Hasmoni, Siti Salwa and Yusoff, Khatijah and Tan, Wen Siang (2005) Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage. The Journal of General and Applied Microbiology, 51 (2). pp. 125-131. ISSN 0022-1260
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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.
|Keyword:||fusion phage, hepatitis B virus, immunoassays, precipitation|
|Faculty or Institute:||Faculty of Biotechnology and Biomolecular Sciences|
|Publisher:||The Microbiology Research Foundation|
|Deposited By:||Erni Suraya Abdul Aziz|
|Deposited On:||16 Apr 2010 04:02|
|Last Modified:||16 Apr 2010 04:04|
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