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Development of a highly sensitive magneto-enzyme lateral flow immunoassay for dengue NS1 detection.

PeerJ (2019-10-04)
Tien V Tran, Ba V Nguyen, Thao T P Nguyen, Tung T Tran, Khanh G Pham, Quang B Le, Binh N Do, Hung N Pham, Chuyen V Nguyen, Duong P H Dinh, Van T Ha, Trang H T Doan, Hoa Q Le
RÉSUMÉ

Dengue infection represents a global health issue of growing importance. Dengue non-structural protein 1 (NS1) plays a central role in the early detection of the disease. The most common method for NS1 detection is testing by lateral flow immunoassays (LFIAs) with varying sensitivity. In this study, we present a highly sensitive magneto-enzyme LFIA for prompt diagnosis of dengue. We have demonstrated the development of a magneto-enzyme LFIA combining super-paramagnetic nanoparticles as labels and Biotin-Streptavidin signal amplification strategy to detect dengue NS1. Factors affecting the test performance including antibody pair, super-paramagnetic nanoparticle size, nitrocellulose membrane type, amounts of detection and capture antibodies, and amounts of Streptavidin-polyHRP were optimized. Analytical sensitivity and cross-reactivity were determined. Clinical performance of the novel assay was evaluated using a panel of 120 clinical sera. This newly developed assay could detect NS1 of all four serotypes of dengue virus (DENV). The limit of detection (LOD) was found to be as low as 0.25 ng ml-1 for DENV-1 and DENV-3, 0.1 ng ml-1 for DENV-2, and 1.0 ng ml-1 for DENV-4. The LOD for DENV-2 was a 50-fold improvement over the best values previously reported. There was an absence of cross-reactivity with Zika NS1, Hepatitis B virus, Hepatitis C virus, and Japanese encephalitis virus. The sensitivity and specificity of the novel assay were 100% when tested on clinical samples. We have successfully developed a magneto-enzyme LFIA, allowing rapid and highly sensitive detection of dengue NS1, which is essential for proper management of patients infected with DENV.

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Anticorps anti-IgG de souris (molécule complète) antibody produced in goat, whole antiserum