Visual and modular detection of pathogen nucleic acids with enzyme–DNA molecular complexes

Abstract

Rapid, visual detection of pathogen nucleic acids has broad applications in infection management. Here we present a modular detection platform, termed enzyme-assisted nanocomplexes for visual identification of nucleic acids (enVision). The system consists of an integrated circuit of enzyme–DNA nanostructures, which function as independent recognition and signaling elements, for direct and versatile detection of pathogen nucleic acids from infected cells. The built-in enzymatic cascades produce a rapid color readout for the naked eye; the assay is thus fast (<2 h), sensitive (<10 amol), and readily quantified with smartphones. When implemented on a configurable microfluidic platform, the technology demonstrates superior programmability to perform versatile computations, for detecting diverse pathogen targets and their virus–host genome integration loci. We further design the enVision platform for molecular-typing of infections in patient endocervical samples. The technology not only improves the clinical inter-subtype differentiation, but also expands the intra-subtype coverage to identify previously undetectable infections.

An ultraportable and versatile point-of-care DNA testing platform

“…Because of our combinatory and integrated approach, the POCKET system was extremely light and small, rendering the POCKET system, to the best of our knowledge, to be one of the smallest and most portable POC DNA testing devices. Compared with other 3D-printed or microfluidic chips, our POCKET system was more versatile for a broader range of possible sample sources and achieved multiplex detections with reduced reagent costs (table S6) (6,9,11,14,17,(27)(28)(29)(30)(31).…”

“…Over the last 5 years, POC DNA analysis research has progressed rapidly not only in clinical cares but also in agriculture, environmental protection, and food safety (3)(4)(5)(6). Two keys to the success of practical applications are the portability and versatility of POCT.…”

Dual-Selective Magnetic Analysis of Extracellular Vesicle Glycans

“…45 Its solution-phase magnetic actuation further improves assay performance and eases probe selection (e.g., lowaffinity binders) to identify weak but true interactions, 23 and could be readily integrated with other technologies to facilitate molecular assay development. 46,47 Such studies will not only improve biomarker characterization but also empower new applications. As demonstrated by the present study, the identification of a conserved EV glycan signature could enable the development of novel glycanbased approaches for ubiquitous EV enrichment from complex biofluids.…”

Single-step multivalent capture assay for nucleic acid detection with dual-affinity regulation using mutation inhibition and allosteric activation

“…The detection of cell-free DNA expression profiles at the genetic level enables broad liquid biopsy approaches in infection diagnostics,17 early childhood wheezing,18 fetal chromosome abnormalities,19 cancer management,20–25 etc. As cf-DNA is more chemically and bioenvironmentally stable than RNA, cf-DNA is particularly informative, suitable as a molecular biomarker 26,27.…”