Comprehensive studies to improve ultrasensitive detection of HIV-1 p24 antigen†

IF 4.1 Q2 CHEMISTRY, ANALYTICAL
Evan Reboli, Ajoke Williams, Ankan Biswas, Tianwei Jia, Ying Luo, Mukesh Kumar and Suri Iyer
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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.

Abstract Image

提高HIV-1 p24抗原†超灵敏检测的综合研究
早期和准确检测HIV-1 p24抗原对于及时诊断和治疗至关重要,特别是在资源有限的环境中,传统方法往往缺乏早期检测的必要灵敏度或价格昂贵。在这里,我们开发了一个通过生物正交TCO/TZ化学功能化的荧光二氧化硅纳米颗粒的逐层信号放大平台。我们评估了不同尺寸的纳米颗粒(25、50和100 nm)和两种染料掺杂纳米颗粒配方,以优化信号强度、检测限和非特异性结合。25 nm的ritc掺杂纳米颗粒表现出优异的性能,实现了7 fg mL−1的超低检测限和高达1 ng mL−1的宽线性范围。与fitc掺杂的纳米颗粒相比,ritc掺杂的纳米颗粒具有增强的亮度和信号强度。进一步的优化表明,使用50 μg的25 nm纳米颗粒获得了最佳的灵敏度,同时最小化了非特异性结合。这种基于纳米颗粒的检测方法明显优于商用ELISA试剂盒,提供了广泛的动态范围和提高的灵敏度。我们的平台提供了一种高度敏感和适应性强的HIV-1 p24抗原检测方法,在即时诊断和其他低丰度生物标志物检测中具有广泛的潜在应用,最终提高疾病的早期检测和治疗可及性。
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CiteScore
2.30
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