Short non-coding RNAs as bacteria species identifiers detected bysurface plasmon resonance enhanced common path interferometry

摘要

Small non-coding RNA sequences have recently been discovered as unique identifiers of certain bacterial species,raising the possibility that they can be used as highly specific Biowarfare Agent detection markers in automated fielddeployable integrated detection systems. Because they are present in high abundance they could allow genomic basedbacterial species identification without the need for pre-assay amplification. Further, a direct detection method wouldobviate the need for chemical labeling, enabling a rapid, efficient, high sensitivity mechanism for bacterial detection.Surface Plasmon Resonance enhanced Common Path Interferometry (SPR-CPI) is a potentially market disruptive, highsensitivity dual technology that allows real-time direct multiplex measurement of biomolecule interactions, includingsmall molecules, nucleic acids, proteins, and microbes. SPR-CPI measures differences in phase shift of reflected S and Ppolarized light under Total Internal Reflection (TIR) conditions at a surface, caused by changes in refractive indexinduced by biomolecular interactions within the evanescent field at the TIR interface. The measurement is performed ona microarray of discrete 2-dimensional areas functionalized with biomolecule capture reagents, allowing simultaneousmeasurement of up to 100 separate analytes. The optical beam encompasses the entire microarray, allowing a solid statedetector system with no scanning requirement. Output consists of simultaneous voltage measurements proportional tothe phase differences resulting from the refractive index changes from each microarray feature, and is automaticallyprocessed and displayed graphically or delivered to a decision making algorithm, enabling a fully automatic detectionsystem capable of rapid detection and quantification of small nucleic acids at extremely sensitive levels. Proof-of-concept experiments on model systems and cell culture samples have demonstrated utility of the system, and efforts arein progress for full development and deployment of the device. The technology has broad applicability as a universaldetection platform for BWA detection, medical diagnostics, and drug discovery research, and represents a new class ofinstrumentation as a rapid, high sensitivity, label-free methodology.