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Fast and accurate detection of nucleic acids is key for pathogen identification. Methods for DNA detection generally rely on fluorescent or colorimetric readout. The development of label-free assays decreases costs and test complexity. We present a novel method combining a one-pot isothermal generation of DNA nanoballs with their detection by electrical impedance. We modified loop-mediated isothermal amplification by using compaction oligonucleotides that self-assemble the amplified target into nanoballs. Next, we use capillary-driven flow to passively pass these nanoballs through a microfluidic impedance cytometer, thus enabling a fully compact system with no moving parts. The movement of individual nanoballs is detected by a change in impedance providing a quantized readout. This approach is flexible for the detection of DNA/RNA of numerous targets (severe acute respiratory syndrome coronavirus 2, HIV, β-lactamase gene, etc.), and we anticipate that its integration into a standalone device would provide an inexpensive (<$5), sensitive (10 target copies), and rapid test (<1 hour).more » « less
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Zhao, Yu; Coelho, Camila; Hughes, Amanda L; Lazar-Stefanita, Luciana; Yang, Sandy; Brooks, Aaron N; Walker, Roy SK; Zhang, Weimin; Lauer, Stephanie; Hernandez, Cindy; et al (, Cell)
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Schindler, Daniel; Walker, Roy SK; Jiang, Shuangying; Brooks, Aaron N; Wang, Yun; Müller, Carolin A; Cockram, Charlotte; Luo, Yisha; García, Alicia; Schraivogel, Daniel; et al (, Cell)
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McCulloch, Laura H; Sambasivam, Vijayan; Hughes, Amanda L; Annaluru, Narayana; Ramalingam, Sivaprakash; Fanfani, Viola; Lobzaev, Evgenii; Mitchell, Leslie A; Cai, Jitong; Anderson, Breeana G; et al (, Cell Genomics)
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Kaminski Strauss, Sivan; Schirman, Dvir; Jona, Ghil; Brooks, Aaron N.; Kunjapur, Aditya M.; Nguyen Ba, Alex N.; Flint, Alice; Solt, Andras; Mershin, Andreas; Dixit, Atray; et al (, PLOS Biology)
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