Isothermal Loop-Mediated Amplification (LAMP) is a nucleic amplification method that has been widely used in diagnostic and field applications due to its simplicity of requiring a constant temperature, use of up to 4 to 6 primers (rendering it highly specific), and capability of amplifying low initial copies of target sequences. The integrative smartphone-based platform, captures images at set time points and measures contact angle over time. The transient change in contact angle detects saturation of LAMP amplicons at the oil-water interface to detect target and nontarget presence. The developed assay has shown endpoint results based on changes in contact angle measurements and Monod mathematical model fitted-constants to differentiate between target and nontarget samples. Furthermore, aqueous LAMP reactions were suspended in pre-heated mineral oil-surfactant mixtures and measured for interfacial tension over time. The developed assay and droplet LAMP platform has been demonstrated to detect targets in the form of whole bacteria, and with promising results for implementation and detection in more biologically complex matrices such as human blood serum and whole blood for modeling of sepsis. The developed integrative smartphone-based platform encompases microreactor droplets and rapid end point detection for possible translation to high-throughput and automated systems such as microfluidic devices, flow cytometry, and microplate instrumentations.

Real-time quantification of droplet isothermal nucleic acid amplification through measurements of interfacial effects

255th National Meeting (2018)

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