基于微流控芯片-巨磁阻抗生物传感器的CYP2C19基因分型研究

批准号:
81300247
项目类别:
青年科学基金项目
资助金额:
23.0 万元
负责人:
洪昌明
依托单位:
学科分类:
H0220.循环系统疾病研究新技术与新方法
结题年份:
2016
批准年份:
2013
项目状态:
已结题
项目参与者:
杨浩、白永怿、祁莉萍、崔华、孙晓楠
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中文摘要
细胞色素P450 2C19 (CYP2C19)存在多个不同的基因型,其基因多态性影响氯吡格雷的代谢,是影响氯吡格雷临床疗效的主要因素之一,对CYP2C19基因进行分型对于指导临床用药具有重要的意义。但目前常用的基因分型技术存在费用高、耗时长等缺点,临床应用受到限制。 本研究拟建立一种基于微流控芯片-巨磁阻抗生物传感器的CYP2C19基因快速分型的新方法。通过携带生物素标记的引物对CYP2C19基因片段进行PCR扩增,使用亲和素修饰的纳米磁球将带有生物素标记的单链片段分离,随后注入固化有捕获探针的微流控芯片内,杂交完成后通过巨磁阻抗生物传感器将捕获结果转化为电磁信号用于判读。该方法可在一个小型芯片上完成所有主要检验步骤,具有快速、方便、灵敏度高、特异性强的优点,能够实现CYP2C19基因分型的快速"床边检测"(point-of-care test)。
英文摘要
Human CYP2C19 gene have several different genotypes. It's polymorphism significantly affect metabolism of clopidogrel. CYP2C19 is one of the most important factors that affected clopidogrel efficacy.Genotyping of CYP2C19 is important to clinical pharmacology. But at present, genotyping technologies have the shortcomings of high cost and time-consuming, which limited its clinical application. Our study intends to establish a new rapid CYP2C19 genotyping technology based on microfluidics and giant magenetoimpedance biosensor. We are going to amplified CYP2C19 gene segment by PCR with whose primer marked by Biotin. Then, we'll use Avidin modified nano magnet particles to purify PCR products by biotin-avidin bind. Next, purified PCR products will be infused into microfluidics which is solidified by capture probe. We'll use giant magnetoinpedence biosensor to read capture results. This new technology can achieve most testing procesures in a chip. It have significant advantages such as rapidness, convenience, high specificity and sensitivity. This new technology can make rapid point-of-care genotyping of CYP2C19 a reality.
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