Isothermal DNA amplification methods have great potential in point-of-care diagnostics aimed at identifying specific strains of infectious diseases. Most isothermal methods, however, require temperatures far above room temperature necessitating the use of a heating element, which complicates the point-of-care assay. We have developed an isothermal ligase chain reaction that achieves turnover without temperature cycling by incorporating a destabilizing lesion into one of the four primers used in the chain reaction. Not only does this strategy appear to be general, as we have demonstrated its applicability to a number of sequences, but the desired replication temperature can also be controlled by probe design. Specifically, we show that incorporating an additional destabilizing element like a mismatch or another abasic lesion into one of the three remaining primers allows the temperature to be tuned in a range that represents ambient temperatures (from 18 - 30 °C). Consequently, with this strategy we have achieved heat-free DNA amplification on the bench top. Our efforts at extending LIDA to other analytes besides DNA will also be discussed.

Achieving room temperature DNA detection with lesion-induced DNA amplification (LIDA)

251st National Meeting (2016)

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