In this work, we demonstrate a rapid diagnostic platform with potential to transform clinical diagnosis of acute febrile illnesses in resource-limited settings. Acute febrile illnesses such as dengue and chikungunya, which pose high burdens of disease in tropical regions, share many nonspecific symptoms and are difficult to diagnose based on clinical history alone in the absence of accessible laboratory diagnostics. Through a unique color-mixing encoding and readout strategy, our platform enabled consistent and accurate multiplexed detection of dengue and chikungunya IgM/IgG antibodies in human clinical samples within 30 min. Our multiplex assay offers several advantages over conventional rapid diagnostic tests deployed in resource-limited settings, including a low sample volume requirement and the ability to concurrently detect four analytes. Our platform is a step toward multiplexed diagnostics that will be transformative for disease management in resourcelimited settings by enabling informed treatment decisions through accessible evidencebased diagnosis.
Background
Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment.
Methods
In this work, a two-colour lateral flow immunoassay (LFA) system was developed to identify infections by Plasmodium spp. and differentiate Plasmodium falciparum infection from the other three human malaria species (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae). To achieve this goal, red and blue colours were encoded to two markers on a single test line of strips, for simultaneous detection of PfHRP2 (red), a marker specific for P. falciparum infection, and pLDH (blue), a pan-specific marker for infections by all species of Plasmodium. The assay performance was first optimized and evaluated with recombinant malarial proteins spiked in washing buffer at various concentrations from 0 to 1000 ng mL−1. The colour profiles developed on the single test line were discriminated and quantified: colour types corresponded to malaria protein species; colour intensities represented protein concentration levels.
Results
The limit of detection (the lowest concentrations of malaria antigens that can be distinguished from blank samples) and the limit of colour discrimination (the limit to differentiate pLDH from PfHRP2) were defined for the two-colour assay from the spiked buffer test, and the two limits were 31.2 ng mL−1 and 7.8 ng mL−1, respectively. To further validate the efficacy of the assay, 25 human whole blood frozen samples were tested and successfully validated against ELISA and microscopy results: 15 samples showed malaria negative; 5 samples showed P. falciparum positive; 5 samples showed P. falciparum negative, but contained other malaria species.
Conclusions
The assay provides a simple method to quickly identify and differentiate infection by different malarial parasites at the point-of-need and overcome the physical limitations of traditional LFAs, improving the multiplexing potential for simultaneous detection of various biomarkers.
Summary
Lithium-sulfur batteries are paid much attention owing to their high specific capacity and energy density. However, their practical applications are impeded by poor electrochemical performance due to the dissolved polysulfides. The concentration of soluble polysulfides has a linear relationship with the internal heat generation. The issue of heat transport inside lithium-sulfur batteries is often overlooked. Here, we designed a functional separator that not only had a high thermal conductivity of 0.65 W m
−1
K
−1
but also alleviated the diffusion of dissolved active materials to the lithium anode, improving the electrochemical performance and safety issue. Lithium-sulfur batteries with the functional separator have a specific capacity of 1,126.4 mAh g
−1
at 0.2 C, and the specific capacity can be remained up to 893.5 mAh g
−1
after 100 cycles. Pouch Cells with high sulfur loading also showed a good electrochemical performance under a lean electrolyte condition of electrolyte/sulfur (E/S) = 3 μL mg
−1
.
Water model experiments were performed in a full-scale, delta-shaped water model tundish, in order to study the removal of inclusions by micro-bubbles. Micro-bubbles were generated using a specially designed ladle shroud with twelve laser-drilled orifices. Gas flow rates, injection positions and multi-port injection were all taken into consideration to create different bubble conditions. Bubbles were recorded using a high speed camera and post-processed with commercial software, Image J. Hollow glass borosilicate microspheres, smaller than 100 μm, were used to simulate inclusions, and detected, in-situ, using a new generation of the Aqueous Particle Sensor, APS III. The results revealed that the effect of microbubbles on inclusion removal depends greatly on the gas injection protocols used. The optimum gas flow rate was an intermediate value, which indicates a minimum particle number density, n p , of about 7.85/ml. This results from the counter-balancing effects of bubble sizes against the total number of bubbles. The highest inclusion removal rate was 80%, when gas was injected through the four ports located closest to the slide gate, at a gas flow rate of 0.2 L/min.
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