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Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage


Citation

Hasmoni, Siti Salwa and Yusoff, Khatijah and Tan, Wen Siang (2005) Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage. Journal of General and Applied Microbiology, 51 (2). pp. 125-131. ISSN 0022-1260; ESSN: 1349-8037

Abstract / Synopsis

The nucleocapsids of hepatitis B virus (HBV) are made of 180 or 240 subunits of core proteins or known as core antigens (HBcAg). A fusion bacteriophage bearing the WSFFSNI sequence that interacts tightly to HBcAg was employed as a diagnostic reagent for the detection of the antigen using the phage-enzyme-linked immunosorbent (phage-ELISA), dot blot and immunoprecipitation assays. The results from phage-ELISA and dot blot assay showed that as low as 10 ng of HBcAg can be detected optimally by 1.0×1012 pfu/ml fusion M13 bacteriophage. The sensitivity of the dot blot assay corresponds with that of the phage-ELISA. HBcAg in HBV positive serum samples can also be detected using the fusion phage via the phage-ELISA and phage-dot blot assay. The phage cross-linked to cyanogen bromide (CNBr) activated agarose can also be used to precipitate HBcAg in bacterial lysate. The optimum amount of phage needed for cross-linking to 1 g of agarose is about 7.0×106 pfu/ml which could also precipitate purified and unpurified HBcAg in bacterial lysate. This study demonstrates the potential of fusion bacteriophage bearing the sequence WSFFSNI as a diagnostic reagent and a ligand for the detection and purification of HBcAg respectively.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.2323/jgam.51.125
Publisher: Applied Microbiology, Molecular and Cellular Biosciences Research Foundation
Keywords: Fusion phage; Hepatitis B virus; Immunoassays; Precipitation
Depositing User: Erni Suraya Abdul Aziz
Date Deposited: 16 Apr 2010 04:02
Last Modified: 25 Apr 2016 04:35
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.2323/jgam.51.125
URI: http://psasir.upm.edu.my/id/eprint/5605
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