Current DNA microarray devices are used primarily for detection of target sequences and require multiple steps prior to application of the amplicons to the microarray and subsequent hybridization of the reporter probes. The sample preparation steps encompass amplification in a thermocycler, desalting the amplified sample, and transfer of the sample to the microarray, either manually or robotically. The proposed research will eliminate these steps by developing a new isothermal on-chip strand displacement amplification (SDA) assay on an electronically active microarray. In addition, molecular beacons or intercalating dyes will be incorporated into the amplification to provide for the first time, real-time detection of target sequences using on-chip anchored SDA. This new assay system will provide an accurate, faster, and simpler method for the analysis of multiplexed biowarfare agents (BWA) and will ensure development of a low cost portable system for detection of BWA and/or emerging infectious diseases.
