Abstract
In this study, we developed a new type of paper-based analytical device (PAD), the three-dimensional (3D) slip-PAD, to detect infectious human norovirus for global healthcare. The 3D configuration of the papers combined with a slip design provides unique features and versatility that overcome the limitations of fluidic manipulation and sensitivity in point-of-care (POC) tests. The assay can be carried out in a single step based on a moveable slip design, making it suitable for unskilled users. The 3D fluidic network developed by layered construction of wax-patterned papers provides different fluidic paths for the sequential delivery of multiple fluids without the need for peripheral equipment. The release and mixing of enhancement reagents on the device improved the sensitivity and detection limit. The assay results could be visualized by naked eye within 10 min, with subsequent amplification of the signal over time (<60 min). The device showed a broad dynamic range of detection and high sensitivity, with a detection limit of 9.5 × 104 copies ml−1 for human norovirus. These results demonstrate that the 3D slip-PAD is a sensitive diagnostic assay for detecting human norovirus infection that is particularly suitable for POC testing in regions where resources are scarce.
Highlights
There is an increasing demand for point-of-care (POC) tests in many fields, including clinical analysis, food safety, and environmental assessment, and many efforts have been made to develop biosensors that are affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free, and deliverable to end-users (ASSURED) that meet World Health Organization criteria[1,2,3,4,5]
We develop a one-step 3D slip-paper-based analytical device (PAD) that embodies features desirable in a POC testing biosensor, including low cost, sensitivity, robustness, rapidity, and ease of use
The device was fabricated by assembling wax-patterned paper layers and was successfully applied to the detection of human norovirus based on immunochromatography coupled with signal amplification
Summary
There is an increasing demand for point-of-care (POC) tests in many fields, including clinical analysis, food safety, and environmental assessment, and many efforts have been made to develop biosensors that are affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free, and deliverable to end-users (ASSURED) that meet World Health Organization criteria[1,2,3,4,5]. A lateral flow assay (LFA) based on PADs is commercially available for a variety of applications that is low cost, rapid, simple to use, equipment-free and mass produced, conforming to ASSURED criteria[9] It has limitations in terms of sensitivity and manipulation of fluid flow. LFA shows considerable potential as a POC device for norovirus detection based on its advantages of rapidity, simplicity, and low cost, it is limited by insufficient sensitivity, as previously stated. To this end, we developed a novel one-step 3D slip-PAD for simple, sensitive, and inexpensive detection of human noroviruses. The manually movable slip design of the 3D slip-PAD enabled the one-step assay without any additional manipulation, making it more accessible to an unskilled user and more suitable for resource-limited settings
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