Portable digital microfluidic serological immunoassays for measles and rubella were developed and evaluated in a remote setting.
We introduce Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes. Specifically, DISCO combines digital microfluidics, laser cell lysis, and artificial intelligence-driven image processing to collect the contents of single cells from heterogeneous populations, followed by analysis of single-cell genomes and transcriptomes by next-generation sequencing, and proteomes by nanoflow liquid chromatography and tandem mass spectrometry. The results described herein confirm the utility of DISCO for sequencing at levels that are equivalent to or enhanced relative to the state of the art, capable of identifying features at the level of single nucleotide variations. The unique levels of selectivity, context, and accountability of DISCO suggest potential utility for deep analysis of any rare cell population with contextual dependencies.
Nanostructured microelectrodes (NMEs) are three-dimensional electrodes that have superb sensitivity for electroanalysis. Here we report the integration of NMEs with the versatile fluid-handling system digital microfluidics (DMF), for eventual application to distributed diagnostics outside of the laboratory. In the new methods reported here, indium tin oxide DMF top plates were modified to include Au NMEs as well as counter and pseudoreference electrodes. The new system was observed to outperform planar sensing electrodes of the type that are typically integrated with DMF. A rubella virus (RV) IgG immunoassay was developed to evaluate the diagnostic potential for the new system, relying on magnetic microparticles coated with RV particles and analysis by differential pulse voltammetry. The limit of detection of the assay (0.07 IU mL(-1)) was >100× below the World Health Organization defined cut-off for rubella immunity. The sensitivity of the integrated device and its small size suggest future utility for distributed diagnostics.
The integration of 3D porous membranes in a digital microfluidic device enables the generation of cell-free plasma from finger-stick volumes of blood with in-line, on-chip analysis.
A digital microfluidic system was developed for portable Zika virus diagnosis, featuring integrated, automated sample extraction and amplification.
Microfluidics has become an important tool that is useful for a wide range of applications. A drawback for microfluidics is that many of the techniques that are commonly used to fabricate devices are not widely accessible, not scalable to high‐volume manufacturing processes, or both. Recently, a number of printing strategies that were originally developed for other applications have been applied to microfluidic device fabrication. These techniques, which include inkjet printing (IJP), screen printing (SP), and solid wax printing (SWP), are proposed to have a transformative effect on the field. Here microfluidics and printing, are introduced and a list of favorite examples is provided that highlights the accessibility and scalability that the combination is bringing to the field.
Background Blood typing, donor compatibility testing, and hematocrit analysis are common tests that are important in many clinical applications, including those found in high-stakes settings such as the trauma center. These tests are typically performed in centralized laboratories with sample batching; the minutes that are lost in this mode can lead to adverse outcomes, especially for critical-care patients. As a step toward providing rapid results at the bedside, we developed a point-of-care hemagglutination system relying on digital microfluidics (DMF) and a unique, automated readout tool, droplet agglutination assessment using digital microfluidics (DAAD). Methods ABO and Rhesus blood grouping, donor crossmatching, and hematocrit assays were developed on a portable DMF platform that allowed for automated sample processing. The result of each assay could be determined by eye or automatically with the DAAD imaging tool. Results DMF-DAAD was applied to 109 samples collected from different sources (including commercial samples, pinpricks from volunteers, and a hospital blood bank), with perfect fidelity to gold-standard results. Some of these tests were carried out by a nonexpert in a hospital trauma center. Proof-of-concept results were also collected from smaller sample sets for donor compatibility testing and hematocrit analysis. Conclusion DMF-DAAD shows promise for delivering rapid, reliable results in a format well suited for a trauma center and other settings where every minute counts.
The Democratic Republic of the Congo (DRC) has a high measles incidence despite elimination efforts and has yet to introduce rubella vaccine. We evaluated the performance of a prototype rapid digital microfluidics powered (DMF) enzyme-linked immunoassay (ELISA) assessing measles and rubella infection, by testing for immunoglobulin M (IgM), and immunity from natural infection or vaccine, by testing immunoglobulin G (IgG), in outbreak settings. Field evaluations were conducted during September 2017, in Kinshasa province, DRC. Blood specimens were collected during an outbreak investigation of suspected measles cases and tested for measles and rubella IgM and IgG using the DMF-ELISA in the field. Simultaneously, a household serosurvey for measles and rubella IgG was conducted in a recently confirmed measles outbreak area. DMF-ELISA results were compared with reference ELISA results tested at DRC’s National Public Health Laboratory and the US Centers for Disease Control and Prevention. Of 157 suspected measles cases, rubella IgM was detected in 54% while measles IgM was detected in 13%. Measles IgG-positive cases were higher among vaccinated persons (87%) than unvaccinated persons (72%). In the recent measles outbreak area, measles IgG seroprevalence was 93% overall, while rubella seroprevalence was lower for children (77%) than women (98%). Compared with reference ELISA, DMF-ELISA sensitivity and specificity were 82% and 78% for measles IgG; 88% and 89% for measles IgM; 85% and 85% for rubella IgG; and 81% and 83% for rubella IgM, respectively. Rubella infection was detected in more than half of persons meeting the suspected measles case definition during a presumed measles outbreak, suggesting substantial unrecognized rubella incidence, and highlighting the need for rubella vaccine introduction into the national schedule. The performance of the DMF-ELISA suggested that this technology can be used to develop rapid diagnostic tests for measles and rubella.
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