Detection of Hepatitis B virus antigen from human blood: SERS immunoassay in a microfluidic system
| Autor: | Jan L. Weyher, Jacek Waluk, Igor Dzięcielewski, Agnieszka Kamińska, Evelin Witkowska, Katarzyna Winkler |
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| Rok vydání: | 2015 |
| Předmět: |
Hepatitis B virus
HBsAg Biomedical Engineering Biophysics Metal Nanoparticles Biosensing Techniques Spectrum Analysis Raman medicine.disease_cause Hepatitis B Antigens Antigen Electrochemistry medicine Humans Immunoassay Detection limit Hepatitis B Surface Antigens Chromatography medicine.diagnostic_test biology Chemistry Substrate (chemistry) General Medicine Microfluidic Analytical Techniques Molecular biology Linear range biology.protein Gold Antibody Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 66:461-467 |
| ISSN: | 0956-5663 |
| Popis: | A highly sensitive immunoassay utilizing surface-enhanced Raman scattering (SERS) has been developed with a new Raman reporter and a unique SERS-active substrate incorporated into a microfluidic device. An appropriately designed Raman reporter, basic fuchsin (FC), gives strong SERS enhancement and has the ability to bind both the antibody and gold nanostructures. The fuchsin-labeled immuno-Au nanoflowers can form a sandwich structure with the antigen and the antibody immobilized on the SERS-active substrate based on Au-Ag coated GaN. Our experimental results indicate that this SERS-active substrate with its strong surface-enhancement factor, high stability and reproducibility plays a crucial role in improving the efficiency of SERS immunoassay. This SERS assay was applied to the detection of Hepatitis B virus antigen (HBsAg) in human blood plasma. A calibration curve was obtained by plotting the intensity of SERS signal of FC band at 1178cm(-1) versus the concentration of antigen. The low detection limit for Hepatitis B virus antigen was estimated to be 0.01IU/mL. The average relative standard deviation (RSD) of this method is less than 10%. This SERS immunoassay gives exact results over a broad linear range, reflecting clinically relevant HBsAg concentrations. It also exhibits high biological specificity for the detection of Hepatitis B virus antigen. |
| Databáze: | OpenAIRE |
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