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

作　　者：姚春艳[1] 王云霞[1] 府伟灵[1] 

机构地区：[1]第三军医大学西南医院检验科,重庆400038

出　　处：《高等学校化学学报》2014年第8期1635-1639,共5页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30970766;81371885)资助~~

摘　　要：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.

关 键 词：滚环扩增 石英晶体传感器 乙型肝炎病毒 

分 类 号：O657[理学—分析化学]