title: Diagnosis of Epstein-Barr virus infection in clinical serum samples by an SPR biosensor assay.
abstract: Label-free affinity biosensors offer a promising platform for the development of a new generation of medical diagnostic technologies. Nevertheless, when such sensors are used in complex biological media, adsorption of non-targeted medium components prevents the specific detection of the analyte. In this work, we introduce for the first time a biosensor assay based on surface plasmon resonance (SPR) capable of diagnosing different stages of Epstein-Barr virus (EBV) infections in clinical serum samples. This was achieved by simultaneous detection of the antibodies against three different antigens present in the virus. To prevent the interference of the fouling from serum during the measurement, the SPR chips were coated by an antifouling layer of a polymer brush of poly[oligo(ethylene glycol) methacrylate] grown by surface-initiated atom transfer radical polymerization. The bioreceptors were then attached via hybridization of complementary oligonucleotides. This allowed the sensor surface to be regenerated after measurement by disrupting the complementary pairs above the oligonucleotides' melting temperature and attaching new bioreceptors. In this way, the same sensing surface could be used repeatedly. The procedure used in this work will serve as a prototype strategy for the development of label-free affinity biosensors for diagnostics in blood serum or plasma samples. This is the first example of detection of marker of a disease in clinical serum samples by an optical affinity biosensor.
authors.affiliation: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic. Electronic address: riedel@imc.cas.cz.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Riedel
authors.affiliation: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic.
authors.forename: Cesar
authors.initials: C
authors.lastname: Rodriguez-Emmenegger
authors.affiliation: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic.
authors.forename: Andres
authors.initials: A
authors.lastname: de los Santos Pereira
authors.affiliation: Vidia s.r.o., Nad Safinou II, no. 365, Vestec, 252 42 Jesenice u Prahy, Czech Republic.
authors.forename: Anna
authors.initials: A
authors.lastname: Bědajánková
authors.affiliation: Vidia s.r.o., Nad Safinou II, no. 365, Vestec, 252 42 Jesenice u Prahy, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Jinoch
authors.affiliation: V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine, Nauki 41, 03028 Kyiv, Ukraine.
authors.forename: Praskovia M
authors.initials: PM
authors.lastname: Boltovets
authors.affiliation: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic.
authors.forename: Eduard
authors.initials: E
authors.lastname: Brynda
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation*; D020031:Epstein-Barr Virus Infections / Q000097:blood* / Q000175:diagnosis* / Q000276:immunology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D000914:Antibodies, Viral
keywords: Antifouling; Epstein–Barr virus infection; Polymer brushes; Real time diagnostics; Surface plasmon resonance biosensor