Detection of HSV type-1 and type-2 IgG using an in vitro PCA based homogeneous immunoassay.
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.
Fry
Scott R
SR
Inverness Medical Innovations Australia Pty Ltd, Department of Research and Development, 532 Seventeen Mile Rocks Road, Sinnamon Park, Brisbane, Qld 4073, Australia.
Li
Jun
J
de las Heras
Rachel
R
McCourt
Jennifer A
JA
Arel
Ece
E
Kachab
Edward H
EH
Hazell
Stuart L
SL
Huang
Chang-Yi
CY
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
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