The ability to detect and quantify HIV RNA in blood is essential to sensitive detection of infections and monitoring viremia throughout treatment. Current options for point-of-care HIV diagnosis (i.e. lateral...
With a nearly 100% mortality rate,
African swine fever (ASF) has
devastated the pork industry in many countries. Without a vaccine
in sight, mitigation rests on rapid diagnosis and immediately depopulating
infected or exposed animals. Unfortunately, current tests require
centralized laboratories with well-trained personnel, take days to
report the results, and thus do not meet the need for such rapid diagnosis.
In response, we developed a portable, sample-to-answer device that
allows for ASF detection at the point of need in <30 min. The device
employs droplet magnetofluidics to automate DNA purification from
blood, tissue, or swab samples and utilizes fast thermal cycling to
perform real-time quantitative polymerase chain reaction (qPCR), all
within an inexpensive disposable cartridge. We evaluated its diagnostic
performance at six farms and slaughter facilities. The device exhibits
high diagnostic accuracy with a positive percent agreement of 92.2%
and a negative percent agreement of 93.6% compared with a lab-based
reference qPCR test.
Supporting InformationThe Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.0c04095. Experimental section: Chip design and fabrication, preparation of ready-to-use microwell array with preloaded antibiotics, preparation of bacteria samples, data analysis of marcAST result, evaluation of rehydration process of the microwell array device, and clinical isolate testing. Figures: Microwell device structure, demonstration of operation procedure with a 30 s step prior to AST, time course of marcAST with ciprofloxacin and E. coli ATCC 25922, MIC determination using benchtop in-tube broth dilution method, ready-to-use microwell array device shelf life, MIC determination of S. aureus ATCC 23922 with gentamicin, and classification of antimicrobial susceptibilities for additional clinical isolates. Tables: Interpretive categories and MIC breakpoints of four antibiotics for Enterobacteriaceae species based CLSI guidelines, and AST profiles of clinical isolates (n = 7).
The rise of highly transmissible SARS‐CoV‐2 variants brings new challenges and concerns with vaccine efficacy, diagnostic sensitivity, and public health responses to end the pandemic. Widespread detection of variants is critical to inform policy decisions to mitigate further spread, and postpandemic multiplexed screening of respiratory viruses will be necessary to properly manage patients presenting with similar respiratory symptoms. In this work, a portable, magnetofluidic cartridge platform for automated polymerase chain reaction testing in <30 min is developed. Cartridges are designed for multiplexed detection of SARS‐CoV‐2 with either identification of variant mutations or screening for Influenza A and B. Moreover, the platform can perform identification of B.1.1.7 and B.1.351 variants and the multiplexed SARS‐CoV‐2/Influenza assay using archived clinical nasopharyngeal swab eluates and saliva samples. This work illustrates a path toward affordable and immediate testing with potential to aid surveillance of viral variants and inform patient treatment.
Quantification of the relative abundance of genetic traits has broad applications for biomarker discovery, diagnostics, and assessing gene expression in biological research. Relative quantification of genes is traditionally done with...
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