研究目的
To develop a dual-color magnetic-quantum dot nanobeads (MQBs) based fluorescent lateral flow immunoassay for the simultaneous point-of-care detection of free and complexed prostate-specific antigen (f-PSA and c-PSA) to improve the diagnostic accuracy of prostate cancer (PCa).
研究成果
The dual-color MQBs-based fluorescent lateral flow immunoassay demonstrated high sensitivity and specificity for the simultaneous detection of f-PSA and c-PSA, with limits of detection of 0.009 ng mL?1 and 0.087 ng mL?1, respectively. The assay showed promising results for the accurate diagnosis of PCa, even in resource-limited settings, due to its easy-to-use format and portable smartphone readout device.
研究不足
The study does not explicitly mention limitations, but potential areas for optimization could include further reducing the limits of detection and improving the specificity and sensitivity of the assay for a wider range of clinical samples.
1：Experimental Design and Method Selection:
The study involved the fabrication of red and green MQBs via polyethyleneimine-mediated electrostatic adsorption of quantum dots onto superparamagnetic Fe3O4 magnetic cores. These MQBs were then conjugated with f-PSA and c-PSA antibodies for immune recognition and detection. A smartphone readout device was developed for imaging and data processing.
2：Sample Selection and Data Sources:
Clinical serum samples from PCa and benign prostatic hyperplasia patients were used to evaluate the clinical feasibility of the assay.
3：List of Experimental Equipment and Materials:
Materials included polyethyleneimine, N-Hydroxysulfosuccinimide sodium salt, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide?HCl, polyclonal goat anti-mouse IgG antibody, bovine serum albumin, carboxyl-functionalized CdSe/ZnS quantum dots, and various antibodies and antigens related to PSA.
4：Experimental Procedures and Operational Workflow:
The MQBs were synthesized and conjugated with antibodies. Lateral flow strips were prepared and used for the simultaneous detection of f-PSA and c-PSA. The smartphone readout device was used to capture and analyze the fluorescence signals.
5：Data Analysis Methods:
The fluorescence signals were analyzed using a custom-designed smartphone application for image recording and data analysis.
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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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