Abstract
The research describes the application of RCA( rolling circle amplification)-based quartz crystal microbalance( QCM) biosensor for the detection of hepatitis B virus( HBV) DNA. RCA is an isothermal amplification technique,which creats long single-stranded products with property of product localization. After amplification,the RCA product is maintained during the assay through the covalent bonding between the capture probes and the gold electrode surface. Using high amplification efficiency of Phi29 DNA polymerase and remarkable precision of Escherichia coli DNA ligase,the detection limit can reach 104 copies /mL. Experimental results show that RCA has significantly enhanced sensitivity for the target strand compared to the single-base mismatch strand. RCA has powerful amplification ability,and QCM has superb mass sensitivity. The combination of these two methods provides a high sensitive RCA-QCM biosensor method,which has the potential to become a successful clinical application.
Published Version
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