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滚环扩增型压电石英晶体传感器检测乙型肝炎病毒
引用本文:姚春艳,王云霞,府伟灵.滚环扩增型压电石英晶体传感器检测乙型肝炎病毒[J].高等学校化学学报,2014,35(8):1635.
作者姓名:姚春艳  王云霞  府伟灵
作者单位:第三军医大学西南医院检验科, 重庆 400038
基金项目: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, Phi29 DNA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104 copies/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.

关 键 词:滚环扩增  石英晶体传感器  乙型肝炎病毒  
收稿时间:2014-06-06

HBV DNA Detection by RCA-QCM Biosensor†
YAO Chunyan,WANG Yunxia,FU Weiling.HBV DNA Detection by RCA-QCM Biosensor†[J].Chemical Research In Chinese Universities,2014,35(8):1635.
Authors:YAO Chunyan  WANG Yunxia  FU Weiling
Affiliation:Department of Laboratory Medicine, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China
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.
Keywords:Rolling circle amplification  Quartz crystal microbalance biosensor  Hepatitis B virus  
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