title: Array-in-well platform-based multiplex assay for the simultaneous detection of anti-HIV- and treponemal-antibodies, and Hepatitis B surface antigen.
abstract: Multiplex assays detecting sets of related clinical analytes simultaneously can save considerable amount of time and resources. Array-in-well (AIW) is a powerful platform for the multiplex detection of different analytes where microarrays can be printed at the bottom of microtiter wells, thus combining the potential of microarrays with the ease of handling microtiter wells. We have developed a single-step AIW assay for the simultaneous screening of HIV, Treponema pallidum subspecies pallidum (causing syphilis) and Hepatitis B virus infections targeting the specific detection of anti-HIV- and treponemal-antibodies and Hepatitis B surface antigen (HBsAg), respectively, using two different fluorescent label technologies i.e. DyLight 633 and europium nanoparticle. Double-antigen assay formats were used for anti-HIV- and treponemal-antibody detection that can simultaneously detect both IgG and IgM, and thus reduce the window period of detection. AIW assay was evaluated with well characterized serum/plasma samples (n=111), and the qualitative results were in near complete agreement with those of the reference assays. The AIW assay exhibited 100% sensitivities for all three analytes, and 100% specificities for anti-HIV antibodies and HBsAg, and 98.6% specificity for treponemal antibodies. The limit of detection of HBsAg in AIW assay was 0.18 ng/ml. This high performing AIW assay has the potential to be used as a multiplex screening test for these three infections.
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland. Electronic address: talshe@utu.fi.
authors.forename: Sheikh M
authors.initials: SM
authors.lastname: Talha
authors.affiliation: VTT Technical Research Centre of Finland Ltd., FI-02044 VTT, Finland.
authors.forename: Petri
authors.initials: P
authors.lastname: Saviranta
authors.affiliation: VTT Technical Research Centre of Finland Ltd., FI-02044 VTT, Finland.
authors.forename: Liisa
authors.initials: L
authors.lastname: Hattara
authors.affiliation: Department of Virology, University of Turku, Turku, Finland.
authors.forename: Tytti
authors.initials: T
authors.lastname: Vuorinen
authors.affiliation: Department of Medical Microbiology and Immunology, University of Turku, Turku, Finland.
authors.forename: Jukka
authors.initials: J
authors.lastname: Hytönen
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi, India; Translational Health Science & Technology Institute, NCR Biotech Science Cluster, Faridabad, India; Department of Paediatrics, Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Navin
authors.initials: N
authors.lastname: Khanna
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Kim
authors.initials: K
authors.lastname: Pettersson
mesh_terms: D000906:Antibodies / Q000032:analysis* / Q000097:blood / Q000276:immunology; D005470:Fluorometry; D006678:HIV / Q000276:immunology*; D015658:HIV Infections / Q000097:blood / Q000276:immunology; D006509:Hepatitis B / Q000097:blood / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D018613:Microscopy, Confocal; D014210:Treponema pallidum / Q000276:immunology*
chemical_list: D000906:Antibodies; D006514:Hepatitis B Surface Antigens
keywords: Array-in-well; Diagnostics; HIV; Hepatitis B virus; Multiplex; Syphilis