Evan Reboli, Ajoke Williams, Ankan Biswas, Tianwei Jia, Ying Luo, Mukesh Kumar and Suri Iyer
{"title":"Comprehensive studies to improve ultrasensitive detection of HIV-1 p24 antigen†","authors":"Evan Reboli, Ajoke Williams, Ankan Biswas, Tianwei Jia, Ying Luo, Mukesh Kumar and Suri Iyer","doi":"10.1039/D5SD00011D","DOIUrl":null,"url":null,"abstract":"<p >Early and accurate detection of HIV-1 p24 antigen is crucial for timely diagnosis and treatment, particularly in resource-limited settings where traditional methods often lack the necessary sensitivity for early-stage detection or is expensive. Here, we developed a layer-by-layer signal amplification platform employing fluorescent silica nanoparticles functionalized <em>via</em> bioorthogonal TCO/TZ chemistry. We evaluated nanoparticles of different sizes (25, 50, and 100 nm) and two dye-doped nanoparticle formulations to optimize signal intensity, detection limits, and nonspecific binding. The 25 nm RITC-doped nanoparticles demonstrated superior performance, achieving an ultra-low detection limit of 7 fg mL<small><sup>−1</sup></small> with a broad linear range up to 1 ng mL<small><sup>−1</sup></small>. Compared to FITC-doped nanoparticles, RITC-doped nanoparticles provided enhanced brightness and signal strength. Further optimization revealed that using 50 μg of 25 nm nanoparticles yielded the best sensitivity while minimizing nonspecific binding. This nanoparticle-based assay significantly outperformed commercial ELISA kits, offering a broad dynamic range and improved sensitivity. Our platform presents a highly sensitive and adaptable approach for HIV-1 p24 antigen detection, with broad potential applications in point-of-care diagnostics and detection of other low-abundance biomarkers, ultimately enhancing early disease detection and treatment accessibility.</p>","PeriodicalId":74786,"journal":{"name":"Sensors & diagnostics","volume":" 7","pages":" 586-595"},"PeriodicalIF":4.1000,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/sd/d5sd00011d?page=search","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sensors & diagnostics","FirstCategoryId":"1085","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2025/sd/d5sd00011d","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
Abstract
Early and accurate detection of HIV-1 p24 antigen is crucial for timely diagnosis and treatment, particularly in resource-limited settings where traditional methods often lack the necessary sensitivity for early-stage detection or is expensive. Here, we developed a layer-by-layer signal amplification platform employing fluorescent silica nanoparticles functionalized via bioorthogonal TCO/TZ chemistry. We evaluated nanoparticles of different sizes (25, 50, and 100 nm) and two dye-doped nanoparticle formulations to optimize signal intensity, detection limits, and nonspecific binding. The 25 nm RITC-doped nanoparticles demonstrated superior performance, achieving an ultra-low detection limit of 7 fg mL−1 with a broad linear range up to 1 ng mL−1. Compared to FITC-doped nanoparticles, RITC-doped nanoparticles provided enhanced brightness and signal strength. Further optimization revealed that using 50 μg of 25 nm nanoparticles yielded the best sensitivity while minimizing nonspecific binding. This nanoparticle-based assay significantly outperformed commercial ELISA kits, offering a broad dynamic range and improved sensitivity. Our platform presents a highly sensitive and adaptable approach for HIV-1 p24 antigen detection, with broad potential applications in point-of-care diagnostics and detection of other low-abundance biomarkers, ultimately enhancing early disease detection and treatment accessibility.