Detection And Precipitation Of Hepatisis B Core Antigen Using A Fusion Bacteriophage.
Hasmoni, Siti Salwa and Yusoff, Khatijah and Wen, Siang Tan (2005) Detection And Precipitation Of Hepatisis B Core Antigen Using A Fusion Bacteriophage. Journal of General and Applied Microbiology, 51 . pp. 125-131. ISSN 0022-1260
The nucleocapsids of hepatisis 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 and dot blot assay showed that as low as 10 ng of HBcAg can be detected optimally by 1.0x1012 pfu/ml fusion m13 bacteriophage. The sensitivity of the dot blot assay correspond 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 7.0x 10 6 pfu/ml which could also prepicipitate purified an 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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