Evaluating the feasibility of a lateral-flow assay for HIV Nef
Date
2024
Authors
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Publisher
University of Delaware
Abstract
Human Immunodeficiency Virus (HIV) pathology deals with lymph nodes, and a delay in anti-retroviral therapy initiation leads to progressive immune system failure. Accurate and prompt diagnosis is critical for initiating effective treatment and improved clinical outcomes. Current HIV detection assays utilize transient biomarkers, resulting in a narrow detection window period. The thesis discusses the development of a competitive lateral flow immunoassay (LFA) for detecting HIV protein Nef, a potentially stable biomarker. Leveraging 40 nm gold nanoparticles as the optical sensors, the critical component of the assay, i.e. the effective concentration of Nef immobilized on the test zone and the strength of the detecting conjugates were optimized. The performance and repeatability of the assay were evaluated with an adequate sample size, determined by power analysis. The mean detection limit for different assay batches was determined to be 12.52 ± 0.29 ng/mL. The rigorous assessment demonstrated a 95% confidence level, i.e., a large effect size was observed when negative and positive test results were compared from the same batch. The repeatability analysis confirms the consistency of the LFA concerning the detection limit within the batch. However, the inter-assay repeatability of the current system is poor. The study is the first report evaluating the feasibility of developing a competitive LFA for detecting Nef. It aims to highlight the potential of LFA for detecting a stable HIV protein as an initial mode of assessment for high-risk individuals and for monitoring the infection. While further refinements are necessary to enhance clinical suitability, this research lays the groundwork for innovative advancements in HIV screening and monitoring.
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Keywords
Competitive, Feasibility, Human Immunodeficiency Virus, Lateral flow immunoassay, Anti-retroviral therapy