AccScience Publishing / ITPS / Volume 5 / Issue 1 / DOI: 10.36922/itps.298
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Highly Sensitive Hepatitis B Virus Identification by Antibody-Aptamer Sandwich Enzyme-Linked Immunosorbent Assay

Huijuan Geng1,2 Subash C.B. Gopinath3,4,5 Wenyan Niu1*
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1 School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China
2 Department of Laboratory Medicine, Baoding People’s Hospital, Baoding 071000, Hebei Province, China
3 Faculty of Chemical Engineering & Technology, Universiti Malaysia Perlis (UniMAP), 02600 Arau, Perlis, Malaysia
4 Institute of Nano Electronic Engineering, Universiti Malaysia Perlis (UniMAP), 01000 Kangar, Perlis, Malaysia
5 Micro System Technology, Centre of Excellence (CoE), Universiti Malaysia Perlis (UniMAP), Pauh Campus, 02600 Arau, Perlis, Malaysia
INNOSC Theranostics and Pharmacological Sciences 2022 , 5(1), 7–14;
Submitted: 14 December 2022 | Accepted: 30 January 2023 | Published: 13 February 2023
© 2023 by the Author(s). Licensee AccScience Publishing, Singapore. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution -Noncommercial 4.0 International License (CC BY-NC 4.0) ( )

Hepatitis B virus (HBV) infection is considered a major global health problem, causing various health issues, including cirrhosis, hepatitis, and liver cancer. The risk of developing such complications increases when the viral load is above 105 copies/mL. Early identification of HBV infection is imperative to preventing the spread of infection to other parts of the body. Although various sensing methods have been developed to identify HBV, researchers are still working toward developing cheap, easy, and sensitive detection methods. We developed a highly sensitive and rapid HBV detection method using nanomaterials on enzyme-linked immunosorbent assay (ELISA). Aptamer-antibody was utilized as the detection probe and immobilized on a zeolite-modified ELISA plate to detect the HBV biomarker hepatitis B surface antigen (HBsAg). To enhance the detection of HBsAg, aptamer and antibody were attached to gold nanoparticle through electrostatic interaction and immobilized on the zeolite-modified ELISA plate through amine linker. This probe-modified ELISA plate detected low levels of HBsAg, with a detection limit of 0.1 ng/mL. Furthermore, serum spiked experiments showed increment of absorbance with increasing HBsAg concentration, but control trials with other biomolecules showed no increment of absorbance, showing the specific and selective detection of HBsAg. This nanomaterial-modified ELISA plate can detect low levels of HBsAg and help in the diagnosis of HBV infection in its early stages.

Enzyme-linked immunosorbent assay
Hepatitis B surface antigen
Hepatitis B virus

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Conflict of interest
Authors have no competing interests in relation to the publication of this article.
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INNOSC Theranostics and Pharmacological Sciences, Electronic ISSN: 2705-0823, Print ISSN: TBA, Published by AccScience Publishing