National Key Laboratory of Nano/Micro Fabrication Technology, Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Lab Chip. 2012 Feb 21;12(4):741-5. doi: 10.1039/c2lc20949g. Epub 2012 Jan 6.
Genotyping of human hepatitis B virus (HBV) can be used to direct clinically effective therapeutic drug-selection. Herein we report that a quick genotyping method for human HBV was established by a specially designed giant magnetoresistive (GMR) biochip combined with magnetic nanoclusters (MNCs), PCR and line probe assay. Magnetic nanoclusters of around 180 nm in diameter were prepared and modified with streptavidin, and resultant streptavidin-modified magnetic nanoclusters were used for capturing biotin-labeled hybrid products on the detection interface of the sensor. The gene fragments of HBV's B and C gene types were obtained by PCR based on a template of B- and C-type plasmids. After gene fragments were hybridized with captured probes, streptavidin-modified magnetic nanoclusters could bind with biotin-conjugated gene fragments, and the resultant hydride products could be quickly detected and distinguished by the GMR sensor, with a detection sensitivity of 200 IU mL(-1) target HBV DNA molecules. The novel method has great potential application in clinical HBV genotyping diagnosis, and can be easily extended to other biomedical applications based on molecular recognition.
乙型肝炎病毒（HBV）的基因分型可用于指导临床有效的治疗药物选择。在此，我们报告了一种快速的乙型肝炎病毒基因分型方法，该方法通过专门设计的巨磁电阻（GMR）生物芯片与磁性纳米簇（MNC）、PCR 和线性探针分析相结合建立。我们制备了直径约 180nm 的磁性纳米簇，并对其进行了链霉亲和素修饰，所得的链霉亲和素修饰的磁性纳米簇用于在传感器的检测界面上捕获生物素标记的杂交产物。基于 B 型和 C 型质粒的模板，通过 PCR 获得 HBV 的 B 和 C 基因类型的基因片段。基因片段与捕获探针杂交后，链霉亲和素修饰的磁性纳米簇可以与生物素化的基因片段结合，所得的氢化物产物可以通过 GMR 传感器快速检测和区分，检测灵敏度为 200IU mL(-1)的靶 HBV DNA 分子。该新方法在临床乙型肝炎病毒基因分型诊断中有很大的应用潜力，并可基于分子识别轻松扩展到其他生物医学应用中。
