研究目的
开发一种基于双色磁性量子点纳米珠(MQBs)的荧光侧向流动免疫分析法,用于同时即时检测游离和复合前列腺特异性抗原(f-PSA和c-PSA),以提高前列腺癌(PCa)的诊断准确性。
研究成果
基于双色MQB的荧光侧向流动免疫分析法对f-PSA和c-PSA的同时检测显示出高灵敏度和特异性,其检测限分别为0.009 ng mL?1和0.087 ng mL?1。该检测方法因其操作简便且配备便携式智能手机读数设备,在资源有限的环境中也能实现对前列腺癌的精准诊断,展现出良好前景。
研究不足
该研究未明确提及局限性,但潜在的优化方向可能包括进一步降低检测限,并提高该检测方法对更广泛临床样本的特异性和灵敏度。
1:实验设计与方法选择:
通过聚乙烯亚胺介导的静电吸附作用,将量子点负载到超顺磁性Fe3O4磁核上制备红绿双色微球(MQBs),随后偶联f-PSA和c-PSA抗体实现免疫识别检测,并开发了智能手机读数装置用于成像与数据处理。
2:样本选择与数据来源:
采用前列腺癌及良性前列腺增生患者的临床血清样本评估该检测方法的临床可行性。
3:实验设备与材料清单:
包括聚乙烯亚胺、N-羟基琥珀酰亚胺钠盐、1-乙基-3-(3-二甲氨基丙基)碳二亚胺盐酸盐、多克隆羊抗小鼠IgG抗体、牛血清白蛋白、羧基功能化CdSe/ZnS量子点,以及与PSA相关的各类抗体抗原。
4:实验流程与操作步骤:
合成MQBs并偶联抗体,制备侧流试纸条用于f-PSA与c-PSA同步检测,通过智能手机读数装置捕获分析荧光信号。
5:数据分析方法:
采用定制化智能手机应用程序进行图像记录与荧光信号数据分析。
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1-ethyl-3-(3-dimethylaminopropyl) carbodiimide?HCl
EDC
Sigma-Aldrich
Used in the conjugation of MQBs with antibodies.
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Bovine serum albumin
BSA
Sigma-Aldrich
Used to block unreacted carboxyl sites in the conjugation process.
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Dual-band emission filter
524/628 nm
Edmund Optical
Used to reduce background noise signals and crosstalk between red and green channels.
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Plano-convex lens
focal length: 20 mm
Edmund Optical
Used for signal collection and image magnification in the smartphone readout device.
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Polyethyleneimine
MW 25 kDa
Sigma-Aldrich
Used for the fabrication of MQBs via electrostatic adsorption of quantum dots onto superparamagnetic Fe3O4 magnetic cores.
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N-Hydroxysulfosuccinimide sodium salt
sulfo-NHS
Sigma-Aldrich
Used in the conjugation of MQBs with antibodies.
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Polyclonal goat anti-mouse IgG antibody
Sigma-Aldrich
Used in the preparation of lateral flow strips.
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Carboxyl-functionalized CdSe/ZnS quantum dots
QD525 and QD625
Meso light Technology Corp.
Used in the fabrication of MQBs for their luminescent properties.
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Free prostate-specific antigen
f-PSA
Lingchao Biotechnology Limited Company
Target analyte for detection in the assay.
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Prostate-specific antigen-α1-antichymotrypsin complex
c-PSA
Fitzgerald Industries International, Inc.
Target analyte for detection in the assay.
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Mouse monoclonal f-PSA, c-PSA, and total PSA antibodies
BiosPacific, Inc.
Used for immune recognition and capture of PSA markers.
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Hi-Flow Plus 90 nitrocellulose membranes
Millipore
Used as the substrate for the lateral flow strips.
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Smartphone
Mix2
Xiaomi
Used as part of the readout device for imaging and data processing.
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UV LED
3 W, 365 nm
Used to excite the MQBs probes captured on the test and control lines of LFIA strips.
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365 nm band-pass filter
Yongxing Optics
Used to filter the UV light source.
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