Development of a Rapid Gold Nanoparticle-Based Lateral Flow Immunoassay for the Detection of Dengue Virus

Abstract

Flavivirus detection in humans and mosquito reservoirs has been an important issue since it can cause a variety of illnesses and could represent a health problem in geographical zones where the vector is endemic. In this work, we designed and characterized a biosensor based on gold nanoparticles (AuNPs) and antibody 4G2 for the detection of dengue virus (DENV) in vitro, obtaining different conjugates (with different antibody concentrations). The AuNP–4G2 conjugates at concentrations of 1, 3, and 6 µg/mL presented an increase in the average hydrodynamic diameter compared to the naked AuNPs. Also, as part of the characterization, differences in the UV-Vis absorbance spectrum and electrophoretic migration were observed between the conjugated AuNPs (with BSA or antibody) and naked AuNPs. Additionally, we used this biosensor (AuNP–4G2 conjugate with 3 µg/mL antibody) in the assembly of a competitive lateral flow assay (LFA) for the development of an alternative test to detect the flavivirus envelope protein in isolated DENV samples as a future tool for dengue detection (and other flaviviruses) in the mosquito vector (Aedesaegypti) for the identification of epidemic risk regions. Functionality tests were performed using Dengue virus 2 isolated solution (TCID50/mL = 4.58 × 103) as a positive sample and PBS buffer as a negative control. The results showed that it is possible to detect Dengue virus in vitro with this gold nanoparticle-based lateral flow assay with an estimated detection limit of 5.12 × 102 PFU. We suggest that this biosensor could be used as an additional detection tool by coupling it to different point-of-care tests (POCT) for the easy detection of other flaviviruses.

A Highly Sensitive Dual-Drive Microfluidic Device for Multiplexed Detection of Respiratory Virus Antigens

“…Colloidal gold immunochromatography assay (GICA) is the most representative POCT method widely used in clinical diagnosis. It has a large market demand due to its rapid detection time, affordable prices, and ease of operation [ 10 ]. However, GICA has been known to give a high false-negative rate due to its low sensitivity [ 5 , 11 ].…”

Europium Nanoparticle-Based Lateral Flow Strip Biosensors for the Detection of Quinoxaline Antibiotics and Their Main Metabolites in Fish Feeds and Tissues

“…LFSBs commonly use gold nanoparticles (AuNPs) as labels. The method is simple and convenient, and results can be observed visually, but low sensitivity and a narrow linear range are considered drawbacks of AuNP-LFSBs [ 18 ]. Therefore, different fluorescence microspheres are employed for LFSB assays, such as quantum dots (QDs) [ 19 ], upconversion nanoparticles (UCNPs) [ 20 ], and europium nanoparticles (EuNPs) [ 21 ].…”

Updates on the Biofunctionalization of Gold Nanoparticles for the Rapid and Sensitive Multiplatform Diagnosis of SARS-CoV-2 Virus and Its Proteins: From Computational Models to Validation in Human Samples

“…Nowadays, AuNPs are used for the ultrasensitive detection of varied targets [29] such as enzymes [30,31], proteins [32], DNA [33,34], cells [35], viruses [36][37][38][39][40][41], and bacteria [42,43]. Moreover, promising therapeutic approaches have just begun using AuNPs as antiviral agents capable of trapping viruses [44][45][46][47], in anticoagulant therapy [48], in photothermal therapy for breast cancer [49], and in the treatment of inflammatory bowel disease [50].…”