Abstract
Rolling circle amplification (RCA) is an isothermal amplification technique for small, circular DNA templates with the unique property of the product localization. This research describes the application of RCA-based quartz crystal microbalance (QCM) biosensor for direct detection of hepatitis B virus (HBV) genomic DNA from clinical samples. The covalent bonding between the capture probes and the gold electrode surface guarantees that the linkage between the capture probes and the amplified RCA products is maintained during the assay. Making use of the high amplification efficiency of Phi29 DNA polymerase and the remarkable precision of Escherichia coli DNA ligase in differentiating mismatched bases at the ligation site, as low as 104copies/mL HBV genomic DNA can be detected. Experimental results show that RCA has significantly enhanced sensitivity for the target strand compared to the single-base mismatch strand. In this paper, with 60min RCA duration, the proposed sensitive detection shows over 10 times amplification of the original signals. The combination of RCA with QCM biosensor has the potential to become a successful clinical application.
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
