Promise and perils of paper-based point-of-care nucleic acid detection for endemic and pandemic pathogens.

From HIV and influenza to emerging pathogens like COVID-19, each new infectious disease outbreak has highlighted the need for massively-scalable testing that can be performed outside centralized laboratory settings at the point-of-care (POC) in order to prevent, track, and monitor endemic and pandemic threats. Nucleic acid amplification tests (NAATs) are highly sensitive and can be developed and scaled within weeks while protein-based rapid tests require months for production. Combining NAATs with paper-based detection platforms are promising due to the manufacturability, scalability, and simplicity of each of these components. Typically, paper-based NAATs consist of three sequential steps: sample collection and preparation, amplification of DNA or RNA from pathogens of interest, and detection. However, these exist within a larger ecosystem of sample collection and interpretation workflow, usability, and manufacturability which can be vastly perturbed during a pandemic emergence. This review aims to explore the challenges of paper-based NAATs covering sample-to-answer procedures along with three main types of clinical samples; blood, urine, and saliva, as well as broader operational, scale up, and regulatory aspects of device development and implementation. To fill the technological gaps in paper-based NAATs, a sample-in-result-out system that incorporates the integrated sample collection, sample preparation, and integrated internal amplification control while also balancing needs of users and manufacturability upfront in the early design process is required.

Portable and power-free serodiagnosis of Chagas disease using magnetic levitating microbeads.

This work describes the detection of anti-T. cruzi antibodies in whole blood solutions using magnetic levitating microbeads (MLµBs). This simple diagnostic method can be easily performed by minimally trained personnel using an inexpensive and portable magnetic stage that requires no electricity. A multiphase test tube containing the MLµBs facilitates the sequential incubation, filtering, and reading of the immunoassays. The diagnostic method starts by adding a blood sample to the top phase of the test tube where the anti-T. cruzi antibodies present in the blood attach to the T. cruzi antigens on the surface of the MLµBs. Shaking the test tube after incubation mixes the top layer with a paramagnetic medium loaded with SiO2 microcrystals. The attachment of SiO2 microcrystals to those MLµBs bound to T. cruzi antibodies decreases their levitation height once the tube is placed between two antialigned permanent magnets. Measuring the levitation height of MLµBs enables the accurate detection and quantification of anti-T. cruzi antibodies in the blood across the clinically relevant range, with a detection limit of 5 µg mL-1. The small size of the test tubes facilitates the simultaneous analysis of over 50 different samples. MLµBs act as partial collimators for non-polarized light, facilitating their visual identification by the naked eye or by projecting incident light on a thin paper screen. A machine-vision algorithm was created to automatically interpret the results of the MLµB tests from a digital image, resulting in a rapid, accurate, and user-friendly assay for Chagas disease that can be used in resource-limited settings.

Wearable and Implantable Epidermal Paper-Based Electronics.

Traditional manufacturing methods and materials used to fabricate epidermal electronics for physiological monitoring, transdermal stimulation, and therapeutics are complex and expensive, preventing their adoption as single-use medical devices. This work describes the fabrication of epidermal, paper-based electronic devices (EPEDs) for wearable and implantable applications by combining the spray-based deposition of silanizing agents, highly conductive nanoparticles, and encapsulating polymers with laser micromachining. EPEDs are inexpensive, stretchable, easy to apply, and disposable by burning. The omniphobic character and fibrous structure of EPEDs make them breathable, mechanically stable upon stretching, and facilitate their use as electrophysiological sensors to record electrocardiograms, electromyograms, and electrooculograms, even under water. EPEDs can also be used to provide thermotherapeutic treatments to joints, map temperature spatially, and as wirelessly powered implantable devices for stimulation and therapeutics. This work makes epidermal electronic devices accessible to high-throughput manufacturing technologies and will enable the fabrication of a variety of wearable medical devices at a low cost.