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About this entry
For the last 6 years the Yager (and Lutz) team has been pushing the limits of NAAT testing in an autonomous disposable device. In the DARPA-funded Multiplexable Autonomous Disposable Nucleic Acid Amplification Test “MAD NAAT” project (2011-2016), our goal was to port NAAT tests for DNA or RNA to a platform as simple to use as a pregnancy test, allowing non-trained personnel to perform the tests in a low-resource setting. The NAAT process used (isothermal strand displacement amplification, or iSDA) was adopted from our collaborators at Epoch Biosciences/ELITech. It is both fast (fluorescence lift-off at about 7 minutes) and sensitive down to 1 copy when performed in tube with well-designed primers. The MAD NAAT device combined enzymatic lysis of cells (using achromopeptidase (ACP)), followed by heating to 95°C to inactivate the enzymes, followed by reconstitution of dried iSDA master mix, then amplification of target sequences at ~49.5°C. Amplification products were detected by binding to Au nanoparticles, and flowing them over a nitrocellulose lateral flow strip that has been striped with capture probes.