TY - JOUR
T1 - A novel electrochemical biosensor based on dynamic polymerase-extending hybridization for E. coli O157:H7 DNA detection
AU - Wang, Lijiang
AU - Liu, Qingjun
AU - Hu, Zhaoying
AU - Zhang, Yuanfan
AU - Wu, Chunsheng
AU - Yang, Mo
AU - Wang, Ping
PY - 2009/5/15
Y1 - 2009/5/15
N2 - A novel biosensor based on single-stranded DNA (ssDNA) probe functionalized aluminum anodized oxide (AAO) nanopore membranes was demonstrated for Escherichia coli O157:H7 DNA detection. An original and dynamic polymerase-extending (PE) DNA hybridization procedure is proposed, where hybridization happens in the existence of Taq DNA polymerase and dNTPs under controlled reaction temperature. The probe strand would be extended as long as the target DNA strand, then the capability to block the ionic flow in the pores has been prominently enhanced by the double strand complex. We have investigated the variation of ionic conductivity during the fabrication of the film and the hybridization using cyclic voltammetry and impedance spectroscopy. The present approach provides low detection limit for DNA (a few hundreds of pmol), rapid label-free and easy-to-use bacteria detection, which holds the potential for future use in various ss-DNA analyses by integrated into a self-contained biochip.
AB - A novel biosensor based on single-stranded DNA (ssDNA) probe functionalized aluminum anodized oxide (AAO) nanopore membranes was demonstrated for Escherichia coli O157:H7 DNA detection. An original and dynamic polymerase-extending (PE) DNA hybridization procedure is proposed, where hybridization happens in the existence of Taq DNA polymerase and dNTPs under controlled reaction temperature. The probe strand would be extended as long as the target DNA strand, then the capability to block the ionic flow in the pores has been prominently enhanced by the double strand complex. We have investigated the variation of ionic conductivity during the fabrication of the film and the hybridization using cyclic voltammetry and impedance spectroscopy. The present approach provides low detection limit for DNA (a few hundreds of pmol), rapid label-free and easy-to-use bacteria detection, which holds the potential for future use in various ss-DNA analyses by integrated into a self-contained biochip.
KW - E. coli O157:H7
KW - Electrochemical biosensor
KW - Nanopore membrane
KW - Polymerase-extending DNA hybridization
UR - http://www.scopus.com/inward/record.url?scp=61649094674&partnerID=8YFLogxK
U2 - 10.1016/j.talanta.2008.12.001
DO - 10.1016/j.talanta.2008.12.001
M3 - Journal article
C2 - 19269406
SN - 0039-9140
VL - 78
SP - 647
EP - 652
JO - Talanta
JF - Talanta
IS - 3
ER -