A New Method for Ultra-sensitive P24 Antigen Assay Based on Near-infrared Fluorescent Microsphere...

Area of Science:

• Immunology
• Biotechnology
• Medical Diagnostics

Background:

• Human Immunodeficiency Virus (HIV) diagnosis relies on detecting viral markers like P24 antigen.
• Current diagnostic methods may lack the desired speed, sensitivity, or quantitative accuracy for early detection and monitoring.
• Development of advanced immunoassay platforms is crucial for improving HIV diagnostics.

Purpose of the Study:

• To develop and validate a novel, highly sensitive, and quantitative method for P24 antigen detection.
• To utilize near-infrared fluorescent microsphere immunochromatography for enhanced diagnostic capabilities.
• To assess the performance of this new method compared to established techniques like ELISA.

Main Methods:

• Preparation of a lateral flow assay test strip with fluorescent microsphere-labeled detection antibodies.
• Optimization of antibody labeling and assay detection conditions.
• Establishment of a working curve and methodological assessment using clinical samples.
• Comparative analysis with quantitative ELISA for P24 antigen detection.

Main Results:

• The developed method demonstrates a low detection limit of 3.4 pg/mL and a broad detection range (3.4 pg/mL to 10 ng/mL).
• High accuracy and precision were observed, with average intra-assay recovery of 99.6% (CV 5.4%-8.6%) and inter-assay recovery of 97.3% (CV 8.5%-11%).
• The fluorescent microsphere immunochromatography method exhibited a higher detection rate and good correlation with ELISA.

Conclusions:

• Near-infrared fluorescent microsphere immunochromatography provides a rapid, highly sensitive, and quantitative approach for P24 antigen detection.
• The method's wide detection range and high performance make it suitable for early clinical diagnosis of HIV.
• This technique is valuable for continuous monitoring of AIDS progression and treatment efficacy.