Electric-Field Assisted Isothermal On-Chip Amplification of DNA

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DESCRIPTION (provided by applicant): Analysis of nucleic acids represents a well established molecular diagnostic test offering a highly specific detection of pathogens causing infectious diseases. Most clinical methods are based on the process of amplifying DNA and/or RNA molecules using polymerase chain reaction (PCR). The proposing team has recently developed another scheme for efficient isothermal amplification of DNA; the ramification amplification (RAM). This isothermal amplification is derived from primer extension, strand displacement, and the propagation of multiple ramification (branching) points. This technology utilizes a capture probe for target DNA or RNA isolation and a circularized RAM probe for subsequent amplification. Isothermal methods of DNA amplification have inherent advantages compared to the PCR method because there is no need to perform thermal cycling and can be easily packaged into a portable detection system. The proposed project combines two technologies, RAM amplification and electronic microarray technology where an electric field is used to assist and accelerate the DNA amplification process. The project is a joint effort in combining two compatible technologies and will leverage earlier efforts in RAM amplification and development of a portable DNA microarray detection system. The main advantage of the proposed approach is that a highly selective and sensitive RAM amplification will be integrated on the electronic microarray which will assure high level of multiplexing and rapid detection. Potential applications are envisioned in portable and/or point of care systems for detection of pathogens, infectious diseases and/or biological agents and emerging pathogens.