All-Serotype Dengue Virus Detection through Multilayered Origami-Based Paper/Polymer Microfluidics

Abstract

The dengue virus (DENV) infection commonly triggers threatening seasonal outbreaks all around the globe (estimated yearly infections are in the order of 100 million, combining all the viral serotypes), testifying the need for early detection to facilitate disease management and patient recovery. The laboratory-based testing procedures for detecting DENV infection early enough are challenged by the need of resourced settings that result in inevitable cost penalty and unwarranted delay in obtaining the test results due to distance-related factors with respect to the patient's location. Recognizing that the introduction of alternative extreme point-of-care technologies for early detection may potentially mitigate this challenge largely, we develop here a multiplex paper/polymer-based detection strip that interfaces with an all-in-one simple portable device, synchronizing the pipeline of nucleic acid isolation, isothermal amplification, and colorimetric analytics as well as readout for detecting all the four serotypes of dengue viruses in around 30 min from about 50 μL of human blood serum with high specificity and sensitivity. Aligned with the mandatory guidelines of the World Health Organization, the ultralow-cost test is ideal for dissemination at different community centers via a user-friendly device interface, not only as a critical surveillance measure in recognizing the potential cocirculation of the infection across regions that are hyperendemic for all four DENV serotypes but also for facilitating effective monitoring of patients infected by any one of the particular viral serotypes as well as timely administration of life-saving measures on need.

Dengue: A review of laboratory diagnostics in the vaccine age

“…Reverse-transcriptase (RT) LAMP has been considered a good candidate for deployment in point-of-care assays, and several adaptations for portable device have been described [120][121][122][123]. Some LAMP assays may be suitable for direct amplification without prior nucleic acid extraction [120,121]. LAMP can be used to differentiate serotypes [124,125] or as a single tube reaction for clinical diagnosis [120,126].…”

Microfluidic systems for infectious disease diagnostics

“…The approach was validated with clinical human blood serum. 494 Seok et al demonstrated a lateral flow type device for RT-LAMP molecular diagnostics of multiple viral RNAs from human serum (DENV, ZIKV, CHIKV). The whole process of nucleic acid testing was integrated on the lab-on-paper strip with an assay time of 60 min.…”

Paper microfluidics with deep learning for portable intelligent nucleic acid amplification tests

“…Relatively large surface-to-volume ratio by the porous nature of paper theoretically enhances substances exchange and reactivity. These merits make μPADs particularly suitable for meeting the need for developing on-site or point-of-care testing (POCT) systems [ 22 , 23 ]. NAATs has been successfully integrated with paper microfluidics [ [24] , [25] , [26] ] which can also be leveraged for detecting SARS-CoV-2.…”