Quick genotyping detection of HBV by giant magnetoresistive biochip combined with PCR and line probe assay
Quick genotyping detection of HBV by giant magnetoresistive biochip combined with PCR and line probe assay
复制标题
巨磁阻生物芯片结合PCR和线探针法快速检测HBV基因型
DOI：
10.1039/c2lc20949g
复制
发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Cui, Daxiang
中科院分区：
文献类型：
--
作者：
Zhi, Xiao;Liu, Qingsheng;Cui, Daxiang
人类B肝炎病毒（HBV）基因分型可用于指导临床有效治疗药物的选择。本文报道了一种利用磁纳米团簇（MNCs）、巨磁阻（GMR）生物芯片、聚合酶链反应（PCR）和线探针技术快速检测HBV基因分型的方法。制备直径约180 nm的磁性纳米团簇，并用链霉亲和素修饰，所得链霉亲和素修饰的磁性纳米团簇用于捕获传感器检测界面上的生物素标记的混合产物。以B型和C型质粒为模板，通过PCR扩增HBV的B和C型基因片段。基因片段与捕获探针杂交后，链霉亲和素修饰的磁性纳米团簇可以与生物素偶联的基因片段结合，生成的氢化物产物可以被GMR传感器快速检测和区分，检测灵敏度为200 IU mL（-1）靶HBV DNA分子。该方法在临床HBV基因分型诊断中具有很大的应用潜力，并可以很容易地扩展到其他基于分子识别的生物医学应用。
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.