov, S. Hydrothermally shrunk alumina nanopores and their application to DNA sensing. Analyst 2006, 131, 1248–1253.
Rai, V.; Hapuarachchi, H.C.; Ng, L.C.; Soh, S.H.; Leo, Y.S.; Toh, C.-S. Ultrasensitive cDNA detection of dengue virus RNA using electrochemical nanoporous membrane-based biosensor. PLoS One 2012, 7, doi:10.1371/journal.pone.0042346.
[12]
Rai, V.; Deng, J.; Toh, C.-S. Electrochemical nanoporous alumina membrane-based label-free DNA biosensor for the detection of Legionella sp. Talanta 2012, 98, 112–117.
[13]
Souza, E.; Nascimento, G.; Santana, N.; Ferreira, D.; Lima, M.; Natividade, E.; Martins, D.; Lima-Filho, J. Label-free electrochemical detection of the specific oligonucleotide sequence of dengue virus type 1 on pencil graphite electrodes. Sensors 2011, 11, 5616–5629.
[14]
Cai, H.; Wang, Y.; He, P.; Fang, Y. Electrochemical detection of DNA hybridization based on silver-enhanced gold nanoparticle label. Anal. Chim. Acta 2002, 469, 165–172.
[15]
De la Escosura-Mu？iz, A.; Merko？i, A. Nanoparticle based enhancement of electrochemical DNA hybridization signal using nanoporous electrodes. Chem. Comm. 2010, 46, 9007–9009.
[16]
Wu, Z.-S.; Jiang, J.-H.; Fu, L.; Shen, G.-L.; Yu, R.-Q. Optical detection of DNA hybridization based on fluorescence quenching of tagged oligonucleotide probes by gold nanoparticles. Anal. Biochem. 2006, 353, 22–29.
[17]
Touahir, L.; Elisabeth, G.; Boukherroub, R.; Gouget-Laemmel, A.C.; Chazalviel, J.-N.; Ozanam, F.; Szunerits, S. Localized surface plasmon-enhanced fluorescence spectroscopy for highly-sensitive real-time detection of DNA hybridization. Biosens. Bioelectron. 2010, 25, 2579–2585.
[18]
Kick, A.; B？nsch, M.; Katzschner, B.; Voigt, J.; Herr, A.; Brabetz, W.; Jung, M.; Sonntag, F.; Klotzbach, U.; Danz, N.; et al. DNA microarrays for hybridization detection by surface Plasmon resonance spectroscopy. Biosens. Bioelectron. 2010, 26, 1543–1547.
[19]
Wang, Q.; Yang, X.; Wang, K. Enhanced surface Plasmon resonance for detection of DNA hybridization based on layer-by-layer assembly films. Sens.Actuators B 2007, 123, 227–232.
[20]
Su, X.; Wu, Y.-J.; Knoll, W. Comparison of surface Plasmon resonance spectroscopy and quartz crystal microbalance techniques for studying DNA assembly and hybridization. Biosens. Bioelectron. 2005, 21, 719–726.
[21]
Li, S.; Li, X.; Zhang, J.; Zhang, Y.; Han, J.; Jiang, L. Quantitative investigation of the influence of gold nanoparticles on the dynamics of DNA hybridization using a programmed multi-channel quartz crystal microbalance system. Colloids Surf. A 2010, 364, 158–162.
[22]
Wang, X.; Smirnov, S. Label-free DNA sensor based on surface charge modulated ionic conductance. ACS Nano 2009, 3, 1004–1010.
[23]
Ito, T.; Hosokawa, K.; Maeda, M. Detection of single-base mismatch at distal end of DNA duplex by electrochemical impedance spectroscopy. Biosens. Bioelectron. 2007, 22, 1816–1819.
[24]
Vlassiouk, I.; Krasnoslobodtsev, A.; Smirnov, S.; Germann, M. “Direct” detection and separation of DNA using nanoporous alumina filters. Langmuir 2004, 20, 9913–9915.
[25]
Vlassiouk, I.; Takmakov, P.; Smirnov, S. Sensing DNA hybridization via ionic conductance through a nanoporous electrode. Langmuir 2005, 21, 4776–4778.
