Enhanced and multiplexed detection via a novel RCA-aptamer based sensing system

ANYL 247

Jeffrey B-H. Tok, tok2@llnl.gov1, Nicholas O. Fischer, fischer29@llnl.gov2, Cheryl E. Dolan, dolan4@llnl.gov1, and Ted M. Tarasow, tarasow2@llnl.gov2. (1) Chemistry & Materials Science Directorate, Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA CA 94551, (2) Chemistry, Materials and Life Sciences Directorate, Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA 94550
To contain and possibly thwart potential domestic bioterrorism, it is highly desirable that low abundance of potential biological threat agents be accurately detected. Largely due to Polymerase Chain Reaction (PCR), significant progress has been made in the area of DNA detection. Recently, with the development of techniques such as “bio-barcodes” and immuno-PCR, giant strides have also been made in protein detection technology. However it still lags behind its DNA counterpart. Herein, the usage of SELEX will first be described to generate DNA aptamers as sensing ligands for the binding domain of the neurotoxin (Tet-C) produced by Clostridium tetani. Second, Rolling Circle Amplification (RCA) technique is being used with a novel aptamer-primer oligomer to enable a significantly enhanced detection sensitivity compared to a singly-labeled aptamer. This biodetection/diagnostic approach is easily amendable to a multiplex format as DNA aptamers against other pathogenic threat agents can easily be displayed on a fixed array, e.g. chip or well-plate, or in a suspended format, e.g individually-encoded beads or wires. Progress in these areas will be discussed.
 

Analytical Approaches: Sensors
8:30 AM-11:40 AM, Wednesday, 13 September 2006 Moscone Center -- Room 123, Oral

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006