title: Detection of HSV type-1 and type-2 IgG using an in vitro PCA based homogeneous immunoassay.
abstract: Enzyme immunoassays (EIAs) are widely used in the clinical laboratory and research institutes for the detection of biologically relevant analytes. Almost all EIAs are heterogeneous in nature and require multiple steps of process. In contrast, homogeneous immunoassays (HA) offer a simplified one-step approach with a number of potential advantages over contemporary heterogeneous EIAs such as higher throughput and greater clinical utility. Utilizing TEM-1 beta-lactamase as a reporter enzyme, we have developed HAs based on in vitro protein fragment complementation (PCA) for the detection of antibodies and potentially be used for antigens or other biomarkers. In this proof-of-principle study we demonstrate the successful in vitro differentiation of anti-herpes simplex virus (HSV) type-1 and type-2 Immunoglobulin G (IgG) in human serum with high sensitivity and specificity.
authors.affiliation: Inverness Medical Innovations Australia Pty Ltd, Department of Research and Development, 532 Seventeen Mile Rocks Road, Sinnamon Park, Brisbane, Qld 4073, Australia.
authors.forename: Scott R
authors.initials: SR
authors.lastname: Fry
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Rachel
authors.initials: R
authors.lastname: de las Heras
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: McCourt
authors.forename: Ece
authors.initials: E
authors.lastname: Arel
authors.forename: Edward H
authors.initials: EH
authors.lastname: Kachab
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Hazell
authors.forename: Chang-Yi
authors.initials: CY
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology; D018259:Herpesvirus 1, Human / Q000276:immunology / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000097:blood*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D015032:Zinc / Q000737:chemistry; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D010455:Peptides; D011993:Recombinant Fusion Proteins; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1; D015032:Zinc