title: Simultaneous determination of CRP and D-dimer in human blood plasma samples with White Light Reflectance Spectroscopy.
abstract: A dual-analyte assay for the simultaneous determination of C-reactive protein (CRP) and D-dimer in human blood plasma based on a white light interference spectroscopy sensing platform is presented. Measurement is accomplished in real-time by scanning the sensing surface, on which distinct antibody areas have been created, with a reflection probe used both for illumination of the surface and collection of the reflected interference spectrum. The composition of the transducer, the sensing surface chemical activation and biofunctionalization procedures were optimized with respect to signal magnitude and repeatability. The assay format involved direct detection of CRP whereas for D-dimer a two-site immunoassay employing a biotinylated reporter antibody and reaction with streptavidin was selected. The assays were sensitive with detection limits of 25ng/mL for both analytes, precise with intra- and inter-assay CV values ranging from 3.6% to 7.7%, and from 4.8% to 9.5%, respectively, for both assays, and accurate with recovery values ranging from 88.5% to 108% for both analytes. Moreover, the values determined for the two analytes in 35 human plasma samples were in excellent agreement with those received for the same samples by standard diagnostic laboratory instrumentation employing commercial kits. The excellent agreement of the results supported the validity of the proposed system for clinical application for the detection of multiple analytes since it was demonstrated that up to seven antibody areas can be created on the sensing surface and successfully interrogated with the developed optical set-up.
authors.affiliation: Immunoassay/Immunosensors Lab, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, NCSR "Demokritos", GR-15310 Aghia Paraskevi, Greece.
authors.forename: Georgios
authors.initials: G
authors.lastname: Koukouvinos
authors.affiliation: Immunoassay/Immunosensors Lab, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, NCSR "Demokritos", GR-15310 Aghia Paraskevi, Greece. Electronic address: ypetrou@rrp.demokritos.gr.
authors.forename: Panagiota
authors.initials: P
authors.lastname: Petrou
authors.affiliation: Optical sensors Lab, Institute of Nanoscience and Nanotechnology, NCSR "Demokritos", GR-15310 Aghia Paraskevi, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Misiakos
authors.affiliation: ThetaMetrisis S.A., Polydefkous 14, GR-12243, Athens, Greece.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Drygiannakis
authors.affiliation: ThetaMetrisis S.A., Polydefkous 14, GR-12243, Athens, Greece.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Raptis
authors.affiliation: Diagnostic Laboratories, "Henri Dunant" Hospital, 115 26 Athens, Greece.
authors.forename: Gerasimos
authors.initials: G
authors.lastname: Stefanitsis
authors.affiliation: Diagnostic Laboratories, "Henri Dunant" Hospital, 115 26 Athens, Greece.
authors.forename: Spyridoula
authors.initials: S
authors.lastname: Martini
authors.affiliation: Diagnostic Laboratories, "Henri Dunant" Hospital, 115 26 Athens, Greece.
authors.forename: Dimitra
authors.initials: D
authors.lastname: Nikita
authors.affiliation: Electronics Engineering Department, TEI of Piraeus, Athens, Greece.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Goustouridis
authors.affiliation: Electronics Engineering Department, TEI of Piraeus, Athens, Greece.
authors.forename: Isabella
authors.initials: I
authors.lastname: Moser
authors.affiliation: Jobst Technologies GmbH, 79108 Freiburg, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Jobst
authors.affiliation: Immunoassay/Immunosensors Lab, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, NCSR "Demokritos", GR-15310 Aghia Paraskevi, Greece.
authors.forename: Sotirios
authors.initials: S
authors.lastname: Kakabakos
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation*; D002097:C-Reactive Protein / Q000032:analysis*; D004867:Equipment Design; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D008027:Light; D057230:Limit of Detection; D015203:Reproducibility of Results; D013057:Spectrum Analysis / Q000295:instrumentation
chemical_list: D055910:Antibodies, Immobilized; D005338:Fibrin Fibrinogen Degradation Products; C036309:fibrin fragment D; D002097:C-Reactive Protein
keywords: C-reactive protein; D-dimer; Dual-analyte determination; Human blood plasma sample; Label-free detection; White light interference spectroscopy