Interfacial Effects Enable Droplet Actuation, Inhibition Relief, and Early Sensing of Polymerase Chain ReactionTechnology #ua15-071
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Background: Quantitative real-time polymerase chain reaction (qPCR) is a method of detecting DNA amplified via the polymerase chain reaction (PCR) in real time, rather than after the completion of the amplification reaction. qPCR is useful in point-of-care diagnostic diagnoses of certain diseases and conditions, however current qPCR methods (e.g. fluorescence detection) still require a relatively high number of amplification cycles to occur before detection and quantification are possible. Therefore improvements that allow for the detection of PCR and quantification of initial target amount to take place at an earlier amplification cycle, and thus at an earlier time, are advantageous because they enable healthcare providers to make faster decisions in point-of-care settings.
Invention: The present invention is a device and associated methods that improve qPCR, especially in point-of-care settings, by allowing detection of PCR amplification and quantification of initial target amount to occur at an earlier amplification cycle. Ultimately this allows for faster results, and thus faster informed medical decision-making in the point-of-care setting. This innovation is possible through the novel utilization of interfacial effects in PCR reaction droplets.
Applications: This device and methods are applicable to improving qPCR generally, and are especially pertinent to the improvement of qPCR as it is used in the point-of-care setting. For instance, this system has been used experimentally to differentiate antibiotic-resistant bacteria from antibiotic-sensitive bacteria in as little as 5 minutes.
Advantages: This technology permits PCR amplification detection and quantification of initial target amount earlier in the PCR reaction, and thus provides healthcare professionals in the point-of-care setting with the ability to make critical decisions more quickly. Detection of PCR amplification and quantification of initial target amount can occur as early as the fifth thermocycle, and sample preparation requirements are also reduced, meaning that detection and quantification can occur within 5 minutes of obtaining a sample. Further, the present technology allows for each thermocycle to occur faster than existing methods, separates contaminant proteins that can slow PCR from PCR reactants, and allows for sub-picogram limits of detection.
Lead Inventors: Jeong-Yeol Yoon, Dustin Harshman