ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2016
·Vol. 408
·期 22
·6115-6121
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IF 3.125
Zhang Yun,Yang Hang,Yu Junping,Wei Hongping
In this study, a system of magnetic beads (MBs) coupled with catalytic fluorescent immunoassay for rapid and sensitive determination of HIV-1 capsid antigen p24 was developed. p24 was captured by antibody immobilized MBs, and the detection antibody was linked to horseradish peroxidase (HRP) through biotin–streptavidin recognition, catalyzing the oxidation of o -phenylenediamine (OPD) and hydrogen peroxide to produce a fluorescent product. This is the first reported utilization of the fluorescence of OPD oxidation product catalyzed by HRP for immunoassay. Optimization of conditions afforded a low detection limit of 0.5 pg/mL (3σ) for p24 with a linear range of 1.4–90.0 pg/mL. The assay exhibited good reproducibility with a relative standard deviation (RSD) of 4.4 %, 4.7 %, and 5.0 % for detecting 1.4 pg/mL, 22.5 pg/mL, and 45.0 pg/mL p24, respectively. The assay can be completed in less than 90 min. Moreover, the proposed method was successfully applied to detect p24 in spiked serum. This method overcomes the interference of MBs to the fluorescence signal and demonstrated higher sensitivity for detection of p24 than conventional ELISA kits. The system could be applied for detecting other antigens with high sensitivity, rapidity, specificity, and simple operation.
A rapid and sensitive biosensing method coupling immunomagnetic separation and catalytic fluorescence for determination of HIV-1 p24 has been developed
研究领域:
医学诊断
·
生物化学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 12
·1-9
·
IF 5.833
Tang Zhuoyun,Wei Zeliang,Huang Ke,Wei Yinhao,Li Dongdong,Yan Shixin,Huang Jin,Geng Jia,Tao Chuanmin,Chen Piaopiao,Ying Binwu
Human immunodeficiency virus (HIV) infection inflicts significant economic and social burdens on many countries worldwide. Given the substantial morbidity and mortality from HIV infection, there is an urgent need for accurate and early detection of the virus. In this study, immunofluorescence and visual techniques are described that detect the HIV-1 p24 antigen, which relied on selective recognition of Ag + /Ag nanoparticles (Ag NPs) and Cu 2+ /Cu + using cadmium telluride quantum dots (CdTe QDs). After the sandwich immunoreactions were accomplished, the alkaline phosphatase (ALP) hydrolyzed L-ascorbic acid 2-phosphate (AAP) to form ascorbic acid (AA) that further reduces Ag + and Cu 2+ to Ag NPs and Cu + , respectively. This method was highly sensitive and selective and could detect as low as 1 pg/mL of p24 antigen by naked eyes and had a good linearity in the concentration range 1–100 pg/mL. When using Ag + and Cu 2+ as media, the limit of detection (LOD) of the new method was 0.3 pg/mL and 0.2 pg/mL, respectively. Compared with clinical electrochemiluminescence immunoassay (ECLIA) results and clinical data, this method demonstrated good consistency for the quantification of HIV-1 p24 antigen in 34 clinical serum samples. In addition, this method could accurately distinguish HIV from other viruses and infections such as hepatitis B virus , systemic lupus erythematosus , hepatitis C virus , Epstein-Barr virus , cytomegalovirus , lipemia , and hemolysis . Therefore, our dual-mode analysis method may provide additional solutions to identify clinical HIV infection. Graphical abstract An immunofluorescence and visualization dual-mode strategy for the detection of p24 antigen was constructed based on immune recognition reaction and a phenomenon that cadmium telluride quantum dots (CdTe QDs) can selectively recognize Ag + /Ag nanoparticles (Ag NPs) and Cu 2+ /Cu + .
研究领域:
病毒学
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免疫学
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
BME Frontiers
·
2025
·
IF 7.7
Yirui Zhang,Jieyu Yan,Wangheng Hou,Qian Gao,Tao Zhang,Yan Gao,Jingwen Li,Kun Han
Objective: Accurate and rapid detection of disease biomarkers is critical for early diagnosis, timely intervention, and effective disease management. Impact Statement: This strategy is exemplified through the development of 2 biosensors for detecting HIV-1 DNA and HIV-1 p24, biomarkers associated with acquired immunodeficiency syndrome (AIDS). Introduction: We propose a general biosensing strategy that leverages a treble signal amplification cascade, demonstrating its versatility and applicability across diverse biomolecular targets. Methods: The method integrates multiple amplification mechanisms to achieve unparalleled sensitivity. Initially, the hybridization of 2 aided probes with the target triggers isothermal amplification facilitated by polymerase and nicking enzyme, providing a robust preliminary signal enhancement. Repeated cycles of primer extension, nicking, and signal primer dissociation then generate multiple signal primers. These primers are further amplified via rolling circle amplification (RCA), resulting in an important secondary signal boost. Finally, the amplified products activate CRISPR-Cas12a-mediated trans-cleavage, achieving a tertiary level of signal enhancement. Results: This cascade amplification approach achieves remarkable sensitivity, with detection limits of 62 aM for nucleic acids and 8.48 pg/ml for proteins, positioning it as a broadly applicable framework. Clinical samples were assayed, which indicates its capability in clinical diagnosis. Conclusion: Beyond HIV detection, the modular design of this strategy allows adaptation for various biomarkers, showcasing its potential as a universal platform for molecular diagnostics in healthcare and research.
研究领域:
生物传感器
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分子诊断
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病毒学
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临床诊断
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生物医学工程
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核酸扩增
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蛋白质检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）