Development of Novel Dengue NS1 Multiplex Lateral Flow Immunoassay to Differentiate Serotypes in Serum of Acute Phase Patients and Infected Mosquitoes

Abstract

Dengue is among the most rapidly spreading arboviral disease in the world. A low-cost, easy to use point-of-care diagnostic tool for the detection and differentiation of dengue virus serotypes could improve clinical management, disease prevention, epidemiological surveillance, and outbreak monitoring, particularly in regions where multiple serotypes co-circulate. Despite widespread deployment, no commercial dengue antigen diagnostic test has proven effective in differentiating among dengue virus serotypes. In the current study, we first established mAb pairs and developed a multiplex lateral flow immunoassay for the simultaneous detection of the dengue viral NS1 antigen and identification of serotype. The proposed system, called Dengue serotype NS1 Multiplex LFIA, provides high sensitivity and specificity. In testing for JEV, ZIKV, YFV, WNV, and CHIKV, the multiplex LFIA gave no indication of cross- reactivity with cell culture supernatants of other flaviviruses or chikungunya virus. In analyzing 187 samples from patients suspected of dengue infection, the detection sensitivity for serotype D1 to D4 was 90.0%, 88.24%, 82.61%, and 83.33% and serotype specificity was 98.74%, 96.13%, 99.39%, and 97.04%, respectively. Our multiplex LFIA can also identify mono- and co-infection of different serotype of dengue viruses in mosquitoes. The proposed Multiplex LFIA provides a simple tool for the rapid detection of dengue serotypes and in the differential diagnosis of fever patients in regions where medical resources are limited and/or multiple DENVs co-circulate.

Simple and field amenable loop-mediated isothermal amplification-lateral flow dipstick assay for detection of west Nile virus in human clinical samples

“…It has so far been utilized for the detection of many viruses consisting of West Nile, Dengue, Japanese encephalitis, Zika virus, Yellow fever and Chikungunya viruses, etc. (Lai et al, 2022;Meia et al, 2018;Wang et al, 2010Wang et al, , 2017Zengguo et al, 2016).…”

Hyphenating paper-based biosensors with smartphones

“…For example, this is particularly useful for febrile illnesses such as dengue fever and yellow fever [53], as early identification can lead to timelier treatment. Due to the highly specific nature of antibodies, a multiplex LFIA can even distinguish between different variants of the same disease, such as different dengue serotypes [54]; this can be useful because different serotypes are associated with the severity of the disease [55]. That said, it can be very challenging to multiplex LFIAs and there are some key points to consider during development.…”

Diagnoses Based on C-Reactive Protein Point-of-Care Tests

“…Lateral flow tests represent a group of analytical devices being commercially available for many analytes including the CRP. The lateral flow tests are simple colorimetric disposable analytical devices historically coming out from thin-layer chromatography and the first introduction into the market as pregnancy tests in the 1980s followed by the introduction of other analytes including biochemical and immunochemical markers, microorganisms and viruses, various toxins, and other substances [96][97][98][99][100]. The color lines formed in the lateral flow tests can be scaled by the naked Despite immunoturbidimetric assays being quite simple and cheap to be performed, they still need an analytical device that excludes them from solitary use.…”