Rapid and sensitive HBsAg immunoassay based on fluorescent nanoparticle labels and time-resolved detection.
The detection of hepatitis B virus in blood specimens is carried out commonly by measuring surface antigen (HBsAg) levels with assays designed for various random access immunoanalysers or rapid near-patient testing. These methods leave much to be desired in performance or throughput. The aim of this study was to develop a nanoparticle label-based rapid and sensitive HBsAg immunoassay and evaluate its performance compared to a well-established reference immunoassay (Enzygnost HBsAg 5.0). The assay developed is based on kinetic format and relies on one-step two-site antibody-antigen interaction. Europium(III)-chelate-doped nanoparticles and microtiter wells were coated with anti-HBsAg monoclonal antibodies specific for discrete epitopes. The adaptation of nanoparticle labels for quantitative HBsAg detection showed improved sensitivity (LLD: 0.028 ng/ml) and dynamics (up to 1000 ng/ml) with reasonably low coefficients of variation (concentration-CV%s 2.8-21.9%). Furthermore, concurrent sample runs with the ELISA reference method showed 100% agreement. The time required for the assay was only 10 min facilitating a rapid and convenient method for hepatitis B screening.
University of Turku, Department of Biotechnology, Tykistökatu 6, 6th floor, 20520 Turku, Finland. antti.valanne@utu.fi
Antti
A
Valanne
Saila
S
Huopalahti
Raija
R
Vainionpää
Timo
T
Lövgren
Harri
H
Härmä
D005063:Europium; D005455:Fluorescent Antibody Technique / Q000379:methods; D006510:Hepatitis B Antibodies / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000276:immunology; D006515:Hepatitis B virus / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007163:Immunosorbent Techniques; D049329:Nanostructures; D036103:Nanotechnology; D012680:Sensitivity and Specificity
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D005063:Europium