Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage
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- Hasmoni Siti Salwa
- Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia
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- Yusoff Khatijah
- Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia
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- Tan Wen Siang
- Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia
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説明
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.
収録刊行物
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- The Journal of General and Applied Microbiology
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The Journal of General and Applied Microbiology 51 (2), 125-131, 2005
公益財団法人 応用微生物学・分子細胞生物学研究奨励会
