HIV molecular detection plays a significant role in early diagnosis and antiretroviral therapy for HIV patients. CRISPR technology has recently emerged as a powerful tool for highly sensitive and specific nucleic acid based molecular detection when used in combination with isothermal amplification. However, it remains a challenge to improve the compatibility of such a multienzyme reaction system for simple and sensitive molecular detection. Inspired by the multicompartment structures in a living cell, we present a nanoporous membrane-separated (compartmentalized), artificial, cascade reaction system to improve the compatibility of a CRISPR-mediated multienzyme reaction. We further integrated the multienzyme cascade reaction system with a microfluidic platform and glucose biosensing technology to develop a bioinspired, CRISPR-mediated cascade reaction (CRISPR-MCR) biosensor, enabling HIV molecular detection by a simple glucose meter, analogous to diabetes home testing. We applied the bioinspired CRISPR-MCR biosensor to detect HIV DNA and HIV RNA, achieving a detection sensitivity of 43 copies and 200 copies per test, respectively. Further, we successfully validated the bioinspired biosensor by testing clinical plasma samples of HIV, demonstrating its great application potential for point-of-care testing of HIV virus and other pathogens at home or in resource-limited settings.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10198471PMC
http://dx.doi.org/10.1021/acsnano.2c12754DOI Listing

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