滚环扩增型压电石英晶体传感器检测乙型肝炎病毒

HBV DNA Detection by RCA-QCM Biosensor

通过捕获探针与纳米金膜之间的共价连接,保证了滚环扩增(RCA)产物始终结合于金膜表面,Phi29DNA聚合酶的高效扩增和Escherichia coli DNA链接酶的高度精确性使检测达到单碱基识别,检测灵敏度达到104copies/mL.实验结果表明,与单碱基错配序列相比,RCA可明显增强检测的灵敏度.该RCA基因传感器操作简单,灵敏度和特异性较高,在乙型肝炎病毒的快速检测方面具有一定的开发潜力.

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 am-plification 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 cap-ture 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. Experimen-tal 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 poten-tial to become a successful clinical application.