Multiplexed, Patterned-Paper Immunoassay for Detection of Malaria and Dengue Fever

Abstract

Multiplex assays detect the presence of more than one analyte in a sample. For diagnostic applications, multiplexed tests save healthcare providers time and resources by performing many assays in parallel, minimizing the amount of sample needed and improving the quality of information acquired regarding the health status of a patient. These advantages are of particular importance for those diseases that present with general, overlapping symptoms, which makes presumptive treatments inaccurate and may put the patient at risk. For example, malaria and dengue fever are febrile illnesses transmitted through mosquito bites, and these common features make it difficult to obtain an accurate diagnosis by symptoms alone. In this manuscript, we describe the development of a multiplexed, patterned paper immunoassay for the detection of biomarkers of malaria and dengue fever: malaria HRP2, malaria pLDH, and dengue NS1 type 2. In areas coendemic for malaria and dengue fever, this assay could be used as a rapid, point-of-care diagnostic to determine the cause of a fever of unknown origin. The reagents required for each paper-based immunoassay are separated spatially within a three-dimensional device architecture, which allows the experimental conditions to be adjusted independently for each assay. We demonstrate the analytical performances of paper-based assays for each biomarker and we show that there is no significant difference in performance between the multiplexed immunoassay and those immunoassays performed in singleplex. Additionally, we spiked individual analytes into lysed human blood to demonstrate specificity in a clinically relevant sample matrix. Our results suggest multiplex paper-based devices can be an essential component of diagnostic assays used at the point-of-care.

Reducing Uncertainty for Acute Febrile Illness in Resource-Limited Settings: The Current Diagnostic Landscape

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Dielectrophoresis‐Based Protein Enrichment for a Highly Sensitive Immunoassay Using Ag/SiO2 Nanorod Arrays

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Diagnosis of communicable diseases using papepr microfluidic platforms

“…A schematic diagram of the paper-based detection device is shown in Figure 10. Deraney et al [63] reported a patterned paper immunoassay for detection of two malaria antigens, malaria HRP2 and malaria pLDH. They developed a three-dimensional paper-based microfluidic device ( Figure 11) capable of detecting both malaria antigens with high sensitivity which can also distinguish between the two malaria species.…”