title: Disposable immunosensor for human cytomegalovirus glycoprotein B detection.
abstract: Combining the advantages of the biosensor field with the problem of detecting Human Cytomegalovirus (HCMV) in human samples, an inexpensive, simple and disposable electrochemical immunosensor for glycoprotein B detection in urine is proposed. Glycoprotein B has been chosen once is the dominant antigen of HCMV. The approach is based on a sandwich-type immunoassay, with the HCMV glycoprotein B sandwiched between the Anti-HCMV antibody adsorbed onto the working electrode, and the Anti-HCMV labeled with gold nanoparticles. Glycoprotein B detection was carried out through electrochemical stripping analysis of silver nanoparticles quantitatively deposited on the immunosensor through catalysis by the nanogold labels. The influence of different steps in the immunosensor construction, namely the silver deposition time, silver enhancer concentration, antibody concentration, BSA concentration and glycoprotein B incubation time, were examined and optimized. The method showed a linear dependence between glycoprotein B concentration and the corresponding anodic stripping peak current, resulting in detection limits of 3.3±1.7ng/mL for samples prepared in buffer and 3.2±0.2ng/mL for urine samples, suggesting that the biological matrix does not interfere with the immunosensor detection capability. Given its mode of preparation, by physical adsorption of the capture antibody in the working electrode, the immunosensor also exhibited an acceptable reproducibility, with a residual standard deviation of 13.5% for samples prepared in buffer and 11.2% for urine samples, thereby presenting a promising development potential for clinical applications.
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, 6200-506 Covilhã, Portugal; Department of Chemistry, University of Beira Interior, 6200-001 Covilhã, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Pires
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, 6200-506 Covilhã, Portugal; Department of Chemistry, University of Beira Interior, 6200-001 Covilhã, Portugal.
authors.forename: H
authors.initials: H
authors.lastname: Silva
authors.affiliation: Analytical Chemistry Department, Faculty of Sciences, University of Burgos, Burgos, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Domínguez-Renedo
authors.affiliation: Analytical Chemistry Department, Faculty of Sciences, University of Burgos, Burgos, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Alonso-Lomillo
authors.affiliation: Analytical Chemistry Department, Faculty of Sciences, University of Burgos, Burgos, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Arcos-Martínez
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, 6200-506 Covilhã, Portugal; Department of Chemistry, University of Beira Interior, 6200-001 Covilhã, Portugal. Electronic address: ccabral@fcsaude.ubi.pt.
authors.forename: A C
authors.initials: AC
authors.lastname: Dias-Cabral
mesh_terms: D000327:Adsorption; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology*; D015374:Biosensing Techniques; D002384:Catalysis; D006046:Gold / Q000737:chemistry; D006801:Humans; D007118:Immunoassay; D053768:Metal Nanoparticles / Q000737:chemistry; D012834:Silver / Q000737:chemistry; D014759:Viral Envelope Proteins / Q000276:immunology / Q000652:urine*
chemical_list: D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; D012834:Silver; D006046:Gold
keywords: Anodic stripping analysis; Glycoprotein B; Human cytomegalovirus; Immunosensor; Screen-printed electrodes