Lee Sang-Myung, Hwang Kyo Seon, Kim Sang Kyung, Kim Tae Song
Department of Chemical Engineering, Kangwon National University, Kangwon-do, Republic of Korea.
Methods Mol Biol. 2012;903:283-93. doi: 10.1007/978-1-61779-937-2_19.
DNA sensors that are capable of detecting specific DNA sequences in a bio-sample have recently been highlighted as a powerful and sensitive approach to detect infectious diseases caused by pathogens such as viruses and bacteria. Generally, DNA samples extracted from biological fluids are amplified by PCR prior to analysis by DNA sensors or directly analyzed by DNA sensors equipped with a signal amplification process. Nanoparticles have recently been used to amplify the sensor signal and have been shown to play an important role in improving the sensitivity of mechanical resonating sensors. This is because the weight of the nanoparticle can increase the change in the resonance response of the mechanical sensor since this signal change is closely related to mass. Here, we introduce an experimental method to detect HBV at femtomolar concentrations using a silica nanoparticle-enhanced microcantilever resonating sensor. This method includes the preparation of detection probe-conjugated silica nanoparticles, immobilization of capture probe on the microcantilever sensor and sandwich type detection of HBV DNA.
能够检测生物样品中特定DNA序列的DNA传感器,最近已成为检测由病毒和细菌等病原体引起的传染病的一种强大且灵敏的方法。一般来说,从生物体液中提取的DNA样品在通过DNA传感器分析之前,先通过聚合酶链式反应(PCR)进行扩增,或者由配备信号放大过程的DNA传感器直接进行分析。纳米颗粒最近已被用于放大传感器信号,并已证明在提高机械共振传感器的灵敏度方面发挥着重要作用。这是因为纳米颗粒的重量可以增加机械传感器共振响应的变化,因为这种信号变化与质量密切相关。在这里,我们介绍一种使用二氧化硅纳米颗粒增强微悬臂梁共振传感器检测飞摩尔浓度乙肝病毒(HBV)的实验方法。该方法包括制备检测探针偶联的二氧化硅纳米颗粒、将捕获探针固定在微悬臂梁传感器上以及对HBV DNA进行夹心型检测。