B1 Non-refereed article in a scientific journal

Europium nanoparticle−based simple to perform dry-reagent immunoassay for the detection of Hepatitis B surface antigen




AuthorsSheikh M. Talha, Teppo Salminen, Etvi Juntunen, Anni Spangar, Chandrasekhar Gurramkonda, Tytti Vuorinen, Navin Khanna, Kim Pettersson

PublisherELSEVIER SCIENCE BV

Publication year2016

JournalJournal of Virological Methods

Journal acronymJ VIROL METHODS

Volume229

First page 66

Last page69

Number of pages4

ISSN0166-0934

eISSN1879-0984

DOIhttps://doi.org/10.1016/j.jviromet.2016.01.001(external)


Abstract

Hepatitis B infection, caused by Hepatitis B virus (HBV), presents a huge global health burden. Serological diagnosis of HBV mainly relies on the detection of hepatitis B surface antigen (HBsAg). Although there are high sensitivity commercial HBsAg enzyme immunoassays (EIAs) available, many low-resource laboratories lacking trained technicians continue to use rapid point-of-care assays with low sensitivities for HBsAg detection, due to their simplicity to operate. We developed a time-resolved fluorometric dry-reagent HBsAg immunoassay which meets the detection limit of high sensitivity EIAs but is simple to operate. To develop the assay, anti-HBsAg monoclonal antibody coated on europium nanoparticles was dried atop of biotinylated anti-HBsAg polyclonal antibody immobilized on streptavidin-coated microtiter wells. To test a sample in dry-reagent assay, serum sample and assay buffer were added to the wells, incubated, washed and europium signals were measured. The assay showed a detection limit of 0.25 ng/ml using HBsAg spiked in serum sample. When evaluated with 24HBV positive and 37 negative serum samples, assay showed 100% sensitivity and specificity. Assay wells are stable for at least 26 weeks when stored at 4 °C, and can tolerate elevated temperatures of up to 35 °C for two weeks. The developed assay has high potential to be used in low-resource laboratories.



Last updated on 2024-26-11 at 20:36