研究目的
To develop a portable, low-cost, and quantitative biosensor for rapid, on-site detection of avian influenza virus (AIV) H7N9 using surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay strips.
研究成果
The SERS-based LFIA strip developed in this study provides a rapid, sensitive, and quantitative method for detecting AIV H7N9, with a detection limit of 0.0018 HAU, which is three orders of magnitude more sensitive than hemagglutination assays. It shows high specificity and reproducibility, making it suitable for on-site diagnostics and potential extension to other pathogens.
研究不足
The stability of the SERS probes may decline over time due to oxidation of the silver layer, and the method requires optimization for other pathogens beyond H7N9.
1:Experimental Design and Method Selection:
A sandwich-type immunoassay was designed using a lateral flow immunoassay strip with SERS probes for ultrasensitive detection. The method involves synthesizing a novel core-shell SERS tag (AuAg4(cid:3)ATP@AgNPs) and immobilizing antibodies on a nitrocellulose membrane.
2:Sample Selection and Data Sources:
Inactivated AIV H7N9 samples were used, with hemagglutination titers measured. Real samples included avian cloacal and throat swabs from poultry.
3:List of Experimental Equipment and Materials:
Materials included chloroauric acid, bovine serum albumin, antibodies, nitrocellulose membranes, and various chemicals. Equipment included a transmission electron microscope, scanning electron microscope, Biodot XYZ3060 dispense platform, Synergy H1 Hybrid Multi-Mode Microplate Reader, Zetasizer Nano ZS, centrifuge, programmable strip cutter, and portable Raman spectrometer.
4:Experimental Procedures and Operational Workflow:
Synthesis of AuAg4(cid:3)ATP@AgNPs, preparation of antibody-labeled probes, assembly of LFIA strips, sample application, migration for 20 minutes, and signal detection using a portable Raman spectrometer.
5:Data Analysis Methods:
Raman peak intensities at 1580 cm?1 were measured and used for quantitative analysis, with calibration curves constructed and limit of detection calculated.
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Scanning Electron Microscope
Carl Zeiss AG
Carl Zeiss
Imaging of immunoassay strips
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Zetasizer Nano ZS
Nano ZS
Malvern Panalytical
Particle size and zeta potential analysis
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Transmission Electron Microscope
PHILIPS TECNAI
PHILIPS
Characterization of nanoparticles
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Biodot XYZ3060 Dispense Platform
XYZ3060
Biodot
Dispensing antibodies and probes onto membranes
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Synergy H1 Hybrid Multi-Mode Microplate Reader
Synergy H1
Bio Tek Instruments
Optical measurements
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Centrifuge
Beckman
Purification and separation of nanoparticles
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Programmable Strip Cutter
HGS201
Shanghai Jiening Biotech
Cutting immunoassay strips
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Portable Raman Spectrometer
Guangzhou Biaoqi Electronic Technology
Quantification of Raman signals
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