Abstract
Dengue is one of the most rapidly spreading mosquito-borne viral diseases in the world. Differential diagnosis is a crucial step for the management of the disease and its epidemiology. Point-of-care testing of blood-borne dengue biomarkers provides an advantageous approach in many health care settings, and the ability to follow more than one biomarker at once could significantly improve the management of the disease. Bead-based multiplex technologies (suspension array) can measure multiple biomarker targets simultaneously by using recognition molecules immobilized on microsphere beads. The overarching objective of our work is to develop a portable detection device for the simultaneous measurement of multiple biomarkers important in dengue diagnosis, monitoring and treatment. Here, we present a bead-based assay for the detection of one of the four serotypes of dengue virus non-structural protein (DENV-NS1) as well as its cognate human IgG. In this system, the fluorescent microspheres containing the classification fluorophore and detection fluorophore are imaged through a microfluidic chip using an infinity-corrected microscope system. Calibration curves were plotted for median fluorescence intensity against known concentrations of DENV-NS1 protein and anti-NS1 human IgG. The limit of quantitation was 7.8 ng/mL and 15.6 ng/mL, respectively. The results of this study demonstrate the feasibility of the multiplex detection of dengue biomarkers and present its analytical performance parameters. The proposed imaging device holds potential for point-of-care testing of biomarkers on a highly portable system, and it may facilitate the diagnosis and prevention of dengue as well as other infectious diseases.
Highlights
Dengue fever is a major public health problem in tropical and sub-tropical areas
Bead set A was coupled with a capture antibody specific to dengue virus (DENV)-NS1, bead set B was coupled with a capture protein specific to anti-DENV-NS1 IgG
After bead coupling, we found that the PBST suspension buffer had a higher bead count recovery than Phosphate buffer saline (PBS) because Tween-20 reduced the attachment of beads to the tube surface
Summary
Dengue fever is a major public health problem in tropical and sub-tropical areas. It is the fastest spreading mosquito-borne viral disease, and it affects an estimated 390 million people in more than 120 countries every year [1,2,3]. Dengue fever is caused by one of four antigenically distinct serotypes of dengue virus (DENV), which is a Flavivirus like Zika virus, West Nile virus, and yellow fever virus. In response to the virus, infected individuals produce immunoglobins (Igs, such as IgG or IgM) against DENV envelope glycoproteins, commonly resulting in the elimination of the virus. In a small number of cases it evolves into a severe illness called dengue haemorrhagic fever, which can result in life-threatening dengue shock syndrome [4]. The early or mild form of the disease shares the same symptomatic manifestation as other tropical infectious diseases, such as malaria, chikungunya, and Zika, which makes molecular testing essential for diagnosis
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